CN1182255C - Flowing Biochip and its usage - Google Patents
Flowing Biochip and its usage Download PDFInfo
- Publication number
- CN1182255C CN1182255C CNB021336229A CN02133622A CN1182255C CN 1182255 C CN1182255 C CN 1182255C CN B021336229 A CNB021336229 A CN B021336229A CN 02133622 A CN02133622 A CN 02133622A CN 1182255 C CN1182255 C CN 1182255C
- Authority
- CN
- China
- Prior art keywords
- biochip
- open
- reactor
- chip
- liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention discloses a flowing Biochip and a use method thereof. At present, the production process of all flowing biochips with unopened reaction vessels as the characteristics is complicated, and the operation and scanning conditions are strict, so the application of the existing flowing biochips is greatly limited. All of the existing biochips with open reaction vessels as the characteristics have the disadvantage that liquid media can not flow in a fixed direction in the detection process, so the service efficiency of the existing biochips with the open reaction vessels as the characteristics is low. The biochip of the present invention is characterized in that the biochip is an open flow reaction vessel with a reaction and operation liquid phase medium, and the reaction and operation liquid phase medium can flow in the fixed direction, reaction results can be directly read by external instruments or naked eyes through an uncovered open structure arranged above a probe array. The biochip provided by the present invention has the advantages of simple structure, function and material combination optimization, low unit reaction vessel cost, convenient operation and easy scanning.
Description
Technical field
The present invention relates to a kind of biochip, particularly a kind of open mobile biochip and using method thereof.
Background technology
Biochip is a kind of qualitative and/or quantitative testing product, its principle is that micro-probe is fixed on the surface of solid phase carriers in addressable mode, make its under the condition of trace detection with biological sample in target molecule generation specific reaction, and then utilize peripheral equipment or naked eyes that the result of specific reaction is read.Fixed probe on the biochip, its kind is a lot of, comprising: various DNA, RNA, protein, polypeptide, various cell, tissue and pharmaceutical cpd etc.The size range of trace probe: the marker area that shows its reaction result usually is less than 1 square millimeter.The biochip scope that has a wide range of applications comprises: genetic expression detection, genescreen, drug screening, medical diagnosis on disease treatment, environmental monitoring and fields such as improvement, judicial expertise.
The core of biochip is the reactor on it.In the present invention, the reactor of biochip is defined as: be fixed with probe array in the biochip, reach other dependency structure that is communicated with it with the place of target compound generation specific reaction when detecting.
Biochip can have reactor of different nature, by the qualitative classification of the characteristic reaction device that it had, it can be defined as dissimilar biochips again.In the present invention, according to the liquid phase medium that is added in the testing process can be in reactor directed flow, reactor is defined as flowing and non-current reactor, and the biochip that is feature with these two kinds of reactors is defined as flow biochip and non-current biochip respectively; Could cover open architecture by the nothing of probe array top according to reaction result is directly read by external instrument or naked eyes, reactor is respectively defined as open and the non-open type reactor, with these two kinds of biochips that reactor is a feature, be respectively defined as open and the non-open type biochip respectively; According to probe array top whether only be only read rank disconnected just have do not have the covering open architecture, reactor is defined as broad sense and non-broad sense open reactive device, and the biochip that is feature with these two kinds of reactors is defined as broad sense and the open chip of non-broad sense respectively.The difference of the structure of the solid phase carrier of root a tree name stationary probe, biochip also is respectively defined as one dimension chip, two-dimentional chip and three-dimensional chip.In the present invention, according to the number n of reactor on the biochip, biochip is defined as single reactor biochip (n=1) and multiple reactor biochip (n is equal to or greater than 2).In the time will showing reactor character, again reactor character is introduced, for example, and how open flow reactor chip, it is defined as: a biochip that includes the open flow reactor more than two or two.
According to definition of the present invention, the present situation of biochip is as follows:
1, non-flowing type biochip
The non-flowing type biochip comprises non-current biochip of non-open type and open non-current chip, and the most widely used at present is open non-current chip.
Using the widest open non-current chip at present is the open non-current chip of single reactor, and an example is to be the chip that substrate is made, non-structure is transformed with the microslide.Its advantage is simple in structure, and point sample and scan operation are all simple and easy to do, and part operation also can be tried hard to recommend outside under the medium flowing state under moving and carry out (for example hydrojet cleaning).The shortcoming reaction is to carry out under first valve state.In addition,, probe array less when the probe kind hour, the situation of peptide chips or protein chip for example, because a reactor only needs the few probe of fixed qty (for example several individual to hundreds of), the single reactor chip just seems that efficient is too low, strengthened and produced and the detection cost, the practical application of biochip has been subjected to considerable restraint in this case.
