CN102866147B - A kind of based on fit micro-fluidic chemical luminous chip and preparation method thereof - Google Patents
A kind of based on fit micro-fluidic chemical luminous chip and preparation method thereof Download PDFInfo
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- CN102866147B CN102866147B CN201210339160.XA CN201210339160A CN102866147B CN 102866147 B CN102866147 B CN 102866147B CN 201210339160 A CN201210339160 A CN 201210339160A CN 102866147 B CN102866147 B CN 102866147B
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Abstract
The invention discloses a kind of based on fit micro-fluidic chemical luminous chip, comprise several sample holes (1), the sample channel (2) identical with sample holes (1) quantity, golden film detection cell (3), outage (4), sample holes (1) connects sample channel (2), each sample channel (2) connects golden film detection cell (3), gold film detection cell (3) is formed by sputtering and electron beam evaporation, and outage (4) derives waste liquid.The present invention achieves integrated on micro-fluidic chemical luminous chip of golden film by sputtering and electron beam evaporation.Compared with chip before, be modified with the fit of sulfydryl and by the direct self assembly of Au-S key on the surface of golden film, thus fit fixing operation can be simplified, improve fixed efficiency, shorten the set time.
Description
Technical field
What the present invention relates to is a kind of based on fit micro-fluidic chemical luminous chip and preparation method thereof.
Background technology
Chemiluminescence refers to that converts chemical energy is the phenomenon of luminous energy by reaction intermediate, reaction product or the additional luminescence reagent in chemical reaction process.Compared with fluorescence analysis etc., chemiluminescence does not have external excitation source background signal to disturb, and disturbs less, has the advantages such as sensitivity is high, the range of linearity is wide; The chemiluminescence analysis of setting up thus has been widely used in the fields such as clinical diagnosis, environmental monitoring, food security.Antibody is the identification molecule the most frequently used in clinical diagnostic applications process of chemiluminescence.At present, antibody mainly relies on monoclonal technigue to be prepared, and the cycle is long, cost is high; And it is larger that antibody is subject to the impact of the testing environment such as pH, temperature factor.The chemiluminescence detector device that domestic clinical detection adopts and the main dependence on import of matched reagent, reagent consumption is large, expensive, and testing cost is high.Therefore, setting up highly sensitive, high specificity, reagent, to consume little, the lower-cost detection method meeting clinical quick detection needs very necessary.
Fit (aptamer), in nineteen ninety reported first, has caused the extensive concern in clinical diagnosis field.Fit by index concentration Fas lignand system evolution (systematic evolution of ligands by exponential enrichment, SELEX) technology screening obtains, be one section can with DNA or the RNA nucleotide sequence of the specific bindings such as albumen, microorganism, organic molecule, also referred to as " chemical antibody ", antibody can be replaced in clinical detection application.Compared with conventional antibodies, fit have: (1) by chemosynthesis, cost is low, manufacturing cycle is short; (2) functional modification is easy to; (3) advantages such as environmental resistance is strong, not easy in inactivation, and period of storage is long.As a kind of novel target body identification molecule, detect in application in clinical diagnosis and there are great potentiality.
Manz proposes micro-total analysis system (Miniaturized Total Analysis System, μ TAS) concept in early 1990s.The Routine Test Lab operations such as the pre-service of clinical diagnosis sample, sample introduction, separation and detection concentrate on the chip of square centimeter level and complete by this system, can realize the microminiaturization of clinical diagnosis detection system, integrated and portability; Namely its core technology is the micro-fluidic chip based on microflow control technique (Microfluidics).Compared with the experimental provision of macro-scale, the micron scale construction of micro-fluidic chip can significantly improve the area of fluid environment and volume ratio, increase reaction efficiency, simplify the operation, shorten analysis time, reduce reagent consumption.
Aptamer technologies, microfluidic chip technology and chemiluminescence detection coupling can significantly reduce costs, improve sensitivity, accelerate analysis speed.At present, micro-fluidic chip fixes fit mode Avidin-Biotin combination, condensation reaction, paramagnetic particle method etc.All there is different defect in these methods: Avidin-Biotin method needs to be fixed in the detection cell of chip by physisorption by Avidin, therefore should not control; Condensation reaction needs to modify detection cell makes it to possess carboxyl, therefore complicated operation; Paramagnetic particle method needs Magnetic control, and equipment is complicated.
Summary of the invention
The present invention is directed to the deficiencies in the prior art and provide a kind of based on fit micro-fluidic chemical luminous chip and preparation method thereof
The present invention adopts following technical scheme:
A kind of based on fit micro-fluidic chemical luminous chip, comprise several sample holes (1), the sample channel (2) identical with sample holes (1) quantity, golden film detection cell (3), outage (4), sample holes (1) connects sample channel (2), each sample channel (2) connects golden film detection cell (3), gold film detection cell (3) is formed by sputtering and electron beam evaporation, and outage (4) derives waste liquid.
