Patent Application
20170008009
The invention relates to a microchannel, a microfluidic chip and a method for processing microparticles in a fluid flow.
For biological assays, chemical tests, chemical synthesis, processing of samples or biological fluids may require processing microparticles. For example, processing microparticles carrying different analytes on their surface may allow for surface-based assays for detecting different types of analytes including (but not limited to) DNA sequences, antigens, lipids, proteins, peptides, hydrocarbons, toxins, chemical compounds or cells. Analysis on microparticles carrying analytes may be performed, for example, by optical or electrochemical monitoring, applying fluorescence, magnetism-based sensing, fluorescence quenching.
Dielectrophoresis relates to the motion of polarizable particles in a non-uniform or asymmetric electric field. In particular, microparticles subjected to an electric field become polarized and make up dipoles aligned to the applied field. In a non-uniform electric field, each half of the dipole experiences unequal Coulomb forces, and a net force is exerted on the microparticle. Depending on dielectric properties including structural, morphological and chemical characteristics, the microparticles respond differently to the applied asymmetric electric field.
According to a first aspect, the invention can be embodied as a microchannel for processing microparticles in a fluid flow. The microchannel comprises a first pair of electrodes and second pair of electrodes. The first pair of electrodes is configured to generate an asymmetric first electric field for sorting the microparticles to provide sorted microparticles. The second pair of electrodes is configured to generate an asymmetric second electric field for trapping at least some of the sorted microparticles.
According to a second aspect, the invention can be embodied as a microfluidic chip comprising a microchannel according to the first aspect of the invention.
According to a third aspect, the invention can be embodied as a method for arranging microparticles in a fluid flow in a microchannel. The method comprises sorting the microparticles by an asymmetric first electric field to provide sorted microparticles and trapping at least some of the sorted microparticles by an asymmetric second electric field.
In the following, exemplary embodiments of the present invention are described with reference to the enclosed figures.
Similar or functionally similar elements in the figures have been allocated the same reference signs if not otherwise indicated.
In the following, moving or deflecting microparticles, in particular polarizable microparticles, by applying an electric field involves the above principles of dielectrophoresis, unless specified otherwise. In particular, an asymmetric electric field as evoked herein (in particular for sorting, trapping and concentrating particles) is such that a microparticle subject to such an electric field will deviate from an average direction of the fluid flow in the microchannel. Typically, asymmetric electric fields involve fringing fields extending through a medium (e.g., a liquid in a microchannel) from one electrode to another, the field being influenced by the shapes and proximity of the electrodes. Such asymmetric electric fields typically have the strongest gradient near the edges of the electrodes.
For example, the elements and devices of the microfluidic chip 10 may be formed by: anisotropic wet etching using silicon substrate and silicon oxide or nitride as mask; thermal oxidation for electrical passivation; patterning a conductive layer (preferably comprising gold, platinum, palladium and/or aluminum) by etching or lift-off; and sealing microfluidic structures preferably using a pre-patterned adhesive film, elastomer, thermoplastic and/or dry-film resist.
Alternatively, the elements and devices of the microfluidic chip 10 may be formed by: providing a substrate (e.g. plastics, FR-4 materials, polydimethylsiloxane (PDMS), preferably silicon) with a passivated layer (e.g. silicon dioxide); patterning electrodes and contacts (preferably comprising gold, platinum, palladium, titanium and/or aluminum) by an etching and/or lift-off process; patterning microfluidic structures by structuring a photosensitive layer (e.g. SU-8 materials, positive photoresist, dry-film resist) or etching a deposited film (e.g. parylene or polyimide); and sealing microfluidic structures using a pre-patterned adhesive film, elastomer, thermoplastic or dry-film resist.
Alternatively, the elements and devices of the microfluidic chip 10 may be formed by: providing a substrate; structuring the microfluidic structures by etching (for silicon or glass), embossing or injection molding (for plastics) and/or soft-lithography (for elastomers); patterning electrodes on a cover layer (preferably comprising glass, silicon, dry-film resist, plastics and/or PDMS); and sealing microfluidic structures by bonding two substrates, e.g. by anodic bonding, adhesive bonding or thermoplastic bonding.