In order to increase the benefit, manufacturer and scientific worker have carried out a lot of improvement to this class chip, for example single open non-current reactor chip are developed into how open non-current reactor chip.The basic structure of the non-current reactor chip of in the market how open is: the open reactive device that forms a few to tens of circles or square on a substrate with height less than the isolated area of 1.0mm.This open reactive device has the liquid zone of adding and reaction zone, but does not have the liquid zone of going out.During detection, the chip plane is arranged essentially parallel to horizontal plane.Owing to there is not the liquid zone of going out, reaction medium cannot be placed in flow state, thereby can not carry out operate continuously, and chip efficient is still treated further raising.In addition, also be owing to there is not a liquid zone of going out, open reactive device isolated area crossed contamination between the adjacent reactor occurs as too low meeting, reads as its too high commonplace chip scanner of present use that can not utilize again.At this problem, we have invented " a kind of biochip of detachable use " (number of patent application 02113540.1), when detecting, load onto a sufficiently high reactor disrupter with anti-cross-contamination, the disrupter of dismantling during scanning is to adapt to the scanner requirement.
Another example of many open non-current reactor organisms chips is " multiple-sample microarray biochip " (number of patent application 01112783.X).Its probe array is fixed on one when being open, reaction when not reacting in the closed reactor that not have import and export.This chip uses the reactor of the airtight original opening of polyester film single face glue material behind application of sample, need remove enclosed material before going out sample, washing, then application of sample, more airtight, react, remove again airtight, finish to detection reaction so repeatedly.
2, mobile biochip
Mobile biochip comprises mobile biochip of non-open type and open mobile biochip.
1) the mobile biochip of non-open type
1. the one dimension non-open type chip that flows
The one dimension chip is the chip that probe stationary is formed on the line style solid phase carrier.Its example comprises kapillary biochip device (notification number is CN 2483395Y) and microchannel (microchannel) biochip.The microchannel chip is to make general size to be on the chip slapper base: width is less than 0.05mm, the degree of depth is less than the microchannel of 0.025mm, again with probe stationary in the microchannel, add sample then and make its probe region that flows through the microchannel, adopt corresponding signal detection system to read reaction result at last.An example of microchannel biochip is the detection biochip (www.caliper.com) of Caliper Technologies Inc. company, this chip is made substrate with slide, adopt photoetching and etching technique on substrate, to etch the microchannel, with probe stationary in the microchannel.The liquid storage tank of a plurality of openings is arranged on the chip, and liquid storage tank is connected by the microchannel.The advantage of microchannel chip be highly sensitive, speed is fast.Its shortcoming is: 1), need etch the microchannel earlier in the production process, and probe on the point, and then carry out the microchannel sealing and make, its complex structure, the suitability for industrialized production difficulty is very big; 2), flow rate of liquid need be with the control of special precision equipment during detection, for example osmosis device etc.; 3), after reaction is finished,, detect, can not directly use the common chip scanner to read the result for some test example such as fluorescent marker because the fixed probe molecule is surperficial within it.
2. the two-dimentional non-open type chip that flows
The two dimension chip, promptly probe array is distributed in the chip that forms on the solid phase carrier of plane.The two dimension non-open type chip that flows, can lift the invention " a kind of probe card and application thereof that includes closed reactor " (number of patent application 0211364.8) that we have applied for is example.The essential characteristic that this two-dimensional closed formula flows through chip is that probe array is fixed on the plane of a closed reactor that import and export arranged, and operations such as reaction can be carried out under the state of medium flowing continuously, all finish until reaction.
2), open mobile biochip
New through looking into, liquid medium can continuous flow react, clean, and reaction result can not have the open architecture of covering by the probe array top and be yet there are no by the open mobile biochip that peripheral equipment directly reads.Some similar having " viral hepatitis differential diagnosis biochip " (patent announcement CN 2438121Y) are arranged, but the essence of the biochip in this patent is multi-ribbon fast diagnosis reagent bar, belong to a kind of improved diagnostic reagent bar combination, its reaction zone is water-absorbing material, for example cellulose acetate membrane or nylon film are with fixedly the canonical biometric chip of microprobe array is different among the present invention.
Although above-mentioned various biochip respectively has many good qualities, produce with use in still have different problems.A main common issue with remains, how each functional zone of chip are being carried out under the condition of economy, technical optimization, utilize liquid transport mechanism or device as far as possible cheaply, realize operation (comprising probe stationary, application of sample, cleaning, scanning etc.) on the simplest, the reliable sheet.
Summary of the invention
The object of the present invention is to provide a kind of new biochip and using method thereof, this chip will overcome the shortcoming of existing various biochips as much as possible, (for example structure of non-open type chip and Operating Complexity, the medium illiquidity of open chip) brings into play advantage (the medium flowable of the chip that for example flows that has various biochips now as much as possible, the constructional simplicity of open non-current chip and scanning simplification), to improve detection sensitivity, reduce the production cost of operating time and reduction chip unit reactor and to detect cost.
Its essential characteristic of the present invention is for containing a kind of open flow reactor.In open flow reactor of the present invention, reaction and operation liquid phase medium can directed flow, and reaction result can cover open architecture and directly be read by external instrument or naked eyes by the nothing of probe array top.
