CN101864360B - Method for preparing microfluidic chip probe array for use in biochip analysis - Google Patents
Method for preparing microfluidic chip probe array for use in biochip analysis Download PDFInfo
- Publication number
- CN101864360B CN101864360B CN 201010193890 CN201010193890A CN101864360B CN 101864360 B CN101864360 B CN 101864360B CN 201010193890 CN201010193890 CN 201010193890 CN 201010193890 A CN201010193890 A CN 201010193890A CN 101864360 B CN101864360 B CN 101864360B
- Authority
- CN
- China
- Prior art keywords
- microchannel
- probe
- mineral oil
- microfluidic chip
- probe array
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Automatic Analysis And Handling Materials Therefor (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for preparing a microfluidic chip probe array for use in biochip analysis, which relates to biochips. The method comprises: introducing solution to a micro channel; arranging storage tanks or small test tubes, which contain mineral oil or other solvents insoluble in water and probe solution, at intervals for supplying liquid, and realizing a liquid drop sequence of 'mineral oil, probe solution, mineral oil, probe solution and mineral oil' in the micro channel by replacing the supplied liquid at the entrance of the micro channel, wherein the mineral oil serves as a carrier current and the probe solution serves as a drops flowing in the carrier current; and after the probe liquid drop sequence flows to a preset position, making the probe liquid drop sequence stop flowing to allow the probes in the liquid drops to emit light automatically or to be immobilized on the inside surface of the micro channel by the reaction or absorption initiated by light, electricity and magnet in the outside, discharging the mineral oil and the probe liquid drops after the immobilization reaction is finished and washing with buffer liquid to finish the whole step.
Description
Technical field
The present invention relates to biochip, particularly relate to a kind of preparation method of the microfluidic chip probe array for analyzing biochips.
Background technology
Biochip is that a large amount of biomolecules or material (as nucleic acid fragment, protein, medicine or acceptor, cell or tissue etc.) are fixed on carrier surface according to the arrangement mode that designs in advance, with this as probe, carry out specific absorption or reaction with determinand in sample, realize the detection to sample message.Thousands of reaction is carried out on chip piece simultaneously, has the ability that large-scale parallel is analyzed.biochip is generally processed at sheet glass, silicon chip, on the solid support materials such as nylon membrane, the making method of probe array mainly contains point sample method (Schena M, Shalon D, Davis R W.et al.Science., 1995, 20:467-470) and in-situ synthesis (Fodor S P A, Read J L, Pirrung M C, et al.Science, 1991, 251:767-773), thereby the point sample method is with point sample instrument, probe points to be fixed at carrier surface again to react the probe mortise at carrier surface, the synthetic oligonucleotide that is mainly used in of original position, utilize polystep reaction, successively the deoxynucleoside acid mono is connected to the probe afterbody, realization is in the extension of carrier surface.These method and technologies requirements are high, tooling cost is high, manufacturing speed is slow, and all need more expensive precision instrument.
Micro-fluidic chip is to utilize various micro-processing technologies to process the microstructure with various functions on chip material (as other materials such as glass, PDMS or PMMA), the functions such as realization response, separation, detection are integrated into the function in laboratory on portable chip to greatest extent.From (Manz A such as early 1990s Manz and Widmer, Graber N, WidemerH M.Sens.Acturators, B, 1990, B1:244) since proposition first, the micro-fluidic chip technology has obtained fast development, expand to all directions from simple chemical analysis, comprise foranalysis of nucleic acids, protein analysis and cell analysis etc.Carry out analyzing biochips on micro-fluidic chip, can effectively strengthen detection signal, improve sensitivity, reduce reagent and sample consumption, shorten analysis time etc. (Chen H, Wang L, Li P C H.Lab Chip, 2008,8:826-829).Make probe array and mainly contain following 3 kinds of forms on micro-fluidic chip:
1) utilize point sample instrument to make highdensity probe array at a slice micro-fluidic chip material surface, and then form complete micro-fluidic chip with another chip block material, whole probe array is by complete being included in wider microchannel or hybridizing (Noerholm M in the chamber, Bruus H, Jakobsen M H, et al.Lab Chip, 2004,4:28-37).
2) use equally low-density probe array in the point sample instrument making, then form micro-fluidic chip, not necessarily all probes all are comprised in and passage, and probe is linear array (Wei C W along passage, Cheng J Y, Huang C T, et al.Nucleic Acids Res., 2005,33:1-11).
3) microchannel of the micro-fluidic chip of utilization energy reversible keying is at the probe array of carrier upper fixed line shape, then, the relative carrier of the passage of micro-fluidic chip is rotated rear conveying sample solution and probe array line generation hybridization (Wang L, Li P C H.J.Agric.Food Chem., 2007,55:10509-10516).Due to the probe non-refractory, therefore in these methods, micro-fluidic chip all can't carry out the high temperature bonding, has brought great difficulty for chip manufacturing, experimental implementation, the system integration and development of new applications; Need on the other hand manual microchannel and the probe array aimed at, a large amount of repetition probes must occur in probe array, be comprised in the microchannel to guarantee required probe, number and the density of probe are extremely restricted, and can't effectively utilize the surface of carrier.
Summary of the invention
The object of the invention is to for the existing shortcoming of aforesaid method, a kind of preparation method of the microfluidic chip probe array for analyzing biochips is provided.
The present invention includes following steps:
1) solution is incorporated in the microchannel;
2) will contain the organic solvent that mineral oil or other do not dissolve each other with water, and the liquid storage tank of probe solution or small test tube are spaced as feed flow, by changing the feed flow of microchannel entrance, realize in the microchannel " organic solvent that organic solvent-probe solution that organic solvent-probe solution that mineral oil or other do not dissolve each other with water-mineral oil or other do not dissolve each other with water-mineral oil or other do not dissolve each other with water-... " sequence of droplets, the organic solvent that its mineral oil in fluid or other do not dissolve each other with water is as current-carrying, and probe solution is as the drop that flows in current-carrying;
3) moving behind the predetermined position in probe drop sequence flows, stop flowing, probe meeting in drop is spontaneous or be fixed to the microchannel internal surface by reaction or absorption under light, electricity, the extraneous initiation of magnetic, after fixation reaction is completed, discharge mineral oil and probe drop, then clean to complete whole step with damping fluid.
In step 1) in, described solution is incorporated in the microchannel, the microchannel entrance can be processed into the prong shape, or connect kapillary as entrance, be inserted in liquid storage tank or small test tube solution is incorporated in the microchannel by entrance; Described microchannel can be glass microchannel, quartzy microchannel, silicon micro-channel or superpolymer microchannel etc., described microchannