CN101241124A - Biological chip substrate and method for making same - Google Patents
Biological chip substrate and method for making same Download PDFInfo
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- CN101241124A CN101241124A CNA2008100577846A CN200810057784A CN101241124A CN 101241124 A CN101241124 A CN 101241124A CN A2008100577846 A CNA2008100577846 A CN A2008100577846A CN 200810057784 A CN200810057784 A CN 200810057784A CN 101241124 A CN101241124 A CN 101241124A
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- glucosan
- aldehyde radical
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- 239000000758 substrate Substances 0.000 title claims abstract description 93
- 238000000034 method Methods 0.000 title claims description 40
- 229920001503 Glucan Polymers 0.000 claims abstract description 65
- 238000005576 amination reaction Methods 0.000 claims abstract description 33
- 238000000018 DNA microarray Methods 0.000 claims abstract description 25
- 125000003563 glycoside group Chemical group 0.000 claims abstract description 5
- TWNIBLMWSKIRAT-VFUOTHLCSA-N levoglucosan Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@H]2CO[C@@H]1O2 TWNIBLMWSKIRAT-VFUOTHLCSA-N 0.000 claims description 27
- FFUAGWLWBBFQJT-UHFFFAOYSA-N hexamethyldisilazane Chemical compound C[Si](C)(C)N[Si](C)(C)C FFUAGWLWBBFQJT-UHFFFAOYSA-N 0.000 claims description 20
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 claims description 14
- 229920002307 Dextran Polymers 0.000 claims description 13
- -1 polysiloxane Polymers 0.000 claims description 13
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 13
- 230000003647 oxidation Effects 0.000 claims description 8
- 238000007254 oxidation reaction Methods 0.000 claims description 8
- 229920001296 polysiloxane Polymers 0.000 claims description 7
- 230000008021 deposition Effects 0.000 claims description 6
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 claims 3
- 238000002360 preparation method Methods 0.000 abstract description 28
- 108020004707 nucleic acids Proteins 0.000 abstract description 7
- 102000039446 nucleic acids Human genes 0.000 abstract description 7
- 150000007523 nucleic acids Chemical class 0.000 abstract description 7
- 125000003172 aldehyde group Chemical group 0.000 abstract description 6
- 229920002521 macromolecule Polymers 0.000 abstract description 5
- 239000000463 material Substances 0.000 abstract description 2
- 230000036186 satiety Effects 0.000 abstract 1
- 235000019627 satiety Nutrition 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 35
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 21
- 239000011159 matrix material Substances 0.000 description 19
- 229920000642 polymer Polymers 0.000 description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 12
- 150000001299 aldehydes Chemical class 0.000 description 12
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 10
- 238000004375 physisorption Methods 0.000 description 10
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- 238000009396 hybridization Methods 0.000 description 7
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- 229920003023 plastic Polymers 0.000 description 6
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- FZHAPNGMFPVSLP-UHFFFAOYSA-N silanamine Chemical compound [SiH3]N FZHAPNGMFPVSLP-UHFFFAOYSA-N 0.000 description 5
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 238000000151 deposition Methods 0.000 description 4
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- OKHIGGWUISQLMG-UHFFFAOYSA-N 3-diethoxysilylpropan-1-amine Chemical compound CCO[SiH](OCC)CCCN OKHIGGWUISQLMG-UHFFFAOYSA-N 0.000 description 1
- AHYFYQKMYMKPKD-UHFFFAOYSA-N 3-ethoxysilylpropan-1-amine Chemical compound CCO[SiH2]CCCN AHYFYQKMYMKPKD-UHFFFAOYSA-N 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 1
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Abstract
The present invention discloses a biochip substrate and preparation thereof. The provide biochip substrate is grafted aldehydized glucan on the surface of amination chip base, wherein, the aldehydized glucan is made by oxidating the o-hydroxy of the glycoside units into aldehyde group. As a macromolecule 3D aldehyde group substrat, the glucan aldehyde group substrate of present invention bonding capacity is large, so the nucleic acid fixation capability is improved, the point shape is satiety; the problem that mostly superchip sample spotting is not easy to fix in common aldehyde group substrate is resolved, the stability of substrate under spotting environment (humidity of which is high) for long time is enhanced, so the present invention is suitable for spotting system of long time spotting superchip. So the glucan aldehyde group substrate of present invention is an excellent biochip substrate material and can apply to preparation of all kinds of biochip substrate.
