TW201710681A - Chip structure for microfluidic transportation and detection of biological specimen comprising a bottom plate, a biochip, a cover plate and a silica gel dry film to greatly simplify the chip structure - Google Patents

Abstract

This invention relates to a chip structure for microfluidic transportation and detection of a biological specimen. The chip structure comprises a bottom plate provided with a chip mounting hole in its top surface; a biochip which is mounted in the chip mounting hole and has a top surface flush with a top surface of the bottom plate as well as implanted with a biological probe on its top surface; a cover plate on which a specimen inlet and a specimen outlet are provided, wherein the specimen inlet and the specimen outlet respectively penetrate through the top surface and the bottom surface of the cover plate; and a silica gel dry film made of a low Young's modulus heat-resistant polymer polymerized by silica gel and aromatic hydrocarbons, a microfluidic channel penetrating through the top surface and the bottom surface of the silica gel dry film is disposed in the silica gel dry film, the silica gel dry film is melted and fixed between the top surface of the bottom plate and the bottom surface of the cover plate by means of hot melting, such that both ends of the microfluidic channel can respectively correspond to the specimen inlet and the specimen outlet and a path of the microfluidic channel can correspond to the chip mounting hole.

Description

Microfluidic transfer and detection of wafer structure of biopsy

The invention relates to a detection structure of a biological sample, in particular to a micro-flow transmission and detection of a wafer structure of a biological sample, in order to greatly simplify the process of the structure of the wafer, so that it can be carried out without the need to pass through extremely precise equipment and extremely complicated reproduction process. The fast volume yields a precise and ideal microfluidic channel.

Generally speaking, biochips can be divided into two types according to functions, namely, a sensing type wafer and a micro processing type wafer, wherein different types of biological probes are implanted on the sensing type wafer, and the biological probe can be a gene. Fragment, short-chain nucleic acid to form a gene wafer, if the bio-probe is a protein, a protein wafer is formed. In recent years, other types of bio-sensing wafers have been developed, such as an enzyme wafer to replace a biological tube in a test tube. Chemical reaction; micro-processed wafers are carried out in a miniaturized space. For wafer applications, pre-processed wafers, capillary electrophoresis (CE) wafers, and many Functional processing wafer (Lab-on-a-Chip) and the like.

Since the present invention has been developed for the application of a biosensing type wafer, only the protein wafer therein will be exemplified as follows. According to FIG. 1 , the protein wafer 10 is immobilized on a substrate 12 by using a protein, an antigen or an antibody as a biological probe to form the protein wafer 10, and the antigen and the antibody on the protein wafer 10 are used. The specific nature of the test, to detect specific protein samples, such as: used to test the concentration of specific antigens and antibodies in certain diseases, as a diagnostic basis for the disease, the production principle is to first use the first antibody 20 as a biological probe, fixed On the top surface of the substrate 12, then, the sample to be tested is added, the specific antigen 21 in the sample to be tested is combined with the first-stage antibody 20, and then the second-stage antibody 22 is added, and the The second-stage antibody 22 binds to the conjugate of the first-stage antibody 20-specific antigen 21; at this time, since one end of the second-stage antibody 22 has previously immobilized the specific fluorescent enzyme 23, the sample to be tested is middle The specific antigen 21 is catalyzed by a specific fluorescent enzyme 23 at the end of the conjugate, and is released or converted into a colored substance, and the surface of the protein wafer 10 is irradiated with a specific fluorescent lamp to be in accordance with the range of fluorescence development. The size and the intensity of the fluorescence are measured, and the concentration of the specific antigen 21 contained in the sample to be tested is measured. Therefore, the protein wafer 10 can detect whether the specific protein sample contains the first level on the wafer 10 with the protein. Antibody 20 binds to antigen 21 and its concentration.

