The method and the device thereof that are used for biomolecule chip micro sample-adding and reaction
Technical field
The present invention relates to a kind of biomolecule chip production method and device thereof, relate in particular to a kind of device that directly on the biomolecule chip, carries out the method for micro sample-adding and immediate reaction and realize this method.
Background technology
The biomolecule chip is a kind of integrated parallel Measurement for Biotechnique that just grew up in recent years, can integrated multiple aglucon on small geometric scale, so just can be simultaneously the multiple index of micro-example be detected.Because biomolecule sample price height requires consumption the least possible, therefore require employed biomolecule reagent of biomolecule chip and sample traceization, this also just requires chip application of sample and reaction unit microminiaturization.What at present, the application of sample of biochip mainly adopted is point sample instrument.Difference according to the point sample mode is divided into two classes, and a class is a contact, at first dip in and get stand-by aglucon with spotting needle, then by the contact chip surface aglucon point on chip; One class is the spray printing formula, draws a spot of aglucon for the treatment of a little with hollow spotting needle earlier, and the mode by similar ink-jet printer is added to aglucon on the chip surface then.The common drawback of this dual mode is that the point sample amount is inhomogeneous, and the surface density of the interior aglucon molecule of a single point distributes also inhomogeneous, and this will have a strong impact on the quality of testing result.Current, what biochip reaction adopted is the W-response mode mostly, exactly whole being immersed in the testing sample solution of chip is reacted.The testing sample solution amount that this method needs is more, and the reaction time is long, and sensitivity is not high.
Another chip application of sample and reaction technology are that fluid channel transports and the microcavity reactor.At present, generally the chip of the fluid channel technology of Shi Yonging is incorporate, be that chip and fluid channel are to be produced on the same block of material, as document 1:Dielectrophoretic cell separation and gene expression profiling onmicroelectronic chip arrays.July 15,2002 Huang Y, Joo S, Duhon M, Heller M, Wallace B, Xu X Anal Chem 2002 Jul 15; 74 (14): described among the 3362-71.The employed fluid channel of this method is disposable use, and this fluid channel is made complicated, and cost is higher, makes to use to be restricted.Carry out biomolecule operation fixing and detection reaction simultaneously and divide in two kinds of devices and carry out for 2 times, therefore, preparation technology is loaded down with trivial details.
Summary of the invention
The objective of the invention is in order to overcome the shortcoming of above-mentioned prior art; For the cost of manufacture that reduces biochip significantly with simplify the technology of the fixing and detection reaction of biomolecule, thereby provide a kind of biochip to make and directly carry out on this biochip that biomolecule is fixed and the method and apparatus of detection reaction immediately.
The object of the present invention is achieved like this:
Biochip preparation method provided by the invention comprises that order is carried out as follows:
(1) base material with chip closely contacts with the microarray template, and compresses fluid channel by external force;
(2) the fine pipe of aglucon molecule by fluid channel is transported to the lip-deep selection area of chip base; By the time after the aglucon molecule is fixed on the chip base;
(3) chip base that is fixed with biomolecule that step (2) is prepared is washed with damping fluid, damping fluid is transported in the zones of different of chip surface by fluid channel; Wash the aglucon molecule that is not fixed on the chip surface;
(4) by on the flexible sheet 13 of band chase 14, covering one deck resilient material diaphragm again, and the two membranes periphery is bonded together, has formed fluid channel 5, the zone that is fixed with the aglucon molecule is together in series by chase 14; Perhaps the switch that forms by the node on the extruding flexible sheet is together in series the zone that is fixed with the aglucon molecule;
(5) biological sample to be detected is transported in each unit on the chip surface that step (4) prepares by fluid channel again, carries out detection reaction immediately;
(6) take off step (5) preparation and the chip that is loaded with the detection reaction result that obtains, use chip detector to detect its reaction result.
Employed aglucon molecular conecentration is 0.001-1mg/ml in the described step (2); The flow velocity of described aglucon molecule in fluid channel is the 0.1-100 mul/min.
Biological sample is the 1-100 mul/min by the flow velocity of fluid channel in the described step (5).
Described chip base material comprises: silicon, glass, metal, plastic or other material or above-mentioned several compound substance, preferred silicon.
This method is combined fluid channel and microarray.Microarray is meant the micro-recesses that is produced on the rubber-like solid material, and it is fixed that the number of microarray further groove comes according to the index that detects, individual or more to hundreds of from one.Fluid channel is meant the pipeline of the small internal diameter that is connected with micro-recesses.