The objective of the invention is to realize by the enforcement following technical proposals:
A kind ofly contain that respond and operate liquid phase medium can directed flow, reaction result can cover the mobile biochip of the open flow reactor that open architecture directly read by external instrument or naked eyes by the nothing of probe array top, it is characterized in that: the basic structure of the open flow reactor that described biochip is contained comprises at least: what liquid phase medium can be by continuous application of sample adds liquid zone (1), liquid phase medium can and be fixed under flow state and not absorb water but the reaction zone (2) of hydrophilic lip-deep probe array effect, liquid phase medium can flow through and leave chip go out liquid zone (3) and the free-pouring isolated area of limits liquid phase medium (6), described open flow reactor add the liquid zone, reaction zone with go out the liquid zone and directly link to each other, or link to each other indirectly by the structure that comprises passage.The isolated area of contained open flow reactor is with respect to the reaction zone plane, between-3.0mm to 3.0mm.
Open mobile chip provided by the invention, the basic structure of contained open flow reactor comprises adding liquid zone (1), reaction zone (2), going out liquid zone (3) and isolated area (6) (see figure 1) at least.Open flow reactor in the described biochip add being characterized as of liquid zone: can reaction and the operation medium carry out application of sample with the continuous flow state.Being characterized as of the reaction zone of the open flow reactor in the described biochip: the surface of stationary probe does not absorb water but the liquid phase medium of possess hydrophilic property, reaction and operation can be under flow state and the effect of immobilization probe array.Open flow reactor in the described biochip go out being characterized as of liquid zone: reaction and operation medium can flow through and leave.Being characterized as of the isolated area of the open flow reactor in the described biochip: limit fluid unrestricted flow.Adding liquid zone, reaction zone and going out the liquid zone of open flow reactor in the biochip of the present invention can directly link to each other, and also can link to each other indirectly by other structure example such as passage.Directly the example of Xiang Lianing as shown in Figure 1.
Biochip among the present invention, be only to read the biochip that the reaction result stage has the open flow reactor of broad sense that does not have the covering open architecture above comprising its probe array, or not only reading the biochip that the reaction result stage has the open flow reactor of non-broad sense that does not have the covering open architecture above comprising its probe array.The example of the open flow reactor of broad sense is seen example 3.
Open mobile chip provided by the invention, can also have following feature: open flow reactor can be the capillary phenomenon mainly utilizing the wetting ability of liquid phase medium deadweight, material surface and/or go out the liquid zone make the liquid phase medium directed flow from flow reactor, can also be mainly to utilize other external force to produce the non-of liquid directed flow from flow reactor.The example of available external force has: mechanical pressure, and the orientation that the electric osmose device provides moves, and the orientation that micro pump provides moves or the like.
It is the unique characteristics of biochip among the present invention that the character of designs such as capillary phenomenon in chip of utilizing the wetting ability of liquid phase medium deadweight, material surface and/or going out the liquid zone is used as promoting the directed flow of liquid phase medium in reactor.In this case, according to the requirement of biochip test project, can optimize liquid phase medium add-on and deadweight, the hydrophilic selection of material in advance and/or go out the design variables such as selection of liquid zone capillary phenomenon, to reach the purpose of effective control flow velocity.For example, (for example add the liquid zone and the reaction zone material is hydrophilic under suitable condition, the wetting ability that goes out the liquid zone is not less than and adds liquid zone and reaction zone), select suitable above-mentioned angle, liquid phase medium can mainly rely on its deadweight, flow into reaction zone to the lowland from adding the liquid zone by height, flow out chip by going out the liquid zone again after the reaction.So, just, can make reaction and operation medium flow through successively adds liquid zone, reaction zone and goes out the liquid zone.
In the open mobile biochip of the present invention, the directed flow of reaction and operation medium both can be that each step is carried out continuously, also can just carry out in individual steps.For example, when the lower or/and reaction medium amount that adds of speed of response seldom the time, corresponding reactions steps (for example the reaction of test sample and probe is or/and contain the reagent of marker and the reaction of target compound etc.) can be hatched by noncurrent mode, and other reactions steps (for example cleaning) is then carried out in the mobile mode.In this example, non-current mode can be considered a special case of flow pattern, and promptly flow velocity is zero.For example, under the hydrophilic/water-absorbent that flows out the district is not too high situation, reduce liquid-flow and help flowing the effect of component in making a concerted effort or/and increase the effect that liquid-flow is unfavorable for mobile component in making a concerted effort, for example make the angle of chip plane and horizontal plane be tending towards 0 and spend, can realize.Thereby becoming again, above-mentioned angle gives the important parameter of different operating step in the operating process with different in flow rate.
Open mobile chip provided by the invention, can contain the above open flow reactor, also can contain two or two the above open flow reactors, can also contain one or more described open flow reactor and one or more other type of reactor.When chip contains described open flow reactor more than two or two, its structure can be the multiple reactor chip of fixed sturcture, also can a plurality of single reactors be linked together, form the built-up type chip by one or more of draw bails such as wedge head, tenon, fastener, suspension member, plug-in unit.For example shown in Fig. 5.
