WO2004072642A1 - A device for application of micro-sample and reaction of a biochip and its method - Google Patents

Abstract

The invention relates to a device for application of micro-sample and reaction of a biochip and assay method thereof, and the device comprises an array of grooves manufactured in a microfluidic channel mould, and through holes at the both ends of the groove. The microfluidic channel mould with grooves is put on a chip, and a micro tube is inserted into the through hole, forming a microfluidic channel with a inlet and a outlet. The device of the present invention can be used to manufacture biochips and perform assay and detection. The method includes that: flowing through the microfluidic channel and entering the cavity, biological solution contacts with the surface of the chip, and the biological molecule is immobilized on the surface; protein bond in each groove can react individually with the solution detected through microfluidic channel, or can react with one or more solution detected by connection of microfluidic chnnels or switches which join all the grooves in series or a set of grooves. Therefore, the chip can be used easily and expediently. It is deserved to mention that the device fabricated by the method could be used repeatedly.

Description

 Device and method for micromolecular sample loading and reaction of biomolecule chip

 The invention relates to a method and a device for preparing a biomolecule chip, and in particular, to a device and method for directly performing micro-sample addition on a biomolecule chip, and directly performing a biomolecule fixing and detection reaction on the biochip. Background technique

 Biomolecule chip is an integrated parallel biological detection technology developed in recent years. It can integrate a variety of ligands on a small geometric scale, so that it can detect multiple indicators of a small amount of samples at the same time. Due to the high price of biomolecule samples and the minimum amount required, it is required to miniaturize the biomolecule reagents and samples used in biomolecule chips, which also requires chip loading and miniaturization of the reaction device. At present, the main application of the biochip is the spotting instrument. Divided into two types according to the different spotting methods, one is contact type, first use the spotting needle to dip the ligand to be used, and then touch the chip surface to place the ligand on the chip; one type is spray printing, first Use a hollow spotting needle to suck a small amount of ligand to be spotted, and then add the ligand to the surface of the chip in a manner similar to an inkjet printer. The common disadvantages of these two methods are the uneven spotting amount and the uneven distribution of the areal density of the ligand molecules in a single spot. This will seriously affect the quality of the test results and is difficult to quantify. At present, most of the biochip reactions use the overall reaction method, that is, the entire chip is immersed in the sample solution to be tested. This method requires a large amount of sample solution, a long reaction time, and low sensitivity.

Another chip loading and reaction technology is the use of microchannel technology. At present, chips commonly used in microchannel technology are integrated, that is, the chip and microchannel are made on the same material, such as reference 1 _: Dielectrophoretic cell separation and gene expression profiling on microelectronic chip arrays. July 15, 2002 Huang Y, Joo S, Duhon M, Heller M, Wallace B, Xu X Anal Chem 2002 Jul 15; 74 (14): 3362-71. The microfluidic channel used in this method is a one-time use. The microfluidic channel is complicated to make and the cost is high, which limits the application. Summary of the invention

 The purpose of the present invention is to overcome the shortcomings of the prior art described above, to greatly reduce the manufacturing cost of biochips and to simplify the process of biomolecule immobilization and detection reactions, so as to provide a set of biochip fabrication and Integrated device and method for directly carrying out biomolecule immobilization and detection reactions on a chip.

The object of the present invention is achieved as follows: The device for micro-sample addition and reaction of a biomolecule chip provided by the present invention includes:

 At least one microchannel template 3 has grooves 2 arranged in an array on the surface, and two openings 10 are formed at both ends of each groove 2 as liquid inlets 7 and 8 respectively; the microchannel template 3 has grooves The surface of 2 is buckled on the chip 1, and a cavity 9 is formed between the groove 2 and the chip 1. The liquid inlet and outlet and the cavity 9 form a microchannel 5 in and out (as shown in FIG. 2a). The liquid inlet 7 and outlet 8 of the device can also be installed with a microtube 12, and the microtube 12 is in communication with the micro pump.