Further, in order to allow the antigen 21 contained in the sample to be sufficiently catalyzed by the fluorescent enzyme to be released or converted into a colored substance, the protein wafer 10 is mounted in a micro flow structure to form A micro flow-through sampling chip structure, as shown in FIG. 2, the biochip structure A includes a bottom plate 31, a protein wafer 10, and a cover 41. Referring to FIG. 3a, the bottom plate 31 is formed by uniformly coating a negative (or positive) photosensitive resin 32 on the top surface of one of the substrates 30 made of glass or polymer plastic; The negative (or positive) photosensitive resin 32 is shielded by a photomask 33 having a microfluidic channel pattern 331, and the negative (or positive) photosensitive resin 32 is developed, and the negative type (or positive) After the photosensitive resin 32 is developed and cured, the undeveloped hardened portion thereof is removed, and a microfluidic channel pattern 321 is formed in the hardened negative (or positive) photosensitive resin 32; The upper surface of the substrate 30 corresponds to the portion of the microfluidic channel pattern 321 and is repeatedly performed. After etching and etching the upper surface of the substrate 30 to a predetermined depth, the hardened negative (or positive) photosensitive resin 32 is removed, that is, a microfluidic channel 311 can be formed on the upper surface of the substrate 30; finally, A wafer mounting hole 312 is formed on the substrate 30 at a position corresponding to the path of the microfluidic channel 311, and the substrate 31 is formed after the protein wafer 10 is mounted in the wafer mounting hole 312.

Referring to FIG. 3b, the cover 41 is formed by drilling at least two through holes on the other substrate 40, wherein the through hole is the sample inlet 411 and the other through hole is the sample outlet 412. That is, the cover plate 41 is formed; finally, as shown in FIG. 2, the top surface of the bottom plate 31 and the bottom surface of the cover plate 41 are bonded or welded to each other, and the sample inlet 411 and the sample are made. The outlets 412 respectively correspond to the two ends of the microfluidic channel 311, and are respectively connected to the microfluidic channel 311, thereby forming a microfluidic structure A for transmitting and detecting the sample.

Thus, referring to FIGS. 1 and 2, when a sample to be tested is pressurized and introduced into the sample inlet 411, the sample to be tested can sequentially flow along the microfluidic channel 311. The egg The top surface of the white matter wafer 10 is such that the specific antigen 21 contained in the sample to be tested can be sufficiently catalyzed by the specific luminescent enzyme 23 on the protein wafer 10 to be released or converted into a colored product, and The protein wafer 10 is irradiated with specific fluorescence, that is, it can be detected whether or not the sample to be tested contains the antigen 21 capable of binding to the first-stage antibody 20 on the protein wafer 10 and its concentration.

However, since the depth of the microfluidic channel 311 is extremely shallow, only tens to hundreds of micrometers, it is obviously impossible to use the above-mentioned etching method or other injection molding methods without using extremely precise equipment and through complicated processes. The ideal microfluidic channel 311 is formed on the top surface of the bottom plate 31. Conversely, through the precision equipment and the complicated reproduction process, it only leads to the manufacturing cost and the market price of the biochip structure A of the microfluidic transmission and detection sample. The main reason for being high.

Although there are also attempts to form a desired microfluidic channel on a conventional silicone dry film, the silicone dry film is assembled between a substrate and a cover to form another type of biochip structure. However, due to the extremely thin thickness of the traditional silicone dry film, it is not only extremely soft but not easy to shape, which leads to the inability to maintain the precision of the microfluidic channel during the microfluidic channel processing. In the process of assembling between the bottom plate and the cover plate, liquid glue must be used, and the adhesive overflow overflows to block the microfluidic channel, so that the structure of the biochip is always ineffective.

According to this, how to design a micro-flow transmission and detection of the wafer structure of the biological sample to greatly reduce the process, and to make accurate on the top surface of the bottom plate 31 without using extremely precise equipment and extremely complicated reproduction process. The ideal microfluidic channel 311 is an important problem to be solved by the present invention.