Microarray can closely contact the cavity that formation seals one by one with smooth chip surface under the effect of external pressure.Need fixing biological aglucon molecule to enter cavity and chip surface reacts by fluid channel, reacted aglucon molecule is discharged by fluid channel again.So just realized carrying out in the zones of different on chip surface micro sample-adding.Then, biological sample to be detected also is to enter each cavity by fluid channel, after the biological aglucon molecular reaction of having fixed on the chip surface, discharges by outlet again.Biochip is used for detecting simultaneously various biomolecules in the biological sample to be measured.Designed the FLOW CONTROL part in this method, make with a biological sample to be measured successively with chip on the multiple biological aglucon molecular reaction that pre-fixes, reduced amount of samples effectively.Can realize connecting between the arbitrary number groove by the FLOW CONTROL part.The fixing zone of aglucon molecule is that strictness limits on the chip, and fixing and reacted surface is washed through damping fluid again, can make the surface go up the fixing of biomolecule and reaction uniformity, has improved the quality of detection effectively.The chip reaction is limited in the tiny area, and under flow state, has quickened the mass transfer rate of biomolecule, has shortened the reaction time effectively, has improved sensitivity.Fixing and the reacted chip of biomolecule is with after the microarray template is separated, use chip detector detection reaction result.Can reuse with the microarray template after chip material separates, repeatedly to carry out the fixing and reaction of biomolecule on the chip.
The isolated plant of biochip preparation method provided by the invention is according to the mode difference that constitutes fluid channel, and this device comprises: two kinds.
At first narrate first kind, this device comprises: be carved with the elastic material sheet 3 of first groove 2 on surface, its first groove 2 is arranged by the array formula, and first groove, 2 two ends have first through hole 10; Wherein the through hole of each groove comprises two fluid channel of one-in-and-one-out; The two ends of also being carved with second groove, 2 ', the second groove 2 ' on one surface of a rigid solid material block 4 have second through hole 11, and the groove one side that is carved with of elastic material sheet 3 is not with groove one side relative fixed together with rigid solid material block 4; Through hole on this rigid solid material block 4 and through hole on the elastic material sheet 3 corresponding and hole of staggering communicates, promptly first groove 2 through hole communicates as solution inlet port with a through hole of second groove 2 ' as taphole, and the outlet of second groove 2 ' communicates with the import of next first groove 2; Solid pieces of material 4 two sides have a testing liquid import 7 and a testing liquid outlet 8, and a switch is installed in the import; Chip 1 closely contact on elastic material sheet 3, the pipe 12 that the plug-in mounting diameter is close in the through hole of solid pieces of material 4, fluid channel is with the formation that is connected of the through hole on the microarray template by pipe 12.
Groove on microarray template under the effect of external pressure closely contacts the cavity that formation seals one by one with smooth chip surface.Need the fixing aglucon molecule can be under the driving of pumping, enter cavity and chip surface reacts by fluid channel, reacted sample be discharged by fluid channel again.So just realized on the zones of different on the chip, carrying out micro sample-adding.Chip behind the application of sample needn't take off from device, can directly carry out detection reaction.Take off fine pipe from microarray, use elastic membrane closed pockets 2 ', groove 2 is together in series, stay next import and an outlet.Biological sample to be detected enters each cavity successively by import, and discharges by outlet after the aglucon molecular reaction fixing on the chip surface again.Reacted chip can take off from device, detects by the reaction result of detecting device to aglucon on the chip and acceptor.
Also comprise an elastic membrane and have an import and an outlet, perhaps have an import, have an outlet in elastic material block one side again in solid pieces of material one side in the solid pieces of material two sides; One switch is installed in its import; Chip behind the application of sample needn't take off from device, can directly carry out detection reaction.Take off fine pipe from microarray, use an elasticity membrane cover on solid pieces of material, seal first groove 2, groove on the elastic material sheet is together in series, relative with solution inlet port opens a hole at the elastic membrane place, as the hole of detecting the liquid import and the solid pieces of material side being opened as detecting taphole.Biological sample to be detected enters each cavity successively by solution inlet port, discharges by taphole after reacting with the biological sample of having fixed on the chip surface again.Reacted chip can take off from device, by detecting device the result is detected.