 Alternatively, a micro-channel template 3 'is further provided, and a through-hole is also formed on the micro-channel template 3'. The position and number of the through-holes are set as required. The surface of the microchannel template 3 'with the groove 2 is buckled on the microchannel template 3, and its through holes communicate with the liquid inlet 7 and the liquid outlet 8 on the microchannel template 3, respectively. The groove 2 communicates the remaining liquid inlet 7 and the liquid outlet 8 on the micro-channel template 3 to form a micro-channel 5 in and out (as shown in FIG. 2b). A micro-tube 12 and a micro-tube 12 can be respectively connected to the liquid inlet 7 and the outlet 8 of the micro-channel template 3 '.

 Alternatively, it also includes two rigid material blocks 4, one of which is provided with a through hole 15 and is mounted on the micro channel template 3, the through hole 15 communicates with the through hole 10 of the micro channel template 3, and the hole size is the same ( (As shown in Fig. 3a); Another piece of rigid material 4 'is installed under the chip 1, and is clamped together with a common clamp as a whole, or at least 2 holes are opened in each piece of rigid material 4, and a magnet is placed in the hole. Block 13; The magnets embedded on two rigid material blocks are of opposite polarity. Under the action of magnetic force, the groove 2 on the microchannel template 3 and the surface of the smooth chip 17 are sandwiched to form a cavity. 9, forming a microchannel 5 in and out. A micro tube 12 can also be installed in the through hole 15 of the rigid material block, and the micro tube 12 is in communication with the micro pump (as shown in Fig. 4a).

 The above device may further include a third rigid material block 4 ", which has an inlet 19 and an outlet 20 thereon. The position and number of the inlet and outlet are set as required. When all points on the chip surface are connected in series When it is up, that is, two through holes are opened in the rigid material block 4 ", one through hole corresponds to the liquid inlet 7 on the microchannel template 3, and the other through hole corresponds to the liquid outlet 8 of the last groove 2; the second The grooved surface of the microchannel template 3 'covers the first rigid material block 4, and the liquid inlet 7 on the microchannel template 3 is connected to the liquid outlet 8 on the microchannel template 3, and the third A rigid material block 4 "covers the second microfluidic template 3 '. A through hole 10 in the microfluidic template 3 communicates with the inlet 19 on the rigid material block; the other outlet 20 communicates with the microfluidic template 3 The liquid outlet 8 of the last groove is connected; the inlet 20 and the outlet 21 of the rigid material block 4 "are installed with microtubes 12 for sample addition to form a flow channel to realize the communication of all the flow channels (as shown in Figure 4b) As shown). After the series connection, the micro pump sends the solution to be tested into the first groove through the through hole 7, and then the solution to be tested will sequentially flow through all the grooves connected in series (ie, the microchannel 5). Outflow.

When it is required to connect the three grooves on the microchannel template 3 in series, that is, the liquid inlet 7 and the liquid outlet 8 are provided at the positions corresponding to the through holes of the first and third grooves of the rigid material block 4 ", On inlet 7 and liquid outlet 8 A microtube 12 is installed to form three grooved serial flow channels.

 The method for manufacturing the integrated biochip provided by the present invention and the method for directly and immediately carrying out the biomolecule fixation and detection reaction on the manufactured biochip include: using the device of the present invention (as shown in FIG. 4)

 1. With the push of a micro pump, different biomolecule solutions enter different grooves through different microchannels, contact the surface of the chip and be fixed on the surface of the chip, and then the buffer is cleaned by the pump. Surfaces and microchannels to remove biomolecules that are not immobilized on the chip surface;

 2. After each point is fixed, when each point needs to react with a separate test solution, in this state, different test solutions can be delivered to different grooves and fixed by pumps. The biomolecules are reacted, and then washed with a buffer solution after the reaction;

 3. Or all the fixed points need to react with the same solution to be tested or group the points on the chip with different solutions to be tested, then connect the grooves in series as required (as shown in Figure 4b). Then, the solution to be tested is transferred into the groove through the liquid inlet, and reacts with the fixed biomolecules in turn, and the reacted liquid is output from the liquid output port, and then washed with buffer solution after the reaction;

 4. Remove the chip after the reaction, and use conventional methods to test the results.

 When the device is restored to the state shown in Figure 4a after testing, it can be reused again.