An object of the present invention is to provide a microfluidic transfer and detection of a wafer structure of a biological sample, the wafer structure comprising a bottom plate, a biochip, a cover plate and a first silicone dry film, wherein the bottom plate is made of glass Or a polymer body made of a polymer, the top surface of the bottom plate is provided with a wafer mounting hole; the biochip is mounted in the wafer mounting hole, and the top surface of the biochip is flush with the top surface of the bottom plate, and The plurality of biological probes are implanted thereon; the cover plate is also a plate body made of glass or polymer plastic, and the cover plate is provided with at least one sample inlet and a sample outlet, the sample inlet and the inspection The body outlets respectively penetrate the top surface and the bottom surface of the cover plate; the first silicone dry film It is made of a low Young's modulus and heat-resistant polymer which is polymerized from aromatic and carbonic acid (Aromatic Hydrocarbon) and has low temperature curing, high temperature melting and high temperature. a cross-linking property, so that at least one microfluidic channel can be accurately opened in the first silicone dry film in a low-temperature curing state, and the micro-fluid channel penetrates the first silicone dry film The top surface and the bottom surface, the depth of the microfluidic channel is equal to the thickness of the first silicone dry film, and the first silicone dry film can be baked by heat, melted at a high temperature, and crosslinked by high temperature (cross linking) The precise cross-linking is fixed between the top surface of the bottom plate and the bottom surface of the cover plate to integrate the three rubbers, and the two ends of the micro-flow channel respectively correspond to the sample inlet and the sample The outlets are respectively connectable to the sample inlet and the sample outlet, and the path of the microfluidic channel can correspond to the wafer mounting hole. In this way, when a biological sample to be tested is pressurized and introduced into the entrance of the sample, the biological sample can sequentially flow through the top surface of the biochip along the microfluidic channel, so that the biological sample is in the biological sample. The specific antigen contained can be sufficiently catalyzed by the specific fluorescing enzyme on the biochip to be released or converted into a colored product, and by irradiating the biochip with specific fluorescence, the organism can be detected. Whether the sample contains an antigen capable of binding to the first-stage antibody on the biochip and its concentration.

Another object of the present invention is that the cover (or the bottom plate) further includes a gate hole and a second silicone dry film, wherein the gate hole extends through the top surface and the bottom surface of the cover plate, and corresponds to the a path of the microfluidic channel; the second silicone dry film is made of a silicone polymer and has a high-temperature melting property, wherein a through hole is respectively penetrated through a position corresponding to the inlet of the sample and the outlet of the sample, the second The silicone dry film is baked by heat, fixed at a high temperature and fixed between the bottom surface of the cover and the top surface of the first silicone dry film (or melted and fixed on the top surface of the bottom plate and the first silicone dry film) Between the bottom surfaces). In this way, when a pressurized air is introduced into the gate hole, the portion of the second silicone dry film corresponding to the gate hole is deformed toward the microfluidic channel to shield the path of the microfluidic channel, thereby The biological sample in the microfluidic channel path stops flowing or slows down the flow rate to enable a sufficient biochemical reaction between the biological sample and the biochip.

For the sake of review, the reviewer can have a further understanding and understanding of the technical, structural features and purposes of the present invention. The embodiments are described in conjunction with the drawings, which are described in detail as follows:

[study]

10‧‧‧protein wafer

12, 30, 40‧‧‧ substrates

20‧‧‧First-level antibody

21‧‧‧ antigen

22‧‧‧second-level antibody

23‧‧‧Fluorescent Enzymes

31‧‧‧floor

311‧‧‧Microflow channel

312‧‧‧ wafer mounting holes

32‧‧‧Photosensitive resin

321, 331‧‧‧ microfluidic channel pattern

33‧‧‧Photomask

41‧‧‧ Cover

411‧‧ ‧ entrance to the body

412‧‧‧Exam export

A‧‧‧Biowafer structure

〔this invention〕

10‧‧‧Biochip

50‧‧‧floor

51‧‧‧First wafer mounting hole

60‧‧‧ cover

61‧‧‧Inspection entrance

62‧‧‧Exam export

63‧‧‧Second wafer mounting hole

64‧‧‧ gate hole

70‧‧‧First silicone dry film

71‧‧‧Microflow channel

80‧‧‧Second silicone dry film

81, 82‧‧‧through holes

B‧‧‧ wafer structure

Figure 1 is a schematic diagram of a biochemical reaction between a conventional biochip and an antigen; Figure 2 is a schematic diagram of the decomposition of a conventional microfluidic transfer and detection biochip structure; and Figures 3a and 3b are conventional microfluidic transmission and detection FIG. 4 is a schematic exploded view of a preferred embodiment of the present invention; FIG. 5 is a schematic view showing the molecular structure of the first silicone dry film; and FIG. 6 is a preferred embodiment shown in FIG. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 7 is a schematic cross-sectional view of a group structure of another preferred embodiment of the present invention.

