CN101597639A - The making method of high-density DNA microarray biochip - Google Patents
The making method of high-density DNA microarray biochip Download PDFInfo
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- CN101597639A CN101597639A CNA2008101145093A CN200810114509A CN101597639A CN 101597639 A CN101597639 A CN 101597639A CN A2008101145093 A CNA2008101145093 A CN A2008101145093A CN 200810114509 A CN200810114509 A CN 200810114509A CN 101597639 A CN101597639 A CN 101597639A
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- dna microarray
- porous alumina
- alumina formwork
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Abstract
A kind of making method of high-density DNA microarray biochip is characterized in that, comprises the steps: step 1: adopt anode oxidation method to make the high-density porous alumina formwork at the aluminum or aluminum alloy film surface; Step 2: porous alumina formwork is peeled off from the aluminum or aluminum alloy film surface; Step 3: porous alumina formwork is transferred to dna microarray bio-chip substrate surface, as the etch mask of dna microarray bio-chip substrate; Step 4: adopt the method for etching that the dna microarray bio-chip substrate is carried out etching; Step 5: remove porous alumina formwork, finish the making of high-density DNA microarray biochip.The present invention its have density height, cost low, simple to operate, be suitable for the advantage that heavy industrialization is made.
Description
Technical field
The present invention relates to dna microarray biochip preparation field, particularly high-density DNA microarray biochip preparation field.
Background technology
The dna microarray biochip is the model that modern biotechnology combines with engineering, the appearance of dna microarray biochip technology has not only promoted development of life science greatly, the feasible mankind can carry out such as complicated explorations such as gene order-checkings, and having promoted the development of engineering greatly, various new technologies continue to bring out and are used.At present institute extensively the biochip preparation method of employing comprise point sample technology, semiconductor processing technology.Yet owing to be subjected to the restriction of technological limits, present biochip technology of preparing can't prepare diameter less than 100nm, and the uniform dna microarray biochip in aperture has restricted single dna molecular and surveyed further developing of contour true tumor technology.
Summary of the invention
The object of the present invention is to provide a kind of making method of high-density DNA microarray biochip, its have density height, cost low, simple to operate, be suitable for the advantage that heavy industrialization is made.
The making method of a kind of high-density DNA microarray biochip of the present invention is characterized in that, comprises the steps:
Step 1: adopt anode oxidation method to make the high-density porous alumina formwork at the aluminum or aluminum alloy film surface;
Step 2: porous alumina formwork is peeled off from the aluminum or aluminum alloy film surface;
Step 3: porous alumina formwork is transferred to dna microarray bio-chip substrate surface, as the etch mask of dna microarray bio-chip substrate;
Step 4: adopt the method for etching that the dna microarray bio-chip substrate is carried out etching;
Step 5: remove porous alumina formwork, finish the making of high-density DNA microarray biochip.
Wherein porous alumina formwork being peeled off from the aluminum or aluminum alloy film surface is to adopt the mechanical-physical stripping means.
Wherein with porous alumina formwork when the aluminum or aluminum alloy film surface is peeled off, control the thickness of porous alumina formwork by the time of control electrolytic reaction.
Wherein porous alumina formwork being transferred to dna microarray bio-chip substrate surface is to adopt the physical bond method.
Wherein the method for the etching that is adopted is the chemical wet etching method.
Wherein the method for the etching that is adopted is the plasma dry lithographic method.
Wherein removing porous alumina formwork, is to utilize strong base solution to remove method.
The wherein said porous alumina formwork that removes is to utilize strong acid solution to remove method.
The wherein said porous alumina formwork that removes is to utilize machinery to remove or the physical removal method.
Description of drawings
For further specifying content of the present invention and characteristics, below in conjunction with drawings and Examples the present invention is done a detailed description, wherein:
Fig. 1 utilizes porous alumina formwork to make the schema of high-density DNA microarray biochip.