 Another device for micromolecular sampling and reaction of a biomolecule chip of the present invention includes:

 A rigid material block 4, a microchannel template 3 and a microchannel template 33.

 The micro-channel template 33 (shown in FIG. 6) is provided with grooves 14, and the pitch of the grooves 14 is equal to the length of the groove 2 on the micro-channel template 3. One end of each of the channels 14 corresponds to a recess. The other hole 6 of the through hole 10 of the groove 2 is extended to the edge; a horizontal groove 11 is also connected between the two grooves 14 corresponding to each groove 2; and the microchannel templates 3 and 33 are bonded together The grooves and grooves face outward, and the holes communicate; the rigid material block 4 is covered on the grooves of the micro-channel template 33, and the four sides are sealed and bonded together, and the chip 1 is covered on the grooves of the micro-channel template 3 A closed microfluidic channel is formed between the microfluidic template and the rigid material block on the surface of the groove 2 (as shown in FIG. 5). In this way, each groove on the microchannel template 3 is connected to two closed microchannels. Each microchannel has a tactile switch 16 and a tactile switch between the two microchannels. 17. Both switches close the microfluidic channel by pressing the elastic material with external force.

 The method for manufacturing the integrated biochip provided by the present invention and directly performing the biomolecule immobilization and detection reaction on the produced biochip, using the device of the present invention (as shown in FIGS. 5 and 6) includes:

 (1) installing the chip material in the device of the present invention, making it in close contact with the microchannel template 3, and pressing the microchannel with an external force;

(2) Transfer the ligand molecules to the selected area on the surface of the chip substrate through the microtube of the microchannel; wait until After the ligand molecule is fixed on the chip substrate;

 (3) Rinse the biomolecule-immobilized chip substrate prepared in step (2) with a buffer solution, and transfer the buffer solution to different areas on the chip surface through a microfluidic channel; wash away the components not fixed on the chip surface Base molecule

 (4) connecting a region in which ligand molecules are fixed in series through a microchannel for tandem; or connecting a region in which ligand molecules are fixed in series by pressing a switch formed by pressing a node on an elastic membrane;

(5) transport the biological sample to be detected through the microfluidic channel to each unit on the surface of the chip prepared in step (4), and perform a detection reaction;

 The micro-channel template includes: made of silicone, rubber or other elastic plastic materials.

The rigid material block is made of plastic, metal, plexiglass and other materials, and has a thickness of 1 mm to 10 dishes. The strip-shaped groove is a groove, which groove strip area from 0.01 ² bandit bandit _^2-1; bandit depth from ΙΟμπι- 1; the number of which at least two strip-shaped grooves, for example, from 1 -500.

 The inner diameter of the through hole 10 can be from 10 μm to 1 leg.

 The inner diameter of the microchannel can be from 10 μm to 1 μm.

 The inner diameter of the groove can be from 10 μm to 1 band.

 The chip base material includes: silicon, glass, metal, plastic, coated silicon wafer and other materials or the above-mentioned composite materials, preferably silicon.

 The advantage of the present invention is that the method provided by the present invention is to use a plurality of tiny grooves arranged in an array on a sheet of elastic material (such as silicone), fasten these grooves to the chip, and select a solution input port and The output port, and a microtube is installed on the solution input port and the output port to form a plurality of closed cavities. Each cavity has one microchannel and one microchannel, and the biomolecule solution enters the cavity through the microchannel. Contact with the chip surface, it is fixed on the chip surface. The proteins fixed in each groove can be individually reacted with the test solution through the microfluidic channel, or all the grooves can be connected in series or grouped in series with one or more of the to-be-measured through the microfluidic plug or switch. The solution was reacted. Makes the use of the chip very flexible and convenient. It is more worth mentioning that the device made by this method can be reused after being washed clean. In addition, the method of the present invention can simultaneously and independently perform multi-point sampling on a chip; the amount of sampling and the size of the points can be controlled. The area on the chip where the ligand biomolecules are fixed is strictly defined by the groove. The surface after fixing and reaction is washed with buffer solution, so that the size of the dots on the chip and the density of the biomolecules within the points are consistent, Effectively improve detection quality. The chip reaction is limited to a small area, and in a flowing state, the mass transfer rate of biomolecules is accelerated, the reaction time is effectively shortened, and the sensitivity is improved.

