Patent Grant
10960397
The present invention relates generally to microfluidic structures, and more specifically, to microfluidic structures and methods for manipulating fluids, fluid components, and reactions.
Microfluidic systems typically involve control of fluid flow through one or more microchannels. One class of systems includes microfluidic “chips” that include very small fluid channels and small reaction/analysis chambers. These systems can be used for analyzing very small amounts of samples and reagents and can control liquid and gas samples on a small scale. Microfluidic chips have found use in both research and production, and are currently used for applications such as genetic analysis, chemical diagnostics, drug screening, and environmental monitoring. Although these systems may allow manipulation of small volumes of fluids, additional methods that allow further control and flexibility are needed.
Microfluidic structures and methods for manipulating fluids, fluid components, and reactions are provided.
In one aspect of the invention, a method is provided. The method comprises providing a microfluidic network comprising a first region and a microfluidic channel in fluid communication with the first region, flowing a first fluid in a first direction in the microfluidic channel, and flowing a second fluid in the first direction in the microfluidic channel. The method also includes partitioning at least a portion of the first fluid at the first region, at least in part through action of the second fluid, so as to form a first droplet of the first fluid at the first region. The method also includes maintaining the droplet at the first region while the second fluid is flowing in the first direction.
In another embodiment, a method comprises providing a microfluidic network comprising at least a first inlet to a microfluidic channel, a first and a second region for forming a first and a second droplet, respectively, the first and second regions in fluid communication with the microfluidic channel, and flowing a first fluid in the microfluidic channel. The method involves partitioning a first portion of the first fluid at the first region, at least in part through action of the second fluid, so as to form the first droplet at the first region, and partitioning a second portion of the first fluid at the second region, at least in part through action of the second fluid, so as to form the second droplet at the second region.
In another embodiment, a microfluidic device comprises a plurality of chamber units positioned in parallel, each chamber unit comprising: a chamber having a chamber inlet and a chamber outlet, a feed channel fluidly connected to a plurality of chamber inlets, a drain channel fluidly connected to a plurality of chamber outlets, a chamber bypass channel extending from the chamber, and a fluid restriction region between the chamber outlet and the drain channel, the fluid restriction region being more restrictive to fluid flow than the chamber.
In another embodiment, a method comprises flowing a fluid containing a plurality of components in a microfluidic system comprising a chamber having a flow direction, a chamber inlet, a chamber outlet, and a chamber bypass channel extending from the chamber between the chamber inlet and the chamber outlet. The method also includes positioning a component in the chamber, the chamber having a cross-sectional area, perpendicular to the flow direction, less than 2 times the largest cross-sectional area of the component perpendicular to the flow direction, and flowing a fluid through the chamber while maintaining the component at its position in the chamber. A portion of the plurality of components may be flowed in the chamber bypass channel.
In another embodiment, a system comprises a microfluidic device comprising an inlet, an outlet, a chamber having a flow direction, and a flow restriction region fluidly connected to the outlet of the chamber, and a plurality of cells generally aligned in the chamber. At least 80% of the cells have a largest cross-sectional area, perpendicular to the flow direction, of between 0.1 and 1.0 times the cross-sectional area of the chamber perpendicular to the flow direction. The flow restriction region is constructed and arranged to allow a fluid but not the cells to pass therethrough.
A method may also comprise providing a microfluidic network comprising at least a first inlet to a microfluidic channel, a first and a second region for positioning a first and a second reactive component, respectively, the first and second regions in fluid communication with the microfluidic channel, wherein the first region is closer in distance to the first inlet than the second region, and flowing a first fluid comprising first and second components in the microfluidic channel. The method may also include positioning the first component at the first region, positioning the second component at the second region, and maintaining the first and second components in the first and second regions, respectively, while a fluid is flowing in the microfluidic channel. In one embodiment, the first and/or second reactive component is a cell. In another embodiment, the first and/or second reactive component is a bead. In some cases, positioning of the first and/or second reactive components does not require use of a fluid immiscible with the first fluid. The method may optionally include flowing a second fluid comprising an associating component in the microfluidic channel, wherein the associating component can interact with the first and/or second reactive components. The associating component may be a binding partner complementary to the first and/or second components. In some embodiments, the microfluidic channel comprises an upstream portion, a downstream portion, and first and second fluid paths extending from the upstream portion and reconnecting at the downstream portion. The first and second fluid paths may have different resistances to flow. In some cases, the first region is positioned within the first fluid path.
Other advantages and novel features of the present invention will become apparent from the following detailed description of various non-limiting embodiments of the invention when considered in conjunction with the accompanying figures. In cases where the present specification and a document incorporated by reference include conflicting and/or inconsistent disclosure, the present specification shall control. If two or more documents incorporated by reference include conflicting and/or inconsistent disclosure with respect to each other, then the document having the later effective date shall control.
Non-limiting embodiments of the present invention will be described by way of example with reference to the accompanying figures, which are schematic and are not intended to be drawn to scale. In the figures, each identical or nearly identical component illustrated is typically represented by a single numeral. For purposes of clarity, not every component is labeled in every figure, nor is every component of each embodiment of the invention shown where illustration is not necessary to allow those of ordinary skill in the art to understand the invention. In the figures:
The present invention relates to microfluidic structures and methods for manipulating fluids, fluid components, and reactions. In one aspect, such structures and methods involve positioning fluid samples, e.g., in the form of droplets, in a carrier fluid (e.g., an oil, which may be immiscible with the fluid sample) in predetermined regions in a microfluidic network. In some embodiments, positioning of the droplets can take place in the order in which they are introduced into the microfluidic network (e.g., sequentially) without significant physical contact between the droplets. Because of the little or no contact between the droplets, coalescence between the droplets can be avoided. Accordingly, in such embodiments, surfactants are not required in either the fluid sample or the carrier fluid to prevent coalescence of the droplets. Positioning of droplets without the use of surfactants is desirable in certain cases where surfactants may negatively interfere with the contents in the fluid sample (e.g., proteins). Structures and methods described herein also enable droplets to be removed sequentially from the predetermined regions to a different region of the fluidic network where they can be further processed.
Once the droplets are positioned at the predetermined regions, they can be stored and/or may undergo manipulation (e.g., diffusion, evaporation, dilution, and precipitation). In some instances, many (e.g., 1000) droplets can be manipulated, sometimes simultaneously. Manipulation of fluid samples can be useful for a variety of applications, including testing for reaction conditions, e.g., in crystallization, and chemical and/or biological assays, including chemical reactions, enzymatic reactions, immuno-based reactions (e.g., antigen-antibody), and cell-based reactions.
It should be understood that while several of the embodiments described herein refer to the positioning and/or manipulation of droplets, the embodiments are also applicable to other components such as cells and beads, which may be contained in a fluid without being in a droplet.
In another aspect, microfluidic structures and methods described herein can allow production of droplets of a precise volume, which can be stored/maintained at precise regions of the device. The droplets can be created at a region (e.g., a storage region) in a self-regulated manner. The method may include, optionally, filling a microfluidic channel with a filling fluid (e.g., oil). The oil can then be flushed out with a first fluid (e.g., an aqueous fluid) to be stored/maintained at a region of the device. This first fluid may be immiscible with the filling fluid. The first fluid can enter a region of the device for storing a droplet, replacing the filling fluid in that region. Next, a second fluid (e.g., a fluid immiscible with the first fluid) may be flowed in the channel, causing partitioning of a portion of the first fluid. Partitioning of the first fluid causes formation of a droplet of the first fluid at the region, while a second portion of the first fluid bypasses the region. In this manner, a plurality of droplets can be generated sequentially down the length of the channel.
In another aspect, microfluidic structures and methods described herein are designed for containing and positioning components in an arrangement such that the components can be manipulated and then tracked even after manipulation. For example, cells may be constrained in an arrangement in microfluidic structures described herein to facilitate tracking during their growth and/or after they multiply. This can allow, for example: 1) cells to be trapped and observed over time; 2) culturing of cells in a manner than allows determination of their identity and lineage; and 3) manipulation of the cells (e.g., by staining or washing) while maintaining the identity and/or position of the cells. Other advantages and applications are described in more detail below.
Certain microfluidic chips described herein may include a micro fluidic network having a region for forming droplets of sample in a carrier fluid (e.g., an oil), and one or more regions (e.g., microreactor regions, microwells, reservoirs, chambers, or portions of a microfluidic channel) in which the droplets can be positioned and reaction conditions within the droplet can be varied. In some embodiments, the droplet formation region is the same as the region in which the droplet is positioned for varying a condition within the droplet. Droplets may be positioned sequentially in regions of the microfluidic network so that upon manipulating and/or performing a chemical and/or biological process within each the droplets, the droplets can be identified at a later time, for example, to determine the particular conditions within the droplets that lead to a favorable outcome (e.g., optimal conditions for forming a product, for crystal growth, etc.).
As used herein, “droplet” means a small portion of a fluid, isolated from other portions of the same fluid. A droplet can have a traditional, rounded shape, or a different shape which can be influenced by its environment. A droplet of a first fluid can be surrounded by different, immiscible fluid, or bounded by a surface of an article, or a gas such as air, or a combination. For example, a droplet of a first fluid can be suspended in (completely surrounded by) a second fluid immiscible with the first fluid. Or a droplet of a first fluid can reside on a surface of a solid article, with portions that are not in contact with the surface exposed to the second fluid or a gas. A droplet can be bounded on multiple sides by one or more surfaces of an article, e.g. the interior of a channel. For example, a portion of a channel completely filled with a first fluid, which resides within a discrete regions of the channel, is a droplet for purposes of the invention.
It should also be understood that any suitable fluid(s) can be used in connection with devices and methods described herein. Where embodiments describe the use of “immiscible” fluids, those of ordinary skill in the art know or can determine by simple experimentation which combination of fluids is immiscible. For instance, solubility parameters of a variety of fluids are available in literature and can be used to determine miscibility/immiscibility. Additionally and/or alternatively, simple experimentation may include, for example, mixing two or more fluids in a container—if the fluids partition after a certain period of time, the fluids are immiscible. Furthermore, it should be understood that where “first” and “second” fluids are described herein, these fluids can have any suitable composition and can be interchangeable in other embodiments. For example, one particular embodiment may describe the use of a “first fluid” that is aqueous and a “second fluid” that is an oil, and a different embodiment may described a “first fluid” as an oil and a “second fluid” that is aqueous. In certain embodiments, first and second fluids can be miscible with one another (e.g., both being aqueous or both being an oil). Gaseous fluids may also be used.
As shown in
It should be understood that any suitable fluid path can be used as a fluid restriction region, which may have a higher hydrodynamic resistance and/or a smaller cross-sectional area for fluid flow than a region immediately upstream or downstream of the fluid constriction region. For instance, fluid restriction region 1024 may be in the form of a narrow fluid path or a channel having the same dimensions as fluid path 1014, but having an obstruction (e.g., posts or a valve) positioned in or at the region. In other embodiments, fluid restriction region 1024 may comprise a porous membrane, a semi-permeable plug (e.g., a gel), a valve, or another structure.
As shown in the embodiment illustrated in
It should be understood that when droplet 1020 is positioned at region 1028, the droplet may plug all or a portion of fluid path 1014 and/or fluid restriction region 1024. For instance, in some cases, the droplet plugs all of such fluid paths such that none of carrier fluid 1021 (or another fluid) flowing in microfluidic channel 1002 passes through fluid restriction region 1024. In other embodiments, the droplet may plug only a portion of such fluid paths such that some fluid passes through fluid restriction region 1024 even though the droplet is positioned at region 1028. The amount of fluid flowing past the positioned droplet may depend on factors such as the dimensions of fluid path portions 1014 and/or 1024, the size of the droplets, the flow rate, etc. As the droplet causes fluid path 1014 to have a higher relative hydrodynamic resistance than fluid path 1018, a second droplet can bypass fluid path 1014 and enter fluid path 1018.
As described above, fluid paths 1014 and 1018 may have different hydrodynamic resistances depending on whether or not a droplet is positioned at region 1028. In the absence of a droplet positioned at region 1028, fluid path 1014 may be configured to have a lower hydrodynamic resistance than fluid path 1018. For example, greater than 50%, greater than 60%, greater than 70%, greater than 80%, or greater than 90% of the fluid flowing in channel 1002 at upstream portion 1006 may flow in fluid path 1014 compared to fluid path 1018. However, when the droplet is positioned and maintained in region 1028, fluid path 1014 may be relatively more restrictive to fluid flow. For example, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the fluid flowing in channel 1002 at upstream portion 1006 may flow in fluid path 1014 compared to that of 1018. In some cases, 100% of the fluid flowing in direction 1012 in microfluidic channel 1002 flows in fluid path 1018 upon positioning of a droplet in region 1028.
As illustrated in the exemplary embodiment of
Accordingly, in some instances, the positioning of the droplets in the microfluidic network can take place without the use of surfactants. In other words, surfactants in either a fluid flowing in channel 1002 (e.g., a carrier fluid) or within the droplets is not required in order to stabilize the droplets and/or prevent the droplets from coalescing with one another during positioning or carrying the droplet in the microfluidic channel, and/or during maintaining the droplets within a predetermined region within the microfluidic network. However, in instances where coalescence is desired (e.g., to allow a reaction between reagents contained in two droplets), the microfluidic network and methods for operating the network can be configured to allow such physical contact and/or coalescence between droplets. These interactions or absence of interactions can be controlled, for example, by varying the volume and/or length of the droplets, as well as the volume and/or length of regions 1028.
In some embodiments, methods for positioning a droplet in a microfluidic network include the steps of providing a microfluidic network comprising a first region (e.g., region 1028 of
In some embodiments, a chemical and/or biological process and/or another manipulation process can be carried out in droplet 1020 of
In some embodiments, droplets that have been positioned at regions of a microfluidic network can be removed or extracted from the regions to a different location in the microfluidic network, where they can be optionally processed, manipulated, and/or collected. As shown in the illustrative embodiments of
In some such embodiments, upstream portion 1006 and downstream portion 1010 of
In some embodiments, sequential positioning of droplets can be performed such that a first droplet is positioned in a first region before a second droplet is positioned in a second region (and, optionally, before third, fourth, fifth droplets, etc. are positioned in their respective regions). As described above, sequential removal of the droplets can be performed such that the first droplet is removed from a region and/or positioned at a different location of the microfluidic network before the second droplet (and, optionally, before third, fourth, fifth droplets, etc. are removed from their respective regions). In other embodiments, removal of the droplets can be performed such that the second droplet is removed and/or positioned at a different location of the microfluidic network before the first droplet.
In some cases, several (e.g., greater than 2, greater than 5, greater than 10, greater than 50, greater than 100, greater than 200, greater than 500, or greater than 1000) droplets can be positioned at regions of the microfluidic network, wherein the droplets are positioned in the regions in the order the droplets are introduced into the microfluidic network. In some cases, removing several droplets positioned at regions of the microfluidic network comprises removing the droplets in the order the droplets were introduced into the microfluidic network (or in the order the droplets were positioned into the regions of the microfluidic network). In other cases, removing several droplets positioned at regions of the microfluidic network comprises removing the droplets in the reverse order the droplets were introduced into the microfluidic network (or in the reverse order the droplets were positioned into the regions of the microfluidic network). Other methods of positioning and removal of droplets are also possible.
The sequential (or predetermined/known order of) removal of droplets from regions of a microfluidic network can allow control over the identification and location of each droplet within the network. This can also allow determination of the contents inside each of the droplets from the time they are formed and/or introduced into the microfluidic network, to the time the droplets are manipulated and/or extracted from the microfluidic network.
A microfluidic network may have any suitable number of microfluidic sections 1001. For instance, the microfluidic network may have greater than or equal to 5, greater than or equal to 10, greater than or equal to 30, greater than or equal to 70, greater than or equal to 100, greater than or equal to 200, greater than or equal to 500, or greater than or equal to 1000 such sections.
In additional, although certain embodiments herein show that sections 1001 can allow positioning of a single droplet in each of the sections, in other embodiments, the sections can be designed such that greater than one droplet (e.g., greater than or equal to 2, greater than or equal to 5, or greater than or equal to 10 droplets) can be positioned at each section.
Furthermore, although only two fluid flow paths 1014 and 1018 are shown extending from channel 1002, in other embodiments, more than two (e.g., greater than or equal to 3, greater than or equal to 5, or greater than or equal to 10) fluid paths may extend from channel 1002. Each extending fluid path may optionally comprise one or more regions (e.g., microwells) for positioning and/or maintaining droplets.
Once droplet 1060 is positioned at region 1028-A, it impedes fluid flow through fluid restriction region 1024-A such that the hydrodynamic resistances of fluid paths 1014-A and 1018-A are altered. This causes the hydrodynamic resistance of portion 1014-A to be higher, and as a result, a greater amount of fluid flows in the direction of 1070 through fluid path portion 1018-A. Accordingly, a second droplet 1062 flowing through microfluidic channel 1002-A and passing upstream portion 1006 now bypasses fluid path portion 1014-A and flows through portion 1018-A. The second droplet, after bypassing region 1028-A, now enters microfluidic channel portion 1002-B. If there is a lower hydrodynamic resistance in fluid path portion 1014-B compared to region 1018-B (e.g., a droplet has not already been positioned in region 1028-B), the droplet can be positioned at this region. Next, a third droplet 1064 can flow through microfluidic channel portion 1002-A in the direction of arrow 1012 and first bypasses region 1028-A due to droplet 1060 already positioned at that region. The droplet can then flow into fluid path portion 1018-A and 1002-B. Since droplet 1062 has already been positioned at region 1028-B, third droplet 1064 bypasses this region and takes the fluid path of least hydrodynamic resistance (fluid path portion 1018-B). Upon entering an empty region such as region 1028-C, the third droplet can now be positioned at that region due to a lower hydrodynamic resistance in fluid path 1014-C compared to that of fluid path portion 1018-C (e.g., prior to any other droplet being positioned at region 1028-C).
Accordingly, a method for positioning droplets in regions of a microfluidic network may include providing a microfluidic network comprising at least a first inlet to a microfluidic channel (e.g., positioned upstream of portion 1006 of
It should be understood that other components may be integrated with fluidic networks described herein in some embodiments of the invention. For example, in some instances, hydrodynamic resistances of fluid paths can be changed dynamically such that the direction of fluid flow (and, therefore, positioning of droplets) can be controlled by the user. In one such embodiment, valves may be positioned at one or more of positions 1070-A, 1070-B, and 1070-C of
As described herein, in some embodiments droplets do not require stabilization (e.g., the use of surfactants or other stabilizing agents) in order to be positioned at predetermined regions within microfluidic networks described herein. This is because in some embodiments, the droplets do not significantly physically contact one another during bypass of one droplet to another. Due to the little or no physical contact between the droplets, the droplets do not have a chance to coalesce with one another. Thus, surfactants or other stabilizing agents are not required to stabilize the droplets from coalescing in some such embodiments.
In some embodiments, the absence of surfactants or other stabilizing agents causes the droplets to wet a surface of the microfluidic network. Even though wetting may occur, the droplets can still be positioned at predetermined regions within the microfluidic network due to, for example, a positive pressure that causes fluid flow to carry these droplets into these regions. As discussed above, the use of droplets and/or a carrier fluid that does not contain a surfactant is advantageous in some embodiments where surfactants may negatively interfere with contents inside the droplets. For example, the droplets may contain proteins, and surfactants are known to denature certain proteins to some extent. However, after manipulation of the droplet and/or carrying out a process such as a chemical and/or biological reaction inside the droplet, surfactants may no longer negatively affect the contents inside the droplet. Accordingly, in such cases, a surfactant or other stabilizing agent can be applied to the droplets after the droplets have been positioned at regions of the microfluidic network. In some embodiments, application of a stabilizing agent to a droplet after manipulation of the droplet and/or carrying out a process inside the droplet can facilitate mobilization of the droplet out of the region in which the droplet is positioned.
It should be understood, however, than in some embodiments, a droplet and/or a carrier fluid may contain a surfactant or other stabilizing agent that stabilizes a droplet prior to positioning of the droplet at a region in the microfluidic network. In some such embodiments, the stabilizing agent does not negatively interfere with contents (e.g., reagents) inside the droplet. Of course, such embodiments will depend on a variety of factors such as the type of stabilizing agent used, the contents inside the droplet, the application, etc.
If desired, after introducing a fluid containing a stabilizing agent to the droplet, the fluid flow may be stopped for a certain amount of time to allow the stabilizing agent to coat the droplet. In other embodiments, however, flow in channel 1002 is not stopped after the stabilizing agent has been introduced. In yet other embodiments, after a droplet has been de-wetted from a surface of the microfluidic network, fluid flowing in the microfluidic network may be replaced by a second fluid (which may or may not contain a stabilizing agent). As shown in the embodiment illustrated in
It should be understood that a fluid containing a stabilizing agent can be introduced into microfluidic network 1090 in any suitable manner. For example, in some embodiments, the stabilizing agent may be introduced by a fluid flowing in the direction of arrow 1012. In other embodiments, region 1028 may be in fluidic communication with another portion of the device extending from region 1028. For instance, above or below region 1028 may be a reservoir, a channel, or other component that can be used to introduce a stabilizing agent or other entity to a droplet in that region.
As shown in
Different types of carrier fluids can be used to carry droplets or components in a microfluidic system. Carrier fluids can be hydrophilic (e.g., aqueous) or hydrophobic (e.g., an oil), and may be chosen depending on the type of droplet being formed or positioned (e.g., aqueous or oil-based) and/or the type of process occurring in the droplet (e.g., crystallization or a chemical reaction). In some cases, a carrier fluid may comprise a fluorocarbon. In some embodiments, the carrier fluid is immiscible with the fluid in the droplet. In other embodiments, the carrier fluid is slightly miscible with the fluid in the droplet. Sometimes, a hydrophobic carrier fluid, which is immiscible with the aqueous fluid defining the droplet, is slightly water soluble. For example, oils such as PDMS and poly(trifluoropropylmethysiloxane) are slightly water soluble. These carrier fluids may be suitable, for example, when fluid communication between the droplet and another fluid is desired. Diffusion of water from a droplet, through the carrier fluid, and into a second droplet is one example of such a case.
As described above, methods for storing and/or extracting droplets in a microfluidic network are provided herein. In some embodiments, the droplets may be stored and/or extracted in sequential order. For example, the droplets may be extracted in the order they are stored or positioned in predetermined regions in the microfluidic network. In other embodiments, the use of valves can allow only certain droplets to be released from regions of the microfluidic system. Advantageously, in some embodiments, such methods do not require the use of surfactants or other stabilizing agents, since the droplets may not come into substantial physical contact with one another in a manner that causes coalescence. This is advantageous in certain cases as surfactants may interfere with contents such as proteins inside the droplet, as is known to those of ordinary skill in the art.
In another aspect of the invention, a method of forming droplets in regions of a microfluidic device is provided. The method can allow production of droplets of a precise volume, which can be stored/maintained at precise regions of the device. The drops can be created at the storage region in a self-regulated manner. The method may include, optionally, filling a microfluidic channel with a filling fluid (e.g., oil). The oil can then be flushed out with a first fluid (e.g., an aqueous fluid) to be stored/maintained at a region of the device. This first fluid may be immiscible with the filling fluid. The first fluid can enter a region of the device for storing a droplet, replacing the filling fluid in that region. Next, a second fluid (e.g., a fluid immiscible with the first fluid) may be flowed in the channel, causing partitioning of a portion of the first fluid (e.g., at least in part through action of the second fluid). Partitioning of the first fluid causes formation of a droplet of the first fluid at the region, while a second portion of the first fluid bypasses the region. In this manner, a plurality of droplets can be generated sequentially down the length of the channel.
Advantageously, the devices and methods described herein may address several problems commonly associated with forming and/or storing droplets: (1) Each droplet of the same volume may be formed (or, the droplets may have different volumes, e.g., depending on the size of the positioning regions) and all of the first fluid (e.g., aqueous phase) may be used with zero or minimal waste, (2) Because the forming and/or positioning of the droplets is done by the serial application of single phase fluids, the process is simple and tolerant to a wide range of flow rates, pressures, fluids, and materials used to form the device. (3) Valves are not required in this device, which may make it easy to manufacture (although valves may be used with the device if desired), (4) Drop generation is robust and simple (e.g., in contrast to certain flow focusing and T-junction methods), (5) The method can be used to position/store reactive components besides (or in addition to) droplets, such cells as beads for use in PCR and ELISA type bioassays.
As shown in this illustrative embodiment, fluid path 1014 has a lower hydrodynamic resistance than fluid path 1018 due to the relatively longer channel length of fluid path 1018. It should be understood, however, that the microfluidic network may have other designs and/or configurations for imparting different hydrodynamic resistances, and such designs and configurations can be determined by those of ordinary skill in the art. For instance, in some embodiments, the length, width, height, and/or shape of the fluid path can be designed to cause one fluid path to have a resistance to fluid flow different from another fluid path. In one particular embodiment, fluid path 1018 has at least one cross-sectional dimension (e.g., a width or height) that is less than a cross-sectional dimension of fluid path 1014. In other embodiments, at least a portion of a fluid path may include an obstruction such as a valve (which may change hydrodynamic resistance dynamically), a semi-permeable plug (e.g., a hydrogel), a membrane, or another structure that can impart and/or change hydrodynamic resistance in that portion.
As shown in
Fluid path 1018, now having a lower resistance to fluid flow, allows second portion 1200-B of the first fluid to continue flowing in the direction of arrows 1013 and 1015, followed by second fluid 1202. As fluid path 1014 includes fluid restriction region 1024, droplet 1200-A cannot flow further down the microfluidic network. Accordingly, droplet 1200-A is positioned within a region 1028 (e.g., a “microwell”) of the microfluidic network. In some embodiments, droplet 1200-A can be maintained at the region even though fluid continues to flow in the microfluidic network (e.g., in the direction of arrow 1012).
The size/volume of droplet 1200-A can vary and may depend and can be determined, at least in part, on the size/volume of region 1028. In some instances, droplets formed at a region have the same volume (or length) as that of the region. In other instances, droplets formed at a region have a different (e.g., a smaller or larger) volume (or length) as that of the region. The droplet may have a volume (or length) that is within, for example, 5%, 10%, 15%, 20%, 25%, or 30% of the volume (or length) of the region in which the droplet is positioned. In other cases, the volume (or length) of a droplet formed in a region is substantially smaller (e.g., less than 50%) than the volume (or length) of the region. For instance, if a region having a volume (or length) of X already contains a droplet having a volume (or length) of Y (e.g., using a method of positioning droplets as described in connection with
Various sizes/volumes of droplets that can be formed/stored are described in more detail below.
As described above, in some embodiments, the positioning/presence of droplet 1200-A at region 1028 of
Accordingly, one method of the invention comprises providing a microfluidic network comprising a first region (e.g., region 1028) and a microfluidic channel (e.g., microfluidic channel 1002) in fluid communication with the first region, flowing a first fluid (e.g., fluid 1200) in a first direction in the microfluidic channel (e.g., in the direction of arrow 1012), flowing a second fluid (e.g., fluid 1202) in the first direction in the microfluidic channel, partitioning at least a portion of the first fluid at the first region so as to form a first droplet (e.g., droplet 1200-A) of the first fluid at the first region, and maintaining the droplet at the first region while fluid is flowing in the first direction. In some instances, forming and/or maintaining of a droplet at a region is independent of flow rate of the first fluid in the microfluidic channel.
The method described in connection with
It should be understood that when droplet 1200-A is formed/positioned at region 1028, the droplet may plug all or a portion of fluid path 1014 and/or fluid restriction region 1024. For instance, in some cases, the droplet plugs all of such fluid paths such that none of the fluid flowing in micro fluidic channel 1002 passes through fluid restriction region 1024. In other embodiments, the droplet may plug only a portion of such fluid paths such that some fluid passes through fluid restriction region 1024 even though the droplet is positioned at region 1028. The amount of fluid flowing past the droplet may depend on factors such as the dimensions of fluid path portions 1014 and/or 1024, the size of the droplets, the flow rate, etc. As long as the droplet causes fluid path 1014 to have a higher relative resistance to fluid flow than fluid path 1018, a second fluid (e.g., fluid 1202) can bypass fluid path 1014 and enter fluid path 1018.
As described above, fluid paths 1014 and 1018 may have different resistances to fluid flow depending on whether or not a droplet has been formed or positioned at region 1028. In the absence of a droplet at region 1028, fluid path 1014 may be configured to have a lower resistance to fluid flow than fluid path 1018. For example, greater than 50%, greater than 60%, greater than 70%, greater than 80%, or greater than 90% of the fluid flowing in channel 1002 at upstream portion 1006 may flow in fluid path 1014 compared to fluid path 1018. However, when a droplet has been formed, positioned and/or maintained at region 1028, fluid path 1014 may have a relatively higher hydrodynamic resistance. For example, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the fluid flowing in channel 1002 at upstream portion 1006 may flow in fluid path 1014 compared to that of 1018. In some cases, 100% of the fluid flowing in direction 1012 in microfluidic channel 1002 flows in fluid path 1018 upon positioning of a droplet at region 1028.
In some embodiments, partitioning of the first fluid into a first portion at region 1028 to cause formation of droplet 1200-A of the first fluid (e.g., a first droplet), can result in the droplet “recoiling” such that it has a slightly smaller volume than that of region 1028. In some instances, this recoiling causes certain subsequent fluids flowing in the microfluidic channel (e.g., a third fluid that is miscible with the first fluid) to not come into contact with the droplet stored at region 1028. The third fluid may be in the form of, for example, a fluid stream or a droplet. If the third fluid is in the form of a second droplet, the second droplet may not come into contact with the first droplet. Accordingly, in certain embodiments, a third fluid (e.g., second droplet or a fluid stream or portion of a fluid stream) does not physically contact the first droplet after forming and/or positioning of a first droplet in region 1028.
In other embodiments, a third fluid (e.g., second droplet or a fluid stream or portion of a fluid stream) comes into physical contact with the first droplet as it bypasses the first droplet, however, due to such minimal contact between the two fluids, the fluids do not coalesce. Accordingly, in some instances, the forming and/or positioning of droplets in the microfluidic network can take place without the use of surfactants. In other words, surfactants in either a fluid flowing in channel 1002 and/or within the droplets is not required in order to stabilize the droplets and/or prevent the droplets or fluids from coalescing with one another during forming, positioning or carrying the droplet in the microfluidic channel, and/or during maintaining the droplets within a predetermined region within the microfluidic network. However, in instances where coalescence is desired (e.g., to allow a reaction between reagents contained in two droplets), the microfluidic network and methods for operating the network can be configured to allow such physical contact and/or coalescence between droplets.
In some embodiments, methods for positioning a droplet in a microfluidic network include the steps of providing a microfluidic network comprising a first region (e.g., region 1028 of
Another method of the invention comprises providing a microfluidic network comprising at least a first inlet to a microfluidic channel (e.g., positioned upstream of portion 1006 of
In some embodiments, a chemical and/or biological process and/or a manipulation process can be carried out in a droplet that is positioned at a region of a microfluidic network (e.g., droplet 1200-A of
Microfluidic network 1000 of
In some embodiments, a fluid of defined volume (e.g., a “slug”) can be introduced into microfluidic networks described herein and portions of the fluid can be partitioned into regions of the fluidic network. For example, a slug having a volume of X may be flowed into a network having a plurality of sections 1001 (and regions 1028, having a volume of Y, where Y is less than X) and the slug can be partitioned at each region 1028. The first region may be filled with the fluid in the amount of volume approximately Y (and forming a droplet of the fluid having a volume of approximately Y at the first region), while the remainder of the slug (X-Y or X minus Y) continues to flow down the network. A second region can then be filled with the fluid in the amount of approximately Y (and forming a droplet of the fluid having a volume of approximately Y at the second region), while the remainder of the slug (X-2Y) continues to flow down the network. This process can continue until the slug has a volume of zero. This method allows all of the fluid of the slug, e.g., a sample fluid, to be used in the microfluidic network with no or minimal waste.