Direct preparation biochip of the present invention and second kind of structure of carrying out the device of detection reaction immediately comprise: fluid channel 5 also comprises the microarray template that is complementary with it; Wherein the microarray template comprises: the elastic material sheet 3 that is carved with at least 2 first grooves 2 on the surface, elastic material sheet 3 has facing up of first groove 2, first through hole 10 is opened at the two ends of each first groove 2 respectively, and the another side of elastic material sheet 3 closely contacts with chip 1 surface; Fluid channel comprises: a resilient material diaphragm 13, on resilient material diaphragm 13, be shaped on chase 14, and a port of its chase 14 is that first through hole, 10 positions of first groove 2 on the corresponding described microarray are provided with another port and extraneous UNICOM; First through hole, 10 corresponding chases 14 of each groove; And the film of corresponding microarray resilient material diaphragm 13 through holes on the resilient material diaphragm 13 is got through pore-forming, on be with chase 14 resilient material diaphragms 13 again, cover one deck elastic material membrane again, and the two membranes periphery is bonded together, formed fluid channel 5 by chase 14, interconnect between the fluid channel 5, at node location, the method for pushing by external force forms first switch 15, second switch 17.
Also comprise a rigid solid material block 4, this solid pieces of material 4 is fixed on the face of resilient material diaphragm 3 with first groove 2, first through hole 10 that open at the two ends of elastic material membrane piece 3 first grooves 2 runs through rigid solid material block 4, and resilient material diaphragm 16 does not have the elastic material membrane of chase and solid pieces of material 4 upper surfaces of microarray are bonded together.
Groove on microarray under the effect of external pressure can closely contact the cavity that formation seals one by one with smooth chip surface.During application of sample, push the pathway closure that makes between fluid channel, just close second switch by external force.Each groove part has and independently passes in and out runner.Under the driving of pumping, different aglucon protein solutions enters into cavity and chip surface reaction, thereby fixes from the teeth outwards.Use buffer solution for cleaning then, remove and be not fixed on proteins on surfaces.So just realized carrying out in the zones of different on chip the purpose of micro sample-adding.When carrying out detection reaction, open the second switch between fluid channel, close first switch on the fluid channel, so just a plurality of cavitys are cascaded, only stay next import, an outlet.Protein solution to be measured enters each cavity successively and is fixed on lip-deep aglucon proteins react along import; After the reaction, use buffer solution for cleaning, silicon chip takes off detection from device.
Described elastic material block, sealing comprise silica gel, rubber or plastic material with elastic membrane, resilient material diaphragm or elastic material membrane.
Described solid pieces of material comprises: materials such as silica gel, rubber, plastics, metal, glass, its thickness 1mm is to 10mm.
Described groove is a strip groove, and its strip groove area is from 0.01mm
2-1mm
2The degree of depth is from 10 μ m-1mm; At least 2 of the numbers of its strip groove for example can be from 2-1000.
Described through hole internal diameter can be from 10 μ m to 1mm.
The internal diameter of described fluid channel can be from 10 μ m to 1mm.
The internal diameter of described chase can be from 10 μ m to 1mm.
The invention has the advantages that:
Because the method that is used for directly preparing the biomolecule chip and carrying out detection reaction immediately of the present invention can and be carried out detection reaction immediately and carry out at same device the preparation biochip, and the fixing zone of aglucon biomolecule is strict the qualification on this chip, fixing and reacted surface is washed through damping fluid again, can make the surface go up the fixing of biomolecule and reaction uniformity, improve the quality that detects effectively.The chip reaction is limited in the tiny area, and under flow state, has quickened the mass transfer rate of biomolecule, has shortened the reaction time effectively, has improved sensitivity.Can reuse with the microarray template after the chip of making separates, carry out the fixing and reaction of biomolecule on the chip.