Description
Technical field
The present invention relates to a kind of bio-chip substrate and preparation method thereof.
Background technology
Now, chip technology has become necessary part in the life science, when characterizing complicated bioprocess and maybe need to measure a large amount of Diagnostic parameters, chip is extensive, the feature of high throughput analysis make chip between analyzing gene group series jump, gene expression profile, protein, react in and be widely adopted.
Substrate is the basis of chip, and the preparation of substrate has very important effect in the development of biochip.
Usually, substrate is divided into three kinds by molecular structure: first kind, simple silanization substrate, amino as carrying out in surface of glass slide on the silylation modification, aldehyde radical, epoxide group, sulfydryl etc., this type substrate exists bigger sterically hindered, and stronger non-special absorption is arranged, and uses more in early days in the chip research; Second kind, slide carries out small numerator modified substrate again behind simple silanization, and this type substrate space surface increases to some extent, has reduced non-special absorption to a certain extent; The third, polymer matrix film, can be divided into molecule does not have the polymer matrix film of reactive group, molecule to contain the polymer matrix film of a reactive group and the polymer matrix film that molecule contains a plurality of reactive groups.Polymer matrix film provides an additional surfaces protective seam, thereby reduces non-specific adsorption, and the polymer matrix film per surface that molecule contains a plurality of reactive groups also provides more active sites, thereby causes more effective sample to be fixed.Polymer matrix film can also be divided into dendritic macromole polymer matrix film, filtration membrane substrate and hydrogel substrate.
Common aldehyde radical substrate belongs to simple silanization substrate, is a kind of one dimension substrate.In the practical application, the fixed amount of common aldehyde radical substrate is lower, and sensitivity is not high; And in a system superchip process, there is stability problem, when promptly (the point sample ambient humidity is big) long-time point sample point sample occurs and has just begun under the point sample environment spot diameter normally, the more and more littler phenomenon of most spot diameters backward, it also is star point phenomenon, cause just having begun behind the chip hybridization region point size of point sample and signal normally, backward overwhelming majority point little to star point down to extracting data, just there is stability problem in Chang Yong simple silanization aldehyde radical substrate in long-time point sample process, and the point sample time is long more, and this problem is serious more.
Summary of the invention
An object of the present invention is to provide a kind of bio-chip substrate.
Bio-chip substrate provided by the present invention is the substrate of aldehyde radical glucosan in the grafting of amination sheet primary surface; Described aldehyde radical glucosan is to be that aldehyde radical obtains with the adjacent hydroxyl oxidize on the glycoside units of glucosan.
Wherein, described aldehyde radical glucosan obtains with periodic acid or water-soluble periodate oxidation glucosan.The dextran polymer molecular structure as shown in Figure 1, after periodic acid or the processing of water-soluble periodate, just the adjacent hydroxyl on all or part glycoside units is interrupted and is become two aldehyde radicals, so dextran polymer molecule basic framework and length after the aldehyde radicalization are constant, be the long chain polymer that a kind of all or part glycoside units contains two aldehyde radicals, as shown in Figure 2.
The optional molecular weight of described glucosan from more than 10000 to the molecule of hundreds of thousands, specifically can be Dextran T 40 or Dextran T 70.
Wherein, amination sheet base can obtain from commercial channels, also can carry out amido modified obtaining to existing methods of employing such as slide, plastics, silicon chips, as adopting amino silane method or physisorption amination method.When described base was slide, described amination sheet base can adopt following physisorption amination method preparation: slide is carried out vapour deposition with hexamethyldisilazane, and then handle with containing amino polysiloxane, obtain the amination slide.
Another object of the present invention provides two kinds of methods that prepare above-mentioned bio-chip substrate.
