CN1208622C - Biological chip with minimized reactor isolation structure height and its preparation method - Google Patents
Biological chip with minimized reactor isolation structure height and its preparation method Download PDFInfo
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- CN1208622C CN1208622C CN 03117397 CN03117397A CN1208622C CN 1208622 C CN1208622 C CN 1208622C CN 03117397 CN03117397 CN 03117397 CN 03117397 A CN03117397 A CN 03117397A CN 1208622 C CN1208622 C CN 1208622C
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Abstract
The present invention discloses a biological chip with the minimized reactor isolation structure and a preparation method thereof, wherein the isolation structure mainly has a high hydrophobic material layer whose contact angle of surface static water is at least 80 DEG; the maximal thickness of the high hydrophobic material layer is 1000 um. Because the present invention realizes the minimization of the isolation structure, the maximization of reactor number and the minimization of sample addition quantity are easily realized.
Description
Technical field:
The present invention relates to a kind of biochip, particularly a kind of biochip and preparation method of reactor isolation structure minimized height, wherein pass through on hydrophilic solid phase carrier substrate in conjunction with high hydrophobic material, form or participate in forming the reactor isolation structure that the control liquid phase medium flows, owing to utilized the high hydrophobic material surface effect different with conventional hydrophobic material, thereby easier realization response device isolation structure minimized height finally reaches the purpose that reduces the biochip test cost.
Background technology:
Biochip, also be called for short chip in the present invention, it 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 testing conditions with biological sample in target molecule generation idiosyncrasy, and then the result of idiosyncrasy discerned.
The most frequently used biochip is polypeptide chip and genetic chip at present.Polypeptide chip is the biochip for preparing on substrate as probe stationary with a plurality of amino acid whose sequential structures (comprising protein).Genetic chip is with sample amplifying nucleic acid to be checked, nucleotide and complementary nucleic acid, nucleotide probe hybridization, forms crossbred, or combines with specific antibody, shows the chip of testing result again with color reaction.
The biochip scope that has a wide range of applications comprises gene 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.The reactor of the biochip among the present invention is meant to be fixed with probe array in the biochip, reaches other dependency structure that is communicated with it with the place of object generation specific reaction when detecting.The probe of the biochip among the present invention comprises that all can be fixed on the material of the biologically active on the solid phase carrier in addressable mode, comprise biotic components such as DNA, polypeptide, protein, cell, tissue.In the present invention, substrate is meant the parts that are used as the solid-support immobilized probe array in the biochip; Probe card is meant the substrate that is fixed with probe array.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).
Among 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 flow reactor and non-current reactor; With flow reactor and non-current reactor is that the biochip of feature is respectively defined as flow biochip and non-current biochip.Among the present invention, in whole testing process, whether open, be defined as open respectively reactor and the non-open type reactor according to reactor probe array top; With this reactor is the biochip of feature, is respectively defined as open and the non-open type biochip.The biochip reaction device has the character of above-mentioned several reactors usually simultaneously.Among the present invention, whole character that these reactors are defined as being had with it are the reactor of common trait, are that the biochip of feature is also similarly defined with this reactor.For example, if above testing process middle probe array, be no obducent open architecture, then the liquid phase medium that is added can be in reactor directed flow, then this reactor is defined as open flow reactor, corresponding biochip is defined as open mobile biochip, or is called for short open mobile chip; Other is analogized in proper order.
The present situation of biochip is as follows at present:
One, no isolation structure biochip
Grow up at first, still widely used biochip is no isolation structure biochip at present, the open non-current chip of single reactor normally, an example is based on microslide, and the chip of other newly-increased structure is made, do not had to activated and point sample.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 media flow state under moving and carry out (for example hydrojet cleaning).The shortcoming of this kind chip be when less, the probe array of 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, and efficient is too low, has strengthened and has produced and detect cost.The practical application of biochip has been subjected to considerable restraint in this case.
Two, the isolation structure biochip is arranged
The object kind is few in test sample, for example less than 100 kinds, thereby in each reactor under the few situation of fixing probe kind, determines the shared average area on the face of substrate stationary probe of each reactor just of chip cost usually.In addition, also need the mobile of liquid phase medium limited in order to reduce the sample addition.Besides, owing to use optical scanner widely, the thickness of reactor isolation structure is subjected to certain limitation, and the isolation structure height that goes to zero is an important channel that reduces the scanister technical requirement.In order to increase the benefit, manufacturer and scientific worker have carried out a lot of improvement to no isolation structure biochip, thereby develop some the isolation structure biochip are arranged.There is the isolation structure biochip can be divided into non-removable and dismountable two classes.