Arbitrary multiple points can be connected in series, making the use of the chip more flexible. By tandem, trace The sample can react with all points, saving the amount of solution to be tested. The method of the invention enables the preparation, reaction, and detection of the biochip to be implemented in the same device, and can also be reused, which effectively reduces the preparation cost of the chip. BRIEF DESCRIPTION OF THE DRAWINGS

 FIG. La is a schematic plan view of a microchannel template structure with microchannels in the device of the present invention

 Figure lb is a cross-sectional view taken along AA 'of Figure la

 Figure 2a is a schematic cross-sectional view of the first composition of the device of the present invention

 Figure 2b is a schematic cross-sectional view of the second composition of the device of the present invention

 Figure 3a is a schematic plan view of the rigid material block structure of the present invention

 Figure 3b is a cross-sectional view taken along AA 'of Figure 3a

 Figure 4a is a schematic cross-sectional view of the third composition in the device of the present invention (representing the structure of the device when the sample is fixed or the detection reaction is performed at each point separately)

 4b is a schematic cross-sectional view of a fourth composition in the device of the present invention (representing a structural diagram when directly preparing a biomolecule chip, the through holes at the two ends of the groove of the microchannel template are staggered with the through holes of the rigid material block to form a micro Flow channel cooperation diagram)

 FIG. 5 is a schematic structural diagram of another microfluidic device according to the present invention.

 Figure 6 is a top view of the microchannel template shown in Figure 5

 The illustration is as follows:

 1 -chip 2 -groove 3 -micro runner template

 3 '-Microchannel template 33-Microchannel template with groove 4-Rigid material block

 4 '-Rigid material block 4 "-Rigid material block 5 -Micro runner

 6-(recessed) port 7-liquid inlet to be tested 8-liquid outlet to be tested

 9-cavity 10-through hole 11-horizontal groove

 12-microtube 13-magnetic block 14-groove

 15— 通 孑 L 16—First touch switch 17—Second touch switch

 18—hole 19—microchannel inlet 20—microchannel outlet

The following describes the present invention in detail with reference to the accompanying drawings and embodiments: See FIGS. 1a and 1b; Take a piece of silicone sheet as the micro-channel template 3, which has twelve grooves 2 formed on the silicone sheet, and the groove area is lrnrn ² 5mm。, a depth of 0. 1mm; the inner diameter of the through hole 10 at both ends of the groove 2 is 0. 5mm.

3a and 3b _{; the} rigid material block 4 is plexiglass, which is opened with a through hole 15 having a pore diameter of 0.5mra, which The position of the through hole 15 corresponds to the through hole 10 of the microchannel template 3; and a hole with an inner diameter of 5 mm is opened on each side of the rigid material block 4, and a magnetic block 13 is placed in the hole.

 The microtube 12 is a polytetrafluoroethylene tube. The material of the chip 1 is silicon. Microchannel inner diameter 0.5

 Referring to FIG. 2a, the surface of the twelve grooves 2 formed on the above-mentioned silicone sheet 3 is buckled on a silicon chip 1 to form a microchannel 5 having a liquid inlet 7 and a liquid outlet 8; the liquid inlet 7 and a A PTFE microtube 12 can also be installed in the liquid outlet 8, and the microtube 12 is in communication with a micro pump.