As shown in
In another aspect of the invention, microfluidic structures and methods described herein are designed for containing and positioning components (e.g., beads, cells, and other reactive or non-reactive components) in an arrangement such that the components can be grown, multiplied and/or manipulated and then tracked even after one or more of these processes has taken place.
As shown in the embodiment illustrated in
Accordingly, a method of the invention comprises providing a microfluidic network comprising at least a first inlet to a microfluidic channel, a first and a second region for positioning a first and a second reactive component, respectively, the first and second regions in fluid communication with the microfluidic channel, wherein the first region is closer in distance to the first inlet than the second region, and flowing a first fluid comprising first and second reactive components in the microfluidic channel. The method may also include positioning the first reactive component at the first region, positioning the second reactive component at the second region, and maintaining the first and second reactive components in the first and second regions, respectively, while a fluid is flowing in the microfluidic channel.
As shown in the embodiment illustrated in
Next, a third fluid 1260 may be flowed in channel 1002 in the direction of arrow 1012, e.g., to introduce yet another reactive component into the microfluidic network. The third fluid may enter both fluid path 1014 (including region 1028) and fluid path 1018. This third fluid may be miscible with the first and/or second fluids and may contain a reactive species 1236 (e.g., an enzyme-linked anti-human antibody). In some cases, reactive species 1236 interacts (e.g., reacts, associates, or binds) with either reactive component 1230 or with reactive species 1234, e.g., as shown in
Optionally, a fourth fluid 1270 (e.g., an oil or a gel) immiscible with the third fluid may be flowed into channel 1002 in the direction of arrow 1012. The fourth fluid may enter region 1028 and, using the methods described above in connection with
In some embodiments, the positioning or presence of a reactive component at region 1028 causes at least a portion of fluid path 1014 to be plugged such it has a higher hydrodynamic resistance, resulting in less fluid flowing past fluid restriction region 1024 (relative to the absence of the component). This may cause fluid path 1014 to have a higher hydrodynamic resistance than fluid path 1018. As a result, when subsequent fluids (including additional reactive components) are flowed in the direction of arrow 1012, at least a portion of the subsequent fluids (and reactive components) may bypass flow path 1014 and enter flow path 1018, which now has a lower hydrodynamic resistance than fluid path 1014. Accordingly, once region 1028 has been “filled” with a reactive component, other regions downstream can be filled with reactive components. This allows a plurality of reactive component to be generated/stored sequentially down the length of the channel.
Reactive components may interact with reactive species by various methods. For example, interaction may be between a corresponding pair of components that exhibit mutual affinity or binding capacity, typically specific or non-specific binding or interaction, including biochemical, physiological, and/or pharmaceutical interactions. Biological binding defines a type of interaction that occurs between pairs of components including proteins, (e.g., adhesion proteins), nucleic acids, glycoproteins, carbohydrates, hormones and the like. Specific examples include cell (e.g., cell surface receptor)/protein, adhesion protein/integrin, antibody/antigen, antibody/hapten, enzyme/substrate, enzyme/inhibitor, enzyme/cofactor, binding protein/substrate, carrier protein/substrate, lectin/carbohydrate, receptor/hormone, receptor/effector, complementary strands of nucleic acid, protein/nucleic acid repressor/inducer, ligand/cell surface receptor, virus/ligand, etc.
By carrying out the methods described above in connection with
In some embodiments, reactive components such as cells can be encapsulated in a droplet (e.g., as part of a water-in-oil emulsions). The droplet may contain one or more reactants, nutrients, or drugs. The droplets may act as, for example, picoliter-scale reaction microvessels. Encapsulation can enable reactions involving, for instance, single molecules, such as in vitro translation of proteins from single genes, or DNA amplification of single genes by polymerase chain reaction (PCR). Droplets also provide an excellent method for studies of single, isolated cells, compounds they secrete, and/or growth rates of single cells.
A method involving the use of droplets as microvessels may require incubation and imaging of droplets. Certain existing methods allow collection and imaging of droplets, however, it is difficult to maintain the identity of the droplets. Using the articles and methods described herein, these challenges can be overcome in certain embodiments since each droplet may be associated with a particular region of the microfluidic device.
In one particular embodiment, cells can be encapsulated in a gel by using methods and articles described herein. By encapsulating cells in a hydrogel functionalized with the required reactants, heterogeneous assays can be performed in a three-dimensional scaffold. Placing hydrogels in a microfluidic network such as the one shown in
In some embodiments, the microfluidic systems and methods described above and below can be used for containing and positioning cells, which can be grown, multiplied and/or manipulated in the microfluidic system. The cells (and their progeny) can be tracked even after one or more of these processes has taken place.
Genetically identical cells show phenotypic variation, for example during differentiation, development, and in response to environmental stress. The mechanisms of phenotypic variation are not fully understood, yet they are central to our understanding of major questions in biology such as differentiation and ageing. Gene expression patterns can be passed on to progeny cells, however, the mechanisms of non-genetic inheritance are not fully understood. To study phenotypic switching and epigenetic mechanisms of gene regulation, the systems and methods described herein can be used to study lineages of single cells and their progeny. For example, in some embodiments, single yeast cells can be trapped in long, thin chambers of a microfluidic device and the cells can be cultured so they grow in a line. This configuration facilitates the following of lineages deriving from a single progenitor cell. Cells can grow and divide in the chambers with typical doubling times, e.g., under constant flow conditions. As described in more detail below, generations of yeast cells can be tracked. The frequency of phenotypic switching of, for example, the GFP (green fluorescent protein)-fusion protein and PH084-GFP, a high affinity phosphate transporter, can also be studied using the methods and devices described herein.
The methods can be applied to yeast cells, and more broadly to any suspension cell including blood cells, stem cells, and other mammalian cells that grow in suspension. This methods are also useful for staining single cells and their lineages: microcolonies deriving from single cells are cultured in the chambers, and then stained. Quantitative imaging can be applied to study correlations between stained entities (protein amount or localization, chromosome position, etc) and single cell genealogy or replicative age. Moreover, the antibodies and DNA probes required for immunofluorescence and fluorescence in situ hybridization (FISH) are expensive; performing these assays in microfluidic devices described herein reduces the volume and thus cost of the required reagents. The methods and devices described herein can also enable imaging suspension cells over time.
At a downstream portion of chamber 1320 is positioned a fluid restriction region 1328, which, in some embodiments, may be in the form of a narrow channel. This fluid restriction region may allow certain components and/or fluids to pass therethrough, while inhibiting other components and/or fluids from passing therethrough, such that certain components and/or fluids are retained in chamber 1320. In some cases, once one or more components are positioned in a chamber, the position of the component(s) can be maintained in the chamber even during subsequent fluid flow in the chamber. This may occur, in some cases, if the component has an average cross-sectional area that is larger than a cross-sectional area of fluid restriction region 1328.
A fluid restriction region may be positioned at an upstream portion, a downstream portion, or in between an upstream and a downstream portion of the chamber. In some cases, a fluid restriction region is positioned between a chamber inlet and a chamber outlet. In other cases, a fluid restriction region is positioned outside of the chamber. In one embodiment, a fluid restriction region is positioned immediately adjacent a chamber outlet.
It should be understood that any suitable structure can be used as fluid restriction region 1328, which may have a higher hydrodynamic resistance and/or a smaller cross-sectional area for fluid flow than a region immediately upstream or downstream of the fluid restriction region. For instance, fluid restriction region 1328 may be in the form of a narrow fluid path or a channel having the same dimensions as chamber 1320, but having an obstruction (e.g., posts or a valve) positioned in or at that region. In other embodiments, fluid restriction region 1328 may comprise a porous membrane, a semi-permeable plug (e.g., a gel), a valve, or another structure. In some cases, a fluid restriction region is a narrow channel portion that does not include a gel or other structure disposed therein; e.g., the size of the opening of the fluid restriction region alone can be used to trap, immobilize, and/or position components in a chamber fluidly connected to the fluid restriction region.
As shown in the illustrative embodiment of
Also included in chambers 1308 and 1309 are chamber bypass channels 1330, which extend from a portion of chamber 1320. One or more chamber bypass channels may extend from an upstream and/or downstream portion of the chamber. In some cases, a chamber bypass channel extends from the chamber between a chamber inlet and a chamber outlet (e.g., as illustrated in
In some embodiments, a chamber bypass channel has a lower hydrodynamic resistance than a chamber (e.g., prior to and/or after a component has been positioned in the chamber). As described in more detail below, this can allow more fluids and/or components to flow into the chamber bypass channel than the chamber. This arrangement may be useful for applications where it is desirable to position only one, or perhaps a few, components in the chamber. In certain embodiments where it is desirable to flow fluids and/or components out of the chamber bypass channel, the chamber bypass channel does not include any fluid restriction regions. This configuration may prevent any fluids and/or components from being immobilized in the chamber bypass channel. In other embodiments, however, a chamber bypass channel may include one or more fluid restriction regions.
As shown in the exemplary embodiment of
A microfluidic device may include, for example, at least 10, at least 25, at least 50, at least 75, at least 100, at least 150, at least 200, at least 500, or at least 1,000 chambers or chamber units, which may be positioned in series and/or in parallel. The number of chambers or chamber units, in some cases, may depend on and/or may be limited by the size of the device, the size of the chamber units, and/or the application. Optionally, one or more chambers or chamber units may be labeled, e.g., using labels 1372, to identify the chambers or chamber units. Accordingly, any suitable number of chambers or chamber units may be included in a device. In addition, a device may include one or more microfluidic systems 1300 which can allow, for example, experiments under conditions or parameters to be performed simultaneously.
Microfluidic system 1300 may also include one or more device inlets (inlets 1346 and 1348) for introducing one or more fluids into the microfluidic system. As shown in
Inlet 1346 may be connected to two channel portions 1362 and 1364, and inlet 1348 may be connected to channel portion 1360. As illustrated, channel portions 1360, 1362, and 1364 may be arranged in a flow focusing arrangement. This arrangement can facilitate focusing of a component within channel portion 1360 towards a center portion of a fluid stream during laminar flow, e.g., when the channel portions intersect at intersection 1366 to form laminar streams of fluid. The fluid can then be introduced into microfluidic channel 1340 via inlet 1368. Other configurations of inlets are also possible.
Fluids flowing in channel portions 1362 and 1364 may be immiscible with a fluid flowing in channel portion 1360, which may allow formation of droplets of the fluid flowing in channel portion 1360 at or near inlet 1368 in some embodiments. In other embodiments, the fluids in channel portions 1360, 1362 and 1364 are miscible.
As described herein, a microfluidic system may be used to position one or more components in one or a series of regions (e.g., “microwells” or “chambers”). A variety of different components can be positioned at a region, including, for example, droplets, cells (e.g., bacterial, yeast, mammalian and stem cells), beads, tissues, and other entities. The number of components positioned at a region may depend on the size of the region. In some cases, a region (or a component) may have a dimension, such as a length, width, and/or height, of less than or equal to about 500 μm, less than or equal to about 250 μm, less than or equal to about 100 μm, less than or equal to about 50 μm, less than or equal to about 25 μm, less than or equal to about 10 μm, less than or equal to about 8 μm, or less than or equal to about 1 μm. The volume of the region (or component) can also vary; for example, the region (or component) may have a volume of less than or equal to about 50 μL, less than or equal to about 10 μL, less than or equal to about 1 μl, less than or equal to about 100 nL, less than or equal to about 10 nL, less than or equal to about 1 nL, less than or equal to about 100 pL, or less than or equal to about 10 pL. In certain embodiments, a region (or a component) may have a length of at least 1 μm, at least 10 μm, at least 25 μm, at least 50 μm, at least 100 μm, at least 200 μm, at least 250 μm, at least 500 μm, or at least 1,000 μm. Long regions may be suitable for positioning a large number of components at the region.
To position components in chamber of microfluidic system 1300, a first fluid stream including a plurality of components (e.g., cells) may be introduced into inlet 1348. Agglomeration of cells may be captured by filter 1350, e.g., a size exclusion filter, while allowing un-agglomerated cells to pass therethrough. A second fluid (e.g., a buffer solution) may be introduced into inlet 1346, and this fluid can flow into channel portions 1362 and 1364. The first fluid including the components may be focused as they pass through inlet 1368 into feed channel 1340. Since feed channel 1340 fluidly connects to a series of chamber inlets, the feed channel can distribute the fluids and the components to various chambers.
The geometry of the channels of the microfluidic system, the flow rate of the first and second fluids, and the concentration (or density) of the components in the first fluid can be chosen so as to allow delivery of a certain number of components into each of the chambers. For example, these parameters can be chosen such that at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the chambers contain a predetermined number of components positioned therein. For instance, in one embodiment, only a single component is delivered to at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the chambers of a microfluidic system. This can be achieved, in part, by designing the chambers to have a cross-sectional area on the order of the cross-sectional area of the component to be positioned. In addition, fluid restriction region 1328 may be designed such that introduction of a single component into the chamber channel substantially blocks or reduces fluid flow through this region. (In other embodiments, the fluid restriction region may be designed such that the introduction of exactly two components, or exactly three components, etc., causes substantial blockage of fluid flow through the fluid restriction region.) This blockage or reduction in fluid flow can result in the chamber being “filled”, causing the chamber to have a higher hydrodynamic resistance than an empty chamber. Thus, fluid and components flowing in channel 1340 may prefer to flow into empty chambers (and/or through a chamber bypass channel) having a lower hydrodynamic resistance. The flow rate and the concentration (or density) of components in the fluids can also be adjusted to control the number of components in each chamber. This control can allow, for example, multiple experiments to be performed in parallel under similar or the exact same conditions.
Accordingly, a method may include flowing a fluid containing a plurality of components in a microfluidic system comprising a chamber having a flow direction, a chamber inlet, a chamber outlet, and a chamber bypass channel extending from the chamber between the chamber inlet and the chamber outlet. The method may also include positioning a component in the chamber, the chamber having a cross-sectional area, perpendicular to the flow direction, less than 2 times the largest cross-sectional area of the component perpendicular to the flow direction. A fluid may be flowed through the chamber while maintaining the component at its position in the chamber, and a portion of the plurality of components may be flowed in the chamber bypass channel.
As described herein, the dimensions of the chamber and fluid restriction region 1328 may be chosen such that a component, or a series of components, can be positioned, maintained, or trapped in the chamber channel without exiting the chamber via fluid restriction region 1328. In some cases, this positioning, maintaining, or trapping take place even while fluid continues to flow through fluid restriction region 1328. The fluid may be flowed at a substantially constant flow rate, at varying flow rates, or flow may take place periodically. This fluid flow may be important for applications that involve treating or manipulating the component(s) after it has been positioned in the chamber channel. For example, a continuous or periodic flow of nutrients may be supplied to cells that are trapped in a chamber channel. These nutrients can facilitate the growth and/or multiplication of the cells. Flow of dyes can allow the cells to be tagged and identified. In other embodiments, components that are positioned in a chamber can be subjected to a stress (e.g., different buffers, nutrients, or exposure to drugs) and the response of different components to the stress can be determined. Other applications are described in more detail below.
Flow of fluid (e.g., media) through the chambers is important for cell culture and also allows the cells to be exposed to controlled environmental changes, as well as immunostaining. Immunofluorescence of suspension cells is generally challenging. For yeast, the cell wall is typically enzymatically removed, and the cells are attached to a substrate using a chemical crosslinker. The cells are then fixed and permeabilized. Other suspension cells are typically centrifuged onto a surface-treated, glass microscope slide before staining. Moreover, immunostaining techniques for suspension cells are typically not compatible with live cell imaging. Using the methods and systems described herein, however, live cells can be positioned and tracked controllably in regions of a microfluidic device, and environmental conditions can be varied, for example, by changing the fluids flowed in the region.
Furthermore, in some embodiments, the dimensions of the chamber can be chosen such that components in the chamber are positioned in a predetermined configuration. For example, the dimensions of the chamber may be chosen such that a series of components are generally aligned in the chamber. As used herein, “generally aligned” means no greater than 20% of the components in a chamber have the same position along the direction of bulk fluid flow in the chamber. The position of the components along the direction of bulk fluid flow can be determined by taking the cross-section of the component, perpendicular to the direction of (bulk) fluid flow in the chamber, which cross-section passes through the center of the component between two opposed points of a surface/surfaces of the component. For example, as shown in the illustrative embodiment of
In some cases, no greater than 15%, 10%, 5%, or 2% of the components in a chamber have the same position along the direction of bulk fluid flow in the chamber. For example, sometimes 100% of the components in a chamber have different positions along the direction of bulk fluid flow in the chamber. When no greater than 5% of the components in a chamber have the same position along the direction of bulk fluid flow in the chamber, the components may be aligned “in a single line” in the chamber, as shown in the embodiment illustrated in
Aligned or generally aligned components can assist in identification of the components in a chamber. For example, if the components are cells derived from one cell, this configuration may be useful for tracking the lineology of the cells. In order to achieve the positioning of aligned or generally aligned components, the chamber may have a cross-sectional area perpendicular to the (bulk) flow direction that is, for example, less than 2 times the largest cross-sectional area of the component perpendicular to the (bulk) flow direction. In some cases, the chamber has a cross-sectional area, perpendicular to the (bulk) flow direction, that is less than 10 times, 7 times, 5 times, or 3 times the cross-sectional area of a component perpendicular to the (bulk) flow direction.
In some embodiments, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the components in a chamber have a largest cross-sectional area, perpendicular to the flow direction, of between 0.1 and 1.0 times (or between 0.3 and 1.0 times, or between 0.5 and 1.0 times) the cross-sectional area of the chamber perpendicular to the flow direction. It should be understood that the cross-sectional area may be static or dynamic, e.g., depending on the materials used to form the chamber. For example, a chamber may comprise an elastomer that allows it to expand upon having a high pressure in the chamber, or a large component positioned in the chamber. If a component completely blocks a portion of the chamber, even causing the chamber to expand, the largest cross-sectional area of the component perpendicular to the flow direction may be the same as (e.g., 1.0 times) the cross-sectional area of the chamber perpendicular to the flow direction.
In one particular embodiment, a system comprises a microfluidic device comprising an inlet, an outlet, a chamber having a (bulk) flow direction, and a flow restriction region fluidly connected to the outlet of the chamber. A plurality of cells may be generally aligned in the chamber, wherein at least 80% of the cells have a largest cross-sectional area, perpendicular to the flow direction, of between 0.1 and 1.0 times the cross-sectional area of the chamber perpendicular to the flow direction. In other embodiments, these dimensions are between 0.3 and 1.0 times, or between 0.5 and 1.0 times the cross-sectional area of the chamber perpendicular to the flow direction. The flow restriction region may be constructed and arranged to allow a fluid but not the cells to pass therethrough, thereby allowing the cells to be maintained in their positions in the chamber.
In certain embodiments, the chamber has a cross-sectional area having a different shape than a cross-sectional area of the component. For instance, the component may be round and the cross-sectional area of the chamber channel may be square. This shape difference can allow the components to be trapped in the chamber, but may allow fluids to pass the component at the corners of the chamber. Thus, a component may have a largest cross-sectional area, perpendicular to the flow direction, of less than 1.0 times, less than 0.8 times, or less than 0.5 times the cross-sectional area of the chamber perpendicular to the flow direction.
A cross-sectional area (or an average cross-sectional area) of a chamber (or a component) perpendicular to the (bulk) flow direction may be, for example, about 10,000 μm2 or less, about 5,000 μm2 or less, about 2,500 μm2 or less, about 1,000 μm2 or less, about 500 μm2 or less, about 250 μm2 or less, about 100 μm2 or less, about 50 μm2 or less, about 30 μm2 or less, about 20 μm2 or less, about 10 μm2 or less, or about 5 μm2 or less. As mentioned above, the cross-sectional area of a chamber may be chosen depending on factors such as the size of the components at or to be flowed in the chamber. The ratio of average cross-sectional areas of the chamber and the components (both measured perpendicular to the (bulk) flow direction) may be, for example, less than 20:1, less than 10:1, less than 5:1, less than 3:1, less than 2:1, or less than 1.5:1. Additionally or alternatively, the cross-sectional area of a chamber may be chosen depending on factors such as the cross-sectional area of a fluid restriction region, the number and type of components, and/or the fluids to be flowed in the system. The ratio of average cross-sectional areas of the chamber and a fluid restriction region (both measured perpendicular to the (bulk) flow direction) may be, for example, greater than 1:1, greater than 2:1, greater than 5:1, greater than 10:1, or greater than 25:1. In some cases, the ratio is between 3:1 and 8:1.
A cross-sectional area (or an average cross-sectional area) of a fluid restriction region (or the average cross-sectional area of the combined fluid paths in the case of a porous membrane or other structure having a plurality of fluid paths) may be, for example, about 5,000 μm2 or less, about 2,500 μm2 or less, about 1,000 μm2 or less, about 500 μm2 or less, about 250 μm2 or less, about 100 μm2 or less, about 50 μm2 or less, about 30 μm2 or less, about 20 μm2 or less, about 10 μm2 or less, or about 5 μm2 or less, or about 1 μm2 or less. In some cases, the cross-sectional area of a fluid restriction region may be chosen depending on factors such as the cross-sectional area of the chamber, the number, type and size of components, and the fluids to be flowed in the system.
In some embodiments, cells are positioned in a chamber and are allowed to multiply. Further analyses may involve comparing a characteristic of a first cell from a first generation to a characteristic of a second cell from a second generation. Additionally or alternatively, the cells can be subjected to a stress and/or a condition, and the response of one or more cells to the stress or condition can be determined. For example, comparative analyses may be performed by determining the response of at least two cells to a stress under controlled conditions.
Understanding how populations of single cells respond to environmental changes provides critical insights into variability in biological response, from differentiation to multi-drug resistance. Imaging populations of single cells is challenging for cells that are not adherent. Suspension cells can be trapped in wells, however, it may be difficult to image non-adherent cells under changing environmental conditions. Budding yeast cells can adhere to treated surfaces (e.g. concanavalin A or agar pads), however, newly budded yeast cells may not adhere under flow conditions. Moreover, yeast cells bud in multiple dimensions (e.g., different focal planes), challenging the analysis of populations of cells over time. Using the devices described herein, individual cells can be placed in an array, cultured, imaged in a single focal plane, and can be subjected to changing conditions. Furthermore, the microfluidic systems described herein can be fabricated on a microscope slide, which can facilitate imaging and viewing of the systems. Accordingly, multiple flow experiments can be performed in parallel for the simultaneous study and/or comparison of different flow conditions or cell types.
It should be understood that a variety of different cells can be used in devices described herein. Non-limiting examples include yeast cells, bacteria, stem cells, and suspension cells. In some embodiments, the cells are round while the chambers are square, allowing for flow past the cells. Other geometries of chambers can also be used.
Certain existing methods for time-lapse microscopy of budding yeast under flow require that the cells are wedged into a chamber much smaller than the height of the cells themselves, so that they do not move during flow; however in such chambers, the cells are subjected to mechanical stress, which may affect the observed response. In addition, certain other existing methods allow cells to grow in a 2D plane, but the cells are randomly distributed in chambers and cannot be subjected to controlled flow conditions, making it difficult to obtain data on many single cell lineages in a single experiment as microcolonies interdigitate as they grow. Using the devices described herein can allow for fixing and staining of populations of single cells without imposing mechanical stress on the cells, while allowing precise positioning and control of the numbers of cells, and manipulation of cells, in chambers.
As shown in the embodiments illustrated in
As shown in the embodiment illustrated in
As described herein, cells (or other components) can be labeled with an identifier (e.g., a dye, a probe, etc.) that identifies a particular portion of the cell and/or a particular process occurring in the cell. For example, cells labeled with GFP may show a certain level of gene expression within the cell. By combining this identification technique with methods described herein (e.g., the multiplication and tracking of cells in a chamber), gene expression of a lineage of cells can be tracked as a function of a cell's replicative age or history. In addition, these techniques may be useful for studying phenomena such as the age effects of DNA repair, how cells of different age respond to stress (e.g., exposure to different growth conditions, to a drug, and/or to a change in temperature (e.g., a heat shock), the stress of being in a densely packed environment, etc.), how gene expression levels depend on the family history of cells, and frequency of cell phenotype switching events. In some cases, these and other phenomena can be studied as a function of a cell's replicative age or history using the systems and methods described herein.
In some cases, certain strains of the GFP-fusion collection showed a bimodal distribution of phenotypes (“ON” vs. “OFF”) after culture from the harsh climate of a freezer. Despite being genetically identical, these cells switch phenotype, and their progeny inherit this phenotype; this provides a model system to study epigenetic mechanisms in gene regulation. The systems and methods described herein can be used to identify colonies and/or conditions in which cells switch. This information may be relevant to the understanding of gene regulation in yeast cells, and more broadly, to the understanding of epigenetic timescales in adaptation and differentiation.
In other experiments, yeast cells were exposed to different concentrations of phosphate. High concentrations of phosphate caused the cells to turn “OFF”, e.g., the cells having low levels of gene expression. Low concentrations of phosphate caused the cells to turn “ON”, e.g., the cells having high levels of gene expression. Intermediate concentrations of phosphate caused the cells to enter into a bistable system where some progeny turned “ON”, and some turned “OFF”, the different levels switching every few generations.
In some embodiments, regions of a fluidic network such as microchannels, microwells, and/or chambers are defined by voids in the structure. A structure can be fabricated of any material suitable for forming a fluidic network. Non-limiting examples of materials include polymers (e.g., polystyrene, polycarbonate, PDMS), glass, and silicon. Those of ordinary skill in the art can readily select a suitable material based upon e.g., its rigidity, its inertness to (i.e., freedom from degradation by) a fluid to be passed through it, its robustness at a temperature at which a particular device is to be used, its hydrophobicity/hydrophilicity, and/or its transparency/opacity to light (i.e., in the ultraviolet and visible regions).
In some instances, a device is comprised of a combination of two or more materials, such as the ones listed above. For instance, the channels of the device may be formed in a first material (e.g., PDMS), and a substrate can be formed in a second material (e.g., glass).
Fluid channels and/or components described herein may have maximum cross-sectional dimensions less than 2 mm, and in some cases, less than 1 mm. In one set of embodiments, all fluid channels containing embodiments of the invention are microfluidic or have a largest cross-sectional dimension of no more than 2 mm or 1 mm. In another embodiment, the fluid channels may be formed in part by a single component (e.g., an etched substrate or molded unit). Of course, larger channels, tubes, chambers, reservoirs, etc. can be used to store fluids in bulk and to deliver fluids to components of the invention. In one set of embodiments, the maximum cross-sectional dimension of the channel(s) or regions described herein are less than about 500 microns, less than about 200 microns, less than about 100 microns, less than about 50 microns, or less than about 25 microns. In some cases the dimensions of the channel may be chosen such that fluid is able to freely flow through the article or substrate. The dimensions of the channel may also be chosen, for example, to allow a certain volumetric or linear flowrate of fluid in the channel. Of course, the number of channels and the shape of the channels can be varied by any method known to those of ordinary skill in the art. In some cases, more than one channel or capillary may be used. For example, two or more channels may be used, where they are positioned inside each other, positioned adjacent to each other, positioned to intersect with each other, etc.
A “channel,” as used herein, means a feature on or in an article (substrate) that at least partially directs the flow of a fluid. The channel can have any cross-sectional shape (circular, oval, triangular, irregular, square or rectangular, or the like) and can be covered or uncovered. In embodiments where it is completely covered, at least one portion of the channel can have a cross-section that is completely enclosed, or the entire channel may be completely enclosed along its entire length with the exception of its inlet(s) and outlet(s). A channel may also have an aspect ratio (length to average cross sectional dimension) of at least 2:1, more typically at least 3:1, 5:1, or 10:1 or more. An open channel generally will include characteristics that facilitate control over fluid transport, e.g., structural characteristics (an elongated indentation) and/or physical or chemical characteristics (hydrophobicity vs. hydrophilicity) or other characteristics that can exert a force (e.g., a containing force) on a fluid. The fluid within the channel may partially or completely fill the channel. In some cases where an open channel is used, the fluid may be held within the channel, for example, using surface tension (i.e., a concave or convex meniscus).
The channels of the device may be hydrophilic or hydrophobic in order to minimize the surface free energy at the interface between a material that flows within the channel and the walls of the channel. For instance, if the formation of aqueous droplets in an oil is desired, the walls of the channel can be made hydrophobic. If the formation of oil droplets in an aqueous fluid is desired, the walls of the channels can be made hydrophilic.
In some cases, the device is fabricated using rapid prototyping and soft lithography. For example, a high resolution laser printer may be used to generate a mask from a CAD file that represents the channels that make up the fluidic network. The mask may be a transparency that may be contacted with a photoresist, for example, SU-8 photoresist, to produce a negative master of the photoresist on a silicon wafer. A positive replica of PDMS may be made by molding the PDMS against the master, a technique known to those skilled in the art. To complete the fluidic network, a flat substrate, e.g., a glass slide, silicon wafer, or a polystyrene surface, may be placed against the PDMS surface and plasma bonded together, or may be fixed to the PDMS using an adhesive. To allow for the introduction and receiving of fluids to and from the network, holes (for example 1 millimeter in diameter) may be formed in the PDMS by using an appropriately sized needle. To allow the fluidic network to communicate with a fluid source, tubing, for example of polyethylene, may be sealed in communication with the holes to form a fluidic connection. To prevent leakage, the connection may be sealed with a sealant or adhesive such as epoxy glue.
In some embodiments, the microfluidic networks described herein can be combined with one or more microfluidic components such as valves, pumps, droplet formation regions (e.g., in the form of a flow-focusing device), membranes, as well as those described in U.S. application Ser. No. 60/925,357, filed Apr. 19, 2007, and entitled “Manipulation of Fluids and Reactions in Microfluidic Systems”, which is incorporated herein by reference in its entirety for all purposes. Any of a number of valves and/or pumps, including peristaltic valves and/or pumps, suitable for use in a fluidic network such as that described herein can be selected by those of ordinary skill in the art including, but not limited to, those described in U.S. Pat. No. 6,767,194, “Valves and Pumps for Microfluidic Systems and Methods for Making Microfluidic Systems”, and U.S. Pat. No. 6,793,753, “Method of Making a Microfabricated Elastomeric Valve,” which are incorporated herein by reference.
The following examples are intended to illustrate certain embodiments of the present invention, but are not to be construed as limiting and do not exemplify the full scope of the invention.
This example shows the design, fabrication, and operation of a microfluidic system for positioning, trapping and storing single cells in chambers according to one embodiment of the invention.
An array of chambers was designed to position, trap and store cells, the chambers having a configuration such as the ones shown in
In this example, the chambers were designed so they were just as wide as a single cell, with dimensions 5 μm high, 5 μm wide, and 400 μm long. Because of this restriction in height and width of the chamber, the cells were constrained to grow in a single line. While the cross-sections of the cells were approximately round, the chambers were approximately square, enabling media to be continually perfused through the chambers even while the cells were positioned in the chamber. The volume of a single chamber was 10,000 μm3 and there was approximately 3,500 μm3 free volume when the chamber was filled with cells. For longer-term culture applications, a porous membrane can be incorporated into the device (e.g., a porous may form one surface of the chamber), which may allow for additional media exchange along the length of the chamber. Examples of semi-permeable membranes are described in more detail in International Patent Apl. Serial No. PCT/US2006/034659, filed Sep. 7, 2006, entitled “Microfluidic Manipulation of Fluids and Reactions”, which is incorporated herein by reference in its entirety for all purposes.
Before the cells reached the chambers, they were passed through a filter that was fabricated on-chip for both media and cell inlets. The smallest pore size of the filter was 5 μm, so aggregates of cells remained trapped in the filter. This filter helped to achieve the flow of only single cells (instead of agglomerations of cells) into the chamber array.