The non-removable biochip of a kind of isolation structure is arranged in the market, it is a kind of how open non-current reactor chip, its basic structure is: on a standard substrate that is of a size of 25mm * 75mm or 26mm * 75mm (wide * height), (surface contact angle is spent between 75 degree 50 with medium hydrophobic material, for example polyester or PVC) form the reactor isolation structure, thus can on substrate, form the open reactive device of a few to tens of circles or square.The isolation structure of this chip is still with thickness difference and flows as the control liquid phase medium.In this type of chip, open reactive device isolated area cross pollution between the adjacent reactor occurs as too low meeting, too highly then the technical requirement of chip scanner is improved as it.
In addition, dismountable biochip of a kind of isolation structure or the maximized biochip of a kind of number of reactors are also arranged at present.But main shortcoming is a complicated operation, or the isolation structure between its reactor can not realize that still thickness minimizes.
In a word, present biochip, isolation structure minimized height in some applications is still a problem to be solved.
Summary of the invention:
The invention provides a kind of biochip and preparation method of reactor isolation structure minimized height, wherein isolation structure mainly is the high hydrophobic material layer that the surface static water contact angle is at least 80 degree, the thickness of high hydrophobic material layer be to the maximum 1000um,, since realized the isolation structure minimized height, can be so that number of reactors maximization and sample addition minimize easier realization.
We are in the process of development blood transfusion detection chip, tumor marker detection chip and hepatitis detection chip, in order to allow the chip of developing except better quality is arranged, and on price, also have more competitive power, so feel that dearly the service efficiency of substrate of one of cost control factor in this type of biochip of improving is the key of dealing with problems than present general Elisa kit.In addition, also feel dearly that when reduce detecting technical requirement to scanner reduces the investment to scanner, reactor isolation structure height is a key factor.Therefore, " isolation structure minimized height and number of reactors maximization " as an important research project, formed chip of the present invention.
In the present invention, term " reactor " comprises probe region, isolated area, has or does not have sample application zone and goes out the liquid zone.Probe region is the zone that is fixed with probe in the reactor in direct or indirect mode; Adding the liquid zone is the zone that adds the required reaction medium of various detections to reaction zone; Go out the liquid zone and be the zone that liquid after reaction is finished and cleansing solution leave reactor; Isolated area is the area of isolation for avoiding cross pollution to make on the border of reactor.Isolation structure is for preventing the mobile structure of controlling liquid that cross pollution designs to take place between the reactor in the isolated area.Isolation mechanism according to different has different isolation structures in the isolated area.
The present invention be with in the biochip on the unit area substrate under the isolation structure minimized height condition number maximum of reactor turn to and pursue one's goal, promptly use the logical scanner of a biochip and a Daepori can detect maximum sample numbers and be purpose.Along with the development of new material and surface engineering technology in recent years, various new materials and functional coating material have also obtained swift and violent development in the application in each field.The isolation structure of biochip of the present invention based on the isolation mechanism of the flow disruption of aqueous solution between hydrophilic material and high-hydrophobic material, mainly is high-hydrophobic material coating or high-hydrophobic material layer.
In the present invention, high hydrophobic material is defined as the material of surface static water contact angle greater than 80 degree, and water wetted material is defined as the material of surface static water contact angle less than 60 degree.For a long time, with a drop of liquid at the contact angle θ of solid matter surface as the moistening quantification test of particular fixed, this point is widely approved.If liquid disperses to form film from the teeth outwards fully, if being liquid bead on 0 material surface, contact angle θ has certain angle, this surface is considered to nonwetting.The most frequently used solid phase carrier substrate material is a glass in the biochip, and its surface static water contact is about 45 degree.
Thereby require in the detection reaction there to be in the biochip of identical reaction environment every probe being fixed with one group of probe, Shang Weiyou only utilizes high hydrophobicity to carry out the report of the isolation between the reactor.In our research, we find high hydrophobicly can be limited in the hydrophilic substrate surface that high hydrophobic material surrounds to aqueous solution effectively independently as the reactor isolation mech isolation test of biochip.Table 1 has been listed the isolation effect of some high hydrophobic material coatings.