A kind of method for preparing above-mentioned bio-chip substrate provided by the present invention is to obtain bio-chip substrate with being grafted to after the glucosan aldehyde radicalization on the amination sheet primary surface.
The another kind of method of the above-mentioned bio-chip substrate of preparation provided by the present invention is to obtain bio-chip substrate with periodic acid or water-soluble periodate oxidation again after being grafted on the amination sheet primary surface after the glucosan aldehyde radicalization.
Wherein, the glucosan aldehyde radical substrate point of sample diameter of back one method preparation can be controlled, and compares the spot diameter size of last method, and can equate also can be bigger.The glucosan aldehyde radical substrate of these two kinds of aldehyde group method preparations can reach identical result by the CONTROL PROCESS condition.In actual applications, can select any method in above-mentioned two kinds of methods to be prepared as required.
Wherein, concrete available periodic acid of described glucosan aldehyde radicalization or water-soluble periodate oxidation glucosan.
Glucosan in the said method specifically can be Dextran T 70 or Dextran T 40.
Wherein, amination sheet base can obtain from commercial channels, also can carry out amido modified obtaining to existing methods of employing such as slide, plastics, silicon chips, as adopting amino silane method or physisorption amination method.When described base was slide, described amination sheet base can adopt following physisorption amination method preparation: slide is carried out vapour deposition with hexamethyldisilazane, and then handle with containing amino polysiloxane, obtain the amination slide.
The biochip that is made by substrate of the present invention also belongs to protection scope of the present invention.
In the glucosan aldehyde radical substrate of the present invention, glucosan itself is a kind of polyphosphazene polymer sugar compounds, has good bio-compatibility, and low-down non-specific adsorption and autofluorescence background are arranged, and makes the sensitivity of sample detection be significantly improved; Glucosan is a hydrophilic macromolecular compounds, and long hydrophilic linking arm has enough suppleness, can make the mode by covalency or high-affinity that the sample molecule that is fixed in substrate surface is fully contacted and combination with target molecule; With it is that skeleton carries out the reactive group modification, Zhi Bei the glucosan aldehyde radical substrate that is rich in aldehyde radical thus, not only biomolecule fixedly sensitivity improve greatly, Signal gradient is obvious, and can realize combination between the molecule faster and balance.So, as a kind of polymer three-dimensional aldehyde radical substrate, its bonding capacity is big, the fixed amount height, and Signal gradient is obvious, not only the nucleic acid crystallized ability has obtained large increase, the point shape is full, and the shape of sample spot remains homogeneous in the long-time sample spot system process, has solved the problem that some system superchip major part sample that common aldehyde radical substrate exists is not fixed, improved the stability of point sample environment subtegulum, convenience point system needs the superchip of long-time point sample.Therefore, glucosan aldehyde radical substrate of the present invention is a kind of biochip base material of function admirable, can be widely used in the preparation of various biochips.
Description of drawings
Fig. 1 is the dextran molecule structure before the modification;
Fig. 2 is the structural representation of glucosan aldehyde radical substrate surface molecule;
Fig. 3 is that glucosan aldehyde radical substrate and common aldehyde radical substrate nucleic acid samples crystallized ability compare scintigram;
Fig. 4 is that glucosan aldehyde radical substrate and common aldehyde radical substrate nucleic acid samples fixed signal value compare histogram;
Fig. 5 is the some system effect contrast figure of glucosan aldehyde radical substrate and common aldehyde radical substrate point system high density people's promoter chip;
Fig. 6 is the crossbreeding effect comparison diagram of glucosan aldehyde radical substrate and common aldehyde radical substrate point system high density people's promoter chip.
Embodiment
The concrete available following several method preparation of the amination sheet base that relates among the present invention.
The sheet base of substrate can be slide or plastics etc., and plastics specifically can be PMMA.Slide sheet base can make its surface amination by the method for amino silaneization or physisorption; Plastic base generally makes its surface amination by the method for amino silaneization.Compare with the amidized surface of glass slide of silane, amidized surface of glass slide of physisorption and the amidized frosting of silane are all more hydrophobic.