Over the time course of the experiments, cells trapped in the filter continued to grow. To avoid dislodged budding cells from the filter and contaminating cell lineages in the chamber array, a flow-focusing junction was included in the device design. During loading, media flowed in from the side channels (e.g., channels portions 1362 and 1364 of
Typical loading efficiencies of single cells in the chambers ranged from about 50-90%, e.g., depending on the flow conditions and the cell density. The majority of the remaining chambers were empty, and in a few instances cases, contained multiple cells. The loading efficiencies can be optimized by varying one or both of the flow rate and cell density. For example, in some cases, a loading flow rate of 95 μL/hr with approximately 107 cells/mL in the loading solution filled at least half of the chambers with a single cell after approximately 5 minutes of loading. In other cases, a concentration of approximately 106 cells/mL was used. Depending on the flow rate and other conditions, an increase of the cell concentration or the loading time may lead to the positioning of multiple cells per chamber.
In chambers of these dimensions used in this experiment, over 100 cells derived from a single cell were captured in each chamber. Chambers may also be made longer to follow more generations. Over the course of 1000-1500 minutes, a relatively constant division time was observed, even for cells positioned adjacent the flow constriction region. This may suggest that the division of cells was not limited by the number of cells in the chamber (or by the dimensions of the chamber).
Once the cells are loaded in the chambers, a buffer was flowed through the chambers at rates of 55 μL/hr. Based on the volume of the device, the media was exchanged on timescales much less than the division time of the cells.
To evaluate growth of cells in the progeny chambers, cells were tracked using time-lapse microscopy. Images were acquired every 7 minutes. The average cell division time was comparable to bulk growth rates.
The chambers facilitated fast and efficient qualitative screening of single cells and their progeny. At the end of an experiment, images of the lines of cells were acquired. The number of single cells in “ON” and “OFF” states was easily determined by simple analysis of the proportion of light to dark channels. The chambers also made it easy to investigate switching frequency for single cells.
For more detailed analysis of cells and their progeny, time-lapse imaging and analysis was performed. It was observed that cells and their closest progeny remained relatively close to each other within the channel, however, recently budded cells sometimes squeezed past each other in the chamber and disrupted the genealogical order of the cells. Furthermore, while the haploid cells typically budded from the same end, they occasionally reoriented in the channels and budded from the other side. The cells and their lineology were tracked manually. However, the cells may also be tracked automatically, e.g., using software that can identify individual cells (e.g., based on differences in fluorescence intensity).
The chambers allowed for growth of cells to densities higher than that which can normally be obtained in traditional laboratory conditions, for example, up to 1×1010 cells/mL. This may facilitate studies of high density cell cultures, such as those found in nature.
With slight modifications in size, the progeny chambers may also be used for the culture of other suspension cells, such as mammalian blood cells or stem cells. By treating the channels with appropriate surface coating(s) (e.g., fibronectin), adherent cells may also be grown in the chambers.
Device fabrication. The microfluidic device used in this example was fabricated as follows. The designs for the chambers were generated in AutoCad. Chrome masks were printed on quartz (HTA Photomask, CA) and soft lithography was used to create polydimethylsiloxane (PDMS) devices. In brief, a SU8 2005 photoresist (MicroChem, Newton, Mass.) was spin-coated onto a silicon wafer (rinsed with methanol and prebaked for 10 minutes at 210° C.) to a final thickness of 5 μm following the protocol described by the manufacturer. A mask was placed on top of the wafer and exposed to UV light for 12 seconds. Exposure of the photoresist to UV light (OAL San Jose, Calif.) crosslinked the exposed pattern, and the non-exposed photoresist was dissolved away using propylene glycol monomethyl ether acetate (PGMEA). The channel height was confirmed to be 5 μm using a scanning profilometer (Stylus). PDMS was mixed with a crosslinker at a ratio of 10:1, and poured onto the master (Sylgard 184 Silicone Elastomer, Dow Coming, Midland, Mich.). Devices were placed in a vacuum to rid the PDMS of air bubbles for at least 5 minutes before baking at 65° C. overnight. A biopsy punch (0.75 mm diameter, Harris Uni-Core, Ted Pella Inc., Redding, Calif.) was used to punch entry and exit holes in the PDMS. The PDMS was then oxygen plasma treated and bonded to a LabTek chamber with a no. 1.5 coverslip bottom (Nunc). The chambers were found to be most stable during the course of scanning the stage and automated image acquisition.
Loading. The microfluidic device was loaded with the yeast cells as follows. Yeast cells (S. cerevisiae) were cultured in YPD at 30° C. to a density of OD600˜0.1 and were washed three times in SD media, and diluted 100-fold. A dilute suspension of the yeast cells and perfusion media were loaded into 1 mL plastic syringes (BD, VWR). Needles (luer-lok, 27½ gauge) were connected to the syringes, and the syringes were inverted to remove all air bubbles. The needles were then connected to PE-20 tubing (VWR/ALA Scientific Instruments Inc., Westbury, N.Y.). Syringe pumps (Kent/Harvard Apparatus) were used to control fluid flow. To begin, a piece of tubing was inserted into the exit port for waste collection, and the cell inlet was blocked with a plug. Plugs were fabricated by holding the tip of a 2″ long piece of PE-20 briefly in a flame, and then placing the melted end of the tubing between two flat objects. This formed a nice handle making the plug easy to hold. The remaining long end of the tubing was cut to make a short stub. With the plug inserted in the cell entry port, media was flowed through the media entry port of the device until the chambers were filled with fluid. The plug was then replaced with the cell tubing. Cells were loaded using flow rates of 95 μL/hr cells and 150 μL/hr media. When about half of the chambers were filled with single cells, the cell tubing was removed from the syringe needle, and the media flow was increased to 45 μL/hr. This flow rate was maintained for the remainder of the experiment.
The cells were stained by connecting a media syringe to a three-way stopcock (Small Parts). A syringe full of fixation medium/staining medium was attached to the third entry of the stopcock at the end of a growth experiment. The staining media was then flowed through the chambers. The chambers were imaged by placing the chambers on the stage of an inverted microscope (Zeiss Axiovert 200M). A timelapse series of images were acquired at different stage positions. Confocal microscopy was performed on a Zeiss microscope.
While several embodiments of the present invention have been described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and/or structures for performing the functions and/or obtaining the results and/or one or more of the advantages described herein, and each of such variations and/or modifications is deemed to be within the scope of the present invention. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and/or configurations will depend upon the specific application or applications for which the teachings of the present invention is/are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, the invention may be practiced otherwise than as specifically described and claimed. The present invention is directed to each individual feature, system, article, material, kit, and/or method described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and/or methods, if such features, systems, articles, materials, kits, and/or methods are not mutually inconsistent, is included within the scope of the present invention.
All definitions, as defined and used herein, should be understood to control over dictionary definitions, definitions in documents incorporated by reference, and/or ordinary meanings of the defined terms.
The indefinite articles “a” and “an,” as used herein in the specification and in the claims, unless clearly indicated to the contrary, should be understood to mean “at least one.”
The phrase “and/or,” as used herein in the specification and in the claims, should be understood to mean “either or both” of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with “and/or” should be construed in the same fashion, i.e., “one or more” of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the “and/or” clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to “A and/or B”, when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.
As used herein in the specification and in the claims, “or” should be understood to have the same meaning as “and/or” as defined above. For example, when separating items in a list, “or” or “and/or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of” or “exactly one of,” or, when used in the claims, “consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (i.e. “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of.” “Consisting essentially of”, when used in the claims, shall have its ordinary meaning as used in the field of patent law.
As used herein in the specification and in the claims, the phrase “at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase “at least one” refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B,” or, equivalently “at least one of A and/or B”) can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.
It should also be understood that, unless clearly indicated to the contrary, in any methods claimed herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited.
In the claims, as well as in the specification above, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” “composed of,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases “consisting of” and “consisting essentially of” shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.
| Number | Name | Date | Kind |
|---|---|---|---|
| 2097692 | Fiegel | Nov 1937 | A |
| 2164172 | Dalton | Jun 1939 | A |
| 2636855 | Schwartz | Apr 1953 | A |
| 2656508 | Coulter | Oct 1953 | A |
| 2692800 | Nichols et al. | Oct 1954 | A |
| 2797149 | Skeggs | Jun 1957 | A |
| 2879141 | Skeggs | Mar 1959 | A |
| 2971700 | Peeps | Feb 1961 | A |
| 3479141 | Smythe et al. | Nov 1969 | A |
| 3608821 | Simm et al. | Sep 1971 | A |
| 3621059 | Bartlett | Nov 1971 | A |
| 3698635 | Sickles | Oct 1972 | A |
| 3784471 | Kaiser | Jan 1974 | A |
| 3816331 | Brown, Jr. et al. | Jun 1974 | A |
| 3828085 | Price et al. | Aug 1974 | A |
| 3930061 | Scharfenberger | Dec 1975 | A |
| 3960187 | Stock et al. | Jun 1976 | A |
| 3980541 | Aine | Sep 1976 | A |
| 3982541 | L'Esperance, Jr. | Sep 1976 | A |
| 4014469 | Sato | Mar 1977 | A |
| 4022575 | Hansen et al. | May 1977 | A |
| 4034966 | Suh et al. | Jul 1977 | A |
| 4059552 | Zweigle et al. | Nov 1977 | A |
| 4091042 | Alexanderson et al. | May 1978 | A |
| 4117550 | Folland et al. | Sep 1978 | A |
| 4130394 | Negersmith | Dec 1978 | A |
| 4210809 | Pelavin | Jul 1980 | A |
| 4253846 | Smythe et al. | Mar 1981 | A |
| 4266721 | Sickles | May 1981 | A |
| 4279345 | Allred | Jul 1981 | A |
| 4297345 | Howarth | Oct 1981 | A |
| 4315754 | Ruzicka et al. | Feb 1982 | A |
| 4378957 | Malkin et al. | Apr 1983 | A |
| 4383767 | Jido | May 1983 | A |
| 4439980 | Biblarz et al. | Apr 1984 | A |
| 4508265 | Jido | Apr 1985 | A |
| 4533634 | Maldonado et al. | Aug 1985 | A |
| 4585209 | Aine et al. | Apr 1986 | A |
| 4618476 | Columbus | Oct 1986 | A |
| 4675285 | Clark et al. | Jun 1987 | A |
| 4676274 | Brown | Jun 1987 | A |
| 4683195 | Mullis et al. | Jul 1987 | A |
| 4683202 | Mullis | Jul 1987 | A |
| 4739044 | Stabinsky | Apr 1988 | A |
| 4757141 | Fung et al. | Jul 1988 | A |
| 4767515 | Scott et al. | Aug 1988 | A |
| 4767929 | Valentine | Aug 1988 | A |
| 4779805 | Jackson et al. | Oct 1988 | A |
| 4795330 | Noakes et al. | Jan 1989 | A |
| 4801086 | Noakes | Jan 1989 | A |
| 4801529 | Perlman | Jan 1989 | A |
| 4829996 | Noakes et al. | May 1989 | A |
| 4853336 | Saros et al. | Aug 1989 | A |
| 4856363 | LaRocca et al. | Aug 1989 | A |
| 4859363 | Davis et al. | Aug 1989 | A |
| 4865444 | Green et al. | Sep 1989 | A |
| 4883750 | Whiteley et al. | Nov 1989 | A |
| 4908112 | Pace | Mar 1990 | A |
| 4931225 | Cheng | Jun 1990 | A |
| 4941959 | Scott | Jul 1990 | A |
| 4962885 | Coffee | Oct 1990 | A |
| 4963498 | Hillman et al. | Oct 1990 | A |
| 4981580 | Auer | Jan 1991 | A |
| 4996004 | Bucheler et al. | Feb 1991 | A |
| 5055390 | Weaver et al. | Oct 1991 | A |
| 5091652 | Mathies et al. | Feb 1992 | A |
| 5096615 | Prescott et al. | Mar 1992 | A |
| 5104813 | Besemer et al. | Apr 1992 | A |
| 5122360 | Harris et al. | Jun 1992 | A |
| 5149625 | Church et al. | Sep 1992 | A |
| 5180662 | Sitkovsky | Jan 1993 | A |
| 5185099 | Delpuech et al. | Feb 1993 | A |
| 5188290 | Gebauer et al. | Feb 1993 | A |
| 5188291 | Cross | Feb 1993 | A |
| 5192659 | Simons | Mar 1993 | A |
| 5204112 | Hope et al. | Apr 1993 | A |
| 5207973 | Harris et al. | May 1993 | A |
| 5241159 | Chatteriee et al. | Aug 1993 | A |
| 5260466 | McGibbon | Nov 1993 | A |
| 5262027 | Scott | Nov 1993 | A |
| 5270163 | Gold et al. | Dec 1993 | A |
| 5296375 | Kricka et al. | Mar 1994 | A |
| 5304487 | Wilding et al. | Apr 1994 | A |
| 5310653 | Hanausek-Walaszek et al. | May 1994 | A |
| 5313009 | Guenkel et al. | May 1994 | A |
| 5333675 | Mullis et al. | Aug 1994 | A |
| 5344594 | Sheridon | Sep 1994 | A |
| 5354670 | Nickoloff et al. | Oct 1994 | A |
| 5376252 | Ekstrom et al. | Dec 1994 | A |
| 5378957 | Kelly | Jan 1995 | A |
| 5397605 | Barbieri et al. | Mar 1995 | A |
| 5399461 | Van et al. | Mar 1995 | A |
| 5399491 | Kacian et al. | Mar 1995 | A |
| 5403617 | Haaland | Apr 1995 | A |
| 5413924 | Kosak et al. | May 1995 | A |
| 5417235 | Wise et al. | May 1995 | A |
| 5427946 | Kricka et al. | Jun 1995 | A |
| 5445934 | Fodor et al. | Aug 1995 | A |
| 5452878 | Gravesen et al. | Sep 1995 | A |
| 5452955 | Lundstrom | Sep 1995 | A |
| 5454472 | Benecke et al. | Oct 1995 | A |
| 5460945 | Springer et al. | Oct 1995 | A |
| 5468613 | Erlich et al. | Nov 1995 | A |
| 5475096 | Gold et al. | Dec 1995 | A |
| 5475610 | Atwood et al. | Dec 1995 | A |
| 5480614 | Kamahori | Jan 1996 | A |
| 5486335 | Wilding et al. | Jan 1996 | A |
| 5498392 | Wilding et al. | Mar 1996 | A |
| 5498523 | Tabor et al. | Mar 1996 | A |
| 5500415 | Dollat et al. | Mar 1996 | A |
| 5503851 | Mank et al. | Apr 1996 | A |
| 5512131 | Kumar et al. | Apr 1996 | A |
| 5516635 | Ekins et al. | May 1996 | A |
| 5518709 | Sutton et al. | May 1996 | A |
| 5523162 | Franz et al. | Jun 1996 | A |
| 5587128 | Wilding et al. | Dec 1996 | A |
| 5589136 | Northrup et al. | Dec 1996 | A |
| 5602756 | Atwood et al. | Feb 1997 | A |
| 5604097 | Brenner | Feb 1997 | A |
| 5610016 | Sato et al. | Mar 1997 | A |
| 5612188 | Shuler et al. | Mar 1997 | A |
| 5616478 | Chetverin et al. | Apr 1997 | A |
| 5617997 | Kobayashi et al. | Apr 1997 | A |
| 5635358 | Wilding et al. | Jun 1997 | A |
| 5636400 | Young | Jun 1997 | A |
| 5641658 | Adams et al. | Jun 1997 | A |
| 5643729 | Taniguchi et al. | Jul 1997 | A |
| 5655517 | Coffee | Aug 1997 | A |
| 5656155 | Norcross et al. | Aug 1997 | A |
| 5656493 | Mullis et al. | Aug 1997 | A |
| 5661222 | Hare | Aug 1997 | A |
| 5662874 | David | Sep 1997 | A |
| 5670325 | Lapidus et al. | Sep 1997 | A |
| 5681600 | Antinone et al. | Oct 1997 | A |
| 5695934 | Brenner | Dec 1997 | A |
| 5726026 | Wilding et al. | Mar 1998 | A |
| 5726404 | Brody | Mar 1998 | A |
| 5733526 | Trevino et al. | Mar 1998 | A |
| 5739036 | Parris | Apr 1998 | A |
| 5744366 | Kricka et al. | Apr 1998 | A |
| 5750988 | Apffel et al. | May 1998 | A |
| 5762775 | DePaoli | Jun 1998 | A |
| 5779868 | Parce et al. | Jul 1998 | A |
| 5783431 | Peterson et al. | Jul 1998 | A |
| 5789206 | Tavtigian et al. | Aug 1998 | A |
| 5840506 | Giordano | Nov 1998 | A |
| 5846719 | Brenner et al. | Dec 1998 | A |
| 5849491 | Radomski et al. | Dec 1998 | A |
| 5851769 | Gray et al. | Dec 1998 | A |
| 5858187 | Ramsey et al. | Jan 1999 | A |
| 5858655 | Arnold | Jan 1999 | A |
| 5858670 | Lam et al. | Jan 1999 | A |
| 5863722 | Brenner | Jan 1999 | A |
| 5868322 | Loucks | Feb 1999 | A |
| 5872010 | Karger et al. | Feb 1999 | A |
| 5876771 | Sizer et al. | Mar 1999 | A |
| 5880071 | Parce et al. | Mar 1999 | A |
| 5882680 | Suzuki et al. | Mar 1999 | A |
| 5882856 | Shuber | Mar 1999 | A |
| 5884846 | Tan | Mar 1999 | A |
| 5887755 | Hood, III | Mar 1999 | A |
| 5888746 | Tabiti et al. | Mar 1999 | A |
| 5888778 | Shuber | Mar 1999 | A |
| 5904933 | Riess et al. | May 1999 | A |
| 5921678 | Desai et al. | Jul 1999 | A |
| 5927852 | Serafin | Jul 1999 | A |
| 5928870 | Lapidus et al. | Jul 1999 | A |
| 5932100 | Yager et al. | Aug 1999 | A |
| 5935331 | Naka et al. | Aug 1999 | A |
| 5942056 | Singh | Aug 1999 | A |
| 5942443 | Parce et al. | Aug 1999 | A |
| 5958203 | Parce et al. | Sep 1999 | A |
| 5972187 | Parce et al. | Oct 1999 | A |
| 5980936 | Krafft et al. | Nov 1999 | A |
| 5989815 | Skolnick et al. | Nov 1999 | A |
| 5989892 | Nishimaki et al. | Nov 1999 | A |
| 5995341 | Tanaka et al. | Nov 1999 | A |
| 5997636 | Gamarnik et al. | Dec 1999 | A |
| 6008003 | Haak-Frendscho et al. | Dec 1999 | A |
| 6023540 | Walt et al. | Feb 2000 | A |
| 6028066 | Unger | Feb 2000 | A |
| 6042709 | Parce et al. | Mar 2000 | A |
| 6045755 | Lebl et al. | Apr 2000 | A |
| 6046056 | Parce et al. | Apr 2000 | A |
| 6048551 | Hilfinger et al. | Apr 2000 | A |
| 6048690 | Heller et al. | Apr 2000 | A |
| 6068199 | Coffee | May 2000 | A |
| 6074879 | Zelmanovic et al. | Jun 2000 | A |
| 6080295 | Parce et al. | Jun 2000 | A |
| 6086740 | Kennedy | Jul 2000 | A |
| 6096495 | Kasai et al. | Aug 2000 | A |
| 6103537 | Ullman et al. | Aug 2000 | A |
| 6105571 | Coffee | Aug 2000 | A |
| 6105877 | Coffee | Aug 2000 | A |
| 6107059 | Hart | Aug 2000 | A |
| 6116516 | Ganan-Calvo | Sep 2000 | A |
| 6118849 | Tanimori et al. | Sep 2000 | A |
| 6119953 | Ganan-Calvo et al. | Sep 2000 | A |
| 6120666 | Jacobson et al. | Sep 2000 | A |
| 6124388 | Takai et al. | Sep 2000 | A |
| 6124439 | Friedman et al. | Sep 2000 | A |
| 6130052 | Van Baren et al. | Oct 2000 | A |
| 6130098 | Handique et al. | Oct 2000 | A |
| 6137214 | Raina | Oct 2000 | A |
| 6138077 | Brenner | Oct 2000 | A |
| 6139303 | Reed et al. | Oct 2000 | A |
| 6140053 | Koster | Oct 2000 | A |
| 6143496 | Brown et al. | Nov 2000 | A |
| 6146828 | Lapidus et al. | Nov 2000 | A |
| 6149789 | Benecke et al. | Nov 2000 | A |
| 6150180 | Parce et al. | Nov 2000 | A |
| 6150516 | Brenner et al. | Nov 2000 | A |
| 6155710 | Nakajima et al. | Dec 2000 | A |
| 6162421 | Ordino et al. | Dec 2000 | A |
| 6165778 | Kedar | Dec 2000 | A |
| 6171796 | An et al. | Jan 2001 | B1 |
| 6171850 | Nagle et al. | Jan 2001 | B1 |
| 6172214 | Brenner | Jan 2001 | B1 |
| 6172218 | Brenner | Jan 2001 | B1 |
| 6174160 | Lee et al. | Jan 2001 | B1 |
| 6174469 | Gañan-Calvo | Jan 2001 | B1 |
| 6177479 | Nakajima | Jan 2001 | B1 |
| 6180372 | Franzen | Jan 2001 | B1 |
| 6184012 | Neri et al. | Feb 2001 | B1 |
| 6187214 | Ganan-Calvo | Feb 2001 | B1 |
| 6189803 | Ganan-Calvo | Feb 2001 | B1 |
| 6196525 | Ganan-Calvo | Mar 2001 | B1 |
| 6197335 | Sherman | Mar 2001 | B1 |
| 6197835 | Ganan-Calvo | Mar 2001 | B1 |
| 6203993 | Shuber et al. | Mar 2001 | B1 |
| 6207372 | Shuber | Mar 2001 | B1 |
| 6207397 | Lynch et al. | Mar 2001 | B1 |
| 6210396 | MacDonald et al. | Apr 2001 | B1 |
| 6210891 | Nyren et al. | Apr 2001 | B1 |
| 6210896 | Chan | Apr 2001 | B1 |
| 6214558 | Shuber et al. | Apr 2001 | B1 |
| 6221654 | Quake et al. | Apr 2001 | B1 |
| 6227466 | Hartman et al. | May 2001 | B1 |
| 6234402 | Ganan-Calvo | May 2001 | B1 |
| 6235383 | Hong et al. | May 2001 | B1 |
| 6235475 | Brenner et al. | May 2001 | B1 |
| 6241159 | Ganan-Calvo et al. | Jun 2001 | B1 |
| 6243373 | Turock | Jun 2001 | B1 |
| 6248378 | Ganan-Calvo | Jun 2001 | B1 |
| 6251661 | Urabe et al. | Jun 2001 | B1 |
| 6252129 | Coffee | Jun 2001 | B1 |
| 6258568 | Nyren | Jul 2001 | B1 |
| 6258858 | Nakajima et al. | Jul 2001 | B1 |
| 6261661 | Ohno et al. | Jul 2001 | B1 |
| 6261797 | Sorge et al. | Jul 2001 | B1 |
| 6263222 | Diab et al. | Jul 2001 | B1 |
| 6266459 | Walt et al. | Jul 2001 | B1 |
| 6267353 | Friedline et al. | Jul 2001 | B1 |
| 6267858 | Parce et al. | Jul 2001 | B1 |
| 6268152 | Fodor et al. | Jul 2001 | B1 |
| 6268165 | O'Brien | Jul 2001 | B1 |
| 6268222 | Chandler et al. | Jul 2001 | B1 |
| 6274320 | Rothberg et al. | Aug 2001 | B1 |
| 6274337 | Parce et al. | Aug 2001 | B1 |
| 6280948 | Guilfoyle et al. | Aug 2001 | B1 |
| 6292756 | Lievois et al. | Sep 2001 | B1 |
| 6294344 | O'Brien | Sep 2001 | B1 |
| 6296020 | McNeely et al. | Oct 2001 | B1 |
| 6296673 | Santarsiero et al. | Oct 2001 | B1 |
| 6299145 | Ganan-Calvo | Oct 2001 | B1 |
| 6301055 | Legrand et al. | Oct 2001 | B1 |
| 6306659 | Parce et al. | Oct 2001 | B1 |
| 6310354 | Hanninen et al. | Oct 2001 | B1 |
| 6310653 | Malcolm, Jr. et al. | Oct 2001 | B1 |
| 6316208 | Roberts et al. | Nov 2001 | B1 |
| 6316213 | O'Brien | Nov 2001 | B1 |
| 6318640 | Coffee | Nov 2001 | B1 |
| 6326145 | Whitcombe et al. | Dec 2001 | B1 |
| 6336463 | Ohta | Jan 2002 | B1 |
| 6344325 | Quake et al. | Feb 2002 | B1 |
| 6352828 | Brenner | Mar 2002 | B1 |
| 6355193 | Stott | Mar 2002 | B1 |
| 6355198 | Kim et al. | Mar 2002 | B1 |
| 6357670 | Ganan-Calvo | Mar 2002 | B2 |
| 6386463 | Ganan-Calvo | May 2002 | B1 |
| 6391559 | Brown et al. | May 2002 | B1 |
| 6394429 | Ganan-Calvo | May 2002 | B2 |
| 6399339 | Wolberg et al. | Jun 2002 | B1 |
| 6399389 | Parce et al. | Jun 2002 | B1 |
| 6403373 | Scanlan et al. | Jun 2002 | B1 |
| 6405936 | Ganan-Calvo | Jun 2002 | B1 |
| 6408878 | Unger et al. | Jun 2002 | B2 |
| 6409832 | Weigl et al. | Jun 2002 | B2 |
| 6429025 | Parce et al. | Aug 2002 | B1 |
| 6429148 | Chu et al. | Aug 2002 | B1 |
| 6432143 | Kubiak et al. | Aug 2002 | B2 |
| 6432148 | Ganan-Calvo | Aug 2002 | B1 |
| 6432630 | Blankenstein | Aug 2002 | B1 |
| 6439103 | Miller | Aug 2002 | B1 |
| 6440706 | Vogelstein et al. | Aug 2002 | B1 |
| 6440760 | Cho et al. | Aug 2002 | B1 |
| 6450139 | Watanabe | Sep 2002 | B1 |
| 6450189 | Ganan-Calvo | Sep 2002 | B1 |
| 6454193 | Busick et al. | Sep 2002 | B1 |
| 6464336 | Sharma | Oct 2002 | B1 |
| 6464886 | Ganan-Calvo | Oct 2002 | B2 |
| 6475441 | Parce et al. | Nov 2002 | B1 |
| 6481648 | Zimmermann | Nov 2002 | B1 |
| 6489103 | Griffiths et al. | Dec 2002 | B1 |
| 6503933 | Moloney et al. | Jan 2003 | B1 |
| 6506609 | Wada et al. | Jan 2003 | B1 |
| 6508988 | Van Dam et al. | Jan 2003 | B1 |
| 6511803 | Church et al. | Jan 2003 | B1 |
| 6520425 | Reneker | Feb 2003 | B1 |
| 6524456 | Ramsey et al. | Feb 2003 | B1 |
| 6540395 | Muhlbauer et al. | Apr 2003 | B2 |
| 6540895 | Spence et al. | Apr 2003 | B1 |
| 6551836 | Chow et al. | Apr 2003 | B1 |
| 6553944 | Allen et al. | Apr 2003 | B1 |
| 6553960 | Yoshikawa et al. | Apr 2003 | B1 |
| 6554202 | Ganan-Calvo | Apr 2003 | B2 |
| 6557334 | Jager | May 2003 | B2 |
| 6557834 | Ganan-Calvo | May 2003 | B2 |
| 6558944 | Parce et al. | May 2003 | B1 |
| 6558960 | Parce et al. | May 2003 | B1 |
| 6560030 | Legrand et al. | May 2003 | B2 |
| 6565010 | Anderson et al. | May 2003 | B2 |
| 6569631 | Pantoliano et al. | May 2003 | B1 |
| 6576420 | Carson et al. | Jun 2003 | B1 |
| 6591852 | McNeely et al. | Jul 2003 | B1 |
| 6592321 | Bonker et al. | Jul 2003 | B2 |
| 6592821 | Wada et al. | Jul 2003 | B1 |
| 6601613 | McNeely et al. | Aug 2003 | B2 |
| 6608726 | Legrand et al. | Aug 2003 | B2 |
| 6610499 | Fulwyler et al. | Aug 2003 | B1 |
| 6614598 | Quake et al. | Sep 2003 | B1 |
| 6627603 | Bibette et al. | Sep 2003 | B1 |
| 6630006 | Santarsiero et al. | Oct 2003 | B2 |
| 6630353 | Parce et al. | Oct 2003 | B1 |
| 6632619 | Harrison et al. | Oct 2003 | B1 |
| 6637463 | Lei et al. | Oct 2003 | B1 |
| 6638749 | Beckman et al. | Oct 2003 | B1 |
| 6645432 | Anderson et al. | Nov 2003 | B1 |
| 6646253 | Rohwer et al. | Nov 2003 | B1 |
| 6653626 | Fischer et al. | Nov 2003 | B2 |
| 6656267 | Newman | Dec 2003 | B2 |
| 6659370 | Inoue | Dec 2003 | B1 |
| 6660252 | Matathia et al. | Dec 2003 | B2 |
| 6670142 | Lau et al. | Dec 2003 | B2 |
| 6679441 | Borra et al. | Jan 2004 | B1 |
| 6680178 | Harris et al. | Jan 2004 | B2 |
| 6682890 | Mack et al. | Jan 2004 | B2 |
| 6717136 | Andersson et al. | Apr 2004 | B2 |
| 6729561 | Hirae et al. | May 2004 | B2 |
| 6738502 | Coleman et al. | May 2004 | B1 |
| 6739036 | Koike et al. | May 2004 | B2 |
| 6744046 | Valaskovic et al. | Jun 2004 | B2 |
| 6752922 | Huang et al. | Jun 2004 | B2 |
| 6753147 | Vogelstein et al. | Jun 2004 | B2 |
| 6766817 | da Silva | Jul 2004 | B2 |
| 6767194 | Jeon et al. | Jul 2004 | B2 |
| 6767704 | Waldman et al. | Jul 2004 | B2 |
| 6790328 | Jacobson et al. | Sep 2004 | B2 |
| 6793753 | Unger et al. | Sep 2004 | B2 |
| 6797056 | David | Sep 2004 | B2 |
| 6800849 | Staats | Oct 2004 | B2 |
| 6806058 | Jesperson et al. | Oct 2004 | B2 |
| 6808382 | Lanfranchi | Oct 2004 | B2 |
| 6808882 | Griffiths et al. | Oct 2004 | B2 |
| 6814980 | Levy et al. | Nov 2004 | B2 |
| 6818395 | Quake et al. | Nov 2004 | B1 |
| 6832787 | Renzi | Dec 2004 | B1 |
| 6833242 | Quake et al. | Dec 2004 | B2 |
| 6841350 | Ogden et al. | Jan 2005 | B2 |
| 6872250 | David et al. | Mar 2005 | B2 |
| 6890487 | Sklar et al. | May 2005 | B1 |
| 6897018 | Yuan et al. | May 2005 | B1 |
| 6905844 | Kim | Jun 2005 | B2 |
| 6918404 | Dias da Silva | Jul 2005 | B2 |
| 6926313 | Renzi | Aug 2005 | B1 |
| 6935768 | Lowe et al. | Aug 2005 | B2 |
| 6936417 | Orntoft | Aug 2005 | B2 |
| 6942978 | O'Brien | Sep 2005 | B1 |
| 6949342 | Golub et al. | Sep 2005 | B2 |
| 6960437 | Enzelberger et al. | Nov 2005 | B2 |
| 6964847 | Englert | Nov 2005 | B1 |
| 6974667 | Horne et al. | Dec 2005 | B2 |