Description of drawings
Fig. 1 a is the microarray stencil plane synoptic diagram in first kind of device of the present invention
Fig. 1 b is the microarray template side view in first kind of device of the present invention
Fig. 2 is the structural drawing of first kind of device of the present invention when being used for directly preparing the biomolecule chip
(through hole at microarray template groove two ends and the through hole at rigid solid material block groove two ends stagger corresponding communicating form fluid channel and cooperate synoptic diagram)
Fig. 3 is the structure drawing of device of first kind of device of the present invention when being used for carrying out detection reaction immediately
Fig. 4 a is microarray (elastic material sheet) the stencil plane synoptic diagram in second kind of device of the present invention
Fig. 4 b is the microarray template side view of Fig. 4 a
Fig. 5 is the structural representation of second kind of device of the present invention
Fig. 6 is second kind of another kind of example structure synoptic diagram of device of the present invention
Fig. 7 is the fluid channel vertical view in second kind of device of the present invention
Drawing is described as follows:
1-chip 2-first groove 3-elastic material block or the diaphragm
4-solid pieces of material 5-fluid channel 6-liquid entrance
7-testing liquid import 8-testing liquid outlet 9-sealing elastic membrane
The 10-first through hole 11-second through hole (not shown in FIG.) 2 '-the second groove
12-pipe 13-resilient material diaphragm 14-chase
The 15-first switch 16-elastic material membrane (not shown in FIG.) 17-second switch
Embodiment
Below in conjunction with drawings and Examples the present invention is described in detail
Embodiment 1
Make first kind device, and the method for carrying out the fixing of 12 kinds of protein thereon and detecting.This device comprises 12 grooves.The groove area is 1mm
2, be 0.1mm deeply, fluid channel internal diameter 0.5mm.Resilient material is a rubber, and solid material is an organic glass, and pipe is a teflon, and the elastic material membrane that is used to seal is a silica gel.Chip material is a silicon.
Make a microarray template by Fig. 1-2; Employing thickness is that the rubber of 1mm is done elastic material block 3, and its above-listed configuration is arranged with 12 first grooves 2, and these first groove, 2 areas are 1mm
2, be 0.1mm deeply; First through hole 10 of an internal diameter 0.5mm is opened as fluid channel 5 (as shown in Figure 1) in these first groove, 2 two ends.Employing thickness is that the organic glass of 5mm is done solid pieces of material 4, is arranged with 12 second grooves 2 ' at plexiglass block 4 above-listed configurations, and this second groove, 2 ' area is 1mm
2, be 0.1mm deeply; Second through hole 11 of an internal diameter 0.5mm is opened at these second groove, 2 ' two ends; Wherein the through hole of each groove comprises two fluid channel 5 of one-in-and-one-out; The face of this sheet rubber 3 band grooves and plexiglass block 4 are not with the face relative fixed of groove not together; Second through hole 11 on this plexiglass block 4 communicates with first through hole 10 on the sheet rubber 3, and first through hole 10 at sheet rubber 3 lip-deep first grooves 2 two ends staggers corresponding with second through hole 11 at plexiglass block 4 second grooves 2 ' two ends, promptly in second through hole 11 of second groove 2 ' communicates as solution inlet port as first through hole 10 of taphole and first groove 2, and the outlet of second groove 2 ' communicates with the import of first groove; Plexiglass block 4 two sides have 2 holes, and one of them hole is testing liquid import 7 when detecting, and the import 7 of this testing liquid installs a switch, and another hole is as the outlet 8 of testing liquid; Chip 1 closely contact is equipped with fine pipe 12 in second through hole 11 of plexiglass block 4 on sheet rubber 3, fluid channel 5 is with the formation (as shown in Figure 2) that is connected of second through hole 11 on the plexiglass block 4 by fine pipe 12.The pipe 12 of fluid channel 5 usefulness of present embodiment is a teflon, and the elastic material membrane 9 that is used to seal is silica gel.Chip 1 material is a silicon.
Use method of the present invention, carry out the fixing and detection of 12 kinds of protein in the device of the foregoing description, its step is as follows:
Under the effect of external pressure (as shown in Figure 2), silicon chip 1 closely contacts with microarray when (1) working, and forms 12 independently cavitys of sealing like this between the silicon chip 1 and first groove 2, and each cavity has two fluid channel 5 of one-in-and-one-out;
(2) under the promotion of micro-ram pump, for example with 12 kinds of protein solutions, perhaps hepatitis B surface antigen, hepatitis B virus e antigen, hepatitis B virus core antigen, hepatitis B surface antibody, hepatitis B e antibody solution 10 microlitres (concentration is 0.1mg/ml) are transported to zones of different on the silicon chip by fluid channel respectively, flow speed control is in 1 mul/min, waits after the aglucon molecule is fixed on the chip 1;
(3) then, carry phosphate buffer to chip 1 surface of above-mentioned steps (2) preparation, wash the biomolecule that is not fixed on the silicon chip surface by fluid channel;
(4) take off the pipe 12 of fluid channel 5 usefulness, use the pellosil 9 of a sealing to cover on fluid channel 5, the zone that is fixed with the aglucon molecule is together in series;
(5) by injecting 100 microlitres patients serum to be checked to testing liquid import 7, be after 10 mul/min are injected reaction by a fluid channel, to flow out with flow velocity through testing liquid outlet 8, re-use phosphate buffer flushing (as shown in Figure 3);
(6) take off chip 1, use detecting device detection reaction result.