 Referring to FIG. 2b, a micro-flow channel template 3 'is further included. The face of the flow channel template 3' with the groove 2 is buckled on the micro-flow channel template 3, and the liquid inlet 7 and the micro-flow of the micro-flow channel template 3 ' The channel template 3 communicates with the liquid outlet 8 to form a microfluidic channel 5 in and out; a microtube 12 can also be installed in the through hole, and the microtube 12 is in communication with the micro pump.

 Referring to FIG. 4a, it also includes two rigid material blocks 4 and 4 '. The rigid material block is a metal material and has a thickness of 2 mm. One of them has a plurality of through holes 15 and is mounted on the micro-channel template 3'. The through-hole 15 communicates with the through-hole 10 of the micro-channel template 3 ', and the hole size is the same; another rigid material block 4' is installed under the chip 1, and is clamped as a whole with a common clamp, or each block The rigid material block 4 is provided with at least two holes, and a magnetic block 13 is placed in the hole; the magnets embedded in the two rigid material blocks have opposite polarities, and the concave of the micro-channel template 3 is under the action of the magnetic force. The slot 2 is sandwiched between the surface of the smooth chip 1 to form a cavity 9, which forms a microchannel 5 in and out; a microtube 12 can also be installed in the through hole, and the microtube 12 is in communication with the micro pump .

 Referring to FIG. 4b, based on the device made in FIG. 4a, the microtube 12 is first removed from the microchannel opening, and then a microchannel template 3 'and a rigid material block 4 "are set on the rigid material block 4". Only one liquid input hole 7 and output hole 8 are opened, and then a microchannel template 3 'with a groove 2 is placed on the rigid material block 4, with the groove 2 facing downward, and the microchannel template 3' on the microchannel template 3 ' The liquid inlet 7 is installed through the liquid outlet 8 of the micro-channel template 3 and communicates with the inlet 19 and the outlet 20 on the rigid material block 4 "to form a micro-channel 5. In this way, each sealed cavity has one inlet and one inlet. Exit the two microchannel inlets and outlets 19, 20. Under the impetus of the 12-channel micropump, different protein solutions enter different grooves through different microchannels, contact the surface of the chip and be fixed on the chip The surface of the chip and the microfluidic channel are then cleaned by pumping the buffer solution to remove the protein molecules that are not fixed on the chip surface. The device is restored to the state shown in Figure 4a, and 12 types of protein fixation can be repeated .

 Refer to the device made in 4a, and fix and react twelve proteins on it.

 (1) As shown in the figure, the silicon wafer 1 is installed under the groove 2 of the micro-channel template 3;

 (2) Under the push of a micro plunger pump, 12 kinds of protein solutions (concentrations of 0.1 mg / ml) were respectively delivered to different areas on the silicon chip through the micro-channel, and the flow rate was controlled at 1 microliter / minute 10 minutes

(3) Then, the phosphate buffer solution is delivered to the surface of the chip 1 prepared in the above step (2) through the microchannel, and the biomolecules not fixed on the surface of the silicon wafer are washed away; It is fixed on the surface of the chip. After washing with buffer solution, the DNA not fixed on the chip surface is discharged.

 Open the switch 17 and close the switch 16 to connect the grooves on the micro-channel template 3 in series. The solution to be tested enters the micro-channel from the inlet through the pump, flows through all the grooves, and is fixed on the chip surface. The DNA is subjected to a hybridization reaction, and finally discharged from the outlet, and then washed with a buffer solution. After the reaction, the chip is removed from the device for detection. The device can be used for the next DNA fixation and reaction.

