Patent Application
20180236444
The present invention relates to a method of manufacture of microfluidic chips having at least one microchannel. Specifically, the present invention relates to manufacturing microfluidic chips having integrated heating elements that are hermetically sealed (electrically isolate) from the fluid in a microchannel. The present invention also relates to microfluidic chips made by the method described herein having hermetically sealed integrated heating elements. The present invention additionally relates to kits comprising such microfluidic chips.
Microfluidic chips for conducting chemical and biological reactions have gained widespread acceptance as a tool used for analytical and research purposes. Provided in a variety of sizes, shapes, and configurations, microfluidic chips are employed for numerous chemical and biological applications.
Microfluidic chips having one or more microchannels formed in silicon, glass, or other substrates are well known in the art. Traditional methods of preparing microfluidic chips are as described in Liu et al. 2000. Proc. Natl. Acad. Sci. USA 97(10):5369-5374 and Anderson et al. 2000. Nucleic Acids Res. 28:e60. Liu et al. describes etching of glass wafers in hydrofluoric acid, followed by sputtering of CR and AU layers and the application of an adhesion layer on top of the metallized layers. The wafers were then spin-coated with a photoresist layer and soft-baked before the photoresist was patterned using UV light through a mask that had the required pattern of microfluidic channels. Following development of the photoresist, the exposed metallized layer was etched off and the channel pattern was chemically etched into the glass. The remaining photoresist and metallized layer was removed, holes to access the microchannels were provided by drilling and the wafer was gleaned before being thermally bonded with a second, blank wafer. Anderson et al. describes the use of a cartridge for use in genetic assays that was prepared by computer-controlled machining as is understood in the art, resulting in a cartridge comprising two 0.25 mm thick silicone layers with a 0.01 mm thick mylar layer between them. Although Anderson et al. used screws and a clamping fixture to hold the layers together, they also provide that ultrasonic welding or adhesives can be used.
In many instances, methods for production of microfluidic chips or “lab-on-a-chip” devices currently use principles and tools of semiconductor process equipment. These methods require patterning metal and electronics on silicon, polymer, and glass substrates. Examples of the integrated microfluidic devices are described in Erickson and Li, “Integrated Microfluidic Devices,” Analytica Chimica Acta 2004, 507:11-26, Zhang and Xing, “Survey and Summary: Miniaturized PCR Chips for Nucleic Acid Amplification and Analysis: Latest Advances and Future Trends,” Nucleic Acids Research June 18, 2007, 35(13):4223-37, Haeberle and Zengerle, “Microfluidic Platforms for Lab-on-a-Chip Applications,” Lab Chip Jul. 27, 2007, 7(9):1094-110, Chen et al., “Total Nucleic Acid Analysis Integrated on Microfluidic Devices,” Lab Chip Aug. 9, 2007, 7:1413-23, Weigl et al., “Towards Non-and Minimally Instrumented, Microfluidics-Based Diagnostic Devices,” Lab Chip Oct. 29, 2008, 8(12):1999-2014, and Sharp et al., Chapter 6: Liquid Flows in Microchannels, in The MEMS Handbook, Ed. M. Gad-el-Hak, CRC Press 2001, each of which are incorporated herein in their entirety.
In some cases, direct electrical contact to the fluidic contents of the channel may be desired for sample manipulation (e.g. for heating, mixing, or impedance-based readout, or other function). In such circumstances, a patterned metal electrode or other circuit element makes intimate direct contact to the fluid. For instance, a capillary electrophoresis chip has high voltage electrodes that are in direct contact with the fluid in a microchannel to provide electrical potential across the microchannel. However, sometimes it is desired to electrically heat the fluid, but avoid electrochemical reactions that would result from direct contact between the fluid in the microchannel and electrical elements.
Fabrication of such microfluidic chips requires bonding of two or more glass or silicon substrates to hermetically seal one or more microchannels that are initially formed in one of the substrates. A metallic layer that serves as a heating element may be integrated into the microfluidic chip. To heat the fluid in a microchannel, the heating element should be in a close proximity to the microchannel which implies that the substrate having the heating element should be relatively thin. For example, Dodge (“Microfluidic devices for heterogeneous assays,” Dissertation, 2003) describes a microfluidic chip comprising two substrates, a channel formed in one of these substrates and hermetically sealed by another, and a metallic layer sputtered over the sealing substrate.
Bonding two glass or silicon substrates by using fusion bonding is known in the art, as previously described. However, fusion bonding often results in void or crack formation due to dust particles, non-planarity and differences in thermal coefficient of expansion are very common. Different approaches have been suggested in the art to bond two substrates by fusion bonding thereby avoiding cracking of the thin substrate.
For example, Stjernström (“Method for fabrication of microfluidic systems in glass,” Journal of Micromechanics and Microengineering, Volume 1, Number 1) describes a method for fabricating integrated microfluidic elements in glass. By employing a matrix of underpinning posts and a thin wall, surrounding etched flow channels, an efficient sealing of glass chips substrates to thin cover glass can be accomplished. However, bonding a thin and a thick substrate still presents a challenge and is rather costly and time consuming.