| 6998232 | Feinstein et al. | Feb 2006 | B1 |
| 7022472 | Robbins et al. | Apr 2006 | B2 |
| 7041481 | Anderson et al. | May 2006 | B2 |
| 7049072 | Seshi | May 2006 | B2 |
| 7056674 | Baker et al. | Jun 2006 | B2 |
| 7057026 | Barnes et al. | Jun 2006 | B2 |
| 7066586 | da Silva | Jun 2006 | B2 |
| 7068874 | Wang et al. | Jun 2006 | B2 |
| 7078180 | Genetta | Jul 2006 | B2 |
| 7081192 | Wang et al. | Jul 2006 | B1 |
| 7081340 | Baker et al. | Jul 2006 | B2 |
| 7090983 | Muramatsu et al. | Aug 2006 | B1 |
| 7115230 | Sundararajan | Oct 2006 | B2 |
| 7118910 | Unger et al. | Oct 2006 | B2 |
| 7129091 | Ismagilov et al. | Oct 2006 | B2 |
| 7138233 | Griffiths et al. | Nov 2006 | B2 |
| 7153700 | Pardee et al. | Dec 2006 | B1 |
| 7156917 | Moriyama et al. | Jan 2007 | B2 |
| 7163801 | Reed | Jan 2007 | B2 |
| 7169560 | Lapidus et al. | Jan 2007 | B2 |
| 7171311 | Dai et al. | Jan 2007 | B2 |
| 7198899 | Schleyer et al. | Apr 2007 | B2 |
| 7204431 | Li et al. | Apr 2007 | B2 |
| 7229770 | Price et al. | Jun 2007 | B1 |
| 7252943 | Griffiths et al. | Aug 2007 | B2 |
| 7267938 | Anderson et al. | Sep 2007 | B2 |
| 7268167 | Higuchi et al. | Sep 2007 | B2 |
| 7282337 | Harris | Oct 2007 | B1 |
| 7291462 | O'Brien et al. | Nov 2007 | B2 |
| 7294503 | Quake et al. | Nov 2007 | B2 |
| 7300765 | Patel | Nov 2007 | B2 |
| 7308364 | Shaughnessy et al. | Dec 2007 | B2 |
| 7314721 | Gure et al. | Jan 2008 | B2 |
| 7316906 | Chiorazzi et al. | Jan 2008 | B2 |
| 7323305 | Leamon et al. | Jan 2008 | B2 |
| 7326529 | Ali et al. | Feb 2008 | B2 |
| 7332280 | Levy et al. | Feb 2008 | B2 |
| 7332590 | Nacht et al. | Feb 2008 | B2 |
| 7341211 | Ganan Calvo et al. | Mar 2008 | B2 |
| 7348142 | Wang | Mar 2008 | B2 |
| 7358231 | McCaffey et al. | Apr 2008 | B1 |
| 7361474 | Siegler | Apr 2008 | B2 |
| 7364862 | Ali et al. | Apr 2008 | B2 |
| 7368255 | Bae et al. | May 2008 | B2 |
| 7378233 | Sidransky et al. | May 2008 | B2 |
| 7378280 | Quake et al. | May 2008 | B2 |
| 7390463 | He et al. | Jun 2008 | B2 |
| 7393634 | Ahuja et al. | Jul 2008 | B1 |
| 7393665 | Brenner | Jul 2008 | B2 |
| 7416851 | Davi et al. | Aug 2008 | B2 |
| 7429467 | Holliger et al. | Sep 2008 | B2 |
| 7432064 | Salceda et al. | Oct 2008 | B2 |
| 7442507 | Polsky et al. | Oct 2008 | B2 |
| 7449303 | Coignet | Nov 2008 | B2 |
| 7468271 | Golovchenko et al. | Dec 2008 | B2 |
| 7473530 | Huttemann | Jan 2009 | B2 |
| 7473531 | Domon et al. | Jan 2009 | B1 |
| 7476506 | Schleyer et al. | Jan 2009 | B2 |
| 7479370 | Coignet | Jan 2009 | B2 |
| 7479371 | Ando et al. | Jan 2009 | B2 |
| 7479376 | Waldman et al. | Jan 2009 | B2 |
| 7482129 | Soyupak et al. | Jan 2009 | B2 |
| 7501244 | Reinhard et al. | Mar 2009 | B2 |
| 7504214 | Erlander et al. | Mar 2009 | B2 |
| 7507532 | Chang et al. | Mar 2009 | B2 |
| 7507541 | Raitano et al. | Mar 2009 | B2 |
| 7510707 | Platica et al. | Mar 2009 | B2 |
| 7510842 | Podust et al. | Mar 2009 | B2 |
| 7514209 | Dai et al. | Apr 2009 | B2 |
| 7514210 | Holliger et al. | Apr 2009 | B2 |
| 7524633 | Sidransky | Apr 2009 | B2 |
| 7527933 | Sahin et al. | May 2009 | B2 |
| 7537897 | Brenner et al. | May 2009 | B2 |
| 7541383 | Fu et al. | Jun 2009 | B2 |
| 7544473 | Brenner | Jun 2009 | B2 |
| 7556776 | Fraden et al. | Jul 2009 | B2 |
| 7582446 | Griffiths et al. | Sep 2009 | B2 |
| 7595195 | Lee et al. | Sep 2009 | B2 |
| 7604938 | Takahashi et al. | Oct 2009 | B2 |
| 7622081 | Chou et al. | Nov 2009 | B2 |
| 7632562 | Nair et al. | Dec 2009 | B2 |
| 7635562 | Harris et al. | Dec 2009 | B2 |
| 7638276 | Griffiths et al. | Dec 2009 | B2 |
| 7655435 | Holliger et al. | Feb 2010 | B2 |
| 7655470 | Ismagilov et al. | Feb 2010 | B2 |
| 7666593 | Lapidus | Feb 2010 | B2 |
| 7691576 | Holliger et al. | Apr 2010 | B2 |
| 7698287 | Becker et al. | Apr 2010 | B2 |
| 7708949 | Stone et al. | May 2010 | B2 |
| 7718578 | Griffiths et al. | May 2010 | B2 |
| 7736890 | Sia et al. | Jun 2010 | B2 |
| 7741130 | Lee, Jr. et al. | Jun 2010 | B2 |
| RE41780 | Anderson et al. | Sep 2010 | E |
| 7814175 | Chang et al. | Oct 2010 | B1 |
| 7824889 | Vogelstein et al. | Nov 2010 | B2 |
| 7888017 | Quake et al. | Feb 2011 | B2 |
| 7897044 | Hoyos et al. | Mar 2011 | B2 |
| 7897341 | Griffiths et al. | Mar 2011 | B2 |
| 7901939 | Ismagliov et al. | Mar 2011 | B2 |
| 7915015 | Vogelstein et al. | Mar 2011 | B2 |
| 7968287 | Griffiths et al. | Jun 2011 | B2 |
| 7990525 | Kanda | Aug 2011 | B2 |
| 8012382 | Kim et al. | Sep 2011 | B2 |
| 8067159 | Brown et al. | Nov 2011 | B2 |
| 8153402 | Holliger et al. | Apr 2012 | B2 |
| 8252539 | Quake et al. | Aug 2012 | B2 |
| 8257925 | Brown et al. | Sep 2012 | B2 |
| 8278071 | Brown et al. | Oct 2012 | B2 |
| 8278711 | Rao et al. | Oct 2012 | B2 |
| 8318434 | Cuppens | Nov 2012 | B2 |
| 8337778 | Stone et al. | Dec 2012 | B2 |
| 8436993 | Kaduchak et al. | May 2013 | B2 |
| 8462269 | Cheng et al. | Jun 2013 | B2 |
| 8528589 | Miller et al. | Sep 2013 | B2 |
| 8535889 | Larson et al. | Sep 2013 | B2 |
| 8592221 | Fraden et al. | Nov 2013 | B2 |
| 8673595 | Nakamura et al. | Mar 2014 | B2 |
| 8715934 | Diehl et al. | May 2014 | B2 |
| 8765485 | Link et al. | Jul 2014 | B2 |
| 8772046 | Fraden et al. | Jul 2014 | B2 |
| 8841071 | Link | Sep 2014 | B2 |
| 8871444 | Griffiths et al. | Oct 2014 | B2 |
| 9029083 | Griffiths et al. | May 2015 | B2 |
| 9029085 | Agresti et al. | May 2015 | B2 |
| 9186643 | Griffiths et al. | Nov 2015 | B2 |
| 9273308 | Link et al. | Mar 2016 | B2 |
| 9364803 | Yurkovetsky et al. | Jun 2016 | B2 |
| 9399797 | Hutchison et al. | Jul 2016 | B2 |
| 9448172 | Griffiths et al. | Sep 2016 | B2 |
| 9816121 | Agresti et al. | Nov 2017 | B2 |
| 9839890 | Griffiths et al. | Dec 2017 | B2 |
| 9857202 | Seki | Jan 2018 | B2 |
| 9919277 | Griffiths et al. | Mar 2018 | B2 |
| 9925501 | Griffiths et al. | Mar 2018 | B2 |
| 9944977 | Link et al. | Apr 2018 | B2 |
| 10151698 | Griffiths et al. | Dec 2018 | B2 |
| 10357772 | Fraden et al. | Jul 2019 | B2 |
| 10526605 | Liu et al. | Jan 2020 | B2 |
| 10584332 | Samuels et al. | Mar 2020 | B2 |
| 10596541 | Weitz et al. | Mar 2020 | B2 |
| 10612081 | Hutchison et al. | Apr 2020 | B2 |
| 10633652 | Link et al. | Apr 2020 | B2 |
| 10639597 | Link et al. | May 2020 | B2 |
| 10639598 | Griffiths et al. | May 2020 | B2 |
| 10675626 | Fraden et al. | Jun 2020 | B2 |
| 20010010338 | Ganan-Calvo | Aug 2001 | A1 |
| 20010020011 | Mathiowitz et al. | Sep 2001 | A1 |
| 20010023078 | Bawendi et al. | Sep 2001 | A1 |
| 20010029983 | Unger et al. | Oct 2001 | A1 |
| 20010034025 | Modlin et al. | Oct 2001 | A1 |
| 20010034031 | Short et al. | Oct 2001 | A1 |
| 20010041343 | Pankowsky | Nov 2001 | A1 |
| 20010041344 | Sepetov et al. | Nov 2001 | A1 |
| 20010041357 | Fouillet et al. | Nov 2001 | A1 |
| 20010042793 | Ganan-Calvo | Nov 2001 | A1 |
| 20010048900 | Bardell et al. | Dec 2001 | A1 |
| 20010050881 | Depaoli et al. | Dec 2001 | A1 |
| 20020004532 | Matathia et al. | Jan 2002 | A1 |
| 20020005354 | Spence et al. | Jan 2002 | A1 |
| 20020008028 | Jacobson et al. | Jan 2002 | A1 |
| 20020012971 | Mehta | Jan 2002 | A1 |
| 20020015997 | Lafferty | Feb 2002 | A1 |
| 20020022038 | Biatry et al. | Feb 2002 | A1 |
| 20020022261 | Anderson et al. | Feb 2002 | A1 |
| 20020033422 | Ganan-Calvo | Mar 2002 | A1 |
| 20020034737 | Drmanac | Mar 2002 | A1 |
| 20020036018 | McNeely et al. | Mar 2002 | A1 |
| 20020036139 | Becker et al. | Mar 2002 | A1 |
| 20020041378 | Peltie et al. | Apr 2002 | A1 |
| 20020058332 | Quake et al. | May 2002 | A1 |
| 20020067800 | Newman et al. | Jun 2002 | A1 |
| 20020085961 | Morin et al. | Jul 2002 | A1 |
| 20020090720 | Mutz et al. | Jul 2002 | A1 |
| 20020106667 | Yamamoto et al. | Aug 2002 | A1 |
| 20020119459 | Griffiths | Aug 2002 | A1 |
| 20020127591 | Wada et al. | Sep 2002 | A1 |
| 20020143437 | Handique et al. | Oct 2002 | A1 |
| 20020155080 | Glenn et al. | Oct 2002 | A1 |
| 20020158027 | Moon et al. | Oct 2002 | A1 |
| 20020164271 | Ho | Nov 2002 | A1 |
| 20020164629 | Quake et al. | Nov 2002 | A1 |
| 20020166582 | O'Connor et al. | Nov 2002 | A1 |
| 20020179849 | Maher | Dec 2002 | A1 |
| 20030008308 | Enzelberger et al. | Jan 2003 | A1 |
| 20030012586 | Iwata et al. | Jan 2003 | A1 |
| 20030015425 | Bohm et al. | Jan 2003 | A1 |
| 20030017305 | Roitman et al. | Jan 2003 | A1 |
| 20030017579 | Corn et al. | Jan 2003 | A1 |
| 20030039169 | Ehrfeld et al. | Feb 2003 | A1 |
| 20030040620 | Langmore et al. | Feb 2003 | A1 |
| 20030059764 | Ravkin et al. | Mar 2003 | A1 |
| 20030061687 | Hansen et al. | Apr 2003 | A1 |
| 20030064414 | Benecky et al. | Apr 2003 | A1 |
| 20030082795 | Shuler et al. | May 2003 | A1 |
| 20030083276 | Li et al. | May 2003 | A1 |
| 20030104372 | Ahmadian et al. | Jun 2003 | A1 |
| 20030108900 | Oliphant et al. | Jun 2003 | A1 |
| 20030124586 | Griffiths et al. | Jul 2003 | A1 |
| 20030143599 | Makarov et al. | Jul 2003 | A1 |
| 20030144260 | Gilon | Jul 2003 | A1 |
| 20030148273 | Dong et al. | Aug 2003 | A1 |
| 20030148544 | Nie et al. | Aug 2003 | A1 |
| 20030181574 | Adam et al. | Sep 2003 | A1 |
| 20030183525 | Elrod et al. | Oct 2003 | A1 |
| 20030207295 | Gunderson et al. | Nov 2003 | A1 |
| 20030219754 | Oleksy et al. | Nov 2003 | A1 |
| 20030224509 | Moon et al. | Dec 2003 | A1 |
| 20030229376 | Sandhu | Dec 2003 | A1 |
| 20030230486 | Chien et al. | Dec 2003 | A1 |
| 20030232356 | Dooley et al. | Dec 2003 | A1 |
| 20040005582 | Shipwash | Jan 2004 | A1 |
| 20040005594 | Holliger et al. | Jan 2004 | A1 |
| 20040018525 | Wirtz et al. | Jan 2004 | A1 |
| 20040027915 | Lowe et al. | Feb 2004 | A1 |
| 20040031688 | Shenderov | Feb 2004 | A1 |
| 20040037739 | McNeely et al. | Feb 2004 | A1 |
| 20040037813 | Simpson et al. | Feb 2004 | A1 |
| 20040041093 | Schultz et al. | Mar 2004 | A1 |
| 20040050946 | Wang et al. | Mar 2004 | A1 |
| 20040053247 | Cordon-Cardo et al. | Mar 2004 | A1 |
| 20040057906 | Hsu et al. | Mar 2004 | A1 |
| 20040058450 | Pamula et al. | Mar 2004 | A1 |
| 20040068019 | Higuchi et al. | Apr 2004 | A1 |
| 20040071781 | Chattopadhyay et al. | Apr 2004 | A1 |
| 20040079881 | Fischer et al. | Apr 2004 | A1 |
| 20040086892 | Crothers et al. | May 2004 | A1 |
| 20040091923 | Reyes et al. | May 2004 | A1 |
| 20040096515 | Bausch et al. | May 2004 | A1 |
| 20040134854 | Higuchi et al. | Jul 2004 | A1 |
| 20040136497 | Meldrum et al. | Jul 2004 | A1 |
| 20040142329 | Erikson et al. | Jul 2004 | A1 |
| 20040146866 | Fu | Jul 2004 | A1 |
| 20040146921 | Eveleigh et al. | Jul 2004 | A1 |
| 20040159633 | Whitesides et al. | Aug 2004 | A1 |
| 20040180346 | Anderson et al. | Sep 2004 | A1 |
| 20040181131 | Maynard et al. | Sep 2004 | A1 |
| 20040181343 | Wigstrom et al. | Sep 2004 | A1 |
| 20040182712 | Basol | Sep 2004 | A1 |
| 20040185484 | Costa et al. | Sep 2004 | A1 |
| 20040188254 | Spaid | Sep 2004 | A1 |
| 20040209299 | Pinter et al. | Oct 2004 | A1 |
| 20040224325 | Knapp et al. | Nov 2004 | A1 |
| 20040224419 | Zheng et al. | Nov 2004 | A1 |
| 20040229349 | Daridon | Nov 2004 | A1 |
| 20040241693 | Ricoul et al. | Dec 2004 | A1 |
| 20040253731 | Holliger et al. | Dec 2004 | A1 |
| 20040258203 | Yamano et al. | Dec 2004 | A1 |
| 20040259083 | Oshima | Dec 2004 | A1 |
| 20050000970 | Kimbara et al. | Jan 2005 | A1 |
| 20050003380 | Cohen et al. | Jan 2005 | A1 |
| 20050008592 | Gardel et al. | Jan 2005 | A1 |
| 20050019776 | Callow et al. | Jan 2005 | A1 |
| 20050032238 | Karp et al. | Feb 2005 | A1 |
| 20050032240 | Lee et al. | Feb 2005 | A1 |
| 20050037392 | Griffiths et al. | Feb 2005 | A1 |
| 20050037397 | Mirkin et al. | Feb 2005 | A1 |
| 20050042639 | Knapp et al. | Feb 2005 | A1 |
| 20050042648 | Griffiths et al. | Feb 2005 | A1 |
| 20050048467 | Sastry et al. | Mar 2005 | A1 |
| 20050064460 | Holliger et al. | Mar 2005 | A1 |
| 20050069920 | Griffiths et al. | Mar 2005 | A1 |
| 20050079501 | Koike et al. | Apr 2005 | A1 |
| 20050079510 | Berka et al. | Apr 2005 | A1 |
| 20050084923 | Mueller et al. | Apr 2005 | A1 |
| 20050087122 | Ismagliov et al. | Apr 2005 | A1 |
| 20050095611 | Chan et al. | May 2005 | A1 |
| 20050100895 | Waldman et al. | May 2005 | A1 |
| 20050103690 | Kawano et al. | May 2005 | A1 |
| 20050123937 | Thorp et al. | Jun 2005 | A1 |
| 20050129582 | Breidford et al. | Jun 2005 | A1 |
| 20050130173 | Leamon et al. | Jun 2005 | A1 |
| 20050152908 | Liew et al. | Jul 2005 | A1 |
| 20050161669 | Jovanovich et al. | Jul 2005 | A1 |
| 20050164239 | Griffiths et al. | Jul 2005 | A1 |
| 20050169797 | Oshima | Aug 2005 | A1 |
| 20050170373 | Monforte | Aug 2005 | A1 |
| 20050170431 | Ibrahim et al. | Aug 2005 | A1 |
| 20050172476 | Stone et al. | Aug 2005 | A1 |
| 20050183995 | Deshpande et al. | Aug 2005 | A1 |
| 20050202429 | Trau et al. | Sep 2005 | A1 |
| 20050202489 | Cho et al. | Sep 2005 | A1 |
| 20050207940 | Butler et al. | Sep 2005 | A1 |
| 20050208495 | Joseph et al. | Sep 2005 | A1 |
| 20050214173 | Facer et al. | Sep 2005 | A1 |
| 20050221339 | Griffiths et al. | Oct 2005 | A1 |
| 20050221341 | Shimkets et al. | Oct 2005 | A1 |
| 20050226742 | Unger et al. | Oct 2005 | A1 |
| 20050227264 | Nobile et al. | Oct 2005 | A1 |
| 20050248066 | Esteban | Nov 2005 | A1 |
| 20050260566 | Fischer et al. | Nov 2005 | A1 |
| 20050272159 | Ismagilov et al. | Dec 2005 | A1 |
| 20050287572 | Mathies et al. | Dec 2005 | A1 |
| 20060003347 | Griffiths et al. | Jan 2006 | A1 |
| 20060003429 | Frost et al. | Jan 2006 | A1 |
| 20060003439 | Ismagilov et al. | Jan 2006 | A1 |
| 20060008824 | Ronaghi et al. | Jan 2006 | A1 |
| 20060035386 | Hattori et al. | Feb 2006 | A1 |
| 20060036348 | Handique et al. | Feb 2006 | A1 |
| 20060040197 | Kabai | Feb 2006 | A1 |
| 20060040297 | Leamon et al. | Feb 2006 | A1 |
| 20060046257 | Pollock et al. | Mar 2006 | A1 |
| 20060051329 | Lee et al. | Mar 2006 | A1 |
| 20060068398 | McMillan | Mar 2006 | A1 |
| 20060078475 | Tai et al. | Apr 2006 | A1 |
| 20060078888 | Griffiths et al. | Apr 2006 | A1 |
| 20060078893 | Griffiths et al. | Apr 2006 | A1 |
| 20060094119 | Ismagilov et al. | May 2006 | A1 |
| 20060100788 | Carrino et al. | May 2006 | A1 |
| 20060108012 | Barrow et al. | May 2006 | A1 |
| 20060110759 | Paris et al. | May 2006 | A1 |
| 20060115821 | Einstein et al. | Jun 2006 | A1 |
| 20060147909 | Rarbach et al. | Jul 2006 | A1 |
| 20060153924 | Griffiths et al. | Jul 2006 | A1 |
| 20060154298 | Griffiths et al. | Jul 2006 | A1 |
| 20060160762 | Zetter et al. | Jul 2006 | A1 |
| 20060163385 | Link et al. | Jul 2006 | A1 |
| 20060169800 | Rosell et al. | Aug 2006 | A1 |
| 20060177832 | Brenner | Aug 2006 | A1 |
| 20060195269 | Yeatman et al. | Aug 2006 | A1 |
| 20060223127 | Yip et al. | Oct 2006 | A1 |
| 20060234254 | An et al. | Oct 2006 | A1 |
| 20060234259 | Rubin et al. | Oct 2006 | A1 |
| 20060234264 | Hardenbol | Oct 2006 | A1 |
| 20060246431 | Balachandran | Nov 2006 | A1 |
| 20060252057 | Raponi et al. | Nov 2006 | A1 |
| 20060257893 | Takahashi et al. | Nov 2006 | A1 |
| 20060258841 | Michl et al. | Nov 2006 | A1 |
| 20060263888 | Fritz et al. | Nov 2006 | A1 |
| 20060269558 | Murphy et al. | Nov 2006 | A1 |
| 20060269934 | Woudenberg et al. | Nov 2006 | A1 |
| 20060269971 | Diamandis | Nov 2006 | A1 |
| 20060281089 | Gibson et al. | Dec 2006 | A1 |
| 20060281098 | Miao et al. | Dec 2006 | A1 |
| 20070003442 | Link et al. | Jan 2007 | A1 |
| 20070009914 | Wallace et al. | Jan 2007 | A1 |
| 20070020617 | Trnovsky et al. | Jan 2007 | A1 |
| 20070026439 | Faulstich et al. | Feb 2007 | A1 |
| 20070045117 | Pamula et al. | Mar 2007 | A1 |
| 20070048744 | Lapidus | Mar 2007 | A1 |
| 20070053896 | Ahmed et al. | Mar 2007 | A1 |
| 20070054119 | Garstecki et al. | Mar 2007 | A1 |
| 20070056853 | Aizenberg et al. | Mar 2007 | A1 |
| 20070065823 | Dressman et al. | Mar 2007 | A1 |
| 20070077572 | Tawfik et al. | Apr 2007 | A1 |
| 20070077579 | Griffiths et al. | Apr 2007 | A1 |
| 20070092914 | Griffiths et al. | Apr 2007 | A1 |
| 20070111303 | Inoue et al. | May 2007 | A1 |
| 20070120899 | Ohnishi et al. | May 2007 | A1 |
| 20070123430 | Pasquier et al. | May 2007 | A1 |
| 20070141593 | Lee et al. | Jun 2007 | A1 |
| 20070154889 | Wang | Jul 2007 | A1 |
| 20070166705 | Milton et al. | Jul 2007 | A1 |
| 20070172873 | Brenner et al. | Jul 2007 | A1 |
| 20070184439 | Guilford et al. | Aug 2007 | A1 |
| 20070184489 | Griffiths et al. | Aug 2007 | A1 |
| 20070195127 | Ahn et al. | Aug 2007 | A1 |
| 20070202525 | Quake et al. | Aug 2007 | A1 |
| 20070213410 | Hastwell et al. | Sep 2007 | A1 |
| 20070241068 | Pamula et al. | Oct 2007 | A1 |
| 20070242105 | Srinivasan et al. | Oct 2007 | A1 |
| 20070243634 | Pamula et al. | Oct 2007 | A1 |
| 20070259351 | Chinitz et al. | Nov 2007 | A1 |
| 20070259368 | An et al. | Nov 2007 | A1 |
| 20070259374 | Griffiths et al. | Nov 2007 | A1 |
| 20070275415 | Srinivasan et al. | Nov 2007 | A1 |
| 20070292869 | Becker et al. | Dec 2007 | A1 |
| 20080003142 | Link et al. | Jan 2008 | A1 |
| 20080003571 | McKernan et al. | Jan 2008 | A1 |
| 20080004436 | Tawfik et al. | Jan 2008 | A1 |
| 20080009005 | Kruk | Jan 2008 | A1 |
| 20080014589 | Link et al. | Jan 2008 | A1 |
| 20080014590 | Dahary et al. | Jan 2008 | A1 |
| 20080020940 | Stedronsky et al. | Jan 2008 | A1 |
| 20080021330 | Hwang et al. | Jan 2008 | A1 |
| 20080023330 | Viovy et al. | Jan 2008 | A1 |
| 20080032413 | Kim et al. | Feb 2008 | A1 |
| 20080038754 | Farias-Eisner et al. | Feb 2008 | A1 |
| 20080044828 | Kwok | Feb 2008 | A1 |
| 20080050378 | Nakamura et al. | Feb 2008 | A1 |
| 20080050723 | Belacel et al. | Feb 2008 | A1 |
| 20080053205 | Pollack et al. | Mar 2008 | A1 |
| 20080057514 | Goldenring | Mar 2008 | A1 |
| 20080058432 | Wang et al. | Mar 2008 | A1 |
| 20080063227 | Rohrseitz | Mar 2008 | A1 |
| 20080064047 | Zetter et al. | Mar 2008 | A1 |
| 20080081330 | Kahvejian | Apr 2008 | A1 |
| 20080081333 | Mori et al. | Apr 2008 | A1 |
| 20080092973 | Lai | Apr 2008 | A1 |
| 20080113340 | Schlegel | May 2008 | A1 |
| 20080118462 | Alani et al. | May 2008 | A1 |
| 20080124726 | Monforte | May 2008 | A1 |
| 20080138806 | Chow et al. | Jun 2008 | A1 |
| 20080166772 | Hollinger et al. | Jul 2008 | A1 |
| 20080166793 | Beer et al. | Jul 2008 | A1 |
| 20080171078 | Gray | Jul 2008 | A1 |
| 20080176211 | Spence et al. | Jul 2008 | A1 |
| 20080176236 | Tsao et al. | Jul 2008 | A1 |
| 20080181850 | Thaxton et al. | Jul 2008 | A1 |
| 20080206756 | Lee et al. | Aug 2008 | A1 |
| 20080216563 | Reed et al. | Sep 2008 | A1 |
| 20080220986 | Gormley et al. | Sep 2008 | A1 |
| 20080222741 | Chinnaiyan | Sep 2008 | A1 |
| 20080234138 | Shaughnessy et al. | Sep 2008 | A1 |
| 20080234139 | Shaughnessy et al. | Sep 2008 | A1 |
| 20080241830 | Vogelstein et al. | Oct 2008 | A1 |
| 20080261295 | Butler et al. | Oct 2008 | A1 |
| 20080268473 | Moses | Oct 2008 | A1 |
| 20080269157 | Srivastava et al. | Oct 2008 | A1 |
| 20080274513 | Shenderov et al. | Nov 2008 | A1 |
| 20080274908 | Chang | Nov 2008 | A1 |
| 20080280285 | Chen et al. | Nov 2008 | A1 |
| 20080280302 | Kebebew | Nov 2008 | A1 |
| 20080286199 | Livingston et al. | Nov 2008 | A1 |
| 20080286801 | Arjol et al. | Nov 2008 | A1 |
| 20080286811 | Moses et al. | Nov 2008 | A1 |
| 20080293578 | Shaugnessy et al. | Nov 2008 | A1 |
| 20080299565 | Schneider et al. | Dec 2008 | A1 |
| 20080305482 | Brentano et al. | Dec 2008 | A1 |
| 20080311570 | Lai | Dec 2008 | A1 |
| 20080311604 | Elting et al. | Dec 2008 | A1 |
| 20090004687 | Mansfield et al. | Jan 2009 | A1 |
| 20090005254 | Griffiths et al. | Jan 2009 | A1 |
| 20090009855 | Nakatsuka et al. | Jan 2009 | A1 |
| 20090012187 | Chu et al. | Jan 2009 | A1 |
| 20090017463 | Bhowmick | Jan 2009 | A1 |
| 20090021728 | Heinz et al. | Jan 2009 | A1 |
| 20090023137 | Van Der Zee et al. | Jan 2009 | A1 |
| 20090026082 | Rothberg et al. | Jan 2009 | A1 |
| 20090029372 | Wewer | Jan 2009 | A1 |
| 20090042737 | Katz et al. | Feb 2009 | A1 |
| 20090053700 | Griffiths et al. | Feb 2009 | A1 |
| 20090053732 | Vermesh et al. | Feb 2009 | A1 |
| 20090060797 | Mathies et al. | Mar 2009 | A1 |
| 20090062144 | Guo | Mar 2009 | A1 |
| 20090068170 | Weitz et al. | Mar 2009 | A1 |
| 20090069194 | Ramakrishnan | Mar 2009 | A1 |
| 20090075265 | Budiman et al. | Mar 2009 | A1 |
| 20090075307 | Fischer et al. | Mar 2009 | A1 |
| 20090075311 | Karl | Mar 2009 | A1 |
| 20090081237 | D'Andrea et al. | Mar 2009 | A1 |
| 20090081685 | Beyer et al. | Mar 2009 | A1 |
| 20090087849 | Malinowski et al. | Apr 2009 | A1 |
| 20090092973 | Erlander et al. | Apr 2009 | A1 |
| 20090098542 | Budiman et al. | Apr 2009 | A1 |
| 20090098543 | Budiman et al. | Apr 2009 | A1 |
| 20090098555 | Roth et al. | Apr 2009 | A1 |
| 20090105959 | Braverman et al. | Apr 2009 | A1 |
| 20090118128 | Liu et al. | May 2009 | A1 |
| 20090124569 | Bergan et al. | May 2009 | A1 |
| 20090127454 | Ritchie et al. | May 2009 | A1 |
| 20090127589 | Rothberg et al. | May 2009 | A1 |
| 20090131353 | Insel et al. | May 2009 | A1 |
| 20090131543 | Weitz et al. | May 2009 | A1 |
| 20090134027 | Jary | May 2009 | A1 |
| 20090191565 | Lapidus et al. | Jul 2009 | A1 |
| 20090197248 | Griffiths et al. | Aug 2009 | A1 |
| 20090197772 | Griffiths et al. | Aug 2009 | A1 |
| 20090215633 | Van Eijk et al. | Aug 2009 | A1 |
| 20090226971 | Beer et al. | Sep 2009 | A1 |
| 20090226972 | Beer et al. | Sep 2009 | A1 |
| 20090233802 | Bignell et al. | Sep 2009 | A1 |
| 20090246788 | Albert et al. | Oct 2009 | A1 |
| 20090317798 | Heid et al. | Dec 2009 | A1 |
| 20090325217 | Luscher | Dec 2009 | A1 |
| 20090325236 | Griffiths et al. | Dec 2009 | A1 |
| 20100003687 | Simen et al. | Jan 2010 | A1 |
| 20100009353 | Barnes et al. | Jan 2010 | A1 |
| 20100015617 | Toyama | Jan 2010 | A1 |
| 20100021984 | Edd et al. | Jan 2010 | A1 |
| 20100022414 | Link et al. | Jan 2010 | A1 |
| 20100035252 | Rothberg et al. | Feb 2010 | A1 |
| 20100075436 | Urdea et al. | Mar 2010 | A1 |
| 20100105112 | Holtze et al. | Apr 2010 | A1 |
| 20100111768 | Banerjee et al. | May 2010 | A1 |
| 20100124759 | Wang et al. | May 2010 | A1 |
| 20100130369 | Shenderov et al. | May 2010 | A1 |
| 20100136544 | Agresti et al. | Jun 2010 | A1 |
| 20100137143 | Rothberg et al. | Jun 2010 | A1 |
| 20100137163 | Link et al. | Jun 2010 | A1 |
| 20100159592 | Holliger et al. | Jun 2010 | A1 |
| 20100172803 | Stone et al. | Jul 2010 | A1 |
| 20100173293 | Woudenberg et al. | Jul 2010 | A1 |
| 20100173394 | Colston, Jr. et al. | Jul 2010 | A1 |
| 20100188073 | Rothberg et al. | Jul 2010 | A1 |
| 20100197507 | Rothberg et al. | Aug 2010 | A1 |
| 20100210479 | Griffiths et al. | Aug 2010 | A1 |
| 20100213628 | Bausch et al. | Aug 2010 | A1 |
| 20100233026 | Ismagliov et al. | Sep 2010 | A1 |
| 20100240101 | Lieberman et al. | Sep 2010 | A1 |
| 20100273173 | Hirai et al. | Oct 2010 | A1 |
| 20100282617 | Rothberg et al. | Nov 2010 | A1 |
| 20100285975 | Mathies et al. | Nov 2010 | A1 |
| 20100300559 | Schultz et al. | Dec 2010 | A1 |
| 20100300895 | Nobile et al. | Dec 2010 | A1 |
| 20100301398 | Rothberg et al. | Dec 2010 | A1 |
| 20100304982 | Hinz et al. | Dec 2010 | A1 |
| 20110000560 | Miller et al. | Jan 2011 | A1 |
| 20110024455 | Bethuy et al. | Feb 2011 | A1 |
| 20110033854 | Drmanac et al. | Feb 2011 | A1 |
| 20110045462 | Fu et al. | Feb 2011 | A1 |
| 20110053151 | Hansen et al. | Mar 2011 | A1 |
| 20110053798 | Hindson et al. | Mar 2011 | A1 |
| 20110059435 | Vogelstein et al. | Mar 2011 | A1 |
| 20110059556 | Strey et al. | Mar 2011 | A1 |
| 20110142734 | Ismagliov et al. | Jun 2011 | A1 |
| 20110151444 | Albers et al. | Jun 2011 | A1 |
| 20110174622 | Ismagilov et al. | Jul 2011 | A1 |
| 20110176966 | Ismagilov et al. | Jul 2011 | A1 |
| 20110177494 | Ismagilov et al. | Jul 2011 | A1 |
| 20110177586 | Ismagilov et al. | Jul 2011 | A1 |
| 20110177609 | Ismagilov et al. | Jul 2011 | A1 |
| 20110188717 | Baudry et al. | Aug 2011 | A1 |
| 20110190146 | Boehm et al. | Aug 2011 | A1 |
| 20110218123 | Weitz et al. | Sep 2011 | A1 |
| 20110223314 | Zhang et al. | Sep 2011 | A1 |
| 20110244455 | Larson et al. | Oct 2011 | A1 |
| 20110250597 | Larson et al. | Oct 2011 | A1 |
| 20110257031 | Bodeau et al. | Oct 2011 | A1 |
| 20110267457 | Weitz et al. | Nov 2011 | A1 |
| 20110275063 | Weitz et al. | Nov 2011 | A1 |
| 20110311978 | Makarewicz, Jr. et al. | Dec 2011 | A1 |
| 20120010098 | Griffiths et al. | Jan 2012 | A1 |
| 20120010107 | Griffiths et al. | Jan 2012 | A1 |
| 20120015382 | Weitz et al. | Jan 2012 | A1 |
| 20120015822 | Weitz et al. | Jan 2012 | A1 |
| 20120021930 | Schoen et al. | Jan 2012 | A1 |
| 20120088691 | Chen et al. | Apr 2012 | A1 |
| 20120122714 | Samuels et al. | May 2012 | A1 |
| 20120167142 | Hey | Jun 2012 | A1 |
| 20120190032 | Ness et al. | Jul 2012 | A1 |
| 20120220494 | Samuels et al. | Aug 2012 | A1 |
| 20120244043 | Leblanc et al. | Sep 2012 | A1 |
| 20120258516 | Schultz et al. | Oct 2012 | A1 |
| 20120288857 | Livak | Nov 2012 | A1 |
| 20120302448 | Hutchison et al. | Nov 2012 | A1 |
| 20130099018 | Miller et al. | Apr 2013 | A1 |
| 20130109577 | Korlach et al. | May 2013 | A1 |
| 20130143745 | Christen et al. | Jun 2013 | A1 |
| 20130157872 | Griffiths et al. | Jun 2013 | A1 |
| 20130178368 | Griffiths et al. | Jul 2013 | A1 |
| 20130178378 | Hatch et al. | Jul 2013 | A1 |
| 20130217601 | Griffiths et al. | Aug 2013 | A1 |
| 20130224751 | Olson et al. | Aug 2013 | A1 |
| 20130244906 | Collins | Sep 2013 | A1 |
| 20130274117 | Church et al. | Oct 2013 | A1 |
| 20130295567 | Link et al. | Nov 2013 | A1 |
| 20130295568 | Link | Nov 2013 | A1 |
| 20130296535 | Church et al. | Nov 2013 | A1 |
| 20140065631 | Froehlich et al. | Mar 2014 | A1 |
| 20140256568 | Link | Sep 2014 | A1 |
| 20140256585 | McCoy | Sep 2014 | A1 |
| 20140274786 | McCoy et al. | Sep 2014 | A1 |
| 20140323317 | Link et al. | Oct 2014 | A1 |
| 20140329239 | Larson et al. | Nov 2014 | A1 |
| 20150018236 | Green et al. | Jan 2015 | A1 |
| 20150038356 | Karlin-Neumann et al. | Feb 2015 | A1 |
| 20150126400 | Watson et al. | May 2015 | A1 |
| 20150184256 | Samuels et al. | Jul 2015 | A1 |
| 20150197790 | Tzonev | Jul 2015 | A1 |
| 20150336072 | Weitz et al. | Nov 2015 | A1 |
| 20160289670 | Samuels et al. | Oct 2016 | A1 |
| 20160304954 | Lin et al. | Oct 2016 | A1 |
| 20170304785 | Link et al. | Oct 2017 | A1 |
| 20180057863 | Larson et al. | Mar 2018 | A1 |
| 20180223348 | Link et al. | Aug 2018 | A1 |
| 20180272294 | Griffiths et al. | Sep 2018 | A1 |
| 20180272296 | Link et al. | Sep 2018 | A1 |
| 20180272299 | Griffiths et al. | Sep 2018 | A1 |