In a preferred embodiment of the present invention, a microfluidic transfer and detection of a wafer structure of a biosample is provided. Referring to FIG. 4, the wafer structure B includes a bottom plate 50, a first biochip 10, a cover plate 60 and a first silicone dry film 70, wherein the bottom plate 50 is a plate made of glass or polymer plastic, and a top wafer mounting hole 51 is defined in a top surface of the bottom plate 50; The wafer 10 is mounted in the first wafer mounting hole 51. The top surface of the first biochip 10 is flush with the top surface of the bottom plate 50, and a plurality of biological probes are implanted thereon; The plate body is made of glass or polymer plastic, and the cover plate 60 is provided with at least one sample inlet 61 and a sample outlet 62. The sample inlet 61 and the sample outlet 62 respectively penetrate the cover plate 60. The top surface and the bottom surface; the first silicone dry film 70 is a low Young's modulus and heat-resistant polymer which is polymerized from silicone and aromatic hydrocarbons (Aromatic Hydrocarbon). Made of, the Young's module value is between 0.01~0.1GPa (Gpa, 1 billion Pascal) or 1 , 500-15,000 lbf / in ² (lbf / in ² , pounds per square inch), equivalent to the hardness and elasticity of rubber materials, and has the characteristics of low temperature curing, high temperature melting and high temperature cross linking. Its molecular structure is shown in Figure 5. In this way, at least one microfluidic channel 71 can be precisely opened in the first silicone dry film 70 in a low temperature curing state, and the microfluidic channel 71 penetrates the top surface and the bottom surface of the first silicone dry film 70. The depth of the microfluidic channel 71 is equal to the thickness of the first silicone dry film 70. The first silicone dry film 70 is baked by heat, melted at a high temperature, and cross-linked by high temperature. The precise cross-linking is fixed between the top surface of the bottom plate 50 and the bottom surface of the cover plate 60 to integrate the three rubbers. The two ends of the micro-flow passage 71 can respectively correspond to the sample inlet 61. The sample outlet 62 is connected to the sample inlet 61 and the sample outlet 62, respectively, and the path of the microfluidic channel 71 corresponds to the first wafer mounting hole 51.

In particular, in the foregoing embodiment, since the first silicone dry film 70 is made of a heat-resistant polymer of a low Young's module obtained by polymerizing a silicone and an aromatic hydrocarbon, it has a low temperature curing. The characteristics of high-temperature melting and high-temperature interlinking, therefore, when the polymer contains a photosensitive material, it can be exposed to a low temperature, in a reticle exposure and development manner, on the first silicone dry film 70. The microfluidic channel 71 is formed accurately; conversely, when the polymer does not contain a photosensitive material, the first silicone dry film 70 can also be cut in a low temperature curing state by laser or knife cutting. The microfluidic channel 71 is formed accurately and rapidly; 嗣, in a low temperature curing state, the first silicone dry film 70 is accurately placed between the bottom plate 50 and the cover plate 60; Bake, the surface of the first silicone dry film 70 is melted and precisely bonded to the bottom plate 50 and the cover 60, and a precise and ideal microfluidic channel 71 is formed between the bottom plate 50 and the cover 60. For transmitting the biological specimen, enabling the biological specimen to interact with the biological The wafer 10 is sufficiently biochemically reacted.

Thus, referring to FIG. 6, a cross-sectional view of the BB section line of the assembled structure of the preferred embodiment shown in FIG. 4, when a biological sample to be tested is pressurized and introduced into the sample inlet 61, The biological sample can sequentially flow through the top surface of the first biochip 10 along the microfluidic channel 71, so that the specific antigen contained in the biological sample can be sufficiently fully on the first biochip 10. By catalyzing a specific fluorescent enzyme, releasing or converting into a colored product, by irradiating the first biochip 10 with a specific fluorescent light, it can be detected whether the biological sample contains the first and the first The antigen bound to the first-stage antibody on the biochip 10 and its concentration.