For example, the microfluidic chip 10 may be plugged to an electrical socket. A fluid can then be pipetted to the inlet port 11 and pulled toward the capillary pump 12 by a capillary force, thereby flowing along the microchannel 13. Alternatively or additionally, the fluid flow F may be generated by a pump or any other device generating an according pressure gradient. In
The fluid may comprise water (distilled, deionized, tap water or water in natural resources), biological buffers such as phosphate-buffered saline (PBS) and Tris-acetate-EDTA (TAE) buffer, human serum, urine and/or saliva. Furthermore, surfactants such as Tween® 20 may be added to the fluid to minimize an aggregation of the microparticles.
In particular, the microparticles are polarizable in an external electric field, i.e. an electric dipole moment is induced at the microparticles by the applied field. A typical size of the microparticles can be in a submillimeter range, preferably from 10−8 m to 10−3 m and even more preferably 10−6 m to 10−4 m. For example, the microparticles comprise beads, microbeads, microspheres. Preferably, the microparticles are suited for capturing other particles, for example biological analytes including cells. The capture of the particles may employ a chemical and/or physical bonding, for example adsorption. Accordingly, the microparticles may have receptors at the surface for capturing smaller particles. The microparticles may have a functionalized surface, e.g. the surface may be amine-terminated, COOH-terminated and/or functionalized with biotin, streptavidin, protein, nucleotides, or oligonucleotides from deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). The microparticles may comprise silica, latex, agarose, one or more polymers, and/or may have a magnetic core. Preferably, the microparticles comprise polystyrene.
In particular, the fluid flow F may follow the principles of microfluidics. Accordingly, typical dimensions of the microchannel 13 can range between 10−7 m to 10−3 m. Further, typical volumes of the fluid, in particular in a microfluidic device, can range from 10−15 L to 10−5 L.
The microfluidic chip 10 further comprises a plurality of electrical contacts 15. In particular, the electrical contacts 15 may be exposed so as to enable an electrical connection to external devices, in particular to a power supply, using sockets, spring-loaded contacts (e.g. Pogo pins), solders, or wirebonds. The electrical contacts 15 can be electrically connected to a concentrating element 16, a sorting element 17, a trapping element 18 and/or further elements. One or more power lines 19 may connect these elements with the respective electrical contact 15.
The capillary pump 12 in particular involves effects of capillarity. The use of the capillary pump 12 could be beneficial in terms of compactness and low cost. Alternatively, microfluidic pumps could be used. Further the capillary pump 13 may eliminate the necessity of microfluidic connectors. A surface tension of the fluid, e.g. water, in the microchannel 13 and adhesive forces between the fluid and inner walls may generate a force that drives the fluid from the inlet port 11 toward the capillary pump 12. The capillary pump 12 may include a plurality of parallel channels in order to increase a capillarity pressure along the microchannel 13. Further, the capillary pump 12 may include a plurality of posts, bars, shapes (e.g. round-shaped or polygonal), etc. arranged in a structure for allowing for a number of parallel flow paths, thereby decreasing a flow resistance. The flow rate can be tuned by changing the wetting properties of the surfaces, hydraulic flow resistance of the microchannels and the viscosity of the liquid. The air vent 14 may be configured for discharging a fluid (e.g. air in the pump) from the capillary pump 12 for eliminating compression of the air in the capillary pump 12.
Generally, capillary pumps can generate lower flow rates and allow for a more precise control of the flow rate compared to external pumps, in particular microfluidic pumps. A non-uniform advancing of the liquid front during the capillary flow can form air bubbles, which may lead to erroneous results in processing of the microparticles and the analysis. Therefore, it could be advantageous to prevent the formation of bubbles in the microchannel 13 as well as undesired sticking of microparticles at the inner walls of the microchannel 13.
The microfluidic chip 10 can comprise a plurality of different elements and devices for sorting and trapping microparticles. For example, a spacing element 21, a decelerating element 22, a partitioning element 23, the concentrating element 16, the sorting element 17 and the trapping element 18 are attached to a first inner wall 24 and/or a second inner wall 25 of the microfluidic channel 13. In particular, the first and second inner walls 24, 25 are parallel to each other. The power lines 19 connect the elements 16-18, 21-23 to one of the electrical contacts 15.