| 20180353913 | Link et al. | Dec 2018 | A1 |
| 20180355350 | Link et al. | Dec 2018 | A1 |
| 20180361346 | Griffiths et al. | Dec 2018 | A1 |
| 20180363050 | Hutchison et al. | Dec 2018 | A1 |
| 20190024261 | Griffiths et al. | Jan 2019 | A1 |
| 20190107489 | Griffiths et al. | Apr 2019 | A1 |
| 20190134581 | Yurkovetsky et al. | May 2019 | A1 |
| 20190316119 | Samuels et al. | Oct 2019 | A1 |
| Number | Date | Country |
|---|---|---|
| 140025 | Jul 1996 | AT |
| 140880 | Aug 1996 | AT |
| 155711 | Aug 1997 | AT |
| 167816 | Jul 1998 | AT |
| 4032078 | Apr 1980 | AU |
| 6415380 | May 1981 | AU |
| 1045983 | Jun 1984 | AU |
| 2177292 | Jan 1993 | AU |
| 4222393 | Nov 1993 | AU |
| 4222593 | Nov 1993 | AU |
| 4222693 | Nov 1993 | AU |
| 4222793 | Nov 1993 | AU |
| 4223593 | Nov 1993 | AU |
| 677197 | Apr 1997 | AU |
| 677781 | May 1997 | AU |
| 680195 | Jul 1997 | AU |
| 2935197 | Jan 1998 | AU |
| 3499097 | Jan 1998 | AU |
| 3501297 | Jan 1998 | AU |
| 1276099 | Jun 1999 | AU |
| 4955799 | Dec 1999 | AU |
| 3961100 | Oct 2000 | AU |
| 4910300 | Nov 2000 | AU |
| 747464 | May 2002 | AU |
| 768399 | Dec 2003 | AU |
| 2004225691 | Jun 2010 | AU |
| 2010224352 | Oct 2010 | AU |
| 1093344 | Jan 1981 | CA |
| 2258481 | Jan 1998 | CA |
| 2520548 | Oct 2004 | CA |
| 563 087 | Jun 1975 | CH |
| 563807 | Jul 1975 | CH |
| 2100685 | Jul 1972 | DE |
| 3042915 | Sep 1981 | DE |
| 43 08 839 | Apr 1997 | DE |
| 69126763 | Feb 1998 | DE |
| 199 61 257 | Jul 2001 | DE |
| 100 15 109 | Oct 2001 | DE |
| 100 41 823 | Mar 2002 | DE |
| 0047130 | Feb 1985 | EP |
| 0402995 | Dec 1990 | EP |
| 0249007 | Mar 1991 | EP |
| 0418635 | Mar 1991 | EP |
| 0476178 | Mar 1992 | EP |
| 0546174 | Jun 1993 | EP |
| 0618001 | Oct 1994 | EP |
| 620432 | Oct 1994 | EP |
| 0637996 | Feb 1995 | EP |
| 0637997 | Feb 1995 | EP |
| 0718038 | Jun 1996 | EP |
| 0540281 | Jul 1996 | EP |
| 0528580 | Dec 1996 | EP |
| 0486351 | Jul 1997 | EP |
| 0895120 | Feb 1999 | EP |
| 1362634 | Nov 2003 | EP |
| 04782399.2 | May 2006 | EP |
| 1741482 | Jan 2007 | EP |
| 2017910 | Jan 2009 | EP |
| 2047910 | Apr 2009 | EP |
| 2127736 | Dec 2009 | EP |
| 2047910 | Jan 2012 | EP |
| 13165665.4 | Nov 2013 | EP |
| 13165667.0 | Nov 2013 | EP |
| 2363205 | Jun 2014 | EP |
| 2534267 | Apr 2018 | EP |
| 2 095 413 | Feb 1997 | ES |
| 2 404 834 | Apr 1979 | FR |
| 2 451 579 | Oct 1980 | FR |
| 2 469 714 | May 1981 | FR |
| 2 470 385 | May 1981 | FR |
| 2 650 657 | Feb 1991 | FR |
| 2 669 028 | May 1992 | FR |
| 2 703 263 | Oct 1994 | FR |
| 1148543 | Apr 1969 | GB |
| 1 446 998 | Aug 1976 | GB |
| 2 005 224 | Apr 1979 | GB |
| 2 047 880 | Dec 1980 | GB |
| 2 062 225 | May 1981 | GB |
| 2 064 114 | Jun 1981 | GB |
| 2097692 | Nov 1982 | GB |
| 2 210 532 | Jun 1989 | GB |
| 922432 | Feb 1993 | IE |
| S5372016 | Jun 1978 | JP |
| S5455495 | May 1979 | JP |
| 55125472 | Sep 1980 | JP |
| S5636053 | Apr 1981 | JP |
| 56-124052 | Sep 1981 | JP |
| 59-49832 | Mar 1984 | JP |
| 59-102163 | Jun 1984 | JP |
| H0665609 | Mar 1994 | JP |
| 6-265447 | Sep 1994 | JP |
| 7-489 | Jan 1995 | JP |
| 8-153669 | Jun 1996 | JP |
| 10-217477 | Aug 1998 | JP |
| 3-232525 | Oct 1998 | JP |
| 2000-271475 | Oct 2000 | JP |
| 2001-301154 | Oct 2001 | JP |
| 2001-517353 | Oct 2001 | JP |
| 2002-085961 | Mar 2002 | JP |
| 2003-501257 | Jan 2003 | JP |
| 2003-502656 | Jan 2003 | JP |
| 2003-222633 | Aug 2003 | JP |
| 2005-037346 | Feb 2005 | JP |
| 2009-265751 | Nov 2009 | JP |
| 2010-198393 | Sep 2010 | JP |
| 2012-204765 | Oct 2012 | JP |
| 2013-143959 | Jul 2013 | JP |
| 2016063824 | Apr 2016 | JP |
| 264353 | May 1996 | NZ |
| 8402000 | May 1984 | WO |
| 9105058 | Apr 1991 | WO |
| 9107772 | May 1991 | WO |
| 9116966 | Nov 1991 | WO |
| 9203734 | Mar 1992 | WO |
| 9221746 | Dec 1992 | WO |
| 9303151 | Feb 1993 | WO |
| 9308278 | Apr 1993 | WO |
| 9322053 | Nov 1993 | WO |
| 9322054 | Nov 1993 | WO |
| 9322055 | Nov 1993 | WO |
| 9322058 | Nov 1993 | WO |
| 9322421 | Nov 1993 | WO |
| 9416332 | Jul 1994 | WO |
| 9423738 | Oct 1994 | WO |
| 9424314 | Oct 1994 | WO |
| 9426766 | Nov 1994 | WO |
| 9800705 | Jan 1995 | WO |
| 9511922 | May 1995 | WO |
| 9519922 | Jul 1995 | WO |
| 9524929 | Sep 1995 | WO |
| 9533447 | Dec 1995 | WO |
| 9634112 | Oct 1996 | WO |
| 9638730 | Dec 1996 | WO |
| 9640062 | Dec 1996 | WO |
| 9640723 | Dec 1996 | WO |
| 9700125 | Jan 1997 | WO |
| 9700442 | Jan 1997 | WO |
| 9704297 | Feb 1997 | WO |
| 9704748 | Feb 1997 | WO |
| 9723140 | Jul 1997 | WO |
| 9728556 | Aug 1997 | WO |
| 9738318 | Oct 1997 | WO |
| 9739814 | Oct 1997 | WO |
| 9740141 | Oct 1997 | WO |
| 9745644 | Dec 1997 | WO |
| 9747763 | Dec 1997 | WO |
| 9800231 | Jan 1998 | WO |
| 9802237 | Jan 1998 | WO |
| 9810267 | Mar 1998 | WO |
| 9813502 | Apr 1998 | WO |
| 9822625 | May 1998 | WO |
| 9823733 | Jun 1998 | WO |
| 9831700 | Jul 1998 | WO |
| 9833001 | Jul 1998 | WO |
| 9834120 | Aug 1998 | WO |
| 9837186 | Aug 1998 | WO |
| 9841869 | Sep 1998 | WO |
| 9852691 | Nov 1998 | WO |
| 9858085 | Dec 1998 | WO |
| 9902671 | Jan 1999 | WO |
| 9922858 | May 1999 | WO |
| 9928020 | Jun 1999 | WO |
| 9931019 | Jun 1999 | WO |
| 9942539 | Aug 1999 | WO |
| 9954730 | Oct 1999 | WO |
| 9961888 | Dec 1999 | WO |
| 0004139 | Jan 2000 | WO |
| 0047322 | Feb 2000 | WO |
| 0052455 | Feb 2000 | WO |
| 0040712 | Jun 2000 | WO |
| 0054735 | Sep 2000 | WO |
| 0061275 | Oct 2000 | WO |
| 0070080 | Nov 2000 | WO |
| 0076673 | Dec 2000 | WO |
| 00078455 | Dec 2000 | WO |
| 0112327 | Feb 2001 | WO |
| 0114589 | Mar 2001 | WO |
| 0118244 | Mar 2001 | WO |
| 0164332 | Sep 2001 | WO |
| 0168257 | Sep 2001 | WO |
| 0169289 | Sep 2001 | WO |
| 0172431 | Oct 2001 | WO |
| 0180283 | Oct 2001 | WO |
| 01089787 | Nov 2001 | WO |
| 0189788 | Nov 2001 | WO |
| 0194635 | Dec 2001 | WO |
| 0216017 | Feb 2002 | WO |
| 0218949 | Mar 2002 | WO |
| 0222869 | Mar 2002 | WO |
| 0223163 | Mar 2002 | WO |
| 0231203 | Apr 2002 | WO |
| 2002036815 | May 2002 | WO |
| 0247665 | Aug 2002 | WO |
| 0260275 | Aug 2002 | WO |
| 0260591 | Aug 2002 | WO |
| 02068104 | Sep 2002 | WO |
| 02078845 | Oct 2002 | WO |
| 02103011 | Dec 2002 | WO |
| 02103363 | Dec 2002 | WO |
| 03011443 | Feb 2003 | WO |
| 03026798 | Apr 2003 | WO |
| 03037302 | May 2003 | WO |
| 03044187 | May 2003 | WO |
| 03078659 | Sep 2003 | WO |
| 2003003015 | Oct 2003 | WO |
| 03099843 | Dec 2003 | WO |
| 2004002627 | Jan 2004 | WO |
| 2004018497 | Mar 2004 | WO |
| 2004024917 | Mar 2004 | WO |
| 2004037374 | May 2004 | WO |
| 2004038363 | May 2004 | WO |
| 2004069849 | Aug 2004 | WO |
| 2004071638 | Aug 2004 | WO |
| 2004074504 | Sep 2004 | WO |
| 2004083443 | Sep 2004 | WO |
| 2004087308 | Oct 2004 | WO |
| 2004088314 | Oct 2004 | WO |
| 2004091763 | Oct 2004 | WO |
| 2004102204 | Nov 2004 | WO |
| 2004103565 | Dec 2004 | WO |
| 2005000970 | Jan 2005 | WO |
| 2005002730 | Jan 2005 | WO |
| 2005003375 | Jan 2005 | WO |
| 200511867 | Feb 2005 | WO |
| 2005021151 | Mar 2005 | WO |
| 2005023427 | Mar 2005 | WO |
| 2005041884 | May 2005 | WO |
| 2005049787 | Jun 2005 | WO |
| 2005103106 | Nov 2005 | WO |
| 2005118138 | Dec 2005 | WO |
| 2005118867 | Dec 2005 | WO |
| 2006002641 | Jan 2006 | WO |
| 2006009657 | Jan 2006 | WO |
| 2006027757 | Mar 2006 | WO |
| 2006038035 | Apr 2006 | WO |
| 2006040551 | Apr 2006 | WO |
| 2006040554 | Apr 2006 | WO |
| 2006078841 | Jul 2006 | WO |
| 2006096571 | Sep 2006 | WO |
| 2006101851 | Sep 2006 | WO |
| 2007012638 | Feb 2007 | WO |
| 2007021343 | Feb 2007 | WO |
| 2007030501 | Mar 2007 | WO |
| 2007081385 | Jul 2007 | WO |
| 2007081387 | Jul 2007 | WO |
| 2007089541 | Aug 2007 | WO |
| 2007114794 | Oct 2007 | WO |
| 2007123744 | Nov 2007 | WO |
| 2007133710 | Nov 2007 | WO |
| 2007138178 | Dec 2007 | WO |
| 2007140015 | Dec 2007 | WO |
| 2008021123 | Feb 2008 | WO |
| 2008063227 | May 2008 | WO |
| 2008097559 | Aug 2008 | WO |
| 2008115626 | Sep 2008 | WO |
| 2008121342 | Oct 2008 | WO |
| 2008130623 | Oct 2008 | WO |
| 2007092473 | Nov 2008 | WO |
| 2008134153 | Nov 2008 | WO |
| 2009015296 | Jan 2009 | WO |
| 2009029229 | Mar 2009 | WO |
| 2009049889 | Apr 2009 | WO |
| 2009059430 | May 2009 | WO |
| 2009085929 | Jul 2009 | WO |
| 2009137415 | Nov 2009 | WO |
| 2009137606 | Nov 2009 | WO |
| 2010009365 | Jan 2010 | WO |
| 2010056728 | May 2010 | WO |
| 2010040006 | Aug 2010 | WO |
| 2010115154 | Oct 2010 | WO |
| 2010151776 | Dec 2010 | WO |
| 2011042564 | Apr 2011 | WO |
| 2011079176 | Jun 2011 | WO |
| 2011100604 | Aug 2011 | WO |
| 2012022976 | Feb 2012 | WO |
| 2012045012 | Apr 2012 | WO |
| 2012047297 | Apr 2012 | WO |
| 2012048341 | Apr 2012 | WO |
| 2012083225 | Jun 2012 | WO |
| 2012167142 | Dec 2012 | WO |
| 201314356 | Jan 2013 | WO |
| 2013165748 | Nov 2013 | WO |
| 2014026031 | Feb 2014 | WO |
| 2014065756 | May 2014 | WO |
| 2014165559 | Oct 2014 | WO |
| 2014204939 | Dec 2014 | WO |
| 2015013681 | Jan 2015 | WO |
| 2015200893 | Dec 2015 | WO |
| 2017117358 | Jul 2017 | WO |
| Entry |
|---|
| Cormack, 1996, FACS-optimized mutants of the green fluorescent protein (GFP), Gene 173(1):33-38. |
| Cortesi, 2002, Production of lipospheres as carriers for bioactive compounds, Biomateials, 23(11): 2283-2294. |
| Courrier, 2004, Reverse water-in-fluorocarbon emulsions and microemulsions obtained with a fluorinated surfactant, Colloids and Surfaces A: Physicochem. Eng. Aspects 244:141-148. |
| Craig, 1995, Fluorescence-based enzymatic assay by capillary electrophoresis laser-induced fluoresence detection for the determinination of a few alpha-galactosidase molecules, Anal. Biochem. 226:147. |
| Creagh, 1993, Structural and catalytic properties of enzymes in reverse micelles, Enzyme Microb Technol 15 (5):383-92. |
| Crosland-Taylor, 1953, A Device for Counting Small Particles suspended in a Fluid through a Tube, Nature 171:37-38. |
| Crowley, 1973, Electrical breakdown of bimolecular lipid membranes as an electromechanical instability, Biophys J. 13 (7):711-724. |
| Cull, 1992, Screening for receptor ligands using large libraries of peptides linked to the C terminus of the lac repressor, PNAS 89:1865-9. |
| Curran, 1998, Strategy-level separations in organic synthesis: from planning to practice. Angew Chem Int Ed, 37:1174-11-96. |
| Czarnik, 1997, Encoding methods for combinatorial chemistry, Curr Opin Chem Biol 1:60-66. |
| Dankwardt, 1995, Combinatorial synthesis of small-molecule libraries using 3-amino-5-hydroxybenzoic acid, 1:113-120. |
| David, 1974, Protein iodination with solid-state lactoperoxidase, Biochemistry 13:1014-1021. |
| Davis, 1987, Multiple emulsions as targetable delivery systems, Meth Enzymol 149:51-64. |
| Davis, 2006, Deterministic hydrodynamics: Taking blood apart, PNAS 103:14779-14784. |
| De Gans, 2004, Inkjet printing of polymers: state of the art and future developments, Advanced materials, 16: 203-213. |
| De Wildt, 2002, Isolation of receptor-ligand pairs by capture of long-lived multivalent interaction complexes, Proceedings of the National Academy of Sciences of the United States, 99, 8530-8535. |
| DelRaso, 1993, In vitro methodologies for enhanced toxicity testing, Toxicol. Lett. 68:91-99. |
| Deng, 2008, Design and analysis of mismatch probes for long oligonucleotide microarrays, BMC Genomics; 9:491, 13 pages. |
| Dickinson, 1994, Emulsions and droplet size control, Wedlock, D.J., Ed., in Controlled Particle Droplet and Bubble Formulation, ButterWorth-Heine-mann, 191-257. |
| DiMatteo, 2008, Genetic conversion of an SMN2 gene to SMN1: A novel approach to the treatment of spinal muscular atrophy, Exp Cell Res. 314(4):878-886. |
| Ding, 2001, Scheduling of microfluidic operations for reconfigurable two-dimensional electrowetting arrays, IEEE Trans CADICS 20(12):1463-1468. |
| Ding, 2003, Direct molecular haplotyping of long-range genomic DNA with M1-PCR, Proc. Natl. Acad. Sci. Usa, 100(33):1449-7453. |
| Dinsmore, 2002, Colioidosomes: Selectively Permeable Capsules Composed of Colloidal Particles, Science 298(5595):1006-1009. |
| Dittrich, 2005, A new embedded process for compartmentalized cell-free protein expression and on-line detection in microfluidic devices, Chembiochem 6(5):811-814. |
| Doi, 1999, STABLE: protein-DNA fusion system for screening of combinatorial protein libraries in vitro, FEBS Lett., 457: 227-230. |
| Doi, 2004, In vitro selection of restriction endonucleases by in vitro compartmentilization, Nucleic Acids Res, 32(12): e95. |
| Doman, 2002, Molecular docking and high-throughput screening for novel inhibitors of protein tyrosine phosphatase-1B, J Med Chem, 45: 2213-2221. |
| Domling, 2000, Multicomponent Reactions with Isocyanides, Angew Chem Int Ed 39(18):3168-3210. |
| Domling, 2002, Recent advances in isocyanide-based multicomponent chemistry, Curr Opin Chem Biol, 6(3):306-13. |
| Dorfman, 2005, Contamination-free continuous flow microfluidic polymerase chain reaction for quantitative and clinical applications, Anal Chem 77:3700-3704. |
| Dove, 2002, Research News Briefs, Nature Biotechnology 20:1213, 1 page. |
| Dower, 1988, High efficiency transformation of E. coli by high voltage electroporation, Nucleic Acids Res 16:6127-6145. |
| Dressman, 2003, Transforming single DNA molecules into fluorescent magnetic particles for detection and enumeration of genetic variations, PNAS 100:8817-22. |
| Dreyfus, 2003, Ordered and disordered patterns in two phase flows in microchannels, Phys Rev Lett 90(14):144505-1-144505-4. |
| Drmanac, 1992, Sequencing by hybridization: towards an automated sequencing of one million M13 clones arrayed on membranes, Elctrophoresis 13:566-573. |
| Du, 2009, SlipChip, Lab Chip, 9, 2286-2292. |
| Dubertret, 2002, In vivo imaging of quantum dots encapsulated in phospholipid micelles, Science, 298: 1759-1762. |
| Duffy, 1998, Rapid Protyping of Microfluidic Systems and Polydimethylsiloxane, Anal Chem 70:474-480. |
| Duggleby, 1995, Analysis of Enzyme Progress Curves by Nonlinear Regression, Pt D. Academic Press 249:61-90. |
| Dumas, 1989, Purification and properties of the phosphotriesterase from Psuedomonas diminuta, J Biol Chem 264:19659-19665. |
| Eckert, 1991, DNA polymerase fidelity and the polymerase chain reaction, Genome Res 1:17-24. |
| Ecole Polytech Federale de Lausanne, 2014, Tracing water channels in cell surface receptors, PhysOrg News (2 pages). |
| Edel, 2002, Microfluidic Routes to the Controlled Production of Nanopaticles, Chemical Communications, 1136-1137. |
| Edris, 2001, Encapsulation of orange oil in a spray dried double emulsion, Nahrung/Food, 45(2):133-137. |
| Effenhauser, 1993, Glass chips for high-speed capillary electrophoresis separations with submicrometer plate heights, Anal Chem 65:2637-2642. |
| Eggers, 1999, Coalescence of Liquid Drops, J Fluid Mech 401:293-310. |
| Ehrig, 1995, Green-fluorescent protein mutants with altered fluorescence excitation spectra, Febs Lett, 367(2):163-66. |
| Eigen, 1980, Hypercycles and compartments: compartments assists—but does not replace—hypercyclic organization of early genetic information, J Theor Biol, 85:407-11. |
| Elghanian, 1997, Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles, Science, 277(5329):1078-1080. |
| Ellington, 1990, In vitro selection of RNA molecules that bind specific ligands, Nature, 346:818-822. |
| Xing, 2011, Novel structurally related compounds reactivate latent HIV-1 in a bcl-2-transduced primary CD4+ T cell model without inducing global T cell activation, Journal of Antimicrobial Chemotherapy, 67(2):398-403. |
| Xu, 2005, Generation of monodisperse particles by using microfluidics: control over size, shape, and composition, Angew. Chem. Int. Ed. 44:724-728. |
| Xu, 2009, Design of 240, 000 orthogonal 25mer DNA barcode probes, PNAS, 106(7) p. 2289-2294. |
| Yamagishi, 1990, Mutational analysis of structure-activity relationships in human tumor necrosis factor-alpha, Protein Eng, 3:713-9. |
| Yamaguchi, 2002, Insulin-loaded biodegradable PLGA microcapsules: initial burst release controlled by hydrophilic additives, Journal of Controlled Release, 81(3): 235-249. |
| Yelamos, 1995, Targeting of non-Ig sequences in place of the V segment by somatic hypermutation. Nature 376(6537):225-9. |
| Yershov, 1996, DNA analysis and diagnostics on oligonucleotide microchips, PNAS 93(10):4913-4918. |
| Yonezawa, 2003, DNA display for in vitro selection of diverse peptide libraries, Nucleic Acids Research, 31(19): e118, 5 pages. |
| Yu, 1997, Specific inhibition of PCR by non-extendable oligonucleotides using a 5′ to 3′ exonuclease-deficient DNA polymerase, Biotechniques 23(4):714-6, 718-20. |
| Yu, 2001, Responsive biomimetic hydrogel valve for microfluidics. Appl. Phys. Lett 78:2589-2591. |
| Yu, 2002, Environmental Carcinogenic Polycyclic Aromatic Hydrocarbons: Photochemisrty and Phototoxicity, J Environ Scie Health C Environ Carcinog Exotoxicol Rev, 20(2), 1-43. |
| Yu, 2007, Quantum dot and silica nanoparticle doped polymer optical fibers, Optics Express 15(16):9989-9994. |
| Zaccolo, 1996, An approach to random mutagenesis of DNA using mixtures of triphosphate derivatives of nucleoside analogues. J Mol Biol 255(4):589-603. |
| Zakrzewski, 1980, Preparation of tritiated dihydrofolic acid of high specific activity, Methods Enzymol, 529-533. |
| Zaug, 1986, The intervening sequence RNA of Tetrahymena is an enzyme, Science 231(4737):470-5. |
| Zaug, 1986, The Tetrahymena intervening sequence ribonucleic acid enzyme is a phosphotransferase and an acid phosphatase, Biochemistry 25(16):4478-82. |
| Zaug, 1986, The Tetrahymena ribozyme acts like an RNA restriction endonuclease, Nature 324(6096):429-33. |
| Zhang, 1993, Substrate specificity of the protein tyrosine phosphatases, PNAS 90: 4446-4450. |
| Zhang, 1999, A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening—Assays, Journal of Biomolecular Screening, 4(2): 67-73. |
| Zhao, 1998, Molecular evolution by staggered extension process (StEP) in vitro recombination. Nat Biotechnol 16(3):258-61. |
| Zhao, 2002, Control and Applications of Immiscible Liquids in Microchannels, J. Am. Chem. Soc, vol. 124:5284-5285. |
| Zheng, 2003, Screening of Protein Crystallization Conditions on a Microfluidic Chip Using Nanoliter-Size Droplets, J Am Chem Soc 125(37):11170-11171. |
| Zheng, 2004, A Droplet-Based, Composite PDMS/Glass Capillary Microfluidic System for Evaluating Protein Crystallization Conditions by Microbatch and Vapor-Diffusion Methods with On-Chip X-Ray Diffraction, Angew. Chem., 116:1-4. |
| Zheng, 2004, Formation of Droplets of Alternating Composition in Microfluidic Channels and Applications to Indexing of Concentrations in Droplet-Based /Assays, Anal. Chem.,76: 4977-4982. |
| Zheng, 2005, A Microiuidic Approach for Screening Submicroliter Volumes against Multiple Reagents by Using Performed Arrays of Nanoliter Plugs in a Three-Phase Liquid/Liquid/Gas Flow, Angew. Chem. Int. Ed., 44(17): 2520-2523. |
| Zhong, 2011 Multiplex digital PCR: breaking the one target per color barrier of quantitative PCR, Lab on a Chip 11(13):2167-2174. |
| Zimmermann, 1974, Dielectric Breakdown of Cell Membranes, Biophys J 14(11):881-889. |
| Zimmermann, 1992, Microscale Production of Hybridomas by Hypo-Osmolar Electrofusion, Hum. Antibod. Hybridomas, 3(1): 14-18. |
| Zimmermann, 2008, Digital PCR: a powerful new tool for noninvasive prenatal diagnosis?, Prenat Diagn 28, 1087-1093. |
| Zubay, 1973, In vitro synthesis of protein in microbial systems, Annu Rev Genet, 7: 267-87. |
| Zubay, 1980, The isolation and properties of CAP, the catabolite gene activator, Methods Enzymol, 65: 856-77. |
| Zuckermann, 1987, Efficient Methods for Attachment of Thiol-Specific Probes to the 3-end of Synthetic Oligodeoxyribonucleotides, Nucleic Acids Res. 15:5305-5321. |
| Pelham, 1976, An efficient mRNA-dependent translation system from reticulocyte lysates, Eur J Biochem 67:247-56. |
| Pelletier, 1999, An in vivo library-versus-library selection of optimized protein-protein interactions, Nature Biotechnology, 17:683-90. |
| Peng, 1998, Controlled Production of Emulsions Using a Crossflow Membrane, Particle & Particle Systems Characterization 15:21-25. |
| Pepe, 2004, Limitations of the odds ratio in gauging the performance of a diagnostic, prognostic, or screening marker, American Journal of Epidemiology 159(9):882-890. |
| Perelson, 1979, Theorectical studies of clonal selection: minimal antibody repertoire size and relaibility of self-non-self discrimination. J Theor Biol 81(4):645-70. |
| Perez-Gilabert, 1992, Application of active-phase plot to the kinetic analysis of lipoxygenase in reverse micelles, Biochemistry J. 288:1011-1015. |
| Petrounia, 2000, Designed evolution of enzymatic properties, Curr Opin Biotechnol, 11:325-330. |
| Pirrung, 1996, A General Method for the Spatially Defined Immobilization of Biomolecules on Glass Surfaces Using Caged' Biotin, Bioconjug Chem 7: 317-321. |
| Ploem, 1993, in Fluorescent and Luminescent Probes for Biological Activity Mason, T. G. Ed., Academic Press, Landon, pp. 1-11. |
| Pluckthun, 2000, In vitro selection and evolution of proteins, Adv Protein Chem, 55: 367-403. |
| Pollack, 1986, Selective chemical catalysis by an antibody, Science 234(4783):1570-3. |
| Pollack, 2002, Electrowetting-based actuation of droplets for integrated microfluidics, Lab Chip 2:96-101. |
| Pons, 2009, Synthesis of Near-Infrared-Emitting, Water-Soluble CdTeSe/CdZnS Core/Shell Quantum Dots, Chemistry of Materials 21(8):1418-1424. |
| Posner, 1996, Engineering specificity for folate into dihydrofolate reductase from Escherichia coli, Biochemistry, 35: 1653-63. |
| Priest, 2006, Generation of Monodisperse Gel Emulsions in a Microfluidic Device, Applied Physics Letters, 88:024106, 3 pages. |
| Qi, 1998, Acid Beta-Glucosidase: Intrinsic Fluorescence and Conformational Changes Induced by Phospholipids and Saposin C, Biochem., 37(33): 11544-11554. |
| Raghuraman, 1994, Emulston Liquid Membranes for Wastewater Treatment: Equillibrium Models for Some Typical Metal-Extractant Systems,Environ. Sci. Technol 28:1090-1098. |
| Ralhan, 2008, Discovery and Verification of Head-and-neck Cancer Biomarkers by Differential Protein Expression Analysis Using iTRAQ Labeling, Multidimensional Liquid Chromatography, and Tandem Mass Spectrometry, Mol Cell Proteomics 7(6):1162-1173. |
| Ramanan, 2016, Algae-bacteria interactions, Biotech ADv 34:14-29. |
| Ramsey, 1999, The burgeoning power of the shrinking laboratory, Nat Biotechnol 17(11):1061-2. |
| Ramstrom, 2002, Drug discovery by dynamic combinatorial libraries, Nat Rev Drug Discov 1:26-36. |
| Rasmussen, 2013, Comparison of HDAC inhibitors in clinical development, Human Vacc Immunother 9(5):993-1001. |
| Raushel, 2000, Phosphotriesterase: an enzyme in search of its natural substrate, Adv Enzymol Relat Areas Mol Biol, 74: 51-93. |
| Rech, 1990, Introduction of a yeast artificial chromosome vector into Sarrachomyeces cervesia by electroporation, Nucleic Acids Res 18:1313. |
| Reyes, 2002, Micro Total Analysis Systems. 1. Introduction, Theory and Technology, Anal Chem 74(12):2623-2636. |
| Riechmann, 1988, Reshaping human antibodies for therapy, Nature, 332:323-327. |
| Riess, 2002, Fluorous micro- and nanophases with a biomedical perspective, Tetrahedron 58(20):4113-4131. |
| Roach, 2005, Controlling nonspecific protein adsorption in a plug-based microfluidic system by controlling inteifacial chemistry using fluorous-phase surfactants, Anal. Chem. 77:785-796. |
| Roberts, 1969, Termination factor for RNA synthesis, Nature, 224: 1168-74. |
| Roberts, 1975, Simian virus 40 DNA directs synthesis of authentic viral polypeptides in a linked transcription-translation cell-free system 72(5):1922-1926. |
| Roberts, 1997, RNA-peptide fusion for the in vitro selection of peptides and proteins, PNAS 94:12297-302. |
| Roberts, 1999, In vitro selection of nucleic acids and proteins: What are we learning, Curr Opin Struct Biol 9(4): 521-9. |
| Roberts, 1999, Totally in vitro protein selection using mRNA-protein fusions and ribosome display. Curr Opin Chem Biol 3(3), 268-73. |
| Roche, 2011, 454 Sequencing System Guidelines for Amplicon Experimental Design, 50 pages. |
| Rodriguez-Antona, 2000, Quantitative RT-PCR measurement of human cytochrome P-450s: application to drug induction studies. Arch. Biochem_ Biophys., 376:109-116. |
| Rolland, 1985, Fluorescence Polarization Assay by Flow Cytometry, J. Immunol. Meth., 76(1): 1-10. |
| Rosenberg, 1975, Inhibition of Human Factor IX by Human Antithrombin, J Biol Chem, 250: 4755-64. |
| Rosenberry, 1975, Acetylcholinesterase, Adv Enzymol Relat Areas Mol Biol, 43: 103-218. |
| Rotman, 1961, Measurement of activities of single molecules of beta-galactosidase, PNAS, 47:1981-91. |
| Rouzioux, 2013, How to best measure HIV reservoirs, Curr Op HIV AIDS 8(3):170-175. |
| Russon et al., Single-nucleotide polymorphism analysis by allele-specific extension of fluorescently labeled nucleotides in a microfluidic flow-through device, Electrophoresis, 24:158-61 (2003). |
| Saarela, 2006, Re-usable multi-inlet PDMS fluidic connector, Sensors Actuators B 114(1):552-57. |
| Sadtler, 1996, Achieving stable, reverse water-in-fluorocarbon emulsions, Angew Chem Int Ed 35(17):1976-1978. |
| Sadtler, 1999, Reverse water-In-fluorocarbon emulsions as a drug delivery system: an in vitro study, Colloids & Surfaces A: Phys Eng Asp 147:309-315. |
| Saiki, 1988, Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase, Science 239(4839):487-91. |
| Sakamoto, 2005, Rapid and simple quantification of bacterial cells by using a microfluidic device, Appl Env Microb 71:2. |
| Sano, 1992, Immuno-PCR: very sensitive antigen-detection by means of sepcific Ab-DNA conjugates, Science 258(5079):120-122. |
| SantaLucia, 1998, A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics, PNAS 95(4):1460-5. |
| Santra, 2006, Fluorescence lifetime measurements to determine the core-shell nanostructure of FITC-doped silica nanoparticles, J Luminescence 117(1):75-82. |
| Sawada, 1996, Synthesis and surfactant properties of novel fluoroalkylated amphiphilic oligomers, Chem Commun 2:179-190. |
| Laird, 2013, Rapid Quantification of the Latent Reservoir for HIV-1 Using a Viral Outgrowth Assay, PLOS PATHOGENS 9(5):e1003398. |
| Lamprecht, 2004, pH-sensitive microsphere delivery increases oral bioavailability of calcitonin, J Control Rel 98(1):1-9. |
| Lancet, 1993, Probability model for molecular recognition in biological receptor repertoirs, PNAS 90(8):3715-9. |