Embodiment
See also shown in Figure 1ly, the making method of a kind of high-density DNA microarray biochip of the present invention comprises the steps:
Step 1: adopt anode oxidation method to make high-density porous alumina formwork 10 at the aluminum or aluminum alloy film surface;
Step 2: porous alumina formwork 10 is peeled off from the aluminum or aluminum alloy film surface, and described peeling off is to adopt the mechanical-physical stripping means, when peeling off, controls the thickness of porous alumina formwork 10 by the time of control electrolytic reaction;
Step 3: porous alumina formwork 10 is transferred to dna microarray bio-chip substrate 11 surfaces, as the etch mask of dna microarray bio-chip substrate 11, described porous alumina formwork 10 is transferred to dna microarray bio-chip substrate 11 surfaces is to adopt the physical bond method;
Step 4: adopt the method for etching that dna microarray bio-chip substrate 11 is carried out etching, the method for the etching of described employing is chemical wet etching method or plasma dry lithographic method;
Step 5: remove porous alumina formwork 10, the described porous alumina formwork 10 that removes, be to utilize strong base solution to remove method or utilize strong acid solution to remove method or utilize machinery to remove or the physical removal method, finish the making of high-density DNA microarray biochip.
Be example to utilize porous alumina formwork to prepare high-density DNA microarray biochip below, the preparation method of the high-density DNA microarray biochip that elaboration the present invention will illustrate.
In conjunction with consulting Fig. 1,
(1) selects high-purity (99.999%) aluminium foil for use, be cut into the substrate that needs size, soaked 3～4 hours with acetone, be that 5% sodium hydroxide solution soaked 90 seconds down at 60 ℃ with massfraction subsequently, behind deionized water rinsing, again aluminium flake is dipped in 5% the salpeter solution 5 minutes, uses deionized water rinsing again; Sulfuric acid (H at 1.2mol/L
2SO
4) or the oxalic acid (C of 0.3mol/L
2H
2O
4) carry out anodic oxidation in the ionogen, the electrode operating voltage is 15～45V, time 2～4h; Aluminium flake is an anode, and platinized platinum is a negative electrode, forms the poroid zone of oxidation of one deck at the aluminium surface energy, and the diameter in hole depends on energising voltage, and oxidated layer thickness depends on conduction time; Utilize cupric chloride as the agent of stripping aluminium, the cupric chloride generation replacement(metathesis)reaction in unreacted aluminium and the Cupric Chloride Solution is with the porous alumina membrane sepn of unoxidized aluminium and oxidation formation; The porous alumina diaphragm that obtains is transferred to the phosphoric acid (H of 0.3mol/L
3PO
4) carry out reaming in the solution and handle, just obtain required porous alumina formwork 10.Above-mentioned operation is all at room temperature carried out, and uses different technology conditions, and we can obtain mean pore size 10～100nm (also can make the aperture increase to 200～300nm) through the reaming processing, hole density 1011～108cm
-2, the serial porous alumina formwork 10 of different structure parameters such as about 30～60 μ m of template thickness.The micropores that form in the porous alumina formwork 10 are parallel to each other and perpendicular to face, arrange by being close to consistent honeycomb six side's imporositys.
(2) porous alumina formwork 10 is peeled off from the aluminium flake surface, described peeling off is to adopt the mechanical-physical stripping means, when peeling off, controls the thickness of porous alumina formwork 10 by the time of control electrolytic reaction;
(3) present commercial dna microarray chip adopts silica-based or silica based glasses is made, the plastics of employing, organism are also arranged as substrate, substrate can be carried out amino or aldehyde group modified in advance, in the present embodiment, adopt silicon as the dna microarray bio-chip substrate, porous alumina formwork 10 is placed into substrate surface, through thermal annealing, 10 of porous alumina formworks stretch naturally, are bonded on the substrate.
(4) the using plasma dry etching technology carries out etching to the dna microarray bio-chip substrate, at this moment, the porous alumina formwork 10 that invests dna microarray bio-chip substrate surface promptly is a blocking layer, the dna microarray bio-chip substrate that is positioned at micropore part below on the porous alumina formwork 10 is not etched owing to being blocked, and the figure of porous alumina formwork 10 has been transferred on the dna microarray bio-chip substrate fully.
(5) the dna microarray biochip that will have a porous alumina formwork 10 is positioned in the NaOH solution, and porous alumina formwork 10 will react and dissolve, and the dna microarray biochip remains.
Be worth pointing out that the described method of above-mentioned steps (5) only is a kind of method that present embodiment adopted, and does not have ubiquity, physical methods such as employing mechanically peel can make porous alumina formwork 10 repeat repeatedly to use, and are applicable to commercialization scale operation.It all is one embodiment of the invention institute employing methods that step (1), (2), (3), (4) and (5) respectively go on foot institute's employing method, the present invention is not had any formality restriction.
In sum, the making method of high-density DNA microarray biochip has the following advantages at least:
1. the making method technical maturity of high-density DNA microarray biochip of the present invention, cost of manufacture is low, adopts the template just can batch making dna microarray biochip, and favorable repeatability is suitable for the scale commercialization and produces.
2. the biochip density height produced of the making method of high-density DNA microarray biochip of the present invention can improve 5～10 times than current methods.
3. the making method of high-density DNA microarray biochip of the present invention adopts the template transfer technology, and the biochip aperture of producing is even, and chip surface can be modified again.
The above; only be embodiments of the invention; be not that the present invention is done any pro forma restriction; every according to the technology of the present invention essence to any simple modification, equivalent variations and modification that above embodiment did; all still belong within the technical solution of the present invention scope, so protection scope of the present invention is when being as the criterion with claims.
Claims (9)
1. the making method of a high-density DNA microarray biochip is characterized in that, comprises the steps:
Step 1: adopt anode oxidation method to make the high-density porous alumina formwork at the aluminum or aluminum alloy film surface;
Step 2: porous alumina formwork is peeled off from the aluminum or aluminum alloy film surface;
Step 3: porous alumina formwork is transferred to dna microarray bio-chip substrate surface, as the etch mask of dna microarray bio-chip substrate;
Step 4: adopt the method for etching that the dna microarray bio-chip substrate is carried out etching;
Step 5: remove porous alumina formwork, finish the making of high-density DNA microarray biochip.
2. the making method of high-density DNA microarray biochip according to claim 1 is characterized in that, wherein porous alumina formwork being peeled off from the aluminum or aluminum alloy film surface is to adopt the mechanical-physical stripping means.
3. the making method of high-density DNA microarray biochip according to claim 1, it is characterized in that, wherein with porous alumina formwork when the aluminum or aluminum alloy film surface is peeled off, control the thickness of porous alumina formwork by the time of control electrolytic reaction.
4. the making method of high-density DNA microarray biochip according to claim 1 is characterized in that, wherein porous alumina formwork being transferred to dna microarray bio-chip substrate surface is to adopt the physical bond method.
5. the making method of high-density DNA microarray biochip according to claim 1 is characterized in that, wherein the method for the etching that is adopted is the chemical wet etching method.
6. the making method of high-density DNA microarray biochip according to claim 1 is characterized in that, wherein the method for the etching that is adopted is the plasma dry lithographic method.
7. the making method of high-density DNA microarray biochip according to claim 1 is characterized in that, wherein removes porous alumina formwork, is to utilize strong base solution to remove method.
8. the making method of high-density DNA microarray biochip according to claim 1 is characterized in that, the wherein said porous alumina formwork that removes is to utilize strong acid solution to remove method.
9. the making method of high-density DNA microarray biochip according to claim 1 is characterized in that, the wherein said porous alumina formwork that removes is to utilize machinery to remove or the physical removal method.
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| Application Number | Priority Date | Filing Date | Title |
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| CNA2008101145093A CN101597639A (en) | 2008-06-06 | 2008-06-06 | The making method of high-density DNA microarray biochip |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106536710A (en) * | 2014-08-08 | 2017-03-22 | 应用材料公司 | Patterned deposition of liquid films for biomedical devices |
| CN109738469A (en) * | 2018-12-29 | 2019-05-10 | 赛纳生物科技(北京)有限公司 | A kind of compactness detection method of FOP surface micro-pit plated film |
| WO2024016656A1 (en) * | 2022-07-22 | 2024-01-25 | 深圳华越再生医学生物科技有限公司 | Alumina film, and preparation method therefor and use thereof |
-
2008
- 2008-06-06 CN CNA2008101145093A patent/CN101597639A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106536710A (en) * | 2014-08-08 | 2017-03-22 | 应用材料公司 | Patterned deposition of liquid films for biomedical devices |
| CN106536710B (en) * | 2014-08-08 | 2018-07-06 | 应用材料公司 | For the patterned deposition of the liquid film of biology device |
| CN109738469A (en) * | 2018-12-29 | 2019-05-10 | 赛纳生物科技(北京)有限公司 | A kind of compactness detection method of FOP surface micro-pit plated film |
| WO2024016656A1 (en) * | 2022-07-22 | 2024-01-25 | 深圳华越再生医学生物科技有限公司 | Alumina film, and preparation method therefor and use thereof |
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Open date: 20091209 |