Enter cavity and silicon chip surface contact reaction by fluid channel, thereby proteinaceous solid is fixed on the silicon chip surface.Re-use buffer solution for cleaning silicon chip surface, cavity and fluid channel, the protein that is not fixed on the silicon chip surface is discharged.
Embodiment 2
Preparation one has 100 independently devices of the present invention of the cavity of sealing.
The elastic material block 3 of present embodiment is a silica gel, comprises on it being carved with 100 first grooves 2.These first groove, 2 areas are 0.1mm
2Or 0.01mm
2Dark is 0.1mm; It is that 10, blocks of solid materials of first through hole 4 of 0.2mm are aluminium that first groove, 2 two ends have internal diameter, comprises on it and is carved with 100 second grooves 2 '.This second groove, 2 ' area is 0.1mm
2, be 0.1mm deeply; Second groove, 2 ' two ends have second through hole 11 that internal diameter is 0.2mm.3 fluted of elastic material blocks and solid material 4 do not have the groove surface relative fixed, wherein first through hole 10,11 dislocation of second through hole communicate, form fluid channel 5, its fluid channel 5 internal diameter 0.2mm, the stainless steel pipe 12 that is inserted with suitable for reading of fluid channel 5, the suitable for reading of stainless steel pipe 12 is liquid-inlet 6, and all the other structures are with embodiment 1.
Comprise also that when detecting one is used to seal the pellosil 9 of usefulness, this pellosil 9 covers on second groove 2 ' of organic glass 4, and gold-plated glass material is as chip 1, and all the other structures are with embodiment 1.
Use the device of present embodiment, carry out the fixing and detection of 1000 kinds of genes, its step is as follows:
(1) when being prepared genetic chip, under the effect of external pressure, silicon chip closely contacts with the microarray template, has just formed 1000 independently cavitys of sealing like this between silicon chip and the groove, and each cavity has two fluid channel of one-in-and-one-out;
(2) under the promotion of micro-ram pump, respectively 1000 kinds of not homotactic dna molecular solution 10 microlitres (concentration is 0.1 μ g/ml) are transported to zones of different on the silicon chip by fluid channel, flow speed control is in 1 mul/min, wait dna molecular fixing after;
(3) then, use the phosphate buffer flushing, wash the molecule that is not fixed on the silicon chip surface;
(4) take off the pipe 12 of fluid channel 5 usefulness, use the pellosil 9 of a sealing to cover on fluid channel 5,1000 grooves are together in series, and the zone that is fixed with dna molecular is together in series;
(5) by inject the to be measured DNA sample of 100 microlitres to testing liquid import 7, be that 10 mul/min are injected, on a fluid channel is flowed through the chip that step (4) makes, after each zone reaction, flow out from testing liquid outlet 8 with flow velocity through degenerative treatments; The flushing of use phosphate buffer;
(7) take off silicon chip, use detecting device detection reaction result.
When detecting: the chip behind the above-mentioned application of sample needn't take off from device, can directly carry out detection reaction.On the microarray template, take off fine pipe 12, use second groove 2 ' on a pellosil 9 sealing organic glass 4,100 grooves on the elastic material sheet 3 are together in series, relative with solution inlet port opens a hole at the elastic membrane place, as testing liquid import 7, export 8 with the hole that the solid pieces of material side is opened as testing liquid, biological sample to be detected enters each cavity successively by testing liquid import 7, discharges by testing liquid outlet 8 after reacting with the biological sample of having fixed on chip 1 surface again.Reacted chip 1 can take off from device, by detecting device the result is detected.
Embodiment 3
Make first kind device, as Figure 1-3; Adopt method of the present invention to carry out the fixing and detection of 100 kinds of protein.
This device comprises 100 grooves.The groove area is 0.06mm
2, be 0.1mm deeply, fluid channel internal diameter 0.2mm.Resilient material is a silica gel, and solid material is an aluminium, and pipe is a stainless steel material, and the elastic material membrane that is used to seal is a silica gel.Chip material is gold-plated glass, and all the other structures are with embodiment 1.