| Landergren, 1988, A ligase mediated gene detection technique, Science 241(4869):1077-80. |
| Lasheras, 1998, Breakup and atomization of a round water jet by a high speed annular air jet, J Fluid Mech 357:351-379. |
| Laufer, 1996, Introduction to Optics and Lasers in Engineering, Cambridge University Press, Cambridge UK:156-162. |
| Leamon, 2003, A massively parallel pictoterplate based platform for discrete picoliter-scale PCR, Electrophoresis 24:3769-3777. |
| Leary, 2000, Application of advanced cytometric and molecular technologies to minimal residual disease monitoring, Proc SPIE 3913:36-44. |
| Lee, 2000, Circulating flows inside a drop under time-periodic non-uniform electric fields, Phys Fuilds 12(8):1899-1910. |
| Lee, 2001, Preparation of silica particles encapsulating retinol using O/W/O multiple emulsions, J Coll Interface Sci 240(1):11:83-89. |
| Lee, 2002, Effective formation of silicone-in-fluorocarbon-in-water double emulsions, J Disp Sci Tech 23(4):491-497. |
| Lee 2002, Investigating the target recognition of DNA cytosine-5 methyltransferase Hhal by library selection using in vitro compartmentalisation (IVC), Nucleic Acids Res 30:4937-4944. |
| Lee, 2004, Special issue on biomedical applications for MEMS and microfluidics, Proc IEEE 92(1):3-5. |
| Lemof, 2003, An AC magnetohydrodynamic microfluidic switch for Micro Total Analysis Systems, Biomed Microdev 5(1):55-60. |
| Leng 2009, Microfluidic crystalizaiton,Lab Chip 9:24-23. |
| Leng, 2010, Agarose droplet microfluidics for highly parallel and efficient single molecule emulsion PCR, Lab Chip 10:2841-2843. |
| Lesley, 1991, Use of in vitro protein synthesis from PCR-generated templates to study interaction of E. coli transcription factors with core RNA polymerase, J Biol Chem 266(4):2632-8. |
| Lesley, 1995, Preparation and use of E. coli S-30 extracts, Methods Mol Biol 37:265-78. |
| Leung, 1989, A method for random mutagenesis of a defined DNA segment using a modified polymerase chain reaction, Technique 1:11-15. |
| Li, 1995, Single-step procedure for labeling DNA strand breaks with fllourescein-or BODIPY-conjugated deoxynucleotides, Cytometry 20:172-180. |
| Li, 1997, Transport, manipulation, and reaction of biological cells on-chip using electrokinetic effects, Anal Chem 69(8):1564-1568. |
| Li, 2005, Multiplexed detection of pathogen DNA with DNA-based fluorescence nanobarcodes, Nat Biotech 23(7):885-889. |
| Li, 2006, Nanoliter microfluidic hybrid method for simultaneous screening and optimization validated with crystallization of membrane proteins, PNAS 103:19243-19248. |
| Li, 2018, Microfluidic fabrication of microparticles for biomedical applications, Chem Soc Rev 47(15):5646-5683. |
| Liao, 1986, Isolation of a thermostable enzyme variant by cloning and selection in a thermophile, PNAS 83:576-80. |
| Lim, 1980, Microencapsulated islets as bioartificial endocrine pancreas, Science 210(4472):908-10. |
| Lin, 2007, Self-assembled combinatorial encoding nanoarrays for multiplexed biosensing, Nano Lett 7(2):507-512. |
| Link, 2004, Geometrically mediated breakup of drops in microfluidic devices, Phys Rev Lettv92(5):054503-1-4. |
| Link, 2006, Electric control droplets in microfluidic devices, Angew Chem Int Ed 45:2556-2560. |
| Lipinski, 2001, Experimental and computational approaches to estimate solubility and permeability in drug discovery, , Adv Drug Deliv Rev 46:3-26. |
| Lipkin, 1988, Biomarkers of increased susceptibility to gastreointestinal cancer: new application to studies of cancer prevention in human subjects, Cancer Res 48:235-245. |
| Liu, 2000, Passive mixing in a three-dimensional serpentine microchannel, J MEMS 9(2):190-197. |
| Liu, 2002, Fabrication and characterization of hydrogel-based microvalves, Mecoelectromech. Syst.11:45-53. |
| Lizardi, 1998, Mutation detection and single-molecule counting using isothermal rolling-circle amplification. Nat Genet 19(3):225-32. |
| Lo, 2007, Digital PCR for the molecular detection of fetal chromosomal aneuploidy, PNAS 104(32):13116-13121. |
| Loakes, 1994, 5-Nitroindole as a universal base analogue, Nucleic Acids Res 22:4039-4043. |
| Loakes, 1997, Stability and structure of DNA oligonucleotides containing non-specific base analogues, J Mol Biol 270:426-435. |
| Lodish, 2000, Structure of Nucleic Acids, Section 4.1 , Molecular Cell Biology, 4th edition, New York, 1-3. |
| Loeker, 2003, FTIR analysis of water in supercritical carbon dioxide microemulsions using monofunctional perfluoropolyether surfanctants, Colloids and Surfaces A: Phys Eng Asp 214:143-150. |
| Loo, 2004, Nanoshell Enabled Photonics-Based Imaging and Therapy of Cancer, Technology in Cancer Research & Treatment 3(1):33-40. |
| Lopez-Herrera, 1995, The electrospraying of viscous and non-viscous semi-insulating liquids: scaling laws, Bull Am Phys Soc 40 (12):2041. |
| Lopez-Herrera, 1999, One-dimensional simulation of the breakup of capillary jets of conducting liquids application to EHD spraying, Aerosol Set 30(7):895-912. |
| Lopez-Herrera, 2003, Coaxial jets generated from electrified Taylor cones, Aerosol Sci 34:535-552. |
| Lorenceau, 2005, Generation of polymerosomes from double-emulsions, Langmuir 21(20):9183-9186. |
| Lorenz, 1991, Isolation and expression of a cDNA encoding Renilla reniformis luciferase, PNAS 88(10):4438-42. |
| Loscertales, 2002, Micro/nano encapsulation via electrified coaxial liquid jets, Science 295(5560):1695-1698. |
| Lowe, 2002, Perfluorochemical respiratory gas carriers: benefits to cell culture systems, J Fluorine Chem 118:19-26. |
| Lu, 2007, Robust fluorescein-doped silica nanoparticles via dense-liquid treatment, Colloids and Surfaces A Phys Eng Asp 303(3):207-210. |
| Luisi, 1987, Activity and conformation of enzymes in reverse micellar solutions, Meth Enzymol 136:188-216. |
| Lund, 1988, Assesment of methods for covalent binding of nucleic acids to magnetic beads, Dynabeads, and the characteristics of the bound nucleic acids in hybridization reactions, Nucleic Acids Res 16(22):10861-10880. |
| Moudrianakis, 1965, Base sequence determination in nucelic acids with the electron microscope 3. Chemistry and microscopy of guanine-labeled DNA, PNAS 53:564-71. |
| Mueth, 1996, Origin of stratification in creaming emulsions, Physical Review Letters 77(3):578-581. |
| Mulbry, 1989, Parathion hydrolase specified by the Flavobacterium opd gene: relationshio between the gene and protein. J Bacteriol, 171: 6740-6746. |
| Mulder, 1993, Characterization of two human monoclonal antibodies reactive with HLA-B12 and HLA-B60, respectively, raised by in vitro secondary immunization of peripheral blood lymphocytes, Hum. Immunol 36(3):186-192. |
| Munson, 1980, Ligand: a versatile computerized approach for characterization of ligand-binding systems, Analytical Biochemistry, 107:220-239. |
| Nakano, 1994, High speed polymerase chain reaction in constant flow, Biosci Biotech and Biochem, 58:349-52. |
| Nakano, 2003, Single-molecule PCR using water-in-oil emulsion, J Biotech, 102:117-124. |
| Nakano, 2005, Single-molecule reverse transcription polymerase chain reaction using water-in-oil emulsion, J Biosci Bioeng 99:293-295. |
| Nametkin, 1992, Cell-free translation in reversed micelles, FEB Letters, 309(3):330-32. |
| Narang, 1979, Improved phosphotriester method for the synthesis of gene fragments, Methods Enzymol, 68:90-98. |
| Neiman, 2011, Decoding a substantial set of samples in parallel by massive sequencing, PLoS ONE 6(3):1-7. |
| Nelson, 1989, Bifunctional oligonucleotide probes synthesized using a novel CPG support are able to detect single base pair mutations, Nucl Acids Res 17(18): 7187-7194. |
| Nemoto, 1997, In vitro virus: bonding of mRNA bearing puromycin at the 3 terminal end to the C-terminal end of its encoded protein on the ribosome in vitro, Federation of European Biochemical Societies, 414:405-8. |
| Ness, 2000, Molecular Breeding: the natural approach to protein design. Adv Protein Chem, 55: 261-292. |
| Ng, 2003, Protein crystallization by capillary counter-diffusion for applied crystallographic structure determination, J. Struct. Biol, 142:218-231. |
| Ng, 2006, Factors affecting flow karyotype resolution, Cytometry, Part A 69A: 1028-1036. |
| Nguyen, 2006, Optical detection for droplet size control in microfluidic droplet-based analysis systems, Sensors and Actuators B 117(2):431-436. |
| Nihant, 1994, Polylactide Microparticles Prepared by Double Emulsion/Evaporation Technique. I. Effect of Primary Emulsion Stability, Pharmaceutical Research, 11(10):1479-1484. |
| Nisisako, 2002, Droplet formation in a microchannel network, Lab Chip 2:24-26. |
| Nisisako, 2002, Formation of droplets using branch channels in a microfluidic circuit, Proceedings of the SICE Annual Conference. International Session Papers 1262-1264. |
| Nisisako, 2005, Controlled formulation of monodisperse double emulsions in a multiple-phase microluidic system, Sot Matter, 1:23-27. |
| Nisisako, 2008, Microstructured Devices for Preparing Controlled Multiple Emulsions. Chem. Eng. Technol 31(8):1091-1098. |
| Nof 2002, Drug-releasing scaffolds fabricated from drug-loaded microspheres, J. Biomed Mater Res 59:349-356. |
| Norman, 1980, Flow Cytometry, Med. Phys., 7(6):609-615. |
| Nygren, 1982, Conjugation of horseradish peroxidase to Fab fragments with different homobifunctional and heterobifunctional cross-linking reagents. A comparative study, J. Histochem. and Cytochem. 30:407-412. |
| Oberholzer, 1995, Enzymatic RNA replication in self-reproducing vesicles: an approach to a minimal cell, Biochem Biophys Res Commun 207(1):250-7. |
| Oberholzer, 1995, Polymerase chain reaction in liposomes, Chem. Biol. 2(10):677-82. |
| Obukowicz, 1988, Secretion and export of IGF-1 in Escerichia coli strain JM101, Mol Gen Genet, 215:19-25. |
| Ogura, 1955, Catalase activity at high concentrations of hydrogen peroxide, Archs Biochem Biophys, 57: 288-300. |
| Oh, 2002, Distribution of Macropores in Silica Particles Prepared by Using Multiple Emulsions, Journal of Colloid and Interface Science, 254(1): 79-86. |
| Oh, 2005, World-to-chip microfluidic interface with built-in valves for multichamber chip-based PCR assays, Lab Chip, 5, 845-850. |
| Okuno, 2003, Recent Advances in Optical Switches Using Silica-based PLC Technology, NTT Technical Review 1(7):20-30. |
| Okushima, 2004, Controlled production of monodisperse double emulsions by two-step droplet breakup in microfluidic devices, Langmuir 20(23): 9905-8. |
| Olsen, 2000, Function-based isolation of novel enzymes from a large library, Nat Bioteoltnol 13(10):1071-4. |
| Omburo, 1992, Characterization of the zinc binding site of bacterial phosphotriesterase, J of Biological Chem, 267:13278-83. |
| Oroskar, 1996, Detection of immobilized amplicons by ELISA-like techniques, Clin. Chem. 42:1547-1555. |
| Ostermeier, 1999, A combinatorial approach to hybrid enzymes independent of DNA homology, Nat Biotechnol, 17(12):1205-9. |
| Ott, 1967, Biological and medical research annual report, Los Alamos Scientific Laboratory, 14 pages. |
| Ouelette, 2003, A new wave of microfluidic devices, Indust Physicist pp. 14-17. |
| Pabit, 2002, Laminar-Flow Fluid Mixer for Fast Fluorescence Kinetics Studies, Biophys J 83:2872-2878. |
| Paddison, 2002, Stable suppression of gene expression by RNAi in mammalian cells, PNAS 99(3):1443-1448. |
| Pain, 1981, Preparation of protein A-peroxidase mono conjugate using a heterobifunctional reagent, and its use in enzyme immunoassays, J Immunol Methods, 40:219-30. |
| Pannacci, 2008, Equilibrium and Nonequilibrium States in Microluidic Double Emulsions Physical Review Leters, 101(16):164502. |
| Park, 2001, Model of Formation of Monodispersed Colloids, J. Phys. Chem. B 105:11630-11635. |
| Park, 2003, Cylindrical compact thermal-cycling device for continuous-flow polymeras chain reaction, Anal Chem, ACS, 75:6029-33. |
| Parker, 2000, Development of high throughput screening assays using fluorescence polarization: nuclear receptor-ligand-binding and kinase/phosphatase assays, J Biomol Screen, 5(2): 77-88. |
| Pasternak, 2013, Cell-associated HIV RNA: a dynmic biomarker of viral persistence, Retrovirology 10:41. |
| Patel, 2003, Formation of Fluorinated Nonionic Surfactant Microemulsions in Hydrfuorocarbon 134a, Journal of Colloid and Interface Science, 258, 345-353. |
| Pedersen, 1998, A method for directed evolution and functional cloning of enzymes, PNAS 95(18):10523-8. |
| Pekin, 2011, Quantitative and sensitive detection of rare mutations using droplet-based microfluidics, Lab on a Chip 11(13):2156-2166. |
| Schatz, 1996, Screening of peptide libraries linked to lac repressor, Meth Enzymol 267:171-91. |
| Schneegass, 2001, Miniaturized flow-through PCR with different template types in a silicone chip thermocycler, Lab on a Chip 1:42-9. |
| Schopman, 2012, Selective packaging of cellular miRNAs in HIV-1 particles, Virus Res 169(2):438-47. |
| Schubert, 2002, Designer Capsules, Nat Med 8:1362. |
| Schweitzer, 2000, Immunoassays with rolling circle DNA amplification, PNAS 97(18):10113-10119. |
| Schweitzer, 2001, Combining nucleic acid amplification and detection. Curr Opin Biotechnol 12(1):21-7. |
| Scott, 1948, The solubility of fluorocarbons, J Am Chem Soc 70:4090-4093. |
| Sedlak, 2013, Viral diagnostics in the era of digital polymerase chain reaction, Diag Microb Inf Dis 75(1):1-4. |
| Seethala, 1997, Homogeneous fluorescence polarization assay for Src-Family tyrosine kinases, Anal Biochem 253(2):210-218. |
| Seiler, 1993, Planar glass chips for capillary electrophoresis: repetitive sample injection, quantitation, and separation efficiency, Anal Chem 65(10):1481-1488. |
| Selwyn, 1965, A simple test for inactivation of an enzyme during assay, Biochim Biophys Acta 105:193-195. |
| Sen, 2012, Development and validation of a new adenosine-independent index of stenosis severity from coronary wave-intensity analysis, Journal of the American College of Cardiology 59(15):1392-1402. |
| Seo, 2007, Microfluidic consecutive flow-focusing droplet generators, Soft Matter 3:986-992. |
| Seong, 2002, Efficient mixing and reactions within microfluidic channels using microbead-supported catalysts, J Am Chem Soc 124(45):13360-1. |
| Seong, 2002, Fabrication of microchambers defined by photopolymerized hydrogels and weirs within microfluidic systems, Anal Chem 74(14):3372-3377. |
| Sepp, 2002, Microbead display by in vitro compartmentalisation: selection for binding using flow cytometry, FEBS Letters 532:455-58. |
| Serpersu, 1985, Reversible and irreversible modification of erythrocyte membrane permeability by electric field, Biochim Biophys Acta 812(3):779-785. |
| Shapiro, 1983, Multistation multiparameter flow cytometry: a critical review and rationale, Cytometry 3: 227-243. |
| Shastry, 2006, Directing droplets using microstructured surfaces, Langmuir 22:6161-6167. |
| Shen, 2006, Eigengene-based linear discriminant model for tumor classification using gene expression microarray data, Bioinformatics 22(21):2635-2642. |
| Shestopalov, 2004, Multi-step synthesis of nanoparticles performed on millisecond time scale in a microfluidic droplet-based system, Royal Soc Chem 4:316-321. |
| Shim, 2007, Using microfluidics to decouple nucleation and growth of protein crystals, Cryst Growth Des 7(11):2192-2194. |
| Shimizu, 1995, Encapsulation of biologically active proteins in a multiple emulsion, Biosci Biotech Biochem 59(3):492-496. |
| Shtern, 1996, Hysteresis in swirling jets, J Fluid Mech 309:1-44. |
| Sia, 2003, Microfluidic devices fabricated in poly(dimethylsiloxane) for biological studies, Electrophoresis 24(21):3563-3576. |
| Siemering, 1996, Mutations that suppress the thermosensitivity of green fluorescent protein, Curr Biol 6:1653-1663. |
| Silva-Cunha, 1998, W/O/W multiple emulsions of insulin containing a protease inhibitor and an absorption enhancer: biological activity after oral administration to normal and diabetic rats, Int J Pharm 169:33-44. |
| Sims, 2000, Immunopolymerase chain reaction using real-time polymerase chain reaction for detection, Anal. Biochem. 281(2):230-2. |
| Sista, 2007, Development of a Digital Microfluidic Lab-on-a-Chip for Automated Immunoassay with Magnetically Responsive Beads, Doctoral Thesis, Florida State University, 128 pages. |
| Sista, 2008, Development of a digital microfluidic platform for point care testing, Lab on a Chip 8:2091-2104. |
| Siwy, 2003, Electro-responsive asymmetric nanopores in polyimide with stable ion-current signal, Appl Phys A: Mat Sci Proc 76:781-785. |
| Slappendel, 1994, Normal cations and abnormal membrane lipids in the red blood cells of dogs with familial stomatocytosis hypertrophic gastritis, Blood 84:904-909. |
| Slob, 1997, Structural identifiability of PBPK models: practical consequences for modeling strategies and study designs, Crit Rev Toxicol. 27(3):261-72. |
| Smith, 1985, The synthesis of oligonucleotides containing an aliphatic amino group at the 5′ terminus: synthesis of fluorescent DNA primers for use in DNA sequence analysis, Nucl Acid Res 13:2399-2412. |
| Smith, 1986, Fluorescence detection in automated DNA sequence analysis, Nature 321:674-679. |
| Smith, 1989 Absolute displacement measurements using modulation of the spectrum of white light in a Michelson interferometer, Applied Optics, 28(16):3339-3342. |
| Smith, 1992, Direct mechanical measurements of the elasticity of single DNA molecules by using magnetic beads, Science 258(5085):1122-1126. |
| Smith, 2010, Highly-multiplexed barcode sequencing: an efficient method for parallel analysis of pooled samples, Nucleic Acids Res 38(13):e142. |
| Smyth, 2000, Markers of apoptosis: methods for elucidating the mechanism of apoptotic cell death from the nervous system, Biotechniques 32:648-665. |
| Sohn, 2000, Capacitance cytometry: Measuring biological cells one by one, PNAS 97(20):10687-10690. |
| Sola, 2014, Fabrication of a microfluidic cell made of thiolene for microarray applications, 18th Int Conf Miniaturized Systems for Chem and Life Sciences, MicroTAS, San Antonio, TX 1719-1721. |
| Somasundaram, 1999, Gain studies of Rhodamine 6G dye doped polymer laser, J Photochem Photobiol 125(1-3):93-98. |
| Song, 2002, Experimental test of scaling of mixing by chaotic advection in droplets moving through microfluidic channels, App Phy Lett 83(22):4664-4666. |
| Song, 2003, A microfluidic system for controlling reaction networks in time, Angew Chem Int Ed 42(7):768-772. |
| Song, 2003, Millisecond kinetics on a microluidic chip using nanoliters of reagents, J Am Chem Soc 125:14613-14619. |
| Song, 2006, Reactions in droplets in microfluidic channels, Angew chem Int ed 45(44):7336-7356. |
| Soni, 2007, Progress toward ultrafast DNA sequencing using solid-state nanopores, Clin Chem 53:1996-2001. |
| Soumillion, 2001, Novel concepts for the selection of catalytic activity. Curr Op Biotech 12:387-394. |
| Spiro, 2000, A bead-based method for multiplexed identification and quantitation of DNA sequences using flow cytometry, Appl Env Micro 66:4258-4265. |
| Sproat, 1987, The synthesis of protected 5′-mercapto-2′,5′-dideoxyribonucleoside-3′-0-phosphorainidites, uses of 5′- mercapto-oligodeoxyribonucleotides, Nucleic Acids Res 15:4837-4848. |
| Heyries, 2011, Megapixel digital PCR, Nat. Methods 8, 649-651. |
| Hildebrand, 1949, Liquid-Liquid Solubility of Perfluoromethylcyclohexane with Benzene, Carbon Tetrachloride, Chlorobenzene, Chloroform and Toluene, J. Am. Chem. Soc, 71:22-25. |
| Hindson, 2011, High-Throughput Droplet Digital PCR System for Absolute Quantitation of DNA Copy Number, Anal. Chem., 83, 8604-8610. |
| Hjelmfelt, 1993, Pattern-Recognition in Coupled Chemical Kinetic Systems, Science, 260(5106):335-337. |
| Ho, 1989, Site-directed mutageneiss by overlap extension using the polymerase chain reaction, Gene, 77(1):51-9. |
| Hochuli, 1987, New metal chelate adsorbent selective for proteins and peptides containing neighbouring histidine residues, J Chromatogr 411: 177-84. |
| Holmes, 1995, Reagents for Combinatorial Organic Synthesis: Development of a New O-Nitrobenzyl Photolabile Linder for Solid Phase Synthesis, J. OrgChem., 60: 2318-2319. |
| Holtze, 2008, Biocompatible surfactants for water-in-fluorocarbon emulsions, Lab Chip, 8, 1632-1639. |
| Hong, 1999, Stereochemical constraints on the substrate specificity of phosphodiesterase, Biochemistry, 38:1159-1165. |
| Hoogenboom, 1997, Designing and optimizing library selection strategies for generating high-affinity antibodies, Trends Biotechnol, 15:62-70. |
| Hopfinger, 1996, Explosive Breakup of a Liquid Jet by a Swirling Coaxial Jet, Physics of Fluids 8(7):1696-1700. |
| Hopman, 1998, Rapid synthesis of biotin-, digoxigenin-, trinitrophenyl-, and fluorochrome-labeled tyramides and their application for in situ hybridization using CARD amplification, J of Histochem and Cytochem, 46(6):771-77. |
| Horton, 1989, Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension, Gene 77(1):61-8. |
| Hosokawa, 1999, Handling of Picoliter Liquid Samples in a Poly(dimethylsiloxane)-Based Microfluidic Device, Analytical Chemistry, 71(20):4781-4785. |
| Hsieh, 2009, Rapid label-free DNA analysis in picoliter microfluidic droplets using FRET probes, Microfluidics and nanofluidics 6(3):391-401. |
| Hsu, 1999, et al., Comparison of process parameters for microencapsulation of plasmid DNA in poly(D, L-lactic-co-glycolic acid microspheres, J Drug Target, 7:313-23. |
| Hua, 2010, Multiplexed Real-Time Polymerase Chain Reaction on a Digital Microfluidic Platform, Analytical Chemistry 32(6):2310-2316. |
| Huang, 1991, Kinetic assay of fluorescein mono-beta-D-galactosidase hydrolysis by beta-galactosidase: a front-face measurement for strongly absorbing fluorogenic substrates, Biochemistry, 30:8530-4. |
| Huang, 1992, A sensitive competitive ELISA for 2,4-dinitrophenol using 3,6-fluorescein diphosphate as a fluorogenic substrate, J Immunol Meth, 149:261. |
| Huang, 2004, Continuous particle separation through deterministic lateral displacement, Science 304(5673):987-990. |
| Huang, 2007, Identification of 8 foodborne pathogens by multicolor combinational probe coding technology in a single real-time PCR, Clin Chem., 53(10):1741-8. |
| Hubert 2003, Data Concordance from a Comparison between Filter Binding and Fluorescence Polarization Assay Formats for Identification of RUOCK-II Inhibitors, J biomol Screen 8(4):399-409. |
| Huebner, 2007, Quantitative detection of protein expression in single cells using droplet microfluidics, Chem Com 12:1218-1220. |
| Hug, 2003, Measurement of the number of molecules of a single mRNA species in a complex mRNA preparation. J Theor Biol.; 221(4):615-24. |
| Hung, 2004, Controlled Droplet Fusion in Microfluidic Devices, MicroTAS 2004, Sep. 26-30, Malmo, Sweden. |
| Hung, 2004, Optimization of Droplet Generation by controlling PDMS Surface Hydrophobicity, 2004 ASME International Mechanical Engineering Congress and RD&D Expo, Nov. 13-19, Anaheim, CA, 47-48. |
| Hutchison, 2005, Cell-free cloning using Phi29 polymerase, PNAS 102(48):17332-17336. |
| Ibrahim, 2003, High-speed cell sorting: fundamentals and recent advances, Curr Opin Biotchnol, 14(1):5-12. |
| Ikeda, 2000, Bioactivation of tegafur to 5-fluorouracil is catalyzed by cytochrome P-450 2A6 in human liver microsomes in vitro, Clin Cancer Res 6(11):4409-4415. |
| Illumina, 2010, Genomic Sequencing, data Sheet, 6 pages. |
| Inai, 1993, Immunohistochemical detection of an enamel protein-related epitope in rat bone at an early stage of osteogenesis, Histochemistry 99(5):335-362. |
| Invitrogen, 2008, Specification sheet for Dynabeads® Oligo (dT)25, http://www.invitrogen.com, 2 pages. |
| Ismagilov, 2003, Integrated Microfluidic Systems, Angew. Chem. Int. Ed 42:4130-4132. |
| Jakobovits, 1993, Analysis of homozygous mutant chimeric mice:deletion of the immunoglobulin heavy-chain joining region blocks B-cell development and antibody production, PNAS USA 90:2551-255. |
| Jakobovits, 1993, Germ-line transmission and expression of a human-derived yeast artificial chromosome, Nature 362:255-258. |
| Janda, 1997, Chemical selection for catalysis in combinatorial antibody libraries, Science, 275:945-948. |
| Jang, 2003, Controllable delivery of non-viral DNA from porous scaffold, J Controlled Release 86(1):157-168. |
| Jarvie, 2007, Amplicon Sequencing, Roche Dx Application Note No. 5 (16 pages). |
| Jermutus, 1998, et al., Recent advances in producing and selecting functional proteins by using cell-free translation, Curr Opin Biotechnol 9(5): 534-48. |
| Jo, 2003, Encapsulation of Bovine Serum Albumin in Temperature-Programmed Shell-in-Shell Structures, Macromol. Rapid Comm 24:957-962. |
| Joerger, 1995, Analyte detection with DNA-labeled antibodies and polymerase chain reaction, Clin. Chem. 41(9):1371-7. |
| Johannsson, 1988, Amplification by Second Enzymes, In ELISA and Other Solid Phase Immunoassays, Kemeny et al (ed.), Chapter 4, pp. 85-106 John Wiley. |
| Johannsson, 1991, Heterogeneous Enzyme Immunoassays, In Principles and Practice of Immunoassay, pp. 295-325 Stockton Press. |
| Johnson, 1993, Human antibody engineering: Current Opinion in Structural Biology, 3:564-571. |
| Johnson, 2002, Protein tyrosine phosphatase 1B inhibitors for diabetes, Nature Review Drug Discovery 1, 696-709. |
| Jones, 1986, Replacing the complementarity-determining regions in a human antibody with those from a mouse, Nature, 321:522-525. |
| Jones, 1997, Quenched BODIPY dye-labeled casein substrates for the assay of protease activity by direct fluorescence measurement, Anal Biochem, 251:144-152. |
| Jones, 1999, Glowing jellyfish, luminescence and a molecule called coelenterazine, Trends Biotechnol. 17(12):477-81. |
| Joo, 1999, Laboratory evolution of peroxide-mediated cytochrome P450 hydroxylaion, Nature 399:670. |
| Joos, 1997, Covalent attachment of hybridizable oligonucleotides to glass supports, Analytical Biochemistry 247:96-101. |
| Branebjerg, 1996, Fast mixing by lamination, MEMS Proc 9th Ann 9:441-446. |
| Braslavsky, 2003, Sequence information can be obtained from single DNA molecules, PNAS 100(7):3960-3964. |
| Breslauer, 2006, Microfluidics based systems biology, Mol Bio Syst 2:97-112. |
| Bringer, 2004, Microfluidic systems for chemical kinetics that rely on chaotic mixing in droplets, Phil Trans A Math Phys Eng Sci 362:1-18. |
| Brown, 1979, Chemical synthesis and cloning of a tyrosine tRNA gene, Methods Enzymol 68:109-151. |