The preparation method of described fit micro-fluidic chemical luminous chip, comprises the steps:
(1) with ethanol and washed with de-ionized water silicon chip, adopt the method for Soft lithograph to make siliceous formpiston, and process with the silylating reagent fluoridized;
(2) making of substrate: by dimethyl silicone polymer (PDMS) and hardening agent according to 10: 1 mass ratio mix, abundant stirring 30min, and carry out vacuum stripping, said mixture is cast on prefabricated siliceous formpiston, and extrusion forming, put into baking oven to dry at 90 DEG C ~ 95 DEG C, complete solidification, the PDMS of solidification is peeled off from formpiston, make the substrate with micro-flow channels, according to chip structure design, offer through hole with tapping and plugging machine in corresponding position, as sample holes, outage;
(3) making of substrate: by PDMS and corresponding hardening agent according to 10: 1 mass ratio be fully uniformly mixed, and carry out vacuum stripping.Planar substrates is made in potpourri cast after degasification, and is placed in oven dry at 90 DEG C ~ 95 DEG C in baking oven;
(4) making of golden film: PDMS substrate is hidden with slide with holes, the corresponding chip detection pond of exposed region, form one deck gold film by sputtering and electron beam evaporation, the size of its thickness is 20-60nm;
(5) passage modification: insert in plasma chamber by the substrate made and substrate, carries out plasma treatment;
(6) bonding encapsulation: the substrate made and substrate are aimed at, makes golden film and detection cell position coincide, carries out the irreversible involution of chip, namely obtain the micro-fluidic chemical luminous chip that golden film is integrated.
The present invention achieves integrated on micro-fluidic chemical luminous chip of golden film by sputtering and electron beam evaporation.Compared with chip before, be modified with the fit of sulfydryl and by the direct self assembly of Au-S key on the surface of golden film, thus fit fixing operation can be simplified, improve fixed efficiency, shorten the set time.Therefore, fit by self assembly different biological molecules on golden film, can be widely used in albumen, bacterium, the separation of organic molecule, detection and screening.In addition, this chip preparation method is simple, and handling ease, is easy to mass production.
Accompanying drawing explanation
Fig. 1 is two sample holes cake core floor map;
Fig. 2 is three sample holes cake core plain film schematic diagram;
Fig. 3 is the making of substrate;
Fig. 4 is the making of substrate;
Fig. 5 is vertical sectional drawing and the detection of chip;
1, sample holes; 2, sample channel; 3, golden film detection cell; 4, outage; 5, prefabricated siliceous formpiston; 6, equal glue; 7, substrate; 8, the substrate of unmodified gold film; 9, with the substrate that glass sheet hides; 10, the substrate of golden film is modified with; 11, photomultiplier;
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
With reference to figure 1, a kind of based on fit micro-fluidic chemical luminous chip, comprise several sample holes 1, the sample channel 2 identical with sample holes 1 quantity, golden film detection cell 3, outage 4, sample holes 1 connects sample channel 2, each sample channel 2 connects golden film detection cell 3, gold film detection cell 3 is formed by sputtering and electron beam evaporation, and outage 4 derives waste liquid.
With reference to figure 1 (or Fig. 2), Fig. 3 and Fig. 4, the preparation method of fit micro-fluidic chemical luminous chip:
(1) with ethanol and washed with de-ionized water silicon chip, adopt the method for Soft lithograph to make siliceous formpiston, and process with the silylating reagent fluoridized.
(2) making of substrate: by dimethyl silicone polymer (PDMS) and hardening agent according to 10: 1 mass ratio mix, fully stir 30min, and carry out vacuum stripping.Said mixture is cast on prefabricated siliceous formpiston, and extrusion forming, put into baking oven and dry at 90 DEG C ~ 95 DEG C, complete solidification.The PDMS of solidification is peeled off from formpiston, makes the substrate with micro-flow channels.According to chip structure design, offer through hole (number of openings is determined according to actual needs) in corresponding position, as sample holes, outage with tapping and plugging machine.
(3) making of substrate: by PDMS and hardening agent according to 10: 1 mass ratio be fully uniformly mixed, and carry out vacuum stripping.Planar substrates is made in potpourri cast after degasification, and is placed in oven dry at 90 DEG C ~ 95 DEG C in baking oven.
(4) making of golden film: PDMS substrate is hidden with slide with holes, the corresponding chip detection pond of exposed region.Form one deck gold film by sputtering and electron beam evaporation, the size of its thickness is 20-60nm.
(5) passage modification: insert in plasma chamber by the substrate made and substrate, carries out plasma treatment.
(6) bonding encapsulation: the substrate made and substrate are aimed at, makes golden film and detection cell position coincide, carries out the irreversible involution of chip, namely obtain the micro-fluidic chemical luminous chip that golden film is integrated.