Described biochip, the water wetted material of the solid phase carrier of its stationary probe comprises: comprise the inorganic hydrophilic material of glass, silicon, comprise the organic hydrophilic material of polypropylene, polyvinyl chloride, nitrocellulose diaphragm; The solid phase material of stationary probe can not be thought water-absorbing material in the reaction zone, and water-absorbing material comprises: nitrocellulose membrane, cellulose acetate film class, natural fiber film (paper) etc.The material of other functional zone, can use same material to make according to function-cost relation, also can use different materials to make to reduce the more expensive solid phase material consumption of price, these materials can be a kind of or its combinations of metal, glass, silicon chip, macromolecular material.For example, when being the solid phase body of stationary probe with preactivated slide, its unit surface cost is higher than general-purpose plastics far away, is satisfying under the prerequisite of function, then can reduce reaction zone activation slide area as far as possible, to reduce the material cost of unit reactor.
In the open mobile biochip of the present invention, there is isolated area to flow, prevent crossed contamination with the limit fluid free movement.In these cases, can also reduce the unit reactor cost.Plane, isolated area height relative response district is between-3.0mm to 3.0mm.The isolated area height is that the connotation of negative value is, one one of reactor reaction zone or all be higher than isolated area, and for example shown in Fig. 5 A.The isolated area height on the occasion of connotation be, one one of reactor reaction zone or all be lower than isolated area, for example shown in Fig. 5 B.The preferred version of isolated area height is between-3.0mm to 3.0mm.The structure that strengthens isolation effect can be arranged on the isolated area, for example, geometry decorative pattern, groove, hole, hydrophobic band or the like.For example, just in case there is liquid to arrive the isolated area top, have on isolated area top or the bottom isolated area along decorative pattern, groove, hole and the water-absorbing material etc. that add liquid zone, reaction zone and go out the water conservancy diversion of liquid zone direction and can strengthen isolation effect, liquid can be drained very soon, can not enter adjacent reactor.The directed flow and avoid the functional structure of crossed contamination between the reactor so chip of the present invention has a good guarantee.
When the isolated area height is negative value, adding the angle of cut place that liquid zone and reaction zone and isolated area form, interrupting appears in the water-wetted surface that adds liquid zone and reaction zone, the surface tension of liquid phase medium makes liquid form the arc with certain altitude and angle in this edge, helps the unrestricted flow of limits liquid phase medium.As this moment chip be in an inclination angle and bigger hydrophilic/water-absorbent arranged or/and go out the liquid zone, then can keep liquid phase medium in reactor along adding liquid zone, reaction zone and going out the directed flow of liquid zone.
Adding liquid zone, reaction zone and going out the liquid zone of open flow reactor in the biochip of the present invention can be formed by various structures respectively, and these structures can be solid and combinations thereof such as rectangle, circle, ellipse, rhombus.The adding liquid zone, reaction zone and go out the liquid zone of open flow reactor in the biochip of the present invention, can with reaction zone on same plane, also can intersect at angle with it not on same plane, to satisfy in the various detections requirements such as control flow velocity, anti-cross-contaminations with reaction zone.In the example of Fig. 4 E, go out bottom surface, liquid zone and reaction zone bottom surface and form one greater than 90 degree, less than 180 angles of spending.When the reaction zone plane kept level, liquid can not flow and hatch like this; When the reaction zone plane becomes one greater than 0 degree angle with go out the plane, liquid zone when outlet becomes one to spend angle greater than 0 with horizontal plane in outlet with horizontal plane, the liquid outflow is easy to carry out flow operation.
In the biochip of the present invention, the wetting ability of controlling liquid phase medium self gravitation, material surface and/or the structure that goes out the capillary phenomenon of liquid zone can be arranged in the described open flow reactor, comprise: add the liquid zone, go out the structure that helps the liquid directed flow (structures such as the ditch of water conservancy diversion layer, flow guide bar, water conservancy diversion rod, diversion rod, water conservancy diversion pin, water conservancy diversion, groove, inclined-plane, taphole etc.) and the material (for example water-absorbing material) of liquid zone, the selection of each functional zone surface hydrophilicity in the kinetics size of the control channel of flow velocity and the reactor in each passage on the chip.For example, when adding the liquid zone, reaction zone and go out the surface of liquid zone, more and more favourable liquid phase medium is capillary when reducing, and just helps liquid phase medium along adding liquid zone, reaction zone and going out the directed flow of liquid zone.
Biochip of the present invention, the dynamic form of its open flow reactor can be the permanent structure that forms by moulding or other irreversible connection technology, also can be the dismantled and assembled structure that forms by the non-irreversible connection that physics and/or chemical process obtain.The material of these structural elements can be a kind of or its combination of the plate made of inorganic materials such as various organic materialss such as plastics, rubber and glass, silicon chip or other material, sheet, film, piece.The physics and/or the chemical process that form non-irreversible connection in the dismantled and assembled structure comprise: mechanical locking device, electromagnetic locking device, reversible agglutinating tackiness agent, elasticity are chimeric or the like.