The isolation effect of some high hydrophobic material coatings of table 1
The hydrophilic substrate surface area of high hydrophobic coating Static Water contact angle coating width coating thickness amount of water 90 degree hang down and flow
Organosilicon coating 95 degree 1mm 30-40um 16mm
220ul-
Nano-spun fabric 155 degree 1mm 400um 16mm
230ul-
Nano silicon oxide CKJ7 160 degree 1mm 20-30um 16mm
230ul-
Wound can be sprayed 85 degree 1mm＜5um 16mm
215ul-
No coated substrate 45 degree--16mm
210ul+
The 90 degree stream that hangs down is meant biochip with respect to the horizontal plane revolved and turn 90 degrees in the table, observes water toward sagging stream, and the positive reaction (+) of flowing is arranged, and does not have the negative reaction (-) of flowing.
Concrete technology contents of the present invention is as follows:
A kind of biochip of reactor isolation structure minimized height, the solid phase carrier that it comprises the reactor isolation structure that contains high hydrophobic material and not hydrophobic solid phase carrier at least or is fixed with probe, it is characterized in that: the surface static water contact angle of described high hydrophobic material is at least 80 degree, described high hydrophobic material isolation structure height is 0.1-1000um, and the distance minimum of high hydrophobic material layer and probe is 0.1mm in the described reactor isolation structure.
The surface static water contact angle preferred version of high hydrophobic material of the present invention is at least 130 degree, more preferably scheme is at least 150 degree.
The preferred version of high hydrophobic material isolation structure height of the present invention is 0.1-60um, more preferably scheme is 0.1-30um, and most preferably scheme is 0.1-10um.
Isolation structure and the solid phase carrier that contains high hydrophobic material of the present invention forms a plurality of open reactive devices.
Reactor isolation structure of the present invention mainly comprises by coating or lacquer or latex or China ink, or the high hydrophobic material coating that forms behind the substrate surface dry solidification of the high hydrophobic material applying coating of its combination in any; Or mainly comprise by high hydrophobic material film or high hydrophobic material sheet or high hydrophobic material powder or high hydrophobic material plate or high hydrophobic material band, or the solid-state high hydrophobic material of its combination in any is by the high hydrophobic material layer of heating and melting or the bonding formation of tackifier.
The high hydrophobic material isolation structure of reactor of the present invention is colourless, and it is transparent or semitransparent that preferred version is, and more preferably scheme is transparent.
Minor increment between the reactor reaction of the present invention pond is longer than the reaction tank width on its extension line, and preferred version is long more than 20%, and more preferably scheme is long more than 50%.
The present invention has to add the liquid zone or/and go out the open mobile chip of liquid zone in reactor.
The present invention except that high hydrophobic material structure, also includes hydrophilic substrate surface in its reactor isolated area.
The structure of the biochip in the invention can also include profiled sheeting except that described substrate and described high hydrophobic material; The part or all of liquid feeding structure that can be provided with reactor in the profiled sheeting is or/and the fluid structure; Reactor adds the liquid zone and/or goes out and can include following one or more the structure of control reaction medium directed flow speed in the liquid zone: water wetted material layer, hydrophobic material layer, absorbent material layer and help to control mobile curb, groove, bar.
The present invention at the ratio of the area of the above high hydrophobic material coating of its substrate and the substrate total area less than 15%.
It is the two-sided biochip that is fixed with probe in the end face and the bottom surface of substrate for the present invention, and the end face of described biochip and the structure on the bottom surface are symmetry or asymmetric.
High hydrophobic material of the present invention is organosilicon polymer, multipolymer or its high Hydrophobic Derivatives based on siloxane bond.
High hydrophobic material of the present invention is a fluororesin, comprise: the high Hydrophobic Derivatives of fluororesin oligomer, polytetrafluoroethylene (PTFE), copolymerization fluororesin (PFEP), polychlorotrifluoroethylene, Kynoar and fluororesin reaches emulsion, nanoscale colloid and film with its preparation.
High hydrophobic material of the present invention is polypropylene, polyacrylate and Hydrophobic Derivatives thereof.
High hydrophobic material of the present invention is that Static Water contact angle minimum is the high dewatering nano material of 150 degree; Containing diameter in the described nano material is the functional nano particulate of 1nm-100nm.