Accordingly, a process for manufacturing a microfluidic chip that allows avoidance of cracks in a thin substrate while performing fusion bonding that results in forming a thin glass layer sealing a microchannel is needed.
Additionally, there is a need for a microfluidic chip having an integrated circuit elements (e.g. a resistors) positioned in close physical proximity to a microchannel, but hermetically sealed (electrically isolated) from the fluid in the microchannel by a high quality transparent insulating layer.
The present invention relates to a microfluidic chip having at least one microchannel and a method for producing the microfluidic chip. According to one aspect of the invention, a microchannel and through holes are formed in a first substrate. A second unpatterned substrate having a first surface and a second surface is provided to be attached to the first substrate. Specifically, the first surface of the second substrate is attached to the first substrate to encapsulate the microchannel formed in the first substrate. Then, the second surface of the second substrate is metallized to provide integrated electrical elements. In one embodiment, the method further comprises thinning the second substrate after the second substrate is attached to the first substrate to allow the circuit elements to be formed in close proximity to the fluidic channels.
In yet another aspect of the invention, a microfluidic chip is provided. Specifically, the microfluidic chip comprises a first substrate having a microchannel formed therein, a second substrate bonded to the first substrate to encapsulate the microchannel, and electrical circuit elements. Specifically, the second substrate has a first surface and a second surface, wherein the first surface of the second substrate is bonded to the first substrate. The electrical elements are formed on the second surface of the second substrate by using a metallization process.
Integrated electrical circuit elements are hermetically sealed and electrically isolated from the fluid the microchannel. The fluid in the microchannel is heated by electrical current in the electrical elements. In one embodiment, fluidic channels are etched using wet or dry (plasma) etching.
In yet another embodiment of the present invention, the first or second substrate is made of glass, silicone, borosilicate glass, pyrex, or polymer. The first and second substrate may be made of the same or different materials. In yet another aspect of the invention, the second substrate is attached to the first substrate by a fusion bond with high temperature anneal. The electrical elements may comprise a platinum layer and gold pads.
The accompanying drawings, which are incorporated herein and form part of the specification, illustrate various embodiments of the subject matter of this disclosure. In the drawings, like reference numbers indicate identical or functionally similar elements.
The present invention has several embodiments and relies on patents, patent applications, and other references for details known in the art. Therefore, when a patent, patent application, or other reference is cited or repeated herein, it should be understood that it is incorporated by reference in its entirety for all purposes as well as for the proposition that is recited.
The present invention relates to microfluidic chips having at least one microchannel(s) where biological reactions can be performed. Microchannels are generally understood in the art to mean a channel having dimensions corresponding to a depth and width of between 1 μm and 1 mm. To control the temperature of the fluid in a microchannel, heating elements may be integrated in the microfluidic chip. More specifically, the present invention relates to a method for manufacturing a microfluidic chip having at least one microchannel and integrated electrical elements that are hermetically isolated from the fluid in the microchannel and which can serve as resistive heating elements.
It is noted that microfluidic chips of the present invention may be made using a variety of techniques used in semi-conductor manufacturing as well as in biological chip and cartridge manufacturing. For instance, reference is made to US Patents and Published Applications U.S. Pat. No. 6,236,447, 2009/0117669, 2007/0111250, 2004/0171043, 2003/0087503, and 2003/0151144, which are incorporated herein in their entirety.
A plurality of microfluidic channels 112 extending across the first substrate 110 can be formed. In step 210, microfluidic channel 112 formation can be preceded by a lithographic or other similar process in order to provide resist patterning for the microfluidic channel 112 on the substrate 110. For instance, the first surface of the substrate 110 can be coated with a photoresist and be patterned with UV light through a mask, wherein the mask provides the pattern of the microfluidic channel 112. Those of skill in the art will readily know and understand those photoresists that would be appropriate for such an application, for instance, an ultraviolet photosensitive organic material, and would otherwise readily understand the process. Those of skill in the art may utilize other alternative processes including molds, etc. for providing the microfluidic channel 112 pattern on the substrate 110.
In one non-limiting embodiment, the resist patterning followed by etching by using wet or dry etching. Wet etching can indicate chemical etching using a liquid, while dry etching may utilize a gas, such as in reactive ion etching or plasma etching. In another embodiment, the microchannel 112 can formed by mechanical abrasion, including sandblasting or powder blasting. Other methods may include ion beam ablation, laser ablations, or some combination thereof. Such methods of channel formation are well known to those of skill in the art, and the appropriate selection of a particular method is within the scope of standard practice for one of skill in the art.
In the next step 230, access holes 114 are created in the substrate 110 to allow the introduction of fluid into the microchannel 112 from an external source. In one embodiment, the access holes 114 are created 230 between the microchannel 112 and the second surface of the substrate 110. In one embodiment, the holes 114 are formed by etching, as described above, or by drilling. Other methods for forming the access holes 114 may include ion beam ablation or laser ablation. Next, in step 240 a second blank (unpatterned) substrate 116 having a first and second surface is attached or bonded to the first substrate 110 to encapsulate microchannel 112 and complete the channel formation process. Such attachment occurs by attaching the second surface of the substrate 116 to the first surface of the substrate 110. The substrate 116 may be formed of quartz, glass, polymer, pyrex, silicon, or borosilicate glass. Substrate 116 may be of any thickness, although typical thicknesses may range from about 100 μm to about 1000 μm. Substrates 110 and 116 can be made of the same or different materials. In one non-limiting embodiment, substrates 110 and 116 are attached to each other. Attachment of substrates 110 and 116 may be accomplished by any means known in the art, including by a thermal or fusion bond.
After the first surface of the first substrate 110 and the second surface of the second substrate 116 are bonded, in the next step 250, the first surface of the second substrate 116 is optionally thinned to allow circuit elements to be formed in close proximity to the microchannel 112. Bonding the first substrate 110 to the second substrate 116 prior to thinning the second substrate 116 allows the bond to be a fusion bond with high temperature anneal. Using the fusion bond for two relatively thick substrates 110 and 116 allows avoiding formation of cracks and voids in the first and second substrates 110 and 116.
In one embodiment, the thinning process is performed by using one or more of wafer lapping, mechanical grinding and/or polishing, wet etching, dry etching, ion beam ablation, laser ablation, or any combinations thereof. In one non-limiting embodiment, the thinned second substrate 116 has a final depth of under about 100 μm. In a further embodiment, the thinned second substrate 116 has a final depth of about 2 μm to about 50 μm, or about 10 μm. In the next step 260, one or more metal layers 118 and 120 are patterned on the first surface of the thinned second substrate 116 using standard processes including, but not limited to, photolithography. In the next step 270, the metal is deposited using techniques known to those of skill in the art, including but not limited to, sputter and lift-off. For instance, U.S. Pat. No. 5,908,319 (which is incorporated by reference herein) describes a method of using photoresists in semiconductor manufacturing which can be equally applied to the present invention. The metal deposited in one or more layers 118 and 120 may be one or more of gold, platinum, aluminum, chrome, titanium, copper, silver, nickel and the like, including alloys and mixtures thereof
In one embodiment, the one or more metal layers formed on the first surface of second substrate 116 may comprise a platinum layer 120 across the first surface of these second substrate 116 and gold pads 118 placed on top of the platinum layer 120 on those portions of substrate 116 outside of the microchannel 112. Circuit elements controlling the temperature of the fluid in the microchannel 112 are based on the patterned metal layers 118 and 120. The metal layers 118 and 120 may therefore be formed from any suitable resistive material that demonstrates good response to temperature and is capable of being used as a heater. Suitable metal layer materials include, but are not limited to, those described above, including platinum, and nickel.
In a further step 280, an optional layer of resin, polymer, or other suitable material may be coated over the metal layers 188 and/or 120. In one embodiment, the coating acts as an insulator. In another embodiment, the coating acts as a physical protectant.
Accordingly, a reaction fluid may be introduced into the microchannel 112 from an external source. The temperature of the fluid in the microchannel 112 is controlled by the metal layer 118 that serves as a resistive heating element. All circuit elements based on the heating elements 118 and gold pads 120 are hermetically isolated from the reaction fluid in the microchannel 112 such that electrochemical reactions in the microchannel 112 can be avoided.
Bonding the first substrate 110 to the second substrate 116 prior to metallization allows the bond to be a fusion bond with high temperature anneal, which will effectively turn the two separate substrates (wafers) 110 and 116 into a single annealed substrate.
One of the advantages of the method as described above is that the quality of isolation between the circuit elements and the fluidic channel could be substantially improved over other methods, allowing for greater device reliability and longevity.
In a further embodiment of the present invention, there is provided kits for biological assays that comprise microfluidic chips as described herein and as prepared by the methods described herein.
The use of the terms “a” and “an” and “the” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.
While the subject matter of this disclosure has been described and shown in considerable detail with reference to certain illustrative embodiments, including various combinations and sub-combinations of features, those skilled in the art will readily appreciate other embodiments and variations and modifications thereof as encompassed within the scope of the present disclosure. Moreover, the descriptions of such embodiments, combinations, and sub-combinations is not intended to convey that the claimed subject matter requires features or combinations of features other than those expressly recited in the claims. Accordingly, the scope of this disclosure is intended to include all modifications and variations encompassed within the spirit and scope of the following appended claims.
This application is the national phase of International Patent Application PCT/US2015/039074 filed Jul. 2, 2015, published as WO 2016/004362, which claims the benefit of U.S. Provisional Patent Application Ser. No. 62/020,102, filed on Jul. 2, 2014, which is incorporated herein by reference in its entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US2015/039074 | 7/2/2015 | WO | 00 |
| Number | Date | Country | |
|---|---|---|---|
| 62020102 | Jul 2014 | US |