The above description is only a preferred embodiment of the present invention, but the present invention is not limited thereto when it is actually applied. According to the person skilled in the relevant art, after considering the technical content of the present invention, if it is desired to more accurately detect whether the biological sample contains a specific antigen and its concentration, a second wafer can be added to the cover 60. a mounting hole 63 and a second biochip 10, wherein the second wafer mounting hole 63 is formed on a bottom surface of the cover 60 and corresponds to a path of the micro flow channel 71; the second biochip 10 is mounted on The bottom surface of the second wafer mounting hole 63 is flush with the bottom surface of the cover plate 60, and a plurality of biological probes are implanted on the bottom surface thereof. As such, when the biosample passes along the microfluidic channel 71 and flows through the bottom surface of the second biochip 10, the specific antigen contained in the biosample can be sufficiently specified on the second biochip 10. Phosphorescent enzymes By releasing or converting into a colored product, 嗣, by irradiating the second biochip 10 with a specific fluorescent light, it can be detected whether the biological sample contains the same energy and the second biochip 10 The primary antibody binds to the antigen and its concentration. Accordingly, by synthesizing the data measured by the first and second biochips 10, it is possible to more accurately determine whether the biological sample contains an antigen capable of binding to the first-order antibody on the biochip 10 and concentration.

In addition, during the actual implementation of the present invention, the first wafer mounting hole 51 can also be designed to penetrate the top surface and the bottom surface of the bottom substrate 50 to adjust the mounting position of the first biochip 10 to ensure The top surface of the first bio-chip 10 can be flush with the top surface of the bottom plate 50. Similarly, the second wafer mounting hole 63 can also be designed to penetrate the top surface and the bottom surface of the cover plate 60. The second biochip 10 is mounted to ensure that the bottom surface of the second biochip 10 is flush with the bottom surface of the cover plate 60, thereby ensuring that the biofilm can be properly biochemically reacted with the biochip 10.

In another preferred embodiment of the present invention, as shown in FIG. 7, in order to suspend or slow the flow rate of the biological sample flowing in the path of the microfluidic channel 71, the biological specimen and the living organism are expected to be A sufficient biochemical reaction can occur between the wafers 10. The cover 60 further includes a gate hole 64 and a second silicone dry film 80. The gate hole 64 extends through the top surface and the bottom surface of the cover plate 60, and corresponds to The second silicone dry film 80 is also made of Silicone Polymer and has a high temperature melting property corresponding to the sample inlet 61 and the sample outlet 62. The second silicone dry film 80 can be baked by heat, and is fixed between the bottom surface of the cover 60 and the top surface of the first silicone dry film 70 by heat baking. In this way, during the detection process, if the detecting person wants to extend the period of biochemical reaction between the biological specimen and the biochip 10, a pressurized air can be introduced into the gate hole 64. At this time, the second silicone rubber is dried. The portion of the film 80 corresponding to the gate hole 64 (as indicated by the broken line in FIG. 7) is deformed toward the microfluidic channel 71 by its own elasticity to shield the path of the microfluidic channel 71, thereby making the micro The biosample in the path of the flow channel 71 stops flowing or slows down the flow rate, enabling the biosample to undergo a sufficient biochemical reaction with the biochip 10.

It is necessary to specifically declare that the second silicone dry film 80 is the same silicone polymer as the first silicone dry film 70, so that the surface of the second silicone dry film 80 can be melted after being baked by heat. On the bottom surface of the cover plate 60 and the top surface of the first silicone dry film 70, Both are the so-called silicone polymers herein. In addition, in other embodiments of the present invention (not shown), the gate hole and the second silicone dry film can also be designed on the bottom plate 50 according to actual needs, wherein the gate hole penetrates the bottom plate 50. a top surface and a bottom surface corresponding to the path of the microfluidic channel 71; a position corresponding to the sample inlet 61 and the sample outlet 62 of the second silicone dry film respectively through a through hole, the second silicone dry The film is baked by heat, and is fixed between the top surface of the bottom plate 50 and the bottom surface of the first silicone dry film 70.