 Using a device with 1000 grooves, to fix and detect 1000 genes, the steps are as follows:

(1) When preparing a gene chip, under the action of external pressure, the silicon wafer is in close contact with the microarray template, so that 1000 independent sealed cavities are formed between the silicon wafer and the groove, and each cavity has one Enter and exit two micro-flow channels;

 (2) Under the impetus of the micro plunger pump, 10 microliters (concentration of 0.1 g / ml) of 1000 kinds of DNA molecule solutions of different sequences were delivered to different regions on the silicon wafer through the microchannel, and the flow rate was controlled at 1 μl / min, after the DNA molecules are fixed;

 (3) Then, rinse with a phosphate buffer solution to wash away molecules that are not fixed on the surface of the silicon wafer;

(4) Remove the microtube 12, use a ± 夬 closed silicone membrane 3, cover the microfluidic channel, and connect 1000 grooves in series so that the regions where the DNA molecules are fixed are connected in series;

 (5) By injecting 100 microliters of the DNA sample to be tested which has undergone denaturation treatment into the liquid inlet 7 to be tested, the sample is injected at a flow rate of 10 microliters / minute, and passed through a microchannel through each of the chips prepared in step (4) After the zone reaction, it flows out from the liquid outlet 8 to be tested; Rinse with phosphate buffer;

(6) Remove the silicon wafer and use the detector to detect the reaction result.

(4) Remove the Teflon microtube, as shown in Figure 4b; install a microchannel template 3 'for series connection with the rigid material block 4, and then install the Teflon microtube in the first 12 holes are formed in series on one hole and the last hole;

 (5) Inject 100 microliters of the test solution into the microchannel with a flow rate of 10 microliters / minute. The test solution flows through the 12 grooves in sequence and flows out through the outlet, and then rinsed with phosphate buffer solution (as shown in Figure 4). .

In this embodiment, a device with 48 grooves is prepared to prepare a 48-point protein chip to react with two solutions to be tested. In order to be able to perform detection of dozens or hundreds of proteins at the same time, the structure of the device is the same as the embodiment 1 of the same. 05mm。 The area of each groove is reduced to 0.3 mm ² and the depth is 0.05 mm. The inner diameter of the microchannel was reduced to 0.3 bandits. Teflon tubing was replaced by stainless steel tubing. Because protein detection results often need to be compared, the 48 grooves in this embodiment are evenly divided into two groups, and 24 grooves in each group are connected in series. The protein fixation procedure was the same as in Example 1. The two solutions to be tested are injected into two sets of grooves connected in series to react with the fixed protein. The results of the reaction are displayed on the same chip and can be easily compared.

 Referring to FIG. 5, a device for making 400 grooves is used for fixing and hybridizing reactions of 400 kinds of DNA fragments: a microchannel template 33 (shown in FIGS. 5 and 6) with microchannels and a block 4 of rigid material are produced. The micro-channel template material in this embodiment is rubber, the rigid material block is plexiglass, and the chip base material is gold-plated glass. The microchannel template 33 is provided with a groove 14. The groove is 0.1 legs wide and 0. lmm deep. One end of the groove 14 has a through hole corresponding to the groove on the microchannel template 3, and the other port 6 of the groove 14 is extended to the edge. There is a groove of the same shape between the two grooves corresponding to each groove. 11 are connected.

Refer to Figure 1 for the microchannel template 3; take a rubber sheet with an area of 20mmx20mm as the microchannel template 3, and the microchannel template 3 with 400 grooves 2 engraved on the surface, each strip groove 2 depth 1mm ^{2。 It is} 0.01mm, the cross-sectional area is 0.1mm2. The 400 pieces are regularly arranged on the rubber sheet 1 in 20 rows, and 20 grooves 2 are arranged in one row. 1mm, 800 条。 The microfluidic template 3 has the groove 2 facing up, and each end of each of the strip-shaped grooves 2 has a through hole 10, the inner diameter of the through hole 10 is 0. 1mm, a total of 800.

 The plexiglass block 4 is covered on the grooves of the microchannel template 33, and the periphery of the two is bonded together. A closed microchannel is formed between the microchannel template and the plexiglass block 4, which can correspond to The holes are connected. The micro-channel template 3 and the micro-channel template 33 are bonded together, and the corresponding holes are communicated. In this way, each groove in the microchannel template 3 is connected to two closed microchannels. Each microfluidic channel is provided with a pressure switch 16. A horizontal groove 11 is provided between the two microfluidic channels, and a pressure switch 17 is provided thereon. Both of these switches close the microfluidic channel by pressing the elastic material with external force.