A chimeric example of elasticity is: reactor is embedded in the chip shaping aperture plate of being made by resilient material or spring piece or spring, by elastic force the two is closely pressed together.An example of reversible agglutinating tackiness agent is: with in type, contain the plastic plate that adds the liquid zone, goes out liquid zone and isolation region structure, by non-setting adhesive with contain the slide of using as reaction zone and be bonded together, be formed with the open mobile chip of a plurality of separate reactors.The example of mechanical locking device, electromagnetic locking device is seen our patent " a kind of biochip of detachable use " (number of patent application 02113540.1).
Also chip provides a kind of special using method according to the present invention in the present invention, its process is: by regulating the angle between chip or wherein a certain structure example such as plane, reaction zone place and the horizontal plane, realize the fluidic directed flow, to adapt to the rate of flow of fluid in all or part of operation, anti-crossed contamination or/and the requirement of other operational condition.The variable range of this angle is the 0-360 degree.
Also chip provides a kind of special being used to regulate the above-mentioned open mobile chip and the recliner of horizontal plane angle according to the present invention in the present invention.This recliner is a kind ofly can adjust reactor planar horizontal angle stationary device then with behind the said chip limit fixed thereon.It is characterized in that: required according to detecting, by machinery or electric device, control said chip reactor plane included angle.On it some additional structures can also be arranged, for example chip fixture apparatus, sample pipetting volume device head stationary installation, chip register, the pin of cover, level-off or the like of preserving moisture.
When above-mentioned open mobile chip and utility appliance are used jointly, according to different samples and reagent, the reactor of different size, angle, the temperature that can regulate speed, exit angle, surging force and the recliner of liquid feeding arrive the best, and its principle is to make to reach detection sensitivity, specificity, detection time best fit.Specifically using method is, behind the adding sample, sample flow is crossed the probe plane of reactor, and flow velocity can be controlled by the inclination angle size of application of sample speed and reactor; When probe molecule with after target molecule combines, add washing lotion by automatic liquid adding device, flush away is binding substance not, then react, wash until reaction and finish from adding the liquid zone or adding other reagent again by automatic liquid adding device, then chip is put into instruments such as chip scanner and got final product sense data, obtain detected result.
The invention has the advantages that:
1) flow biochip relatively with present non-open type, the biochip simple structure among the present invention, easy to manufacture, easy to use, scanner there is not particular requirement.
2) with the non-current biochip of present non-open type relatively, but the biochip among the present invention can control sample injection rate and operate continuously thereby have the higher and speed of detection sensitivity advantage faster according to reaction needed,
3) with the open biochip of present single reactor relatively, biochip among the present invention is owing to can there be a plurality of reactors, and each functional zone can be carried out relatively independent cost control and realize optimum combination in selection and making on the chip, thereby when probe array is not very big, when for example the probe array distribution area was less than 8 square centimeters, production cost and use cost were all lower.
Description of drawings
Fig. 1: the open gravity flow reactor organisms of a kind of list of the present invention chip synoptic diagram
Fig. 2: the present invention is open from flow reactor liquid feeding district synoptic diagram
Fig. 3: used herein open from flow reactor fluid district synoptic diagram
Fig. 4: the present invention is open from flow reactor isolated area synoptic diagram
Fig. 5: single open gravity flow combination of reactors chip synoptic diagram
Fig. 6: a kind of open by locating slot from the flow reactor synoptic diagram
Fig. 7: single 8 open gravity flow reactor chip synoptic diagram
Fig. 8: open from flow reactor recliner synoptic diagram
Mark among the figure
1 for add liquid zone 2 for reaction zone 3 be liquid zone 4 for fluid hole 5 for liquid filling hole 6 for isolated area 7 for being that sump pit 11 is the chip operation face for isolated area 9 highly for locating slot 10 highly for positive number for the isolated area 8 of negative
Embodiment
Below in conjunction with some examples of implementation, the invention will be further described.
Reactor isolated area height is 0.5mm in the chip, the adding liquid zone 1, reaction zone 2, go out liquid zone 3 and directly link to each other of each reactor, and add liquid zone 1, reaction zone 2 bottom surfaces at grade, have fluid hole 4 going out liquid zone 3, going out liquid zone 3 and fluid hole 4 inwalls have thieving paper.Chip places on the thieving paper.Behind the liquid feeding, liquid is by gravity and surface affinity effect, and through adding liquid zone 1, reaction zone 2, going out liquid zone 3 inflow thieving papers, this moment, thieving paper and thieving paper aperture became control flow velocity principal element.
Reactor isolated area height is 0.5mm in the chip, the adding liquid zone 1, reaction zone 2, go out liquid zone 3 and directly link to each other of each reactor, and add liquid zone 1, reaction zone 2 and go out 3 bottom surfaces, liquid zone at grade.Each reactor side has tongue-and-groove, the tenon of a concave, convex respectively, and the tenon of 3 reactor sides is inserted the tongue-and-groove of other reactor respectively, has promptly formed one four combination of reactors chip (Fig. 5).