A width W of the microchannel 13 can be 10−6 m to 10−2 m, preferably 10−5 m to 10−3 m. Distances D between element sets can be 10−5 m to 10−2 m, preferably 10−4 m to 10−3 m. A minimum distance C of the power lines 19 from the inner walls 24, 25 of the microchannel 13 can be 10−5 m to 10−2 m, preferably 5.10−5 m to 10−3 m. Distance D may prevent an electrical cross-talk between the different elements for concentrating, sorting and trapping. Similarly, distance C may prevent an electrical cross-talk between the power lines 19 and electrodes inside the microchannel 13. In particular, such electrical cross-talks may influence the sorting and trapping elements 17, 18 by coupling through the substrate, sealing layer, air and the liquid.
Microparticles which come into contact with the inner walls 24, 25 can lead to an aggregation of microparticles because the velocity of the liquid decreases towards the inner walls 24, 25 under the laminar flow regime. Such aggregation of microparticles can impair the fluid flow F and operation of the microfluidic chip 10. In
The concentrating element 16 can comprise a pair of linearly shaped electrodes. Generally, the microparticles moving in the fluid along the microchannel 13 are randomly distributed over the width W. The electrodes of the concentrating element 16 preferably generate an asymmetric electric field that concentrates the microparticles with respect to the width W of the microchannel 13, i.e. drives them into a column 26 with a smaller width than the width W of the microchannel 13. In other words, a spatial distribution of the microparticles is locally limited to the column 26 in terms of a w-direction, i.e. a direction parallel to the width W of the microchannel 13. For example, the microparticles passing through the electric field of the concentrating element 16 may experience a force perpendicular to the fluid flow direction F and/or in a direction parallel to the linear extension of the electrodes. Preferably, the microparticles are deflected toward one of the inner walls 24, 25 (without touching the inner walls 24, 25) in order to provide a space as wide (i.e. in the w-direction) as possible for a sorting process of the microparticles. In
The electrodes of the concentrating element 16 may be arranged parallel to each other. In particular, one of the electrodes can extend between the first and second inner walls 24, 25 whereas the other electrode extend from the second inner wall 25 into the microchannel 13 without reaching the first inner wall 24 in order not to move the microparticles so far as to touch the first inner wall 24. The gap between the electrode and the first inner wall 24 can be 10−6 m to 10−3 m, preferably 10−5 m to 10−4 m. After being concentrated by the concentrating element 16, the microparticles move inside the column 26 along the fluid flow F, preferably a laminar flow.
The sorting element 17 can comprise a pair of linearly shaped electrodes for generating an asymmetric electric field that selectively moves the microparticles depending on their properties and thereby provide sorted microparticles. In particular, the properties of the microparticles can comprise size, permittivity, polarizability, porosity and/or material. As a result, the microparticles passing through the electric field of the sorting element 17 can be shifted into different positions in terms of w-direction. For example, microparticles having a diameter of 10 μm can be displaced further in the w-direction than microparticles having a diameter of 5 μm since the dielectrophoretic force increases with increasing microparticle size. Further, the sorting element 17 can be configured to divide the microparticles into a plurality of particles groups by selectively moving them depending on their properties.
The partitioning element 23 is configured to prohibit the sorted microparticles and/or the particle groups from intermixing, i.e. to prevent the microparticles of one particle group joining the microparticles from another particle group, particularly while they are trapped on the trapping element 18. Preferably, the partitioning element 23 is linearly shaped along the microchannel 13 and arranged parallel to each other between the inner walls 24, 25. For example, the partitioning element 23 comprises a pair of electrodes for generating a repulsive electric field and/or a solid barrier.
The trapping element 18 is configured to trap the sorted microparticles in specified areas. Trapping can refer to retaining, arresting, positioning, localizing microparticles at one or more defined positions. Trapping microparticles may facilitate imaging and/or processing microparticles, e.g. for an analysis, or increasing their concentration. For example, the trapping element 18 can comprise a plurality of linearly shaped electrodes in an interdigitated arrangement as shown in
The spacing element 21 and the concentrating element 16a are connected to the power lines 19. The spacing element 21 is attached to the first inner wall 24 and extends from the first inner wall 24 into the microchannel 13. Preferably, the spacing element 21 comprises a pair of linearly shaped electrodes arranged parallel to each other. The spacing element 21 intersects the first inner wall 24 at an angle of intersection A21. The angle of intersection A21 is preferably an acute angle, i.e. at most 89°. The spacing element 21 can generate a repulsive electric field for repelling the microparticles in order to push the microparticles away from the first inner wall 24. A length of the electrodes of the spacing element 21 can be 10−6 m to 10−3 m, preferably 10−5 m to 10−4 m.
The concentrating element 16a can comprise a pair of electrodes extending between the first and second inner walls 24, 25. In
The second electrode 32 can comprise two sections 33, 34 that are linearly shaped and connected to each other inside the microchannel 13. A first section 33 can extend from the second inner wall 25 into the microchannel 13 and is arranged parallel to the first electrode 31. A second section 34 can extend from the first inner wall 24 into the microchannel 13. The second section 34 can be inclined at an angle A34 to the first inner wall 24. Preferably, the angle A34 is greater than the angle A31. As a result, a gap between the first and second electrodes 31, 32 can widen from the second inner wall 25 toward the first inner wall 24 in order not to move the microparticles so far as to touch the first inner wall 24.
The pair of electrodes 31, 32 of the concentrating element 16a is configured to generate an electric field for concentrating the microparticles in the fluid flow F in the manner described above. Preferably, the microparticles passing through the electric field of the concentrating element 16a move toward the first inner wall 24 without coming into contact with the inner wall 24.
The concentrating element 16b can comprise a first electrode 41 and a second electrode 42, with the first electrode 41 arranged upstream of the second electrode 42 with respect to the fluid flow direction F. Preferably, the first electrode 41 is shorter than the second electrode 42. The first and second electrodes 41, 42 can extend between the inner walls 24, 25. The first and second electrodes 41, 42 can be linearly shaped. The first electrode 41 can be inclined at an angle A41 to the first inner wall 24. The second electrode 42 can be inclined at an angle A42 to the first inner wall 24. The angles A41, A42 are preferably an obtuse angle and more preferably 120°-150°. Preferably, the angle A41 is greater than the angle A42 so that a gap between the first and second electrodes 41, 42 widens from the second inner wall 25 to the first inner wall 24. In particular, a position in the w-direction to which the microparticles are concentrated by the electric field of the concentrating element 16b may be tuned, for example by adjusting an amplitude and/or frequency of the applied signal and/or changing electrical properties of the microparticles and/or the fluid, so that the location of column 26 can be adjusted
For all embodiments of the concentrating elements 16, 16a, 16b, the linearly shaped electrodes may have a width of 10−6 m to 10−4 m, preferably 10−5 m to 5.10−5 m. The gap between two electrodes of the pair of electrodes may be 10−6 m to 10−4 m, preferably 5.10−6 m to 5.10−5 m.
In
Some metals, such as Au and Pd, can be less hydrophilic than the surface of the microchannel 13, which is typically glass or SiO2. Such inhomogeneity in the hydrophilicity can lead to a non-uniform fluid flow during the capillary flow of the liquid. A non-uniform fluid flow can result in an instability such as bubble that can impair the operation of the microfluidic chip 10. The decelerating element 22 can contribute to a uniformity of the fluid flow through the microchannel 13 by slowing down a part of the initial fluid flow that moves along the second inner wall 25. In particular, the decelerating element 22 slows down the corresponding portion of the fluid flow during an initial filling process of the microchannel 13.
For example, a length L51 of the electrodes 51 grows in the fluid flow direction F, and a length L52 of the electrodes 52 decreases in the fluid flow direction F. Here, the lengths L51, L52 can refer to a spatial extension of the respective electrodes 51, 52 in the w-direction. A width W53 of the gap 53 may be constant. As a result, a position of the gap 53 moves from near the second inner wall 25 toward the first inner wall 24 in the fluid flow direction F. For example, the lengths L51, L52 change in a manner that the position of the gap 53 changes linearly from the second inner wall 25 toward the first inner wall 24 along the direction of the fluid flow F.
In particular, the electrodes are configured to generate an electric field that moves the microparticles M toward the position of the respective gap 53. After passing through the electric field of the concentrating element 16c, the microparticles M are positioned inside a column 55.
A further decelerating element 63 extends from the second inner wall 25 into the microchannel 13. The decelerating element 63 can comprise a pair of linearly shaped metallic bodies arranged parallel to each other, as shown in
The electrodes 61, 62 of the sorting element 17 are configured to generate an electric field, in particular an asymmetric electric field, for sorting the microparticles M in the fluid flow F, thereby providing sorted microparticles. For example, the microparticles M can comprise a first group of microparticles M1 and a second group of microparticles M2, with the microparticles of the first group of microparticles M1 being smaller than those of the second group of microparticles M2. In the electric field generated by the sorting element 17, microparticles M2 may be forced to move toward the second inner wall 25, whereas the microparticles M1 are less affected or not affected. As a result, the microparticles M2 are positioned in a central part of the microchannel 13 with respect to the w-direction, and the microparticles M1 stay in a position close to the first inner wall 24.
Further, the microparticles M in the fluid flow F can comprise more than two randomly mixed groups of microparticles. The sorting element 17 may then be suited for dividing the microparticles M into N respective groups of M1-MN depending on their properties. Amplitude and/or frequency of the applied signal and/or the extension of the sorting element 17 in the w-direction, and/or the tapered gap between the electrodes 61, 62 can be tuned to adjust the position of the microparticles M1 in microchannel 13 with respect to the w-direction.
Two intermediate power lines 73, 74 that are connected to the power lines 19 may be arranged outside of the microchannel 13, with a first intermediate power line 73 being close to the first inner wall 24 and a second intermediate power line 74 being close to the second inner wall 25. For example, the intermediate power lines 73, 74 are arranged parallel to the first and second inner walls 24, 25.
A plurality of first electrodes 73a-73d may be formed extending from the first intermediate power line 73 into the microchannel 13. The first electrodes 73a-73d may be arranged parallel to one another and inclined at an angle A73 to the first inner wall 24. Preferably, the angle A73 is an obtuse angle, more preferably 120°-150°. In particular, the first electrodes 73a-73d can be arranged parallel to the concentrating element 16. The first electrodes 73a-73d can extend toward the second inner wall 25 such that that the first electrodes 73a-73d end in a boundary line 77 inside the microchannel 13. The boundary line 77 can be inclined at an angle A77 to the first inner wall 24. The angle A77 can be an acute angle, preferably 5°-30°.
A plurality of second electrodes 74a-74d may be formed extending between the first and second intermediate power lines 73, 74 across the microchannel 13. The first electrodes 74a-74d may be arranged parallel to one another and inclined at an angle A74 to the second inner wall 25. The angle A74 is preferably an obtuse angle, more preferably 120°-150°.
A plurality of first branches 75a-75c may be formed being connected to the first electrodes 73a-73c, respectively. In particular, the first branches 75a-75c can be connected to the respective first electrodes 73a-73c at a position where the first electrodes 73a-73c intersect the first inner wall 24. A further first branch 75p may be formed being connected to one of the electrodes of the concentrating element 16. The first branches 75a-75p may be linearly shaped and arranged parallel to one another. In particular, the first branches 75a-75p can be inclined at an angle A75 to the first inner wall 24. Preferably, the first branches 75a-75p are arranged parallel to the second electrodes 74a-74d. Preferably, the first branches 75a-75d extend toward the second inner wall 25 as far as to the boundary line 77. The first branches 75a-75p may be electrically conductive, in particular metallic electrodes.
A plurality of second branches 76a-76d may be formed being connected to the second electrodes 74a-74d, respectively. In particular, the second branches 76a-76d can be connected to the respective second electrodes 74a-74d at a position where the second electrodes 74a-74d intersects the boundary line 77. The second branches 76a-76d may be electrically conductive, in particular metallic electrodes. The second branches 76a-76d may be linearly shaped and arranged parallel to one another. In particular, the second branches 76a-76d can be inclined at an angle A76 to the second inner wall 25. Preferably, the second branches 76a-76d are arranged parallel to the first electrodes 73a-73d.
The first branch 75p with the second electrode 74a builds the sorting element 71a. The first branches 75a-75c build with the second electrodes 74b-74d, respectively, the sorting elements 71b-71d. Each of the sorting elements 71a-71d is configured to generate an electric field for selectively moving the microparticles depending on their properties.
The second branches 76a-76d build with the first electrodes 73a-73d, respectively, the concentrating elements 72a-72d. Each of the concentrating elements 72a-72d is configured to generate an electric field for concentrating the microparticles in the w-direction.
In total, a cascade of concentrating elements 72a-72d and sorting elements 71a-71d is provided. For example, a part of the microparticles M that passes the electric field of the sorting elements 71a-71c without being affected can be concentrated by the concentrating elements 72a -72c for being sorted by the following sorting elements 71b-71d. Preferably, sorting of the microparticles can be performed gradually since the microparticles with different properties react differently to the electric fields of the sorting elements 71a-71d. In this manner, a sorting efficacy can be increased and the risk of having unsorted bigger particles ending up in the region of smaller particles can be minimized
In particular, smaller microparticles, e.g. spherical particles with a diameter of 3-5 μm, can be deflected toward the first inner wall 24 by the concentrating elements 72a-72d. Bigger microparticles, e.g. spherical particles with a diameter of 8-10 μm, can be deflected toward the second inner wall 25 by the sorting elements 71a-71d. Unsorted bigger microparticles, i.e. a part of the bigger microparticles that is not affected by the preceding sorting element 71a-71c, can be deflected toward the first inner wall 24 by the concentrating element 72a-72c and sorted by the following sorting element 71b-71d. As a result, the bigger microparticles move in a column near the second inner wall 25, while the smaller microparticles move in a column near the first inner wall 24. The number of sorting and concentrating elements can be adjusted according to the dimensions of the particles, area reserved for these elements and the required efficacy.
In this embodiment, the sorting elements 71a-71d and the concentrating elements 72a-72d are electrically coupled to one another by being connected to both intermediate lines 73, 74. In order to increase the Dielectrophoresis forces for the concentrating elements 72a-72d, a concentrator gap between the second branches 76a-76d and the respective first electrodes 73a-73d is smaller than a sorter gap between the first branches 75p, 75a-75c and the respective second electrodes 74a-74d. A ratio of the concentrator gap to the sorter gap can be, for example, 0.1 to 0.9, preferably 0.3 to 0.7, with the concentrator and sorter gaps being 10−6 m-10−4 m.
The sorting element 80 comprises two intermediate power lines 81, 82. A plurality of electrodes 83 is connected to the first intermediate power line 81 and extend toward the second intermediate power line 82 across the microchannel 13. A further plurality of electrodes 84 is connected to the second intermediate power line 82 and extend toward the first intermediate power line 81 across the microchannel 13. The electrodes 83, 84 are arranged parallel to one another in an interdigitated arrangement. Each electrode 83, 84 includes a plurality of plates 85 connected to another by a wire 86. The plates 85 are, for example, rectangular-shaped. The electrodes 83, 84 are arranged parallel to one another with a constant distance between each neighboring electrodes 83, 84.
For example, the microparticles M approach the sorting element 80 in a column near the first inner wall 24 after being concentrated by an upstream concentrating element. A first part M1 of the microparticles M that are sensitive to the asymmetric electric field of the electrodes 83, 84 can be gradually deflected toward the second inner wall 25 by the cascade of the electrodes 83, 84. A second part M2 of the microparticles M that are less or not affected by the electric fields of the electrodes 83, 84 can thereby be separated from the first part M1.
A dimension of the sorting element 80 and parameters of the applied electric field including frequency and amplitude can be tuned to define a position of the column the microparticles of the first part M1 move along as well as to keep the microparticles of the second part M2 unaffected. This embodiment can reduce the probability of microparticles not being deflected by a sorting element.
The sorting element 90 comprises two intermediate power lines 91, 92 that are connected to the power lines 19. N first electrodes 931-93N are connected to the first intermediate power line 91 and extend toward the second intermediate power line 92 across the microchannel 13. N second electrodes 941-94N are connected to the second intermediate power line 92 and extend toward the first intermediate power line 91 across the microchannel 13. The electrodes 93, 94 are arranged parallel to one another in an interdigitated arrangement. The first electrodes 931-93N build with the second electrodes 941-94N, respectively, N pairs of neighboring electrodes 951-95N.
A plurality of plates 96 are attached to each of the electrodes 931-93N. A further plurality of plates 97 are attached to each of the second electrodes 941-94N. In particular, the plates 96, 97 are shaped thus that the plates 96 and the plates 97 complement one another, i.e. the plates 96 and the plate 97 geometrically add to form another, for example rectangular or circular, shape. In
The plates 96, 97 can be formed between two neighboring electrodes 931-93N and 941-94N, respectively. In
In particular, the sorting element 90 does not require a concentrating element. The microparticles M entering the electric field of the electrodes 931-93N and 941-94N are gradually deflected toward one of the inner walls 24, 25. A direction of the deflection depends on properties, for example size, of the microparticles M. A dimension of the sorting element 90 and parameters of the applied electric field including frequency and amplitude can be tuned to achieve, for example, that microparticles having a diameter of 8-10 μm may be deflected toward the second inner wall 25, and microparticles having a diameter of 3-5 μm may be deflected toward the first inner wall 24. The sorting element 90 provides a continuous sorting and reduces a probability of the microparticles M not to be sorted.
The partitioning element 23 can comprise a pair of linearly shaped electrodes 103, 104 arranged at the center of the microchannel 13 between the first and the second inner walls 24, 25. Preferably, the partitioning element 23 extends in a direction parallel to the fluid flow direction F.
The partitioning element 23 is configured to generate an electric field for repelling the microparticles M. Sorted microparticles are preferably not able to penetrate or move across the partitioning element 23. The partitioning element 23 is thus configured to prevent the sorted particles from intermixing.
The trapping element 18 comprises a plurality of electrodes 106-109 in an interdigitated arrangement. A first intermediate power line 101 and a second intermediate power line 102 are arranged outside of the microchannel 13 and parallel to the first and second inner walls 24, 25. The electrodes 106 are connected to the first intermediate power line 101 and extend from the first intermediate power line 101 into the microchannel 13 toward the partitioning element 23. The electrodes 108 are connected to the second intermediate power line 102 and extend from the second intermediate power line 102 into the microchannel 13 toward the partitioning element 23. Electrodes 107 extend from a third intermediate power line 103 at the center of the microchannel 13 outward and through the first inner wall 24. Electrodes 109 extend from a third intermediate power line 104 at the center of the microchannel 13 outward and through the second inner wall 25. Each of the electrodes 106-109 can have a width of 10−7 m to 10−3 m, preferably 10-6 m to 10−4 m. The electrodes 106-109 are spaced from one another constantly at a distance of 10−7 m to 10−3 m, preferably 10−6 m to 10−4 m. Different respective spacings between 106, 107 and 108, 109 may allow for generating different dielectrophoretic forces on sorted microparticles M.
Connection wires 105 extending across the microchannel 13 in the w-direction may be formed to connect the intermediate power lines 101, 102 to the power lines 19. Preferably, a distance D101 between the connection wires 105 and the nearest electrodes 106-109 is larger than the spacing between the electrodes 106-109 in order to prevent undesired bead accumulation on the connection wires 105. For example, the distance D101 can be 10−4 m-10−3 m.
The electrodes 106-109 are configured to generate an electric field, in which sorted microparticles can be trapped. Trapped microparticles can be imaged for an analysis.
The sorting element 121 can comprise a pair of linearly shaped electrodes 124, 125 that extend across the microfluidic channel 13 crossing the inner walls 24, 25. A first electrode 124 is inclined at an angle A124 to the first inner wall 24. A second electrode 125 is inclined at an angle A125 to the first inner wall 24. The first electrode 124 is arranged upstream of the second electrode 125 with respect to the fluid flow direction F and is, in particular, shorter than the second electrode 125. For example, the first angle A124 is greater than the second angle A125 so that a gap between the electrodes 124, 125 widens from the first inner wall 24 toward the second inner wall 25.
The sorting element 121 may be configured to divide microparticles, in particular microparticles that are concentrated by a preceding concentrating element, in a plurality of groups. In particular, the sorting element 121 is configured to generate an asymmetric electric field. The microparticles M passing through this asymmetric electric field can be deflected in the w-direction. In particular, microparticles having different material, surface chemistry, topological and/or electrical properties may be differently deflected in the asymmetric electric field of the sorting element 121.