The above description is only a few embodiments of the present invention, and the technical features of the present invention are not limited thereto, and those skilled in the relevant art, after considering the technical content of the present invention, can easily think about the equivalent changes. They should not depart from the scope of protection of the present invention.

10‧‧‧Biochip

50‧‧‧floor

51‧‧‧First wafer mounting hole

60‧‧‧ cover

61‧‧‧Inspection entrance

62‧‧‧Exam export

70‧‧‧First silicone dry film

71‧‧‧Microflow channel

B‧‧‧ wafer structure

Claims (8)

A microfluidic transmission and detection of a wafer structure of a biological sample, comprising: a bottom plate having a first wafer mounting hole on a top surface thereof; a first biochip mounted in the first wafer mounting hole, the top surface of the substrate It is flush with the top surface of the bottom plate, and a plurality of biological probes are implanted on the top surface thereof; a cover plate is provided with at least one sample inlet and one sample outlet, and the sample inlet and the sample outlet are Passing through the top surface and the bottom surface of the cover plate respectively; and a first dry film of silicone rubber, which is made of a heat-resistant polymer of low Young's module which is polymerized with silicone and aromatic hydrocarbons, and the Young's module The value is between 0.01 and 0.1 GPa, and has the characteristics of low temperature curing, high temperature melting and high temperature interlinking. In the low temperature curing state, at least one microfluidic channel is opened therein, and the microfluidic channel penetrates the first silicone The top surface and the bottom surface of the dry film, the depth of the micro flow channel is equal to the thickness of the first silicone dry film, the first silicone dry film is baked by heat, melted and crosslinked to the top surface of the bottom plate and Between the bottom surfaces of the cover plates, so that the three are integrated into one body, and Ends of the flow channel are corresponding to the sample inlet and the sample outlet, and respectively with the inlet and a sample outlet communicating this sample, the path of the microfluidic channel of the wafer corresponding to the first mounting hole. The wafer structure of claim 1, wherein the cover further comprises: a second wafer mounting hole formed on a bottom surface of the cover and corresponding to the path of the micro flow channel; A second biochip is mounted in the second wafer mounting hole, the bottom surface of which is flush with the bottom surface of the cover plate, and a plurality of biological probes are implanted on the bottom surface thereof. The wafer structure of claim 2, wherein the first wafer mounting hole extends through a top surface and a bottom surface of the bottom plate. The wafer structure of claim 3, wherein the second wafer mounting hole extends through the top surface and the bottom surface of the cover. The wafer structure according to claim 4, wherein when the first silicone dry film contains a photosensitive material, the film can be exposed and developed in a low temperature curing state on the first silicone dry film. Forming the microfluidic channel. The wafer structure according to claim 4, wherein when the first silicone dry film does not contain a photosensitive material, the first silicone can be dried in a low temperature curing state by laser or cutter cutting. The microfluidic channel is formed on the membrane. The wafer structure of claim 1, 2, 3, 4, 5 or 6, wherein the cover plate further comprises: a gate hole extending through the top surface and the bottom surface of the cover plate, and corresponding to the micro flow And a second silicone dry film, which is made of a silicone polymer and has a high-temperature melting property, wherein a through hole is respectively penetrated through a position corresponding to the inlet of the sample and the outlet of the sample, and the second The silicone dry film is baked by heat, and is fixed between the bottom surface of the cover and the top surface of the first silicone dry film. The wafer structure of claim 1, 2, 3, 4, 5 or 6, wherein the substrate further comprises: a gate hole penetrating through the top surface and the bottom surface of the bottom plate and corresponding to the path of the microfluidic channel; and a second silicone dry film made of a silicone polymer having high temperature melting characteristics corresponding to The sample inlet and the outlet of the sample respectively penetrate through a through hole, and the second silicone dry film is baked by heat, and is fixed between the top surface of the bottom plate and the bottom surface of the first silicone dry film.