The preparation method of the biochip of a kind of reactor isolation structure minimized height of the present invention comprises the steps:
A, provide and to be attached to on-chip, as to contain high hydrophobic material flowing material (bag Chinese juniper solution, coating, paint, ink, gel, emulsion, aerogel);
B, described flowing material is coated in the position that needs to form high hydrophobic isolation structure on the substrate;
C, drying are or/and add hardening agent cured coated material to form the solid-state coating of covering.
Curing reaction wherein, comprise oxidative polymerization type (for example containing the lacquer of drying oil modification etc.), solvent evaporates type (for example the varnish that solidifies of air drying etc.), hardening agent curing type, heat curing, biological enzyme type (for example lacquer etc.), or the like.
The another kind of preparation method of the biochip of reactor isolation structure minimized height of the present invention comprises the steps:
A, provide can be attached to on-chip, contain high hydrophobic material, in type solid matter (comprising sheet, film, plate, band, powder);
B, with described solid matter with gluing or/and hot sticking method is combined in the position that needs to form isolation structure on the substrate.
The ultimate principle of technical scheme of the present invention is as follows:
High hydrophobic material isolation structure of the present invention and solid phase carrier form a plurality of open reactive devices, and the distance minimum of high hydrophobic material layer and probe is the minimum 0.3mm of being of 0.1mm, preferred version.This is because chip is different with the other products of stationary probe at random, and the former has many probes and latter's probe only.Because chip requires the reaction environment of each bar probe identical, according to experiment of the present invention, the repeatability that this minor increment helps detecting.
Reactor isolation structure of the present invention mainly is by curable high hydrophobic material coating, comprise that coating or lacquer or latex or China ink or its combination in any are coated in the high hydrophobic material coating that forms behind the substrate surface dry solidification, or mainly be by with the high hydrophobic material of the form of film, sheet, powder, plate, band by heating and melting or the bonding high hydrophobic material layer that forms of tackifier.Wherein, high hydrophobic material coating is defined as with physics, chemistry or other method the overlayer with high hydrophobic function of a layer thickness between 0.1um-1mm in substrate surface formation.In the present invention, high hydrophobic material coating is meant the material that can form high hydrophobic material coating.
On the other hand, in biochip according to the present invention, the high hydrophobic material in the described reactor isolation structure is the colourless material that produces minimum scan transponding when detecting, and preferred version is transparent or semitransparent material, and more preferably scheme is a material transparent.According to experimental verification, use especially more preferably scheme of preferred version, the existence of isolation structure can obviously not increase background and disturb.
The present invention is in described reactor isolation structure, and the minor increment between the described reactor reaction pond is longer than the reaction tank width on its extension line, and preferred version is long more than 20%, and more preferably scheme is long more than 50%.The isolated area of broad helps making the minimizing possibility of the cross pollution between reactor.
Except that high hydrophobic material structure, also include hydrophilic substrate surface in the biochip provided by the invention, its reactor isolated area.According to experimental verification of the present invention, this hydrophilic substrate surface helps being detained low amounts of water solution, thereby reduces cross contamination risk.
In biochip according to the present invention, the material of substrate comprise all can with less average area form the biochip reaction device, the surface static water contact angle is less than the materials of 60 degree, for example inorganic material such as glass, silicon and silicon compound etc., organic high molecular polymers such as polypropylene, Polyvinylchloride, polystyrene, nylon and cellulose nitrate, and surface coverage has the organic material of metals such as gold, silver, metallic compound.
The advantage of biochip of the present invention is: owing to realized the isolation structure minimized height, and can be so that number of reactors maximization and/or sample addition minimize easier realization.
Accompanying drawing and drawing explanation thereof:
The used high hydrophobic material of Fig. 1 the present invention is the basic block diagram of the multiple reactor biochip of fluent material
The used high hydrophobic material of Fig. 2 the present invention is the basic block diagram of the multiple reactor biochip of solid phase material
The basic block diagram of the open mobile biochip of Fig. 3 multiple reactor of the present invention
The basic block diagram of Fig. 4 multiple reactor biochip of the present invention
The basic block diagram of Fig. 5 two-sided biochip of the present invention
1, substrate 2, isolation structure 3, reaction tank 4, probe 5, add liquid zone 6, go out the liquid zone
Embodiment:
The substrate that uses in the embodiment of the invention is glass substrate, is of a size of 75 * 25 * 1mm, and its surface active groups is amino.Slide is a microscope slide, and the surface of glass slide amination is with reference to work " biomolecule mobilization technology and application " (Beijing, Chemical Industry Press, 1998) such as Jiang Zhonghua.