The present invention achieves integrated on micro-fluidic chemical luminous chip of golden film by sputtering and electron beam evaporation.Compared with chip before, be modified with the fit of sulfydryl and by the direct self assembly of Au-S key on the surface of golden film, thus fit fixing operation can be simplified, improve fixed efficiency, shorten the set time.Therefore, fit by self assembly different biological molecules on golden film, can be widely used in albumen, bacterium, the separation of organic molecule, detection and screening.In addition, this chip preparation method is simple, and handling ease, is easy to mass production.
Should be understood that, for those of ordinary skills, can be improved according to the above description or convert, and all these improve and convert the protection domain that all should belong to claims of the present invention.
Claims (1)
1. one kind based on fit micro-fluidic chemical luminous chip, it is characterized in that, comprise several sample holes (1), the sample channel (2) identical with sample holes (1) quantity, golden film detection cell (3), outage (4), sample holes (1) connects sample channel (2), each sample channel (2) connects golden film detection cell (3), gold film detection cell (3) is formed by sputtering and electron beam evaporation, outage (4) derives waste liquid
The preparation method of this is fit micro-fluidic chemical luminous chip, comprises the steps:
(1) with ethanol and washed with de-ionized water silicon chip, adopt the method for Soft lithograph to make siliceous formpiston, and process with the silylating reagent fluoridized;
(2) making of substrate: by dimethyl silicone polymer (PDMS) and hardening agent according to 10: 1 mass ratio mix, abundant stirring 30min, and carry out vacuum stripping, said mixture is cast on prefabricated siliceous formpiston, and extrusion forming, put into baking oven to dry at 90 DEG C ~ 95 DEG C, complete solidification, the PDMS of solidification is peeled off from formpiston, make the substrate with micro-flow channels, according to chip structure design, offer through hole with tapping and plugging machine in corresponding position, as sample holes, outage;
(3) making of substrate: by PDMS and corresponding hardening agent according to 10: 1 mass ratio be fully uniformly mixed, and carry out vacuum stripping; Planar substrates is made in potpourri cast after degasification, and is placed in oven dry at 90 DEG C ~ 95 DEG C in baking oven;
(4) making of golden film: PDMS substrate is hidden with slide with holes, the corresponding chip detection pond of exposed region, form one deck gold film by sputtering and electron beam evaporation, the size of its thickness is 20-60nm;
(5) passage modification: insert in plasma chamber by the substrate made and substrate, carries out plasma treatment;
(6) bonding encapsulation: the substrate made and substrate are aimed at, makes golden film and detection cell position coincide, carries out the irreversible involution of chip, namely obtain the micro-fluidic chemical luminous chip that golden film is integrated.
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| CN201210339160.XA CN102866147B (en) | 2012-09-05 | 2012-09-05 | A kind of based on fit micro-fluidic chemical luminous chip and preparation method thereof |
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| CN201210339160.XA CN102866147B (en) | 2012-09-05 | 2012-09-05 | A kind of based on fit micro-fluidic chemical luminous chip and preparation method thereof |
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| CN102866147B true CN102866147B (en) | 2015-08-26 |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103439493A (en) * | 2013-08-08 | 2013-12-11 | 南京大渊生物技术工程有限责任公司 | Aptamer percolated biochip and preparation method thereof |
| CN107469879A (en) * | 2017-07-31 | 2017-12-15 | 深圳大学 | A kind of micro-fluidic chip and preparation method thereof and detection method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1472526A (en) * | 2002-07-31 | 2004-02-04 | 中国科学院生态环境研究中心 | Tunnel capillary electrophoretic chemiluminescence testing microfluid control chip |
| CN101221168A (en) * | 2008-01-08 | 2008-07-16 | 东南大学 | Microfluidic chip based on microsphere biological detection |
| CN101817495A (en) * | 2010-03-25 | 2010-09-01 | 湖南大学 | Micro fluid control chip and preparation method and application thereof |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1472526A (en) * | 2002-07-31 | 2004-02-04 | 中国科学院生态环境研究中心 | Tunnel capillary electrophoretic chemiluminescence testing microfluid control chip |
| CN101221168A (en) * | 2008-01-08 | 2008-07-16 | 东南大学 | Microfluidic chip based on microsphere biological detection |
| CN101817495A (en) * | 2010-03-25 | 2010-09-01 | 湖南大学 | Micro fluid control chip and preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Rare cell chemiluminescence detection based on aptamer-specific capture in microfluidic channels;wu liu等;《Biosensors and Bioelectronics》;20110803;第28卷(第1期);摘要，第439页右栏第1段至第440页左栏第2段，第441页左栏第2段 * |
| 电化学发光DNA和适体生物传感器的研究;李延;《中国博士学位论文全文数据库信息科技辑》;20090615(第6期);第30页最后两段 * |
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