| Bru, 1991, Product inhibition of alpha-chymotrypsin in reverse micelles. Eur J Biochem 199(1):95-103. |
| Bru, 1993, Catalytic activity of elastase in reverse micelles, Biochem Mol Bio Int, 31(4):685-92. |
| Brummelkamp, 2002, A system for stable expression of short interfering RNAs in mammalian cells, Science 296(5567):550-3. |
| Buican, 1987, Automated single-cell manipulation and sorting by light trapping, Appl Optics 26(24):5311-5316. |
| Burbaum, 1998, Miniaturization technologies in HTS, Drug Disc Today 3:313-322. |
| Burns, 1996, Microfabricated structures for integrated DNA analysis, PNAS 93:5556-5561. |
| Burns, 1998, An integrated nanoliter DNA analysis device, Science 282:484-487. |
| Burns, 2002, The intensification of rapid reactions in multiphase systems using slug flow in capillaries, Lab on a Chip 1:10-15. |
| Byrnes, 1982, Sensitive fluorogenic substrates for the detection of trypsin-like proteases and pancreatic elastase, Anal Biochem 126:447. |
| Cahill, 1991, Polymerase chain reaction and Q beta replicase amplification, Clin Chem 37(9):1482-5. |
| Caldwell, 1991, Limits of diffusion in the hydrolysis of substrates by the phosphodiesterase from Pseudomonas diminuta, Biochem 30:7438-7444. |
| Calvert, 2001, Inkjet printing for materials and devices, Chem Mater 13:3299-3305. |
| Caruccio, 2009, Nextura technology for NGS DNA library preparation: simulaneous fragmentation and tagging by in vitro transposition, Epibio Newsletter. |
| Caruthers, 1985, Gene synthesis machines: DNA chemistry and its uses, Science 230:281-285. |
| Cavalli, 2010, Nanosponge formulations as oxygen delivery systems, Int J Pharmaceutics 402:254-257. |
| Chakrabarti, 1994, Production of RNA by a polymerase protein encapsulated within phospholipid vesicles, J Mol Evol 39(6):555-9. |
| Chamberlain, 1973, Characterization of T7-specific ribonucleic acid polymerase. 1. General properties of the enzymatic reaction and the template specificity of the enzyme, J Biol Chem 248:2235-44. |
| Chan, 2003, Size-Controlled Growth of CdSe Nanocrystals in Microfluidic Reactors, Nano Lett 3(2):199-201. |
| Chan, 2008, New trends in immunoassays, Adv Biochem Engin/Biotech 109:123-154. |
| Chang, 1987, Recycling of NAD(P) by multienzyme systems immobilized by microencapsulation in artifical cells, Methods Enzymol, 136(67):67-82. |
| Chang, 2008, Controlled double emulsification utilizing 3D PDMS microchannels, Journal of Micromechanics and Microengineering 18:1-8. |
| Chao, 2004, Control of Concentration and Volume Gradients in Microfluidic Droplet Arrays for Protein Crystallization Screening, 26th Annual International Conference of the IEEE Engineering in Medicine and Biology Society, San Francisco, California Sep. 1-5. |
| Chao, 2004, Droplet Arrays in Microfluidic Channels for Combinatorial Screening Assays, Hilton Head: A Solid State Sensor, Actuator and Microsystems Workshop, Hilton Head Island, South Carolina, Jun. 6-10. |
| Chapman, 1994, In vitro selection of catalytic RNAs, Curr. op. Struct. Biol., 4:618-22. |
| Chayen, 1999, Crystallization with oils: a new dimension in macromolecular crystal growth Journal of Crystal Growth, 196:434-441. |
| Chen, 2001, Capturing a Photoexcited Molecular Structure Through Time-Domain X-ray Absorption Fine Structure, Science 292(5515):262-264. |
| Chen, 2003, Microfluidic Switch for Embryo and Cell Sorting the 12th International Conference on Solid State Sensors, Actuators, and Microsystems, Boston, MA, Transducers, 1: 659-662. |
| Chen-Goodspeed, 2001, Enhancement, relaxation, and reversal of the stereoselectivity for phosphotriesterase by rational evolution of active site residues, Biochemistry, 40: 1332-1339. |
| Chen-Goodspeed, 2001, Structural Determinants of the substrate and stereochemical specificity of phosphotriesterase, Biochemistry, 40(5):1325-31. |
| Cheng, 2003, Electro flow focusing inmicrofluidic devices, Microfluidics Poster, presented at DBAS, Frontiers in Nanoscience, 1 page. |
| Cheng, 2006, Nanotechnologies for biomolecular detection and medical diagnostics, Current Opinion in Chemical Biology, 10:11-19. |
| Chetverin, 1995, Replicable RNA vectors: prospects for cell-free gene amplification, expression, and cloning, Prog Nucleic Acid Res Mol Biol, 51:225-70. |
| Chiang, 1993, Expression and purification of general transcription factors by FLAG epitope-tagging and peptide elution, Pept Res, 6:62-64. |
| Chiba, 1997, Controlled protein delivery from biodegradable tyrosino-containing poly(anhydride-co-imide) microspheres, Biomaterials, 18(13):893-901. |
| Chiou, 2001, A closed-cycle capillary polymerase chain reaction machine, Analytical Chemistry, American Chamical Society, 73:2018-21. |
| Chiu, 1999, Chemical transformations in individual ultrasmall biomimetic containers, Science, 283:1892-1895. |
| Chou, 1998, A microfabricated device for sizing and sorting DNA molecules 96:11-13. |
| Clackson, 1994, In vitro selection from protein and peptide libraries, Trends Biotechnol, 12:173-84. |
| Clausell-Tormos, 2008, Droplet-based microfluidic platforms for the encapsulation and screening of Mammalian cells and multicellular organisms, Chem Biol 15(5):427-437. |
| Cohen, 1991, Controlled delivery systems for proteins based on poly(lactic/glycolic acid) microspheres, Pharm Res, 8(6):713-720. |
| Collins, 2003, Optimization of Shear Driven Droplet Generation in a Microluidic Device, ASME International Mechanical Engineering Congress and R&D Expo, Washington, 4 pages. |
| Collins, 2004, Microlluidic flow transducer based on the measurements of electrical admittance, Lab on a Chip, 4:7-10. |
| Compton, 1991, Nucleic acid sequence-based amplification, Nature, 350(6313):91-2. |
| Cook, 2007, Use and misuse of receiver operating characteristic curve in risk prediction, Circulation 115(7):928-35. |
| Cooper, 2000, The Central Role of Enzymes as Biological Catalysts, The Cell: A Molecular Approach, 2nd Edition, pp. 1-6. |
| Giusti, 1993, Synthesis and characterization of 5′ fluorescent dye labeled oligonucleotides, Genome Res 2:223-227. |
| Glass, 1995, Development of primer sets designed for use with the PCR to amlify conserved genes from filamentous ascomycetes, Applied and Environmental Microbiology, vol. 6, pp. 1323-1330. |
| Gold, 1995, Diversity of Oligonucleotide Functions Annu Rev Biochem, 64: 763-97. |
| Gong, 2015, Simple method to prepare oligonucleotide conjugated antibodies and its applicaiotn in multiplex protein detection in single cells, Bioconjugate Chm 27(1):271-225. |
| Goodall, 1998, Operation of Mixed-Culture Immobilized Cell Reactors for the Metabolism of Meta- and Para-Nitrobenzoate by Comamonas Sp. JS46 and Comamonas Sp. JS47, Biotechnology and Bioengineering, 59 (1): 21-27. |
| Gordon, 1999, Solid phase synthesis—designer linkers for combinatorial chemistry: a review, J. Chem. Technol. Biotechnol., 74(9):835-851. |
| Grasland-Mongrain, 2003, Droplet coalescence in microfluidic devices, 30 pages, From internet: http://www.eleves.ens.fr/home/grasland/rapports/stage4.pdf. |
| Gray, 1987, High speed crhomosome sorting, Science 238(4825):323-329. |
| Green, 1992, Selection of a Ribozyme That Functions as a Superior Template in a Self Copying Reaction, Science, 258: 1910-5. |
| Gregoriadis, 1976, Enzyme entrapment in liposomes, Methods Enzymol 44:218-227. |
| Griffiths, 2000, Man-made enzymes-from design to in vitro compartmentalisation, Curr Opin Biotechnol 11:338-353. |
| Griffiths, 2003, Directed evolution of an extremely fast phosphotriesterase by in vitro compartmentalization, EMBO J, 22:24-25. |
| Griffiths, 2006, Miniaturising the laboratory in emulsion droplets, Trend Biotech 24(9):395-402. |
| Grinwood, 2004, The DNA sequence and biology of human chromosome 19, Nature 428:529-535. |
| Grothues, 1993, PCR amplification of megabase DNA with tagged random primers (T-PCR), Nucl. Acids Res vol. 21(5):1321-1322. |
| Grund, 2010, Analysis of biomarker data:logs, odds, ratios and ROC curves, Curr Opin HIV AIDS 5(6):473-479. |
| Guatelli, 1990, Isothermal, in vitro amplification of nucleic acids by a multienzyme reaction modeled after retroviral replication, PNAS, 87(5):1874-8. |
| Guixe, 1998, Ligand-Induced Conformational Transitions in Escherichia coli Phosphofructokinase 2: Evidence for an Allosteric Site for MgATP2n, Biochem., 37: 13269-12375. |
| Gupta, 1991, A general method for the synthesis of 3′-sulfhydryl and phosphate group containing oligonucleotides, Nucl Acids Res 19 (11): 3019-3026. |
| Haber, 1993, Activity and spectroscopic properties of bovine liver catalase in sodium bis(2-ethylhexyl) sulfosuccinate/isooctane reverse micelles, Eur J Biochem 217(2): 567-73. |
| Habig, 1981, Assays for differentiation of glutathione S-transferases, Methods in Enzymology, 77: 398-405. |
| Hadd, 1997, Microchip Device for Performing Enzyme Assays, Anal. Chem 69(17): 3407-3412. |
| Haeberle, 2007, Microfluidic platforms for lab-on-a-chip applications, Lab on a Chip 7:1081-1220. |
| Hagar, 1992, The effect of endotoxemia on concanavalin A induced alterations in cytoplasmic free calcium in rat spleen cells as determined with Fluo-3, Cell Calcium 13:123-130. |
| Hai, 2004, Investigation on the release of fluorescent markers from the w/o/w emulsions by fluorescence-activated cell sorter, J Control Release, 96(3): 393-402. |
| Haies, 1981, Morphometric study of rat lung cells. I. Numerical and dimensional characteristics of parenchymal cell population, Am. Rev. Respir. Dis. 123:533-54. |
| Hall, 2003, The EBG system of E. coli: origin and evolution of a novel beta-galactosidase for the metabolism of lactose, Genetica 118(2-3):143-56. |
| Hamady, 2008, Error-correcting barcoded primers for pyrosequencing hundreds of samples in multiplex. Nature Nethods vol. 5, No. 3, p. 235-237. |
| Han, 2001, Quantum-dot-tagged Microbeads for Multiplexed Optical Coding of Biomolecules, Nat Biotech 19(7): 631-635. |
| Handen, 2002, High-throughput screening—challenges for the future, Drug Discov World, 47-50. |
| Handique, 2001, On-Chip Thermopneumatic Pressure for Discrete Drop Pumping, Analytical Chemistry, 73:1831-1838. |
| Hanes, 1997, In vitro selection and evolution of functional proteins by using ribosome display, PNAS 94:4937-42. |
| Hanes, 1998, Degradation of porous poly(anhydide-co-imide) microspheres and implication for controlled macromolecule delivery, Biomaterials, 19(1-3): 163-172. |
| Hansen 2002, A robust and scalable microfluidic metering method that allows protein crystal growth by free interface diffusion, PNAS 99(26):16531-16536. |
| Harada, 1993, Monoclonal antibody G6K12 specific for membrane-associated differentiation marker of human stratified squamous epithelia and squamous cell carcinoma, J. Oral Pathol. Med 22(4):145-152. |
| Harder, 1994, Characterization and kinetic analysis of the intracellular domain of human protein tyrosine phosphatase beta (HPTP beta) using synthetic phosphopeptides, Biochem J 298 (Pt 2): 395-401. |
| Harries, 2006, A Numerical Model for Segmented Flow in a Microreactor, Int J of Heat and Mass Transfer, 46:3313-3322. |
| Harris, 2008, Single-molecule DNA sequencing of a viral genome, Science 320(5872):106-109. |
| Harrison, 1993, Micromachining a miniaturized capillary electrophoresis-based chemical analysis system on a chip, Science 261(5123):895-897. |
| Hasina, 2003, Plasminogen activator inhibitor-2: a molecular biomarker for head and neck cancer progression, Cancer Research 63:555-559. |
| Haynes, 2012, Digital PCR: A Technology Primer, Principles of Digital PCR and Measurement Issues: The certification of Cytomegalovirus Standard Reference Material (SRM 2366) as a model for future SRMs, National Institute of Standards and Tecnology, San Diego, CA, 4 pages. |
| Hayward, 2006, Dewetting Instability during the Formation of Polymersomes from BloceCopolymer-Stabilized Double Emulsions, Langmuir, 22(10): 4457-4461. |
| He, 2005, Selective encapsulation of single cells and subcellular organelles into picoliter- and femtoliter-volume droplets, Anal Chem 77(6):1539-1544. |
| Head, 2014, Library construction for next generation sequencing, Biotech Rap Disp 56(2):61. |
| Heim, 1996, Engineering Green Fluorescent Protein for Improved Brightness, Longer Wavelengths and Fluorescence Response Energy Transfer, Carr. Biol, 6(2): 178-182. |
| Hellman, 2009, Differential tissue-specific protein markers of vaginal carcinoma, Br J Cancer, 100(8): 1303-131. |
| Henrich, 2012, Low-level detection and quantitation of cellular HIV-1 DNA and 2-ILTR circles using droplet dPCR, J Virol Meth 186(1-2):68-72. |
| Hergenrother, 2000, Small-Molecule Microarrays: Covalent Attachment and Screening of Alcohol-Containing Small Molecules on Glass Slides, J. Am. Chem. Soc, 122: 7849-7850. |
| Hermankova, 2003, Analysis of human immunodeficiency virus type 1 gene expression in lately infected reseting CD4 T lymphocytes in vivo, J Virology 77(13):7383-7392. |
| Herzer, 2001, DNA Purification, in Molecular Biology Problem Solver: A Laboratory Guide, Edited by Alan S. Gerstein, Ch.1. |
| Squires, 2005, Microfluidics: fluid physics at the nanoliter scale, Rev Mod Phys 77:977-1026. |
| Stauber, 1993, Rapid generation of monoclonal antibody-secreting hybridomas against African horse sickness virus by in vitro immunization and the fusion/cloning technique, J Immunol Meth 161(2):157-168. |
| Stemmer, 1994, DNA shuffling by random fragmentation and reassembly: in vitro recombination for molecular evolution. PNAS 91(22):10747-51. |
| Stemmer, 1994, Rapid evolution of a protein in vitro by DNA shuffling, Nature 370(6488):389-91. |
| Stober, 1998, Controlled growth of monodisperse silica spheres in the micron size range, J Colloid Interface Sci 26(1):62-69. |
| Stofko, 1992, A single step purification for recombinant proteins, Febs Lett 302:274-278. |
| Stone, 2004, Engineering flows in small devices: microfluidics toward a lab-on-a-chip, Ann Rev Fluid Mech 36:381-441. |
| Strizhkov, 2000, PCR amplification on a microarray of gel-immobilized oligonucleotides: Detection of bacterial toxin-and drug-resistant genes and their mutations, BioTechniques 29(4):844-857. |
| Strommenger, 2003, Multiplex PCR assay for simultaneous detection of nine clinicly relevant antibiotic resistance genes in S aureus, J Clin Microb 41(9):4089-4094. |
| Stroock, 2002, Chaotic mixer for microchannels, Science 295(5555):647-651. |
| Studer, 1997, Fluorous synthesis: a fluorous-phase strategy for improving separation efficiency in organic synthesis, Science 275:823-826. |
| Sugiura, 2001, Interfacial tension driven monodispersed droplet formation from mtcrofabricated channel array, Langmuir 17:5562-5566. |
| Sugiura, 2002, Effect of channel structure on microchannel emuisification, Langmuir 18:5708-5712. |
| Sundberg, 1995, Spatially-addressable immobilisation of macromolecules on solid supports, J Am Chem Soc 117:12050-12057. |
| Sung, 2005, Chip-based microfluidic devices coupled with electrospray ionization-mass spectrometry, Electrophoresis 26:1783-1791. |
| Sutcliffe, 1986, Dynamics of UV laser ablation of organic polymer surfaces, J Appl Phys 60(9):3315-3322. |
| Suzuki, 1996, Random mutagenesis of thermus aquaticus DNA polmerase I: concordance of immutable sites in vivo with the crystal structure, PNAS 93:96701-9675. |
| Suzuki, 2013, A novel guidewire approach for handling acute-angle bifurcations, J Inv Cardiol 25(1):48-54. |
| Syed, 2009, Next-generation sequencing library preparation: simultaneous fragmentation and tagging using in vitro transposition, Nat Meth 6:1-2. |
| Takayama, 1999, Patterning cells and their environmnets using multiple laminar fluid flows in cappillary networks, PNAS 96:5545-5548. |
| Takeuchi, 2005, An axisymmetric flow-focusing microfluidic device, Adv Mater 17(8):1067-1072. |
| Taly, 2007, Droplets as microreactors for high-throughput biology, Chembiochem 8(3):263-272. |
| Tan, 2003, Controlled fission of droplet emulsions in bifurcating microfluidic channels, 12th Int Conf SSAM 28-31. |
| Tan, 2003, Microfluidic liposome generation from monodisperse droplet emulsion, Summer Bioeng Conf, Florida, 2 pages. |
| Tan, 2003, Monodisperse droplet emulsions in co-flow microfluidic channels, Micro TAS, 2 pages. |
| Tan, 2004, Design of microluidic channel geometries for the control of droplet volume, chemical concentration, and sorting, Lab Chip 4(4):292-298. |
| Tang, 2009, A multi-color fast-switching microfluidic droplet dye laser, Lab Chip 9:2767-2771. |
| Taniguchi, 2002, Chemical reactions in microdroplets by electrostatic manipulation of droplets in liquid media, Lab Chip 2:19-23. |
| Tawfik, 1998, Man-made cell-like compartments for molecular evolution, Nat Biotech 7(16):652-56. |
| Taylor, 1934, The formation of emulsions in definable field of flow, Proc R Soc London A 146(858):501-523. |
| Taylor, 1991, Characterization of chemisorbed monolayers by surface potential measurments, J Phys D Appl Phys 24:1443. |
| Tencza, 2000, Development of a fluorescence polarization-based diagnostic assay for equine infectious anemia virus, J Clin Microbiol 38(5):1854-185. |
| Terray, 2002, Fabrication of linear colloidal structures for microfluidic applications, Applied Phys Lett 81(9):1555-1557. |
| Terray, 2002, Microlluidic control using colloidal devices, Science 296(5574):1841-1844. |
| Tewhey, 2009, Microdroplet based PCR environment for large scale targeted sequence, Nat Biotech 27(11):1025-1031. |
| Theberge, 2010, Microdroplets in microfluidics: an evolving platform for discoveries in chemistry and biology, Angew Chem Int Ed 49(34):5846-5868. |
| Thompson, 1983, Introduction to Lithography, ACS Symp Ser 219:1-13. |
| Thorsen, 2001, Dynamic pattern formation in a vesicle-generating microfluidic device, Phys Rev Lett 86(18):4163-4166. |
| Thorsen, 2002, Microfluidic large-scale integration, Science 298:580-584. |
| Thorsen, 2003, Microfluidic technologies for highthroughput screening applications, California Institute of Technology. |
| Tice, 2003, Formation of droplets and mixing in multiphase microfluidics at low values of the reynolds and the capillary numbers, Langmuir 19:9127-9133. |
| Tice, 2004, Effects of viscosity on droplet formation and mixing in microfluidic channels, Analytica Chimica Acta 507:73-77. |
| Titomanlio, 1990, Capillary experiments of flow induced crystallization of HDPE, AIChe J 36(1):13-18. |
| Tleugabulova, 2004, Evaluating formation and growth mechanisms of silica particles using fluorescence anisotropy decay analysis, Langmuir 20(14):5924-5932. |
| Tokatlidis, 1995, Nascent chains: folding and chaperone infraction during elongation on ribosomes, Philos Trans R Soc Lond B Biol Sci, 348:89-95. |
| Tokeshi, 2002, Continuous-flow chemical processing on a microchip by combining microunit operations and a multiphase flow network, Anal Chem 74(7):1565-1571. |
| Tokumitsu, 1999, Preparation of gadopentetic acid-loaded chitosan microparticles for gadolinium neutron-capture therapy of cancer by a novel emulsion-droplet coalescence technique, Chem Pharm Bull 47(6):838-842. |
| Tonelli et al., 2002, Perfluoropolyether functional oligomers: unusual reactivity in organic chemistry, Journal of fluorine Chemistry, 118; 107-121. |
| Trolier-McKinstry, 2004, Thin Film Piezoelectric for MEMS, Journal of Electroceramics 12:7-17. |
| Tsuchiya, 2007, On-chip polymerase chain reaction microdevice employing a magnetic droplet-manipulation system, Sens Actuators B 130:583-588. |
| Lunderberg, 1995, Solid-phase technology: magnetic beads to improve nucleic acid detection and analysis, Biotech Ann Rev 1:373-401. |
| Lundstrom, 2002, Breakthrough in cancer therapy: Encapsulation of drugs and viruses, Curr Drug Disc 19-23. |
| Lyne, 2002, Structure-based virtual screening: an overview, Drug Disc Tod 7(20):1047-1055. |
| Ma, 1993, In vitro protein engineering using synthetic tRNA(Ala) with different anticodons, Biochemistry 32(31):7939-45. |
| Mackenzie, 1985, IABS Symposium on Reduction of Animal Usage in the Development and Control of Biological Products, London, UK, 16 pages. |
| Mackenzie, 1986, The application of flow microfluorimetry to biomedical research and diagnosis: a review, Dev Biol Stand 64:181-193. |
| Maclean, 1999, Glossary of terms used in combinatorial chemistry, Pure Appl. Chem. 71(12):2349-2365. |
| Magdassi, 1984, Multiple Emulsions: HLB Shift Caused by Emulsifier Migration to External Interface, J. Colloid Interface Sci 97:374-379. |
| Mahajan, 1998, Bcl-2 and Bax Interactions in Mitochondria Probed with Green Florescent Protein and Fluorescence Resonance Energy Transfer, Nat. Biotechnol. 16(6): 547-552. |
| Mahjoob, 2008, Rapid microfluidic thermal cycler for polymerase chain reaction nucleic acid amplification. Int J HeatMass Transfer;51:2109-22. |
| Manafi, 2000, New developments in chromogenic and fluorogenic culture media, 2000, International Journal of Food Microbiology, 60, 205-218. |
| Manley, 1983, In vitro transcription: whole cell extract, Methods Enzymol, 101:568-82. |
| Manz, 1991, Micromachining of monocrystalline silicon and glass for chemical analysis systems A look into next century's technology or just a fashionable craze, Trends in Analytical Chemistry 10(5):144-149. |
| Mao, 1991, Substrate effects on the enzymatic activity of alphachymotrypsin in reverse micelles, Biochem Biophys Res Commun, 178(3):1105-12. |
| Mao, 1992, Kinetic behaviour of alpha-chymotrypsin in reverse micelles: a stopped-flow study, Eur J Biochem 208(1):165-70. |
| Mardis, 2008, The impact of next-generation sequencing technology on genetics, Trends Genet 24:133-141. |
| Margulies, 2005, Genome sequencing in microfabricated high-density picolitre reactors, Nature 437(7057):376-380. |
| Marks, 1992, Bypassing immunization: building high affinity human antibodies by chain shuffling, BioTechnol 10:779-783. |
| Marques, 1996, Porous Flow within Concentric Cylinders, Bull Am Phys Soc Div Fluid Dyn 41:1768, 1 page. |
| Maruno, 1991, Fluorine containing optical adhesives for optical communications systems, J. Appl. Polymer. Sci. 42:2141-2148. |
| Mason, 1997, Shear Rupturing of Droplets in Complex Fluids, Langmuir, 13(17):4600-4613. |
| Mastrobattista, 2005, High-throughput screening of enzyme libraries: in vitro evolution of a beta-galactosidase by fluorescence-activated sorting of double emulsions, Chem. Biol. 12(12): 1291-1300. |
| Masui, 1998, Probing of DNA-Binding Sites of Escherichia coli RecA Protein Utilizing 1-anilinonaphthalene-8-Sulfonic Acid, Biochem 37(35):12133-12143. |
| Matayoshi, 1990, Novel fluorogenic substrates for assaying retroviral proteases by resonance energy transfer, Science 247:954. |
| Matsubara, 2003, Detection of Single Nucleotide Substitution by Competitive Allele-Specific Short Oligonucleotide Hybridization (CASSOH) With Ummunochromatographic Strip, Human Mutation 22:166-172. |
| Mattheakis, 1994, An in vitro polysome display system for identifying ligands from very large peptide libraries, PNAS 91:9022-6. |
| Mayr, 2008, The Future of High-Throughput Screening, JBiomol Screen 13:443-448. |
| Mazutis, 2009, Droplet-Based Microfluidic Systems for High-Throughput Single DNA Molecule Isothermal Amplification and Analysis, Anal Chem 81(12):4813-4821. |
| Mazutis, 2009, Multi-step microfluidic droplet processing: kinetic analysis of an in vitro translated enzyme, Lab Chip 9:2902-2908. |
| McDonald, 2000, Fabrication of microfluidic systems in poly(dimethylsiloxane), Electrophoresis 21(1):27-40. |
| McDonald, 2002, Poly(dimethylsiloxane) as a material for fabricating microfluidic devices, Account Chem. Res. 35:491-499. |
| Melton, 1984, Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter, Nucl. Acids Res. 12(18):7035-7056. |
| Mendel, 1995, Site-Directed Mutagenesis with an Expanded Genetic Code, Annu Rev Biophys Biomol Struct, 24:435-62. |
| Mendieta, 1996, Complementary sequence correlations with applications to reflectometry studies, Instrumentation and Development 3(6):37-46. |
| Metzker, 2010, Sequencing Technologies—the next generation, Nature Reviews, vol. 11, pp. 31-46. |
| Meylan, 1995, Atom/fragment contribution method for estimating octanol-water partition coefficients, J Pharm Sci. 84(1):83-92. |
| Michalatos-Beloin, 1996, Molecular haplotyping of genetic markers 10 kb apart by allele-specific long-range PCR, Nucleic Acids Research, 24:4841-4843. |
| Miele, 1983, Autocatalytic replication of a recombinant RNA, J Mol Biol, 171:281-95. |
| Milstein, 1983, Hybrid hybridomas and their use in immunohistochemistry, Nature 305:537-540. |
| Mindlin, 1936, A force at a point of a semi-infinite solid, Physics, 7:195-202. |
| Minshuil, 1999, Protein evolution by molecular breeding, Curr Opin Chem Biol 3(3): 284-90. |
| Miroux, 1996, Over-production of proteins in Escherichia coli: mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels, J of Mol Biol 260(3):289-98. |
| Miyawaki, 1997, Fluorescent Indicators for Ca2+ Based on Green Fluorescent Proteins and Calmodulin, Nature, 388:382-887. |
| Mize, 1989, Dual-enzyme cascade—an amplified method for the detection of alkaline phosphatase, Anal Biochem 179(2): 229-35. |
| Mock, 1985, A fluorometric assay for the biotin-avidin interaction based on displacement of the fluorescent probe 2-anilinonaphthalene-6-sulfonic acid, Anal Biochem, 151:178-81. |
| Moldavan, 1934, Photo-electric technique for the counting of microscopical cells, Science 80:188-189. |
| Monie, 2005, A Novel Assay Allows Genotyping of the Latent Reservoir for Human Imnunodefi ciency Virus Type 1 in the Resting CD4+ T Cells of Viremic Patients, Journal of Virology, 79(8):5185-5202. |
| Montigiani, 1996, Alanine substitutions in calmodulin-binding peptides result in unexpected affinity enhancement, J Mol Biol, 258:6-13. |
| Moore, 1995, Exploration by lamp light, Nature, 374:766-7. |
| Morrison, 1984, Chimeric human antibody molecules: mouse antigen-binding domains with human constant region domains, PNAS 81:6851-6855. |
| Joyce,1994, In vitro Evolution of Nucleic Acids, Cum Opp. Structural Biol, 4: 331-336. |
| Kadir, 1990, Haem binding to horse spleen ferritin, Febs Lett, 276: 81-4. |
| Kallen, 1966, The mechanism of the condensation of formaldehyde with tetrahydrofolic acid, J. Biol. Chem., 241:5851-63. |
| Kambara, 1988, Optimization of Parameters in a DNA Sequenator Using Fluorescence Detection, Nature Biotechnology 6:816-821. |