Wherein, the used silylating reagent of amino silaneization can be selected amino silane reagent such as 3-aminopropyl triethoxysilane, 3-aminopropyl diethoxy silane, 3-aminopropyl-Ethoxysilane for use; Physisorption can be with containing amino polysiloxane, as Poly[dimethylsiloxane-co-(3-aminopropyl) methylsiloxane] etc.
Following mask body division:
1) method of amino silane slide is as follows:
With surface of glass slide Piranha (dense H
2SO
4: 30%H
2O
2, V/V=7: 3) solution soaks more than the 2h or uses the chromic acid solution soaked overnight, uses washed with de-ionized water, drying then; Or will carry out plasma treatment 2min after the drying of slide washed with de-ionized water; React in the ethanolic solution with above-mentioned slide and 1% (volumn concentration) the 3-TSL 8330 (APTES) after handling; Ethanol cleans the back and dries the slide that promptly gets the surface amino groups silanization.
The method of 2) physisorption amination slide is as follows:
Clean slide; Take out after clean beaker is put in 120 ℃ of baking ovens baking 30min, drip several HMDS (hexamethyldisilazane) rapidly and cover clean surface plate immediately, open surface plate after 10 seconds, the slide that cleans up is put into beaker and covered surface plate immediately, make surface of glass slide carry out HMDS vapour deposition 30min; The preparation 0.5% (volumn concentration) Poly[dimethylsiloxane-co-(3-aminopropyl) methylsiloxane] dichloromethane solution; Slide after handling is made the polysiloxane of surface of glass slide physisorption last layer band amino by czochralski method in above-mentioned dichloromethane solution, thereby realize the amination of slide.
3) method of amino silane plastics following (is example with PMMA):
After PMMA sheet base cleans, dries, carry out surperficial ozone treatment 1h or Plasma and handle 30min.The aqueous phase solution of preparation 1% (volumn concentration) 3-TSL 8330 (APTES), the PMMA sheet base after will handling is immediately put into and is wherein reacted 2h, thereby makes PMMA sheet primary surface amino silaneization.
The preparation of embodiment 1, glucosan aldehyde radical substrate
Elder generation's degree of depth aldehyde radical dextran molecule is grafted to it amination sheet primary surface again, and concrete method is as follows:
1, the preparation of aldehyde radical glucosan
Take by weighing the 5.68g sodium metaperiodate and be dissolved in the 100ml water, add the glucosan T-40 of 0.02g, cover 30 ℃ of reactions of light 3h after the dissolving; After 4 ℃ of dialysis are clean, promptly get aldehyde radical glucosan macromolecule concentrate, 4 ℃ of preservations.
2, the preparation of amination sheet base
With surface of glass slide Piranha (dense H
2SO
4: 30%H
2O
2, V/V=7: 3) solution soaks 2h, uses washed with de-ionized water, drying then; React 2h in the ethanolic solution with above-mentioned slide and 1% (volumn concentration) the 3-TSL 8330 (APTES) after handling; Reaction back ethanol cleans the back and dries the slide that promptly gets the surface amino groups silanization.
3, the preparation of glucosan aldehyde radical substrate
1) in the 600ml phosphate buffer, adds the aldehyde radical glucosan macromolecule concentrate that 50ml step 1 obtains, and add reductive agent NaBH
3CN 0.08g adds the amination slide reaction 30min that step 2 makes then.
2) clean, dry, promptly get glucosan aldehyde radical substrate.
The preparation of embodiment 2, glucosan aldehyde radical substrate
Shallow degree aldehyde radical dextran molecule is grafted to it amination sheet primary surface again, and then carries out the sodium metaperiodate deep oxidation earlier, and concrete grammar is as follows:
1, the preparation of aldehyde radical glucosan
Take by weighing the 5.68g sodium metaperiodate and be dissolved in the 100ml water, add the glucosan T-70 of 0.02g, cover 30 ℃ of reactions of light 1h after the dissolving; After 4 ℃ of dialysis are clean, promptly get aldehyde radical glucosan macromolecule concentrate, 4 ℃ of preservations.