For example, the microparticles M may include three groups of microparticles M1-M3 differing from one another in particle size. The microparticles of the first group M1 may be smaller than the rest of the microparticles of the other groups M2, M3. The microparticles of the third group M3 may be bigger than the microparticles of the other groups M1, M2. The electric field of the sorting element 121 may be configured such that microparticles are deflected farther toward the second inner wall 25 with increasing particle size. As a result, the microparticles M1-M3 may be positioned in different columns in the microchannel 13 as illustrated in
The described sorting mechanism may be advantageous in particular for multiplexed bioassays employing beads with different sizes corresponding to different receptors on their surfaces. Different receptors may be configured for capturing different analytes, respectively, for detection and analysis.
The trapping element 122 is configured to generate an electric field for retaining the sorted microparticles at defined positions. For example, the trapping element 122 can be suited for trapping the microparticles of each group of microparticles M1-M3 in a trapping area 126-128, respectively.
The partitioning element 123 can comprise two physical walls, in particular linearly shaped solid bodies, which the microparticles M cannot penetrate. The partitioning element 123 thus prevents the sorted microparticles M1-M3 from intermixing, in particular from entering the trapping area of other groups of microparticles. Alternatively, the partitioning element 123 may comprise at least one pair of electrodes configured to generate an electric field for repelling the microparticles M. Preferably, the partitioning elements 123 may partition the microchannel 13 in equal widths, thereby allowing for a uniform fluid flow.
The microfluidic chip 10 can comprise a first electrical contact 131 for operating the concentrating element 16, a second electrical contact 132 for operating the sorting element 17, a third electrical contact 133 for operating the trapping element 18 and a fourth electrical contact 134 that is a grounded contact. Preferably, the electrical contacts 131-134 are connected to an alternate current (AC) power supply. Additionally, a function generator and/or control unit may be connected to at least one of the electrical contacts 131-134 that operates and/or controls the elements and devices of the microfluidic chip. For example, the applied signal can be sinusoidal or square wave or a combination thereof
Preferably, the elements and devices of the microfluidic chip 10 are operable and/or controllable via the electrical contacts 131-134. The sorting element 17, trapping element 18, partitioning element 23 spacing element 21 and/or concentrating element 16 may require to be electrically connected to the ground 134 and one of the other electrical contacts 131-133. Preferably, a size of electrical contacts is as small as possible in order to save manufacturing cost and required volume. Accordingly, the number of electrical contacts can be reduced by providing one single ground contact for all elements and devices of the microfluidic chip 10.
In an embodiment, the concentrating element 16, sorting element 17 and trapping element 18 may be connected to two electrical contacts each, resulting in six contacts in total. The total number of contacts may be reduced to four by sharing the ground contact and having independent counter contacts for each element.
Electrical signals applied to the electrical contacts 131-134 may have a peak-to-peak amplitude of 10−1 V to 103 V, preferably 1 V to 100 V. A frequency applied to the electrical contacts 131-134 may be 104 Hz-107 Hz, preferably 105 Hz-3.106 Hz.
In a preferable further embodiment, as shown in
Unless specified otherwise, the angles A31-A125 described above refer to angles of intersection formed by the respective electrode and the corresponding inner wall 24 or 25 inside the microchannel 13 and opening in the fluid flow direction F. In particular, the angles A31-A125 are indicated in the respective
The suggested microfluidic chip, microchannel or method may allow for processing microparticles, in particular beads carrying analytes, via deterministic displacement based on dielectrophoretic forces. Accordingly, the microparticles may be grouped, separated and localized in specific areas using the suggested microfluidic chip, microchannel or method. In particular, the suggested microfluidic chip, microchannel or method may prevent the microparticles from being located at unwanted positions, aggregating, adhering to surfaces of the microfluidic chip or microchannel and sedimenting. The suggested microfluidic chip or microchannel may be implemented in a microfluidic device. The suggested method may be performed using the suggested microfluidic channel or the suggested microchannel, optionally implemented in a microfluidic device.
The descriptions of the various embodiments of the present invention have been presented for purposes of illustration, but are not intended to be exhaustive or limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The terminology used herein was chosen to best explain the principles of the embodiments, the practical application or technical improvement over technologies found in the marketplace, or to enable others of ordinary skill in the art to understand the embodiments disclosed herein.
More generally, while the present invention has been described with reference to certain embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the scope of the present invention. In addition, many modifications may be made to adapt a particular situation to the teachings of the present invention without departing from its scope. Therefore, it is intended that the present invention not be limited to the particular embodiments disclosed, but that the present invention will include all embodiments falling within the scope of the appended claims.