Embodiment 1: the multiple reactor biochip
The basic structure of the biochip in this example as shown in Figure 1.
In this example, substrate for use is the glass material substrate, used high hydrophobic material is fluent material, is respectively " the hydrophobic latex paint of superelevation " (morning twilight chemical design institute, Chinese Chengdu provides), " organic silicon water-proofing coating " (morning twilight chemical design institute, Chinese Chengdu provides) and " polyacrylic acid grease coating material " (morning twilight chemical design institute, Chinese Chengdu provides).
In the present embodiment, substrate 1 is of a size of 75 * 25 * 1mm (length * wide * height), high hydrophobic material evenly spreads upon in the on-chip isolated area, operation instruction by the supplier is solidified after drying at room temperature, form the coating isolation structure 2 shown in Fig. 1, form 16 reaction tanks 3 with substrate, reaction tank is of a size of 3mm * 3mm.On this surface of substrate, except that reaction tank, all be covered with high hydrophobic material coating, parameters such as coating thickness see Table 2.
After chip is made, the probe region 4 internal fixation probes of the reaction tank 3 that surrounds in high hydrophobic material coating, probe region 4 is 100um with the minor increment of high hydrophobic isolation structure 2.Three kinds of probes are respectively HCV antigen, HIV
1+2Antigen and syphilis antigen.Every kind of probe is 3 points, forms one 3 * 3 probe array.Chip is standby with bovine serum albumin(BSA) sealing back.
The biochip master data of four kinds of different isolated materials of table 2
The high hydrophobic material Static Water of chip contact angle coating thickness color and probe minor increment
The hydrophobic latex paint of A superelevation＞colourless, the translucent 100um of 120 degree 20-40um
The B organic silicon water-proofing coating～translucent 100um of 95 degree 20-40um
C polyacrylic acid grease coating material～colourless, the translucent 100um of 85 degree 20-40um
D do not have～45 degree---
In the present embodiment, No. 1 sample is a HCV antibody positive serum, and No. 2 samples are HIV
1+2The antibody positive human serum, No. 3 samples are syphilis antibody positive human serum, No. 4 positive testers of sample (HCV antibody, HIV
1+2The potpourri of antibody and syphilis antibody positive serum tester), No. 5 negative testers of sample (HCV antibody, HIV
1+2The serum tester that antibody and syphilis antibody are all negative).All samples all are to determine through using the classical open chip of single reactor to detect in advance under the equivalent responses condition.Get each one of above-mentioned 4 kinds of chip A, B, C, D during experiment, 1,2, No. 3 sample is respectively done 3 holes on each piece, and 2 holes are respectively done in the contrast of yin and yang attribute blood plasma, and blank is done 1 hole, and the result gets same probe result's average.During experiment, the application of sample amount is 10ul, and the each addition of cleansing solution is 15ul, and the label addition is 10ul.Reaction conditions is identical with the open chip of single reactor that uses classics, experimental result such as table 3.
The testing result of the mobile biochip of table 3
Sample
HCV antibody HIV1+2 antibody Syphilis antibody
A B C D A B C D A B C D
No. 1++++--------
No. 2----++++----
No. 3--------++++
No. 4++++++++++++
No. 5------------
Embodiment 2: the multiple reactor biochip
The basic structure of the biochip in this example as shown in Figure 2.
In this example, substrate for use is the glass material substrate, used high hydrophobic material is solid phase material, (morning twilight chemical design institute, Chinese Chengdu provides to be respectively " teflon Pressure sensitive adhesive tape ", Static Water order of contact 117 degree) and " nano-spun fabric " (Chinese Zhoushan nano material tomorrow company provides, and Static Water order of contact 155 is spent).
In the present embodiment, substrate 1 is of a size of 75 * 25 * 1mm (length * wide * height), high hydrophobic material is bonded in respectively in the on-chip isolated area after by perforate, after drying at room temperature, form the high hydrophobic isolation structure 2 shown in Fig. 2, form 16 reaction tanks 3 with substrate 1, reaction tank is of a size of 4.5mm * 4.5mm.High hydrophobic material layer thickness is respectively: teflon Pressure sensitive adhesive tape 100um (color is a black); Nano-spun fabric 400um.