2, the preparation of amination sheet base
Clean slide; Take out after clean beaker is put in 120 ℃ of baking ovens baking 30min, drip several HMDS (hexamethyldisilazane) rapidly and cover clean surface plate immediately, open surface plate after 10 seconds, the slide that cleans up is put into beaker and covered surface plate immediately, make surface of glass slide carry out HMDS vapour deposition 30min; The preparation 0.5% (volumn concentration) Poly[dimethylsiloxane-co-(3-aminopropyl) methylsiloxane] dichloromethane solution; Slide after handling is made the polysiloxane of surface of glass slide physisorption last layer band amino by czochralski method in above-mentioned dichloromethane solution, thereby realize the amination of slide.
3, the aldehyde radical glucosan is grafted on the amination sheet base
(1) (1) is described in the step 3 with embodiment 1.
(2) clean, dry.
4, further oxidation glucosan
In the 600mL deionized water, add the 12.8g sodium metaperiodate and be mixed with sodium periodate solution, high molecular slide on the surface grafting of step 3 acquisition is immersed in this sodium periodate solution react 3h.
Clean, dry, promptly get glucosan aldehyde radical substrate.
The crystallized ability of embodiment 3, glucosan aldehyde radical substrate and common aldehyde radical substrate relatively
The used common aldehyde radical substrate of present embodiment is according to document (" Colloids and Surfaces B:Biointerfaces " Volume 36, Issues 3-4,1 August 2004, Pages 178) in the method preparation mentioned, be baking was crosslinked after slide carry out amino silaneization earlier, then with the glutaraldehyde prepared in reaction.
The glucosan aldehyde radical substrate that present embodiment is used is by embodiment 1 described method preparation.
The preparation process of biochip is as follows:
1) be mixed with 35mer Oligo sample P BH (sequence the is as shown in table 1) solution of 3 kinds of variable concentrations that contain 50%DMSO respectively with 100%DMSO, every kind of sample takes out 10uL and shifts and add in 384 orifice plates; Preparation 50%DMSO solution also takes out 10uL and shifts in adding 384 orifice plates as blank (BC) contrast; QC (positive quality control of point sample) is that an end has the HEX mark, the oligonucleotide probe that the other end is amido modified is used to observe chip point sample and fixing efficient, and its sequence is as shown in table 1, be mixed with the solution that contains 50%DMSO with 100%DMSO, take out 10uL and shift in adding 384 orifice plates.
The title of table 1, sample and sequence
| The sample title | Sequence |
| PBH | NH2-5’-(T)15-TCGGATTCGACAACACCCGT |
| QC | NH2-5’-(T)15-GTG?CAA?CTC?ACT?CGA?CTG-3’-HEX |
| Arab-HEX | HEX-5’-ACTGGACTTGACGGGTGTTGTCGAATCCGA |
2) by automatic spot sample device ready sampling liquid is put glucosan aldehyde radical substrate surface and common aldehyde radical substrate surface respectively.Concrete dot matrix way as shown in Figure 3 (wherein, A is a glucosan aldehyde radical substrate of the present invention, B is common aldehyde radical substrate): every chip comprises 4 dot matrix, each dot matrix comprises 30 points, the dot matrix of each dot matrix is designed to 5 row *, 6 row, sample repetition form is 1*6, and dot spacing is 400um, and 5 positions of sample on the chip dot matrix are followed successively by from top to bottom: 2.5uM QC, BC, 2.5uM PBH, 5.0uM PBH, 10uM PBH.
3) spend the night 37 ℃ of wet boxes of the chip behind the point sample fixing;
4) use NaBH
4The solution sealing is cleaned subsequently, is dried;
5) add chip hybridization liquid (mainly containing SSC, Denhardt ' s, SDS in the chip hybridization liquid) 15ul that contains the Arab-HEX that fluorescence HEX modifies by cover plate to each dot matrix, in 40 ℃-60 ℃ reaction 2h;
6) reaction back chip respectively in two kinds of washing lotions 42 ℃ respectively wash 2min, washing lotion I:0.3 * SSC/0.1%SDS, washing lotion II:0.06 * SSC.At last, slide centrifuge dripping.