| Kamensky, 1965, Spectrophotometer: new instrument for ultrarapid cell analysis, Science 150(3696):630-631. |
| Kanouni, 2002, Preparation of a stable double emulsion (W1/0/W2): role of the interfacial films on the stability of the system, Adv. Collid. Interf. Sci., 99(3): 229-254. |
| Karapatis, 1998, Direct rapid tooling:a review of current research, Rapid Prototyping Journal, 4(2):77-89. |
| Katanaev, 1995, Viral Q beta RNA as a high expression vector for mRNA translation in a cell-free system, Febs Lett, 359:89-92. |
| Katsura, 2001, Indirect micromanipulation of single molecules in water-in-oil emulsion, Electrophoresis, 22:289-93. |
| Kawakatsu, 1997, Regular-sized cell creation in microchannel emulsification by visual microprocessing method, Journal of the American Oil ChemistS Society, 74:317-21. |
| Keana, 1990, New reagents for photoaffinity labeling: synthesis and photolysis of functionalized perfluorophenyl azides, J. Org. Chem.55(11):3640-3647. |
| Keefe, 2001, Functional proteins from a random-sequence library, Nature, 410: 715-718. |
| Keij, 1994, High-speed photodamage cell sorting: An evaluation of the ZAPPER prototype, Methods in cell biology, 42:371-358. |
| Kelly, 2005, Detection of Vascular Adhesion Molecule-1 Expression Using a Novel Multimodal Nanoparticle, Circulation Research 96:327-336. |
| Kelly, 2007, Miniaturizing chemistry and biology in microdroplets, Chem Commun 18:1773-1788. |
| Kerker, 1983, Elastic and inelastic light scattering in flow cytometry, Cytometry, 4:1-10. |
| Khandjian, 1986, UV crosslinking of RNA to nylon membrane enhances hybridization signals, Mol. Bio. Rep. 11:107-115. |
| Kheir, 2012, Oxygen Gas-Filled Microparticles Provide Intravenous Oxygen Delivery, Science Translational Medicine 4(140):140ra88 (10 pages). |
| Kim, 2003, Type II quantum dots: CdTe/CdSe (core/shell) and CdSe/ZnTe (core/shell) heterostructures, J. Am Chem Soc. 125:11466-11467. |
| Kim, 2004, Comparative study on sustained release of human growth hormone from semi-crystalline poly(L-lactic acid) and amorphous poly(D,L-lactic-co-glycolic acid) microspheres: morphological effect on protein release, Journal of Controlled Release, 98(1):115-125. |
| Kircher, 2010, High-throughput DNA sequencing—concepts and limitations, Bioessays 32(6):524-536. |
| Kiss, 2008, High-throughput quantitative polymerase chain reaction in picoliter droplets, Anal. Chem 80:8975-8981. |
| Kitagawa, 1995, Manipulation of a single cell with microcapillary tubing based on its electrophoretic mobility, Electrophoresis 16:1364-1368. |
| Klug, 1994, All you wanted to know about selex, Molecular Biology Reports, 20:97-107. |
| Klug, 1995, Gene Regulatory Proteins and Their Interaction with DNA, Ann NY Acad Sci, 758: 143-60. |
| Klug, 1995, Protein motifs 5. Zinc fingers, FASEB J 9(8):597-604. |
| Knaak, 1995, Development of partition coefficients, Vmax and Km values, and allometric relationships, Toxicol Lett. 79(I-3):87-98. |
| Knight, 1998, Hydrodynamic Focusing on a Silicon Chip: Mixing Nanoliters in Microseconds, Physical Review Left 80(17):3863-3866. |
| Koeller, 2001, Enzymes for chemical synthesis, Nature 409:232-240. |
| Kohler, 1975, Continuous cultures of fused cells secreting antibody of predefined specificity, Nature, 256:495-7. |
| Kojima, 2005, PCR amplification from single DNA molecules on magnetic beads in emulsion: application for high-throughput screening of transcription factor targets. Nucleic Acids Res. 33:e150, 9 pages. |
| Kolb, 1995, Cotranslational folding of proteins, Biochem Cell Biol, 73:1217-20. |
| Komatsu, 2001, Roles of cytochromes P450 1A2, 2A6, and 2C8 in 5-fluorouracil formation rom tegafur, an anticancer prodrug, in human liver microsomes. Drug Met. Disp., 28:1457-1463. |
| Kopp, 1998, Chemical amplification: continuous flow PCR on a chip, Science, 280:1046-48. |
| Koster, 2008, Drop-based microfluidic devices for encapsulation of single cells, Lab on a Chip 8:1110-1115. |
| Kowalczykowski, 1994, Biochemistry of homologous recombination in Escherichia coli, Microbiol Rev 58(3):401-65. |
| Kozbor, 1984, A human hybrid myeloma for production of human monoclonal antibodies, J. Immunol., 133:3001-3005. |
| Krafft, 1991, Synthesis and preliminary data on the biocompatibility and emulsifying properties of perfluoroalkylated phosphoramidates as injectable surfactants, Eur. J. Med. Chem., 26:545-550. |
| Krafft, 2001, Fluorocarbons and fluorinated amphiphiles in drug delivery and biomedical research, Adv Rev Drug Disc 47:209-228. |
| Krafft, 2003, Emulsions and microemulsions with a fluorocarbon phase, Colloid and Interface Science 8(3):251-258. |
| Kralj, 2005, Surfactant-enhanced liquid-liquid extraction in microfluidic channels with inline electric-field enhanced coalescence, Lab Chip 5:531-535. |
| Kricka, 1996, Micromachining: a new direction for clinical analyzers, Pure and Applied Chemistry 68(10):1831-1836. |
| Kricka, 2003, Microchip PCR, Anal Bioanal Chem 377(5):820-825. |
| Kritikou, 2005, “It's cheaper in the Picolab,” Nature, September, vol. 6, 1 page. |
| Krumdiek, 1980, Solid-phase synthesis of pteroylpolyglutamates, Methods Enzymol, 524-29. |
| Kruth, 2003, Lasers and materials in selective laser sintering, Assembly Automation, 23(4):357-371. |
| Kumagai, 1994, Ablation of polymer films by a femtosecond high-peak-power Ti:sapphire laser at 798 nm, Applied Physics Letters, 65(14):1850-1852. |
| Kumar, 1989, Activity and kinetic characteristics of glutathione reductase in vitro in reverse micellar waterpool, Biochem Biophys Acta, 996(1-2):1-6. |
| Kumaresan, 2008, High-throughput single copy DNA amplification and cell analysis in engineered nanoliter droplets, Anal Chem, 80:3522-3529. |
| Lage, 2003, Whole genome analysis of genetic alterations in small DNA samples using hyperbranched strand displacement amplification and array-CGH, Genome Res 13:294-307. |
| Ellman, 1991, Biosynthetic method for introducing unnatural amino acids site-specifically into proteins, Methods Enzymol, 202:301-36. |
| Endo, 1996, Autocatalytic decomposition of cobalt complexes as an indicator system for the determination of trace amounts of cobalt and effectors, Analyst 121:391-394. |
| Endo, 1998, Kinetic determination of trace cobalt by visual autocatalytic indication, Talanta 47:349-353. |
| Engl, 2005, Droplet Traffic at a Simple Junction at Low Capillary Numbers Physical Review Letters, vol. 95, 208304, 1 page. |
| Eow, 2002, Electrocoalesce-separators for the separation of aqueous drops from a flowing dielectric viscous liquid, Separation and Purification Tech 29:63-77. |
| Eow, 2002, Electrostatic and hydrodynamic separation of aqueous drops in a flowing viscous oil, Chemical Eng Proc 41:649-657. |
| Eow, 2002, Electrostatic enhancement of coalescence of water droplets in oil: a review of the technology, Chemical Engineeing Journal 85:357-368. |
| Eow, 2003, Motion, deformation and break-up of aqueous drops in oils under high electric field strengths, Chemical Eng Proc 42:259-272. |
| Eow, 2003, The behavior of a liquid-liquid interface and drop-interface coalescence under the influence of an electric field, Colloids and Surfaces A: Physiochern. Eng. Aspects 215:101-123. |
| Eriksson, 2013, Comparative analysis of measures of viral reservoirs in HIV-1 eradication studies, PLoS Pathogens 9(2):e1003174, 17 pages. |
| Faca, 2008, A mouse to human search for plasma proteome changes associated with pancreatic tumor development, PLoS Med 5(6):e123:0953-0967. |
| Fahy, 1991, Self-sustained sequence replication (3SR): an isothermal transcription-based amplification system alternative to PCR, PCR Methods Appl 1:25-33. |
| Fan, 1994, Micromachining of capillary electrophoresis injectors and separators on glass chips and evaluation of flow at capillary intersections, Anal Chem 66:177-184. |
| Fan, 2007, Detection of Aneuploidy with Digital PCR, available at https://arxiv.org/ftp/arxiv/papers /0705/0705.1 030.pdf, 16 pages. |
| Fastrez 1997, In vivo versus in vitro screening or selection for catalytic activity in enzymes and abzymes, Mol Biotechnol 7(1)37-55. |
| Fettinger, 1993, Stacked modules for micro flow systems in chemical analysis: concept and studies using an enlarged model, Sens Actuat B. 17:19-25. |
| Fiedler, 1998, Dielectrophoretic sorting of particles and cells in a microsystem, Anal Chem 70(9):1909-1915. |
| Field, 1988, Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cervisiae by use of an epitope addition method. Mol Cell Biol, 8: 2159-2165. |
| Fields, 1989, A novel genetic system to detect protein-protein interactions, Nature 340(6230):245-6. |
| Filella, 1994, TAG-72, CA 19.9 and CEA as tumor markers in gastric cancer, Acta Oncol. 33(7):747-751. |
| Finch, 1993, Encapsulation and controlled release, Spec Publ R Soc Chem, 138:35, 12 pages. |
| Fingas, 1997, Studies of Water-In-Oil Emulsions: Stability Studies, Environment Canada, Proceedings of the Twentieth Arctic Marine Oilspill Program Technical Seminer, 1-20. |
| Fire, 1995, Rolling replication of short DNA circles, PNAS 92(10):4641-5. |
| Firestine, 2000, Using an AraC-based three hybrid system to detect biocatalysts in vivo, Nat Biotechnol 18: 544-547. |
| Fisher, 2004, Cell Encapsulation on a Microfluidic Platform, The Eighth International Conference on Miniaturised Systems for Chemistry and Life Scieces, MicroTAS, Malmo, Sweden. |
| Fletcher, 2002, Micro reactors: principles and applications in organic synthesis, Tetrahedron 58:4735-4757. |
| Fluri, 1996, Integrated capillary electrophoresis devices with an efficient postcolumn reactor in planar quartz and glass chips, Anal Chem 68:4285-4290. |
| Fornusek, 1986, Polymeric microspheres as diagnostic tools for cell surface marker tracing, Crit Rev Ther Drug Carrier Syst, 2:137-74. |
| Fowler, 2002, Enhancement of Mixing by Droplet-Based Microfluidics, Int Conf MEMS 97-100. |
| Frenz, 2008, Reliable microfluidic on-chip incubation of droplets in delay-lines, Lab on a Chip 9:1344-1348. |
| Fu, 1999, A microfabricated fluorescence-activated cell sorter, Nature Biotechnology, 17(11):1109-1111. |
| Fu, 2002, An Integrated Microfabricated Cell Sorter, Anal. Chem., 74: 2451-2457. |
| Fulton, 1997, Advanced multiplexed analysis with the FlowMetrix system, Clin Chem 43:1749-1756. |
| Fulwyler, 1965, Electronic Separation of Biological Cells by Volume, Science 150(3698):910-911. |
| Galan, 2010, A 454 multiplex sequencing method for rapid and reliable genotyping of highly polymorphic genes in large-scale studies., BMC Genomics 11(296):1-15. |
| Gallarate, 1999, On the stability of ascorbic acid in emulsified systems for topical and cosmetic use, Int J Pharm 188(2):233-241. |
| Ganan-Calvo, 1998, Generation of Steady Liquid Microthreads and Micron-Sized Monodisperse Sprays and Gas Streams, Phys Rev Lett 80(2):285-288. |
| Ganan-Calvo, 2001, Perfectly Monodisperse Microbubbling by Capillary Flow Focusing, Phys Rev Lett 87(27): 274501-1-4. |
| Garcia-Ruiz, 1994, Investigation on protein crystal growth by the gel acupuncture method, Acta, Cryst., D50, 99. pp. 484-490. |
| Garcia-Ruiz, 2001, A super-saturation wave of protein crystallization, J. Crystal Growth, 232:149-155. |
| Garstecki, 2004, Formation of monodisperse bubbles in a microhuidic flow-focusing device, Appl Phys Lett 85(13):2649-2651. |
| Gasperlin, 1994, The structure elucidation of semisolid w/o emulsion systems containing silicone surfactant, Intl J Pharm, 107:51-6. |
| Gasperlin, 2000, Viscosity prediction of lipophillic semisolid emulsion systems by neural network modeling, Intl J Pharm, 196:37-50. |
| Gelderblom, 2008, Viral complemntation allows HIV-1 replication without integration, Retrovirology 5:60. |
| Georgiou, 1997, Display of heterologous proteins on the surface of microorganisms: from the screenign of combinatiorial libraires to live recombinant vaccines. Nat Biotechnol 15(1), 29-34. |
| Georgiou, 2000, Analysis of large libraries of protein mutants using flow cytometry, Adv Protein Chem, 55: 293-315. |
| Gerdts, 2004, A Synthetic Reaction NetWork: Chemical Amplification Using Nonequilibrium Autocatalytic Reactions Coupled in Time, J. Am. Chem. Soc 126:6327-6331. |
| Ghadessy, 2001, Directed Evolution of Polymerase Function by Compartmentalized Self-Replication, PNSAS 98(8): 4552-4557. |
| Gibbs, 1989, Detection of single DNA base differences by competitive oligonucleotide priming, Nucleic Acids Res. 17(7): 2437-48. |
| Gilliland, 1990, Analysis of cytokine mRNA and DNA: Detection and quantitation by competitive polymerase chain reaction, PNAS, 87(7):2725-9. |
| Adang, 2001, The contribution of combinatorial chemistry to lead generation: an interim analysis, Curr Med Chem 8:985-998. |
| Affholter 1999, Engineering a Revolution, Chemistry in Britain 48-51. |
| Agrawal, 1990, Site-specific functionalization of oligodeoxynucleotides for non-radioactive labelling, Tetrahedron Let 31:1543-1546. |
| Aharoni, 2005, High-Throughput screens and selections of enzyme-encoding genes, Curr Opin Chem Biol, 9(2):210-6. |
| Ahn, 2006, Dielectrophoretic manipulation of drops for high-speed microluidic sorting devices, Applied Phys Lett 88:024104. |
| Akasheh, 2004, Development of piezoelectric micromachined ultrasonic transducers, Sensors and Actuators A Physical, 111:275-287. |
| Allen, 2000, High throughput fluorescence polarization: a homogeneous alternative to radioligand binding for cell surface receptors J Biomol Screen. 5(2):63-69. |
| Ammar, 2003, UV/Vis absorption and fluorescence spectroscopic study of novel symmetrical biscoumarin dyes, Dyes and Pigments 57:259-265. |
| Amstutz, 2001, In vitro display technologies: novel developments and applications. Curr Opin Biotech 12:400-405. |
| Anarbaev, 1998, Klenow fragment and DNA polymerase alpha-primase fromserva calf thymus in water-in-oil rnicroemulsions, Biochim Biophy Acta 1384:315-324. |
| Anderson, 1983, Preparation of a cell-free protein-synthesizing system from wheat germ, Methods Enz 101:635-644. |
| Anderson, 1993, Restriction endonucleases and modification methylases, Curr Op Struct Biol 3:24-30. |
| Ando, 1999, PLGA microspheres containing plasmid DNA: preservation of supercoiled DNA via cryopreparation and carbohydrate stabilization, J Pharm Sci 88(1):126-130. |
| Angell, 1983, Silicon micromechanical devices, Scientific Am 248:44-55. |
| Anhuf, 2003, Determination of SMN1 and SMN2 copy number using TaqMan technology, Hum Mutat 22(1):74-78. |
| Anna, 2003, Formation of dispersions using flow focusing in microchannels, Appl Phys Lett82(3):364-366. |
| Armstrong, 1996, Multiple-Component condensation strategies for combinatorial library synthesis, Acc Chem Res 29(3):123-131. |
| Ashkin, 1987, Optical trapping and manipulation of single cells using infrared laser beams, Nature 330:769-771. |
| Ashkin, 1987, Optical trapping and manipulation of viruses and bacteria, Science 235(4795):1517-20. |
| Auroux, 2002, Micro Total Analysis Systems 2: Analytical standard operations and applications, Anal Chem 74(12):2637-2652. |
| Baccarani, 1977, Escherichia coli dihydrofolate reductase: isolation and characterization of two isozymes, Biochemistry 16(16):3566-72. |
| Bagwe, 2001, Improved drug delivery using microemulsions: rationale, recent progress, and new horizons, Crit Rev Ther Drug Carr Sys 18(1):77-140. |
| Baker, 2010, Clever PCR: more genotyping, smaller volumes, Nat Meth 7:351-356. |
| Ballantyne, 1973, Selective area metallization by electron-beam controlled direct metallic deposition, J Vac Sci Tech 10:1094. |
| Barany, 1991, Genetic disease detection and DNA amplification using cloned thermostable ligase, PNAS 88(1):189-93. |
| Barany, 1991, The ligase chain reaction in a PCR World, PCR Meth App 1(1):5-16. |
| Baret, 2009, Fluorescence-activated droplet sorting (FADS): efficient microfluidic cell sorting based on enzymatic activity, Lab Chip 9:1850-1858. |
| Baret, 2009, Kinetic aspects of emulsion stabilization by surfactants: a microfluidic analysis, Langmuir 25:6088-6093. |
| Baroud, 2004, Multiphase flows in microfluidics, Physique 5:547-555. |
| Bauer, 1999, Advances in cell separation: recent developments in counterflow centrifugal elutriation and continuous flow cell separation, J Chromot 722:55-69. |
| Beebe, 2000, Functional hydrogel structures for autonomous flow control inside microfluidic channels, Nature 404:588-590. |
| Beer, 2007, On-chip, real-time, single-copy polymerase chain reaction in picoliter droplets, Anal Chem 79(22):8471-8475. |
| Beer, 2008, On-chip single-copy real-time reverse transcription PCR in isolated picoliter droplets, Anal Chem 80(6):1854-1858. |
| Bein, 1999, Efficient assays for combinatorial methods for the eiscovery of catalysts, Agnew Chem Int Ed 38:3:323-26. |
| Benichou, 2002, Double emulsions stabilized by new molecular recognition hybrids of natural polymers, Polym Adv Tech 13:1019-1031. |
| Benner, 1994, Expanding the genetic lexicon, Trends Biotech 12:158-63. |
| Benning, 2000, The binding of substrate analogs to phosphotriesterase. J Biol Chem 275:30556-30560. |
| Berman, 1987, An agarose gel electrophoresis assay for the detection of DNA-binding activities in yeast cell extracts, Meth Enz 155:528-37. |
| Bernath, 2004, In Vitro Compartmentalization by double emulsions: sorting and gene enrichment by FACS Anal Biochem 325:151-157. |
| Bernath, 2005, Directed evolution of protein inhibitors of DNA-nucleases by in vitro compartmentalization (IVC) and nano-droplet delivery, J Mol Biol 345(5):1015-26. |
| Betlach, 1976, A restriction endonuclease analysis of the bacterial plasmid controlling the EcoRI restriction and modification of DNA, Fed Proc 35:2037-2043. |
| Bibette, 1999, Emulsions: basic principles, Rep Prog Phys 62:969-1033. |
| Bico, 2002, Rise of liquids and bubbles in angular capillary tubes, J Colloid & Interface Sc 247:162-166. |
| Bico, 2002, Self-Propelling Slugs, J Fluid Mech 467:101-127. |
| Binder, 2009, Mismatch and G-stack modulated probe signals on SNP microarrays, PLoS One, 4(11):e7862. |
| Binladen, 2007, The use of coded PCR primers enables high-throughput sequencing of multiple homolog amplification products by 454 parallel sequencing, PLoSOne 2(2):e197. |
| Blanchet, 1993, Laser Ablation and the Production of Polymer Films, Science, 262(5134):719-721. |
| Boder, 1997, Yeast surface display for screening combinatorial polypeptide libraries, Nat Biotech 15(6):553-7. |
| Bosque, 2009, Induction of HIV-1 latency and reactivation in primary memory CD4+ T cells, Blood, 113(1):58-65. |
| Bougueleret, 1984, Characterization of the gene coding for the EcoRV restriction and modification system of E coli, Nucleic Acids Res 12(8):3659-76. |
| Tuzel, 2006, Region Covariance: A Fast Descriptor for Detection and Classification, European Conference on computer Vision (ECCV), 14 pages. |
| Umbanhowar, 2000, Monodisperse Emulsion Generation via Drop Break Off in a Coflowing Stream, Langmuir 16(2):347-351. |
| Unger, 2000, Monolithic microfabricated valves and pumps by multylayer soft lithography, Science 288(5463):113-116. |
| Utada, 2005, Monodisperse double emulsions generated from a microcapillary device, Science, 308:537-541. |
| Vainshtein, 1996, Peptide rescue of an N-terminal truncation of the stoffel fragment of Taq DNA polymerase, Protein Science, 5:1785-92. |
| Van der Sluis, 2013, Dendritic Cell-induced Activation of Latent HIV-1 Provirus in Actively Proliferating Primary T Lymphocytes, PLOS Pathog. 9(3): 16 pages. |
| Van Dilla, 1968, The fluorescent cell photometer: a new method for the rapid measurement of biological cells stained with fluorescent dyes, Annual Report of the Los Alamos Scientific Laboratory of the University of California (Los Alamos, NM), Biological and Medical Research Groupp (H-4) of the Health Division, Compiled by D. G. Ott, pp. 100-105. |
| Van Dilla, 1969, Cell Microfluorometry: A Method for Rapid Fluorescence Measurement, Science 163(3872):1213-1214. |
| Vanhooke, 1996, Three-dimensional structure of the zinc-containing phosphotrieesterase with the bound substrate analog diethy 4-methylbenzylphosphonate, Biochemistry 35:6020-6025. |
| Varga, 1991, Mechanism of allergic cross-reactions-I. Multispecific binding of ligands to a mouse monoclonal anti-DNP IgE antibody. Mol Immunol 28(6), 641-54. |
| Vary, 1987, A homogeneous nucleic acid hybridization assay based on strand displacement, Nucl Acids Res 15(17):6883-6897. |
| Venkateswaran, 1992, Production of Anti-Fibroblast Growth Factor Receptor Monoclonal Antibodies by In Vitro Immunization, Hybirdoma, 11(6):729-739. |
| Verhoeyen, 1988, Reshaping human antibodies: grafting an antilysozyme activity, Science, 239:1534-1536. |
| Vogelstein, 1999, Digital PCR, PNAS 96(16):9236-9241. |
| Voss, 1993, Kinetic measurements of molecular interactions by spectrofluorometry, J Mol Recognit, 6:51-58. |
| Wahler, 2001, Novel methods for biocatalyst screening. Curr Opin Chem Biol, 5: 152-158. |
| Walde, 1988, Structure and activity of trypsin in reverse micelles, Eur J Biochem, 173(2):401-9. |
| Walde, 1993, Spectroscopic and kinetic studies of lipases solubilized in reverse micelles, Biochemistry, 32(15):4029-34. |
| Walde, 1994, Oparin's reactions revisited: enzymatic synthesis of poly(adenylic acid) in micelles and self-reproducing vesicles. J Am Chem Soc, 116: 7541-7547. |
| Walker, 1992, Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system, PNAS 89(1):392-6. |
| Walker, 1992, Strand displacement amplification—an isothermal, in vitro DNA amplification technique, Nucleic Acid Res, 20(7):1691-6. |
| Wang, 1989, Quantitation of mRNA by the polymerase chain reaction. Proc natl Acad Sci USA 86(24), 9717-21. |
| Wang, 1990, Design and synthesis of new fluorogenic HIV protease substrates based on resonance energy transfer, Tetrahedron Lett., 31:6493. |
| Wang, 2002, Preparation of Titania Particles Utilizing the Insoluble Phase Interface in a MicroChannel Reactor, Chemical Communications 14:1462-1463. |
| Wang, 2008, DEP actuated nanoliter droplet dispensing using feedback control, Lab on a Chip 9:901-909. |
| Wang, 2010, Quantifying EGFR Alterations in the Lung Cancer Genome with Nanofluidic Digital PCR Arrays, Clinical Chemistry 56:4. |
| Warburton, 1993, Microcapsules for Multiple Emulsions, Encapsulation and Controlled Release, Spec Publ R Soc Chem, 35-51. |
| Wasserman, 1989, Structure and reactivity of allyl-siloxane monolayers formed by reaction of allcyltrichlorosilanes on silicon substrates, Langmuir 5:1074-1087. |
| Weaver, 2010, Taking qPCR to a higher level: Analysis of CNV reveals the power of high throughput qPCR to enhance quantitative resolution, Methods 50, 271-276. |
| Weil, 1979, Selective and accurate initiation of transcription at the Ad2 major late promotor in a soluble system dependent on purified RNA polymerase II and DNA, Cell, 18(2):469-84. |
| Werle, 1994, Convenient single-step, one tube purification of PCR products for direct sequencing, Nucl Acids Res 22(20):4354-4355. |
| Wetmur, 2005, Molecular haplotyping by linking emulsion PCR: analysis of paraoxonase 1 haplotypes and phenotypes, Nucleic Acids Res 33(8):2615-2619. |
| White, 2009, Digital PCR provides sensitive and absolute calibration for high throughput sequencing, BMC Genomics 10:116. |
| Wick, 1996, Enzyme-containing liposomes can endogenously produce membrane-constituting lipids, Chem Biol 3(4):277-85. |
| Wiggins, 2004, Foundations of chaotic mixing, Philos Transact A Math Phys Eng Sci 362(1818):937-70. |
| Williams, 1979, Methotrexate, a high-affinity pseudosubstrate of dihydrofolate reductase, Biochemistry, 18(12):2567-73. |
| Williams, 2006, Amplification of complex gene libraries by emulsion PCR, Nature Methods 3(7):545-550. |
| Wilson, 1999, In vitro selection of functional nucleic acids, Ann. Rev. Biochem. 68: 611-647. |
| Wittrup, 2001, Protein engineering by cell-surface display. Curr Opin Biotechnology, 12: 395-399. |
| Wittwer, 1989, Automated polymerase chain reaction in capillary tubes with hot air, Nucleic Acids Res., 17(11) 4353-4357. |
| Wittwer 1990, Minimizing the Time Required for DNA Amplification by Efficient Heat Transfer to Small Samples, Anal. Biochem., 186, 328-331. |
| Wolff, 2003, Integrating advanced functionality in a microfabricated high-throughput fluorescent-activated cell sorter, Lab Chip, 3(1): 22-27. |
| Woolley, 1994, Ultra-high-speed DNA fragment separations using microfabricated capillary array electrophoresis chips, Proc. Natl. Acad. Sci. USA, 91, 11348-11352. |
| Woolley, 1996, Functional Integration of PCR Amplification and Capillary Electrophoresis in a Microfabricated DNA Analysis Device, Anal. Chem. 68, 4081-4086. |
| Wronski, 2002, Two-color, fluorescence-based microplate assay for apoptosis detection. Biotechniques, 32:666-668. |
| Wu, 1989, The ligation amplification reaction (LAR)-amplification of specific DNA sequences using sequential rounds of template-dependent ligation, Genomics 4(4):560-9. |
| Wyatt, 1991, Synthesis and purification of large amounts of RNA oligonucleotides, Biotechniques 11(6):764-9. |
| Xia, 1998, Soft Lithography, Angew. Chem. Int. Ed. 37:550-575. |
| Xia, 1998, Soft Lithography, Ann. Rev. Mat. Sci. 28:153-184. |
| Xiao, 2007, Rapid DNA mapping by fluorescent single molecule detection, Nucleic Acids Research 35:1-12. |
| Abate, 2011, Synthesis of monidisperse microparticles from non-Newtonian polymer solutions with microfluidic devices, Adv Mat 23(15):1757-1760. |
| Dickinson, 1992, Interfacial interactions and the stability of oil-in-water emulsions, Pure Appl Chem 64(11):1721-1724. |
| Luft, 20001, Detection of integrated papillomavirus sequences by ligation-mediaated PCR (DIPS-PCR) and molecular characterization in cervical cancer cells, In J Cancer 92:9-17. |
| Rogers, 2005, Closing bacterial genoimc sequence gaps with adaptor-PCR, BioTechniques 39(1):1-3. |
| Number | Date | Country | |
|---|---|---|---|
| 20200330993 A1 | Oct 2020 | US |
| Number | Date | Country | |
|---|---|---|---|
| 60925357 | Apr 2007 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 16400401 | May 2019 | US |
| Child | 16916532 | US | |
| Parent | 16105283 | Aug 2018 | US |
| Child | 16400401 | US | |
| Parent | 15415156 | Jan 2017 | US |
| Child | 16105283 | US | |
| Parent | 14737865 | Jun 2015 | US |
| Child | 15415156 | US | |
| Parent | 14070953 | Nov 2013 | US |
| Child | 14737865 | US | |
| Parent | 12595107 | US | |
| Child | 14070953 | US |