After chip is made, the fixing probe identical of probe region in reaction tank beyond the high hydrophobic material coating 0.5mm 4 with embodiment one.Every kind of probe is 3 points, forms one 3 * 3 probe array.Chip is standby with bovine serum albumin(BSA) sealing back.
In the present embodiment, specimen in use, liquid volume added and reaction conditions are identical with embodiment one, and experimental result is also identical with embodiment one.
Embodiment 3: the open mobile biochip of multiple reactor
The basic structure of the biochip in this example as shown in Figure 3.The left side is a vertical view among the figure, the right side is the a-a cut-open view, dotted portion is an activation slide substrate 1 in the middle of it, substrate is of a size of 75mm * 5mm * 1mm (length * wide * height), substrate is fixed on the plastic molding board that liquid feeding structure and fluid structure are arranged, the profiled sheeting overall dimensions are 80mm * 25mm * 1mm (length * wide * height), and liquid feeding structure and fluid structure on it are the groove of wide 2mm, and the water wetted material of water conservancy diversion is arranged in the groove.On-chip reaction tank periphery is the super-hydrophobic isolation structure 2 that one deck superelevation hydrophobic " nano silicon oxide CDS7 " of applying (Chinese Zhoushan nano material tomorrow company provide) forms.By supplier's requirement, in containing the solution of tackifier, after powerful the dispersion, promptly evenly spread upon in the on-chip isolated area, after drying at room temperature, solidify, form the approaching transparent coating shown in Fig. 3, profiled sheeting and substrate form 16 reaction tanks 3, and the reaction tank diameter is 3.0mm.As shown in Figure 3, the liquid zone of adding 5 is arranged on each reactor and go out liquid zone 6.On this surface of substrate, at reaction tank 3, add liquid zone 5 and go out the high hydrophobic isolation structure 2 that scribbles wide 2mm beyond the liquid zone 6, coating thickness 20um-30um.Sample flows to out the liquid zone through reaction zone after add the liquid zone and add.After chip is made, the probe that probe region 4 internal fixation in reaction tank beyond the high hydrophobic material coating 0.5mm and embodiment one are identical.Every kind of probe is 3 points, forms one 3 * 3 probe array.Chip is standby with bovine serum albumin(BSA) sealing back.
In the present embodiment, specimen in use is identical with embodiment one, and the application of sample amount is 10ul, and cleansing solution adding speed is about 30ul/ minute, and the label addition is 10ul.Reaction conditions with use common open mobile chip identical, experimental result is identical with embodiment one.
Embodiment 4: the multiple reactor biochip
The basic structure of the biochip in this example as shown in Figure 4.
In this example, substrate for use is the glass material substrate, and used high hydrophobic material is respectively " organic silicon water-proofing coating " (morning twilight chemical design institute, Chinese Chengdu provides) and a kind of automobile, and (wound can be sprayed from spraying paint, HD-036, B-92P, black pearl, Chinese Shanghai opens abundant industry development company)." organic silicon water-proofing coating " reaches " wound can be sprayed " and evenly smears respectively or be injected in the on-chip isolated area, solidifies after drying at room temperature, forms the coating of the approaching transparent or black shown in Fig. 4, forms 16 reaction tanks 3 with substrate.Coating thickness is respectively 20-40um and less than 5um, width is respectively 1.5mm.Be substrate surface between the high hydrophobic coating 2.
After chip is made, the probe that the probe region internal fixation in reaction tank beyond the high hydrophobic material coating 0.5mm and embodiment one are identical.Every kind of probe is 3 points, forms one 3 * 3 probe array.Chip is standby with bovine serum albumin(BSA) sealing back.
In the present embodiment, chip specimen in use, liquid volume added and the reaction conditions of three kinds of different materials isolation structures are identical with embodiment one, and experimental result is also all identical with embodiment one.
Embodiment 5: two-sided biochip
The basic structure of the biochip in the present embodiment as shown in Figure 5.
Embodiment one is identical.
Claims (18)
1, a kind of biochip of reactor isolation structure minimized height, the solid phase carrier that it comprises the reactor isolation structure that contains high hydrophobic material and not hydrophobic solid phase carrier at least or is fixed with probe, it is characterized in that: the surface static water contact angle of described high hydrophobic material is at least 80 degree, described high hydrophobic material isolation structure height is 0.1-1000um, and the distance minimum of high hydrophobic material layer and probe is 0.1mm in the described reactor isolation structure.
2, the biochip of a kind of reactor isolation structure minimized height according to claim 1 is characterized in that: the surface static water contact angle of described high hydrophobic material is at least 130 degree.
3, the biochip of a kind of reactor isolation structure minimized height according to claim 1 and 2 is characterized in that: described high hydrophobic material isolation structure height is 0.1-60um.
4, the biochip of a kind of reactor isolation structure minimized height according to claim 3 is characterized in that: described isolation structure and the solid phase carrier that contains high hydrophobic material forms a plurality of open reactive devices.
5, the biochip of a kind of reactor isolation structure minimized height according to claim 4, it is characterized in that: described reactor isolation structure mainly comprises by coating or lacquer or latex or China ink, or the high hydrophobic material coating that forms behind the substrate surface dry solidification of the high hydrophobic material applying coating of its combination in any; Or mainly comprise by high hydrophobic material film or high hydrophobic material sheet or high hydrophobic material powder or high hydrophobic material plate or high hydrophobic material band, or the solid-state high hydrophobic material of its combination in any is by the high hydrophobic material layer of heating and melting and the bonding formation of tackifier.
6, the biochip of a kind of reactor isolation structure minimized height according to claim 4 is characterized in that: the high hydrophobic material isolation structure of described reactor is colourless.
7, the biochip of a kind of reactor isolation structure minimized height according to claim 4 is characterized in that: the minor increment between the described reactor reaction pond is longer more than 20% than the reaction tank width on its extension line.
8, the biochip of a kind of reactor isolation structure minimized height according to claim 4 is characterized in that: described biochip is that liquid feeding zone/and go out the open mobile chip of liquid zone is arranged in reactor.
9, the biochip of a kind of reactor isolation structure minimized height according to claim 4 is characterized in that: described biochip except that high hydrophobic material structure, also includes hydrophilic substrate surface in its reactor isolated area.
10, the biochip of a kind of reactor isolation structure minimized height according to claim 4 is characterized in that: the structure of described biochip also includes profiled sheeting except that described substrate and described high hydrophobic material; The part or all of liquid feeding structure that is provided with reactor in the profiled sheeting is or/and the fluid structure; Reactor adds the liquid zone and/or goes out and includes following one or more the structure of control reaction medium directed flow speed in the liquid zone: water wetted material layer, hydrophobic material layer, absorbent material layer and help to control mobile curb, groove, bar.
11, the biochip of a kind of reactor isolation structure minimized height according to claim 4 is characterized in that: the area of the above high hydrophobic material coating of its substrate and the ratio of the substrate total area are less than 15%.
12, the biochip of a kind of reactor isolation structure minimized height according to claim 4, it is characterized in that: described biochip is the two-sided biochip that is fixed with probe in the end face of substrate and bottom surface, and the end face of described biochip and the structure on the bottom surface are symmetry or asymmetric.
13, the biochip of a kind of reactor isolation structure minimized height according to claim 5 is characterized in that: described high hydrophobic material is organosilicon polymer, multipolymer or its high Hydrophobic Derivatives based on siloxane bond.
14, the biochip of a kind of reactor isolation structure minimized height according to claim 5 is characterized in that: described high hydrophobic material is a fluororesin.
15, the biochip of a kind of reactor isolation structure minimized height according to claim 5 is characterized in that: described high hydrophobic material is polypropylene, polypropylene fat or its Hydrophobic Derivatives.
16, the biochip of a kind of reactor isolation structure minimized height according to claim 5 is characterized in that: described high hydrophobic material is that Static Water contact angle minimum is the high dewatering nano material of 150 degree; Containing diameter in the described nano material is the functional nano particulate of 1nm-100nm.
17, the preparation method of the biochip of a kind of reactor isolation structure minimized height according to claim 1 is characterized in that: comprise the steps:
A, provide and to be attached to flowing material on-chip, that contain high hydrophobic material;
B, described flowing material is coated in the position that needs to form high hydrophobic isolation structure on the substrate;
C, drying are or/and add hardening agent cured coated material to form the solid-state coating of covering.
18, the preparation method of the biochip of a kind of reactor isolation structure minimized height according to claim 1 is characterized in that: comprise the steps:
A, provide and to be attached to on-chip, that contain high hydrophobic material, in type solid matter;
B, with described solid matter with gluing or/and hot sticking method is combined in the position that needs to form isolation structure on the substrate.