 Under the action of external pressure, the chip 1 is in close contact with the microchannel template 3, so that 400 independent cavities are formed between the chip and the groove. Each cavity has one microchannel and one microchannel. After the switch 17 is turned off, under the push of the micro pump, 400 kinds of DNA solutions enter the cavity through the microfluidic channel and contact the surface of the silicon wafer, so that the DNA

7

Claims

Rights request  A device for micro-sample loading and reaction of a biomolecule chip, comprising: a chip (1) and a microchannel (5); characterized in that it further comprises at least one microchannel template (3), wherein Grooves (2) arranged in an array are formed on the surface of the microchannel template (3), and two through holes (10) are opened at both ends of each groove (2), which are respectively used as a liquid inlet (7) and an outlet (8); The grooved surface of the microfluidic template (3) is buckled on the chip (1) to form a microfluidic channel (5).  2. The device for micro-sample loading and reaction of a biomolecule chip according to claim 1, further comprising a microchannel template (3,), the microchannel template (3 ') having a groove The surface is buckled on the micro-channel template (3), and the liquid inlet (7) of the micro-channel template (3 ') communicates with the liquid outlet (8) of the micro-channel template (3) to form a one-in-one micro-flow Road (5).  3. The device for micromolecular sampling and reaction of a biomolecule chip according to claim 1, further comprising two pieces of rigid material (4), one of which is provided with a through hole (15), and is mounted on On the micro-channel template (3), the through-hole (15) communicates with the through-hole (10) of the micro-channel template (3), and the hole size is the same; another rigid material block (4 ') is mounted on the chip ( 1) Below, the whole is clamped and integrated with a general jig.  4. The device for micro-sample loading and reaction of a biomolecule chip according to claim 3, characterized in that: the two rigid material blocks (4) and (4 ') are provided with at least 2 holes, A magnet block (13) is placed in the hole; the magnets embedded in the two rigid material blocks have opposite polarities. Under the action of the magnetic force, the groove (2) on the micro-channel template (3) and the chip (1) The tight contact is sandwiched as a whole to form a micro-flow channel (5) which enters and exits.  5. The device for micromolecular sampling and reaction of a biomolecule chip according to claim 3, further comprising a third rigid material block (4 ") having an inlet (19) and an outlet opening thereon. (20); covering the grooved surface of the second microchannel template (3 ') on the first rigid material block (4), and the liquid inlet (7) on the microchannel template (3') Installed in communication with the liquid outlet (8) on the microchannel template (3), a third rigid material block (4 ") covers the second microchannel template (3 '), and its inlet (19) corresponds to the micro The liquid inlet (7) and the outlet (20) on the flow channel template (3) correspond to the liquid outlet (8) of the last groove (2) on the micro flow channel template (3), forming a micro flow channel with one in and one out (5).  6. The device for micro-sample loading and reaction of a biomolecule chip according to any one of claims 1, 2, 3, 4 or 5, characterized in that: the inlet and outlet of the microfluidic channel (5) A microtube (12) is also installed in the (12) Communicate with the micro pump.  7. A device for micro-sample loading and reaction of a biomolecule chip, comprising: a chip (1) and a microfluidic channel (5); characterized in that: further comprising: A piece of rigid material supporting the role (4) _; At least one micro-channel template (3) has grooves (2) engraved on its surface, and the grooves (2) are arranged in an array, each Through-holes (10) are respectively opened at two ends of each groove (2); the grooved surface of the micro-fluid channel template (33) is in close contact with the surface of the chip (1);  Another micro-channel template (33) is provided with grooves (14), and the pitch of the grooves (14) is the length of the grooves (2) on the micro-channel template (3), and each groove ( One end of 14) corresponds to the through hole (10) of the groove (2); one end of the groove (14) has a through hole corresponding to the groove on the microchannel template (3), and the other port of the groove (6 ) Extended to the edge; two grooves (14) corresponding to each groove (2) are connected by a groove (11); the periphery of the micro-channel template (3) and (33) is sealed and bonded together, The grooves and grooves face outwards and the holes communicate; the rigid material block (4) is covered on the grooves of the microchannel template (33), and the periphery is sealed and bonded together, between the microchannel template and the rigid material block A closed microfluidic channel is formed. Each microfluidic channel has a pressure switch (16), a horizontal groove (11) between the two microfluidic channels, and a pressure switch (17).  