Under the effect of external pressure, silicon chip closely contacts with microarray, has just formed 100 independently cavitys of sealing like this between silicon chip and the groove.Each cavity has two fluid channel of one-in-and-one-out.Under the promotion of micro-ram pump, 100 kinds of protein solutions enter cavity and silicon chip surface contact reaction by fluid channel, thereby proteinaceous solid is fixed on the silicon chip surface.Re-use buffer solution for cleaning silicon chip surface, cavity and fluid channel, the protein that is not fixed on the silicon chip surface is discharged.Take off the stainless steel pipe, use pellosil to seal organic groove on glass, 100 grooves are together in series.Under the promotion of micro-ram pump, solution to be measured enters cavity and the proteins react that is fixed on the silicon chip surface from the import of plexiglass block side.Solution to be measured flows through 100 cavitys successively, discharges from outlet at last, uses buffer solution for cleaning then.The silicon chip that has reacted takes off detection from microarray.
Embodiment 4
Make second kind of device (shown in Fig. 4,5,7), comprising: fluid channel 5 and the microarray template that is complementary with it; Wherein the microarray template comprises: an area be the sheet rubber of 20mm * 20mm as elastic material block 3, be carved with the elastic material block 3 of 100 first grooves 2 on its surface, first groove, 2 degree of depth of each bar shaped are that 0.01mm, sectional area are 0.1mm
2100 are divided into 10 rows and are arranged in regularly on the sheet rubber 1, and a row has 10 first grooves 2.Elastic material block 3 has facing up of first groove 2, and first through hole 10 is opened at the two ends of each bar shaped first groove 2 respectively, and these first through hole, 10 internal diameters are 0.1mm, totally 200.The another side of block rubber 3 closely contacts with chip 1 surface; Fluid channel is produced on the pellosil 13, on pellosil 13, be shaped on wide 0.1mm, the chase 14 of dark 0.1mm, a port of its chase 14, first through hole, 10 positions that are first groove 2 on the corresponding microarray are provided with, and the another port 6 of chase 14 extends to pellosil 13 edges and extraneous UNICOM; First through hole, 10 corresponding chases 14 of each groove; And the film of corresponding microarray pellosil 13 through holes on the pellosil 13 is got through pore-forming, on band chase 14 pellosils 13, cover one deck plastic foil 16 more again, and pellosil 13 and plastic foil 16 two membranes peripheries are bonded together, formed fluid channel 5 by chase 14, have wide 0.1mm between the fluid channel 5, the chase of dark 0.1mm interconnects it, at node location, the method for pushing by external force forms first switch 15, second switch 17.
Embodiment 5
Also comprising on the plexiglass block 4 as rigid solid soon 4 on the basis of embodiment 4, this solid pieces of material 4 is the plexiglass block of 25mm * 25mm; One area is the sheet rubber 3 of 20mm * 20mm, and this sheet rubber 3 coheres at 3 bar shaped first grooves 2 that are carved with degree of depth 0.01mm on sheet rubber 1 on the plexiglass block 4, and totally 100, each bar shaped first groove 2 sectional area is 0.1mm
2100 are divided into 10 rows and are arranged in regularly on the sheet rubber 3, and a row has 10 first grooves 2.First through hole 10 is opened at the two ends of 100 first grooves 2 on the elastic material sheet 3 of this substrate respectively, and these first through hole, 10 internal diameters are 0.1mm, totally 200.Fluid channel is produced on the pellosil 13, has wide 0.1mm on pellosil 13, dark 0.1mm chase 14 (shown in Fig. 4,6,7).
Under the effect of external pressure, silicon chip closely contacts with microarray, forms 100 cavitys.During application of sample, close the second switch II between fluid channel, each groove part has and independently passes in and out runner.Under the driving of ram pump, different protein solutions enters into cavity and silicon chip surface reaction, thereby fixes from the teeth outwards.Use buffer solution for cleaning then, remove and be not fixed on proteins on surfaces.Open the second switch 17 between fluid channel, close first switch 15 on the fluid channel, so just 100 cavitys are cascaded, only stay 6, one outlets of next import.Protein solution to be measured enters each cavity successively and is fixed on the proteins on surfaces reaction along import 6.After the reaction, use buffer solution for cleaning.Silicon chip takes off detection from device.