7) chip scanning and data processing.Brilliant core is used in data analysis
TMLuxscan-10K/B (Boao Biological Co., Ltd) scans slide, and sweep parameter is all set Laser/PMT=90/650.
The scan image of two kinds of chips as shown in Figure 3, (wherein, A is a glucosan aldehyde radical substrate of the present invention to signal intensity histogram relatively as shown in Figure 4; B is common aldehyde radical substrate).
The result shows, the nucleic acid samples of 3 kinds of variable concentrations, fixed amount on the chip that glucosan aldehyde radical substrate of the present invention is made significantly increases than the fixed amount of common aldehyde radical substrate, corresponding signal intensity has 2-5 times of raising in various degree respectively, and the nucleic acid samples crystallized ability that shows this glucosan aldehyde radical substrate is than common aldehyde radical substrate height.
Embodiment 4, the application of glucosan aldehyde radical substrate in the high density nucleic acid chip
The glucosan aldehyde radical substrate that present embodiment is used is by embodiment 1 described method preparation.
Used common aldehyde radical substrate identical with described in the embodiment 3.
The preparation process of biochip is as follows:
1) with the Oligo sample of 33.5K kind 46Mer～55Mer (available from Operon company, people's promoter h sapiens V4.0.2 datasheet, Cat#852102) be mixed with the solution that contains 50%DMSO respectively with 100%DMSO, every kind of sample takes out 10ul and shifts in adding 384 orifice plates;
2) by automatic spot sample device with ready sampling liquid according to parallel glucosan aldehyde radical substrate of the present invention and the common aldehyde radical substrate surface selected of identical dot matrix way.Every chip comprises 48 dot matrix, and each dot matrix comprises 697 points, and dot spacing is 160um.Parallel some system effect as shown in Figure 5.
3) spend the night 37 ℃ of wet boxes of the chip behind the point sample fixing;
4) use NaBH
4The solution sealing is cleaned subsequently, is dried;
5) adding the chip hybridization liquid that contains fluorescently-labeled target by cover plate to each dot matrix spends the night in 40 ℃ of-60 ℃ of reactions;
6) reaction back chip cleans (washing lotion I:0.3 * SSC/0.1%SDS, washing lotion II:0.06 * SSC with two kinds of washing lotions respectively.), centrifugal, drying;
7) chip scanning and data processing.The parallel detection result as shown in Figure 6.
In this experiment, when making chip respectively with glucosan aldehyde radical substrate of the present invention and common aldehyde radical substrate, used probe and hybridization solution are all identical, mainly contain formamide, SSC, Denhardt ' s, SDS in the chip hybridization liquid, and corresponding reaction conditions etc. are also all identical.
Use brilliant core
TMChip behind Luxscan-10K/B (Boao Biological Co., Ltd) the scanning point sample, sweep parameter is all set Laser/PMT=90/900.Scanning result shows, in a system high density oligo chip processes, for common aldehyde radical substrate, when just beginning point sample, the spot diameter of common aldehyde radical substrate is at 120-125um, when each lattice point system during to about 1/3 place spot diameter begin to diminish, the spot diameter that diminishes is (Fig. 5 A, wherein zone C and region D are enlarged drawing) between 50-80um; For glucosan aldehyde radical substrate of the present invention, in whole long-time point sample process (the point sample ambient humidity is big), spot diameter does not almost change, always at 135-140um (Fig. 5 B, wherein zone C and region D are enlarged drawing).
Use brilliant core
TMChip after Luxscan-10K/B (Boao Biological Co., Ltd) the scanning hybridization, sweep parameter is all set Laser/PMT=90/900.Scanning result shows, the chip that common aldehyde radical substrate is made, except indivedual spot diameter sizes are about 120-125um, little and fuzzy (especially region D) (Fig. 6 A of boundary of the diameter of most of point, wherein zone C and region D are enlarged drawing), cause extracting data difficulty, inaccurate even can't extract data; By contrast, the diameter that glucosan aldehyde radical substrate of the present invention is had a few is almost equal, is 135-140um, and boundary is very obvious, is easy to extract data and data analysis (Fig. 6 B, wherein zone C and region D are enlarged drawing).