8. The device for micro-sample loading and reaction of a biomolecule chip according to any one of claims 1, 2, 3, 4, 5, 6, or 7, characterized in that: said microchannel template (3 ), (3 '), or (33), including those made of silicone, rubber, plastic, or other flexible materials.  9. The device for micro-sample loading and reaction of a biomolecule chip according to any one of claims 1, 2, 3, 4, 5, 6, or 7, characterized in that: said rigid material block material (4 ), Including those made of plastic, metal, or glass, with a thickness of 1 to 10 mm. 10. The device for micro-sample loading and reaction of a biomolecule chip according to any one of claims 1, 2, 3, 4, 5, 6, or 7, characterized in that: the micro-channel template (3 ) Or (3 ') The grooves (2) are strip grooves, the area of the strip grooves is from 0.01 mm ^{2 to 1} mm ² _{; the} depth is from 10 μm to 1 mm; the number of strip grooves is at least 2 -500.  11. The device for micro-sample loading and reaction of a biomolecule chip according to any one of claims 1, 2, 3, 4, 5, or 7, characterized in that: the groove on the micro-channel template (2) The inner diameters of the through holes (10) at both ends and the through holes (10) of the rigid material block material (4) are both 10 μm to 1 mm.  12. The device for micro-sample loading and reaction of a biomolecule chip according to claim 7, characterized in that: the inner diameter of the groove is from 10 μm to 1 μm.  13. A method for applying micromolecular sampling and reaction of a biomolecule chip by using any one of the devices according to claim 1, 2, 3, 4, 5, or 6, characterized in that it comprises the following steps:  (a) Driven by a micro-pump or external force, different biomolecular solutions enter different grooves through different microfluidic channels, contact the surface of the chip and be fixed on the surface of the chip, and then transport the buffer through the pump. The liquid cleans the chip surface and the microfluidic channel to remove the biomolecules that are not fixed on the chip surface; (b) Each point on the surface of the chip after being fixed, when each point is required to react with a separate test solution, In this state, different solutions to be tested are sent to different grooves through the pump to react with the fixed biomolecules, and then the buffer is washed after the reaction;  (C) Or all the points fixed on the chip surface need to react with the same test solution or group the points on the chip with different test solutions, you need to connect the grooves in series as required, and then The solution to be tested is transferred into the groove through the liquid inlet, and reacts with the fixed biomolecules in turn. The liquid after the reaction is output from the liquid output port, and then washed with buffer solution after the reaction;  (d) Remove the chip after the reaction, and perform the result detection by a conventional method.  14. A method for applying a micromolecular biochip chip sample and reaction using the device according to claim 7, characterized in that it comprises the following steps in order: (1) installing the chip material in the device according to claim 6 so that it is in close contact with the microfluidic channel template (3), and pressing the microfluidic channel by external force;  (2) transport the ligand molecule to a selected area on the chip surface through the microtube of the microchannel; wait until the ligand molecule is fixed on the chip substrate;  (3) washing the biomolecule-immobilized chip prepared in step (2) with a buffer solution, and transferring the buffer solution to different regions of the chip surface through a microchannel; washing away the ligand not fixed on the chip surface molecule; (4) connecting a region in which ligand molecules are fixed in series through a tandem microchannel template (3 ′); or in series by pressing a pressure switch on an elastic diaphragm;  (5) transport the biological sample to be detected through the microchannel to each unit on the surface of the chip prepared in step (4), and perform a detection reaction;  (6) Remove the chip carrying the detection reaction result obtained in step (5), and use a chip detector to detect the reaction result.