The liquid feeding plot structure of this biochip such as Fig. 2 D, 2H, go out structure such as 3B, the 3E of liquid zone, add the liquid zone and be made up of a liquid filling hole, reaction zone is a square planar, going out the liquid zone is a trapezoidal and quadrate combination, becomes one 25 degree angles by reaction zone with horizontal plane to liquid outlet.During assembling, using pressure sensitive adhesive on isolated area a transparent film above the reaction zone.This chip level placed add sample and hatch, hatch finish after, again this chip is placed to add the liquid zone height, to go out the liquid zone low and become miter angle to wash with horizontal plane.Having under the situation of certain inclination angle, institute adds liquid and is easy to flow out the liquid zone.After total overall reaction is finished, tear transparent film off, promptly available scanner scans.Film on the open chip reaction zone of this part can the protective reaction district when chip does not use, and reaction zone is worked under certain pressure and prevents crossed contamination, when needs are hatched, also has moisture-keeping function.
The liquid feeding plot structure of this biochip such as Fig. 2 D, 2H go out structure such as 3B, the 3D of liquid zone, add the liquid zone and are made up of a liquid filling hole, and reaction zone is a square planar, and going out the liquid zone is a trapezoidal and quadrate combination, with reaction zone on same plane.During assembling, using pressure sensitive adhesive on isolated area a transparent film above the reaction zone.During use, this chip level is placed, and reaction and washings add liquid filling hole having under certain external force effect, and institute adds liquid after reaction zone forms certain height, by going out the liquid zone outflow.After total overall reaction is finished, tear transparent film off, promptly available scanner scans.
Embodiment 5, a kind of open 8 preparations from flow reactor c-hepatitis antibody detection chip
One with 8 holes, 75mm * 25mm * 1.5mm polyphenyl alkene plastics plate, span 5mm, the half slot of a 3mm diameter, dark 0.6mm is done as adding the liquid zone in the hole in liquid feeding side, connect the reach through hole of a 3 * 3mm down, the activation slide of assembling one 3 * 3mm, thick 1mm in each reach through hole, the slit with adhesive closure slide and reactor limit forms a dark 0.6mm reaction zone, the slide bottom surface is lower than plastic plate bottom surface 0.1mm, the benchmark during as scanning.The below of reaction zone meets a wide 2mm, and dark 0.6mm goes out the liquid zone.Go up NS3, NS4, NS5, the CORE antigen of hepatitis C virus (HCV) on each slide with 2 * 2 array point, the diameter of each point is 0.2mm, and dot center is apart from 1mm, apart from pool side 1mm.With the skim-milk sealing, make the c-hepatitis antibody detection chip of one 8 reaction tank again.
Embodiment 6, a kind of preparation of the open 8 reactors blood transfusion detection chip that flows
One with 8 holes, 75mm * 25mm * 1.5mm polyphenyl alkene plastics plate, pitch of holes 5mm, the half slot of a 3mm diameter, dark 0.6mm is done as adding the liquid zone adding sample prescription in the hole, connect the reach through hole of a 5 * 3mm down, the activation slide of one 5 * 3mm, thick 1mm in the assembling in each reach through hole with the slit of adhesive closure slide and pool side, makes to form a dark 0.6mm reaction zone, the slide bottom surface is lower than plastic plate bottom surface 0.1mm, the benchmark during as scanning.The below of reaction tank meets a wide 2mm, and dark 0.6mm goes out the liquid zone.On each slide, go up NS3, NS4, NS5, CORE fused antigen, HIV 1+2 antigen, HTLV antigen and the syphilis antigen of hepatitis B surface antibody, hepatitis C virus with 3 * 2 array point, the diameter of each point is 0.2mm, dot center is apart from 1mm, apart from pool side 1mm.With the skim-milk sealing, make the blood transfusion detection chip of one 8 anti-reaction tank again.