Priority Applications (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03117397 CN1208622C (en) | 2003-03-04 | 2003-03-04 | Biological chip with minimized reactor isolation structure height and its preparation method |
| PCT/CN2004/000169 WO2004083863A1 (en) | 2003-03-04 | 2004-03-04 | An integrating analysis chip with minimized reactors and its application |
| EP04717012A EP1605264B1 (en) | 2003-03-04 | 2004-03-04 | An integrating analysis chip with minimized reactors and its application |
| US10/547,681 US20060182655A1 (en) | 2003-03-04 | 2004-03-04 | Integrating analysis chip with minimized reactors and its application |
| JP2006504196A JP2006519384A (en) | 2003-03-04 | 2004-03-04 | Highly integrated analysis chip with extremely small height reactor and its application |
| CN200480000654.9A CN1697976B (en) | 2003-03-04 | 2004-03-04 | High integration analysis chip of minimized height for reactor and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03117397 CN1208622C (en) | 2003-03-04 | 2003-03-04 | Biological chip with minimized reactor isolation structure height and its preparation method |
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| Publication Number | Publication Date |
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| CN1434296A CN1434296A (en) | 2003-08-06 |
| CN1208622C true CN1208622C (en) | 2005-06-29 |
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| CN 03117397 Expired - Fee Related CN1208622C (en) | 2003-03-04 | 2003-03-04 | Biological chip with minimized reactor isolation structure height and its preparation method |
| CN200480000654.9A Expired - Fee Related CN1697976B (en) | 2003-03-04 | 2004-03-04 | High integration analysis chip of minimized height for reactor and application |
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| Application Number | Title | Priority Date | Filing Date |
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| CN200480000654.9A Expired - Fee Related CN1697976B (en) | 2003-03-04 | 2004-03-04 | High integration analysis chip of minimized height for reactor and application |
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| CN (2) | CN1208622C (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1253585C (en) * | 2003-09-30 | 2006-04-26 | 成都夸常科技有限公司 | Capillary chamber chip, capillary chip and chip element and device |
| WO2005095961A1 (en) * | 2004-03-17 | 2005-10-13 | Matsushita Electric Industrial Co., Ltd. | Process for producing biochip, probe solution, and biochip |
| CN1648671B (en) * | 2005-02-06 | 2012-09-26 | 成都夸常医学工业有限公司 | Detecting method for multiple reactor analytic chip and analytic chip and detector |
| CA2712572C (en) * | 2008-01-18 | 2016-09-13 | National University Corporation University Of Toyama | Reaction device, reaction method and method of synthesizing cdna |
| CN105861309B (en) * | 2016-04-14 | 2018-05-11 | 清华大学 | A kind of super-hydrophobic micro-pit array chip and preparation method and application |
| CN118080035A (en) * | 2018-12-13 | 2024-05-28 | 迪亚莱博（张家港）生物科技有限公司 | Preparation method of bedside diagnosis micro-fluidic chip |
| CN115652440B (en) * | 2022-12-27 | 2023-04-14 | 北京百奥纳芯生物科技有限公司 | Low-density aldehyde-based substrate and preparation method of low-density biochip |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN2280908Y (en) * | 1996-07-22 | 1998-05-06 | 高劲松 | Spot immunizing or spot-cross-breeding preparation case |
| AU8908198A (en) * | 1997-08-15 | 1999-03-08 | Hyseq, Inc. | Methods and compositions for detection or quantification of nucleic acid species |
| CN1159454C (en) * | 1999-05-21 | 2004-07-28 | 陆祖宏 | Compound micro-array chip preparing process and the micro-array chip using the same |
| CN1109243C (en) * | 2000-02-04 | 2003-05-21 | 吴昌 | Stereo pool type medium wafer used for preparing biochip |
| CN1343790A (en) * | 2000-09-19 | 2002-04-10 | 河南省生物技术开发中心 | Biochip and integrated biochip |
| CN1138145C (en) * | 2001-04-27 | 2004-02-11 | 上海晶泰生物技术有限公司 | Multiple-sample microarray biochip |
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2003
- 2003-03-04 CN CN 03117397 patent/CN1208622C/en not_active Expired - Fee Related
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Also Published As
| Publication number | Publication date |
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| CN1697976B (en) | 2010-12-22 |
| CN1697976A (en) | 2005-11-16 |
| CN1434296A (en) | 2003-08-06 |
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