Claims (10)
1, a kind of bio-chip substrate is the substrate of aldehyde radical glucosan in the grafting of amination sheet primary surface; Described aldehyde radical glucosan is to be that aldehyde radical obtains with the adjacent hydroxyl oxidize on the glycoside units of glucosan.
2, substrate according to claim 1 is characterized in that: described aldehyde radical glucosan obtains with periodic acid or water-soluble periodate oxidation glucosan.
3, substrate according to claim 1 and 2 is characterized in that: described glucosan is Dextran T 40 or Dextran T 70.
4, substrate according to claim 1 and 2, it is characterized in that: described amination sheet base is prepared as follows: slide is carried out vapour deposition with hexamethyldisilazane, and then handle with containing amino polysiloxane, obtain the amination slide.
5, a kind of method for preparing the described bio-chip substrate of claim 1 is to obtain bio-chip substrate with being grafted to after the glucosan aldehyde radicalization on the amination sheet primary surface.
6, a kind of method for preparing the described bio-chip substrate of claim 1 is to obtain bio-chip substrate with periodic acid or water-soluble periodate oxidation again after being grafted on the amination sheet primary surface after the glucosan aldehyde radicalization.
7, according to claim 5 or 6 described methods, it is characterized in that: described glucosan aldehyde radicalization is with periodic acid or water-soluble periodate oxidation glucosan.
8, according to claim 5 or 6 described methods, it is characterized in that: described glucosan is Dextran T 40 or Dextran T 70.
9, according to claim 5 or 6 described methods, it is characterized in that: described amination sheet base is prepared as follows: slide is carried out vapour deposition with hexamethyldisilazane, and then handle with containing amino polysiloxane, obtain the amination slide.
10, the biochip that makes by arbitrary described substrate in the claim 1 to 4.
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| CNA2008100577846A CN101241124A (en) | 2008-02-18 | 2008-02-18 | Biological chip substrate and method for making same |
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Cited By (5)
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| WO2009135388A1 (en) * | 2008-05-09 | 2009-11-12 | Zhiping Liu | A molecule detecting system |
| CN101857900A (en) * | 2010-05-06 | 2010-10-13 | 东华大学 | Method for preparing double-epoxy modified biochip substrate |
| CN101633742B (en) * | 2009-09-01 | 2012-05-23 | 博奥生物有限公司 | Amino plastic substrate, and preparation method and application thereof |
| CN102520151A (en) * | 2011-12-09 | 2012-06-27 | 东南大学 | Method for producing quasi three dimensional biological chip |
| CN102276863B (en) * | 2009-09-01 | 2013-03-20 | 博奥生物有限公司 | Amino plastic substrate and preparation method and application thereof |
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| CN1381590A (en) * | 2001-04-13 | 2002-11-27 | 缪金明 | Simple and fast technique for detecting single nucleotide polymorphism (SNP) by high-temp hybridized chip |
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| CN1381590A (en) * | 2001-04-13 | 2002-11-27 | 缪金明 | Simple and fast technique for detecting single nucleotide polymorphism (SNP) by high-temp hybridized chip |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009135388A1 (en) * | 2008-05-09 | 2009-11-12 | Zhiping Liu | A molecule detecting system |
| CN101633742B (en) * | 2009-09-01 | 2012-05-23 | 博奥生物有限公司 | Amino plastic substrate, and preparation method and application thereof |
| CN102276863B (en) * | 2009-09-01 | 2013-03-20 | 博奥生物有限公司 | Amino plastic substrate and preparation method and application thereof |
| CN101857900A (en) * | 2010-05-06 | 2010-10-13 | 东华大学 | Method for preparing double-epoxy modified biochip substrate |
| CN101857900B (en) * | 2010-05-06 | 2013-01-02 | 东华大学 | Method for preparing double-epoxy modified biochip substrate |
| CN102520151A (en) * | 2011-12-09 | 2012-06-27 | 东南大学 | Method for producing quasi three dimensional biological chip |
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