This example is used the biochip of embodiment 5, and chip reactor plane sample application zone side height during detection, to go out the liquid zone side low, becomes 30 degree angles.8 reaction tanks slowly add 8 each 20ul of different plasma samples that detected with the RIBA method of dilution in 1: 100 separately, each reaction tank adds goat anti-human igg 100ul, the unconjugated traget antibody of 0.01M phosphate buffered saline buffer 300ul flush away of the unconjugated impurity of 0.01M phosphate buffered saline buffer 300ul flush away, rhodamine mark more successively, at last with drying up after the dehydrated alcohol 100ul washing, chip scanner (GMS 418 ARRAY SCANNER) is read, the computer special software is handled, and obtains table one result:
Table one, the third liver detection chip and RIBA be detected result relatively
The open chip detection result of serum sample RIBA result
NS3 NS4 NS5 CORE NS3 NS4 NS5 CORE
1 -- -- - -- 88 114 86 99
2 -- -- + -- 124 88 15415 116
3 + + - -- 15498 16214 111 132
4 ++ -- - ++ 18146 106 128 19553
5 ++++ -- ++ -- 22051 115 18734 93
6 -- + - - 91 15426 86 129
7 -- ++ ++ - 113 19712 19246 127
8 ++++ ++ ++ ++ 22183 18956 21179 20132
This example is used the biochip of embodiment 6, chip reactor plane sample application zone side height during detection, it is low to go out the liquid zone side, become miter angle, 8 reactors slowly add 8 each 100ul of different plasma samples that detected with the RIBA method of dilution in 1: 100 separately, each reactor adds the unconjugated impurity of 0.01M phosphate buffered saline buffer 300ul flush away more successively, the anti-hepatitis b surface specific antibody (HbsAb) of rhodamine mark, third LS antigen (HCV), the HIV specific antigen, the HTLV specific antigen, each 100ul of syphilis specific antigen, 0.01M the unconjugated traget antibody of phosphate buffered saline buffer 300ul flush away, antigen, at last with drying up after the dehydrated alcohol 100ul washing, chip scanner (GMS 418 ARRAYSCANNER) is read, the computer special software is handled, and obtains table 2 result:
Table 2, blood transfusion detection chip and RIBA comparative result
The open chip detection result of serum sample RIBA result
HbsAb HCV HIV HTLV syphilis HbsAb HCV HIV HTLV syphilis
1 - - - - + 88 114 86 99 15895
2 - - + - - 124 88 15415 116 113
3 + + - - - 15498 16214 111 132 105
4 ++ - - ++ - 18146 106 128 19553 116
5 ++++ - ++ - - 22051 115 18734 93 98
6 - + - - + 91 15426 86 129 16027
7 - ++ ++ - - 113 19712 19246 127 121
8 ++++ ++ ++ ++ - 22183 18956 21179 20132 94
Claims (12)
1, a kind ofly contain that respond and operate liquid phase medium can directed flow, reaction result can cover the mobile biochip of the open flow reactor that open architecture directly read by external instrument or naked eyes by the nothing of probe array top, it is characterized in that: the basic structure of the open flow reactor that described biochip is contained comprises: what liquid phase medium can be by continuous application of sample adds liquid zone (1), liquid phase medium can and be fixed under flow state and not absorb water but the reaction zone (2) of hydrophilic lip-deep probe display effect, liquid phase medium can flow through and leave chip go out liquid zone (3) and the free-pouring isolated area of limits liquid phase medium (6), described open flow reactor add the liquid zone, reaction zone with go out the liquid zone and directly link to each other, or it is continuous indirectly by the structure that comprises passage, the isolated area of contained open flow reactor, with respect to the reaction zone plane, height is at-3.0mm to 3.0mm.
2, mobile biochip according to claim 1, it is characterized in that: be to contain its probe display top only the reaction result rank are disconnected to have the biochip that does not have the open flow reactor of broad sense that covers open architecture reading, or contain its probe array top and not only have the biochip that does not have the open flow reactor of non-broad sense that covers open architecture reading the reaction result stage.
3, mobile biochip according to claim 1, it is characterized in that: be that the capillary phenomenon that contains the wetting ability of mainly utilizing liquid phase medium deadweight, material surface and/or go out the liquid zone makes the open biochip from flow reactor of liquid phase medium directed flow, or contain the open non-biochip that mainly utilizes other external force to produce the liquid directed flow from flow reactor.
4, mobile biochip according to claim 1, it is characterized in that: be the single reactor biochip that contains a described open flow reactor, or contain the how open flow reactor biochip of two or two the above open flow reactors, or contain the multiple reactor biochip of one or more described open flow reactor and one or more other type of reactor, the multiple reactor chip that above-mentioned how open flow reactor biochip is a fixed sturcture, the latter is by the wedge head, tenon, fastener, the contrary built-up type chip that forms that connects of suspension member or plug-in unit.
5, according to claim 1,2,3 or 4 arbitrary described mobile biochips, it is characterized in that: the reaction zone of contained open reactive device in order to stationary probe do not absorb water but hydrophilic solid phase carrier comprises glass, silicon inorganic hydrophilic material, polypropylene, polyvinyl chloride, nitrocellulose diaphragm are at interior organic hydrophilic material; The material of other functional zone uses same material to make, and perhaps uses a kind of or its combination of metal, glass, silicon chip, macromolecular material.
6, according to claim 1,2,3 or 4 arbitrary described mobile biochips, it is characterized in that: how much decorative patterns, ditch, groove, hole, hole or hydrophobic band structures strengthening isolation effect are arranged on the isolated area of contained open flow reactor.
7, according to claim 1,2,3 or 4 arbitrary described mobile biochips, it is characterized in that: the adding liquid zone, reaction zone and go out the liquid zone of contained open flow reactor, formed by various structures respectively, these structures are rectangle, circle, ellipse, rhombus or other solids and combination thereof.
8, mobile biochip according to claim 7, it is characterized in that: the described structure that is beneficial to the liquid directed flow comprises water conservancy diversion layer, baffle, water conservancy diversion rod, diversion rod, water conservancy diversion pin, the ditch of water conservancy diversion, groove, inclined-plane, taphole, and described material is a water-absorbing material.
9, according to claim 1,2,3 or 4 arbitrary described mobile biochips, it is characterized in that: the dynamic form of its reactor is the permanent structure that forms by moulding or other irreversible connection technology; Or the dismantled and assembled structure that forms by the non-irreversible connection that physics and/or chemical process obtain; Described dismantled and assembled structure comprises: mechanical locking device or electromagnetic locking device, maybe can separate agglutinating glued construction or elasticity embedded structure.
10, a kind of using method of mobile biochip according to claim 1, it is characterized in that: its process is for passing through to regulate the angle of cut between chip or wherein a certain structure example such as plane, reaction zone place and the horizontal plane, independently realize or with external force with realizing the fluidic directed flow, to adapt to the rate of flow of fluid in all or part of operation, anti-crossed contamination/or the requirement of other operational condition, the variable range of this angle of cut is the 0-360 degree.
11, a kind of using method of mobile biochip according to claim 10, it is characterized in that: a kind of recliner that is used to regulate the described mobile chip and the horizontal plane angle of cut, it is the device of a kind of fixed angle or adjustable angle, by machinery or electric device, control the horizontal angle of cut of above-mentioned reaction zone planar, on it some additional structures can also be arranged.
12, a kind of using method of mobile biochip according to claim 11 is characterized in that: described additional structure comprises chip fixture apparatus, liquid fillers outlet device for positioning and securing, chip register, the cover of preserving moisture.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021336229A CN1182255C (en) | 2002-08-19 | 2002-08-19 | Flowing Biochip and its usage |
| PCT/CN2003/000055 WO2003106999A1 (en) | 2002-06-01 | 2003-01-22 | Biochip with maximization of the reactor number |
| EP03701458A EP1519191A4 (en) | 2002-06-12 | 2003-01-22 | Biochip with maximization of the reactor number |
| US10/517,452 US20060057580A1 (en) | 2002-06-12 | 2003-01-22 | Biochip with maximization of the reactor number |
| AU2003203334A AU2003203334A1 (en) | 2002-06-12 | 2003-01-22 | Biochip with maximization of the reactor number |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021336229A CN1182255C (en) | 2002-08-19 | 2002-08-19 | Flowing Biochip and its usage |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1403589A CN1403589A (en) | 2003-03-19 |
| CN1182255C true CN1182255C (en) | 2004-12-29 |
Family
ID=4747302
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB021336229A Expired - Fee Related CN1182255C (en) | 2002-06-01 | 2002-08-19 | Flowing Biochip and its usage |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1182255C (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101831384A (en) * | 2009-10-30 | 2010-09-15 | 中国人民解放军第三军医大学第一附属医院 | Biochip reaction device and method for detecting target molecules by using device |
| TWI530683B (en) * | 2014-10-16 | 2016-04-21 | 國立清華大學 | A fabric-base biochemical detecting device and the fabricating method thereof |
| CN108709985B (en) * | 2018-09-06 | 2018-12-18 | 湖南乐准智芯生物科技有限公司 | A kind of biochip reaction device topples over drainage structure |
| CN113976192B (en) * | 2021-08-23 | 2023-08-15 | 上海汉原生物科技有限公司 | Microsphere marking microfluidic chip and microsphere protein marking method |
-
2002
- 2002-08-19 CN CNB021336229A patent/CN1182255C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN1403589A (en) | 2003-03-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US7238521B2 (en) | Microarray hybridization device having bubble-fracturing elements | |
| CN101529227B (en) | A microarray system and a process for producing microarrays | |
| US7390464B2 (en) | Fluidic circuits for sample preparation including bio-discs and methods relating thereto | |
| JP5396857B2 (en) | Analysis chip and analysis method | |
| US20090220948A1 (en) | Methods and Device for Transmitting, Enclosing and Analysing Fluid Samples | |
| US20070280859A1 (en) | Fluidic circuits for sample preparation including bio-discs and methods relating thereto | |
| CN102224260A (en) | Kits and devices for detecting analytes | |
| US20190217290A1 (en) | Encoded Chip Based Micro-Array, Preparation Method Thereof and Application | |
| US20060078472A1 (en) | Microchannel chip system and detection chip | |
| Geissler et al. | Microfluidic patterning of miniaturized DNA arrays on plastic substrates | |
| CN1182255C (en) | Flowing Biochip and its usage | |
| JP4857882B2 (en) | Sample solution agitation method | |
| JP2006519384A5 (en) | ||
| JP5092405B2 (en) | Selective binding substance immobilization carrier | |
| CN2596360Y (en) | Flowing biological chip | |
| CN1697976B (en) | High integration analysis chip of minimized height for reactor and application | |
| JP2007171144A (en) | Microarray having cover | |
| JP2007171030A (en) | Selectively-bonding substance immobilizing base material | |
| CN1409110A (en) | Two-dimensional optic coder produced by micro processing and use | |
| WO2005073363A1 (en) | An analysis chip, an apparatus comprising the chip, the chip test kit and the measurement method thereof | |
| JP5145752B2 (en) | Analysis chip | |
| CN2575676Y (en) | Vehicles & boats satellite positioning radio monitor | |
| CN1432655A (en) | Multilateral biochip | |
| CN1208449C (en) | Biochip with small area reactor | |
| US8445201B2 (en) | Hybridization device, methods, and system using mixing beads |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20041229 Termination date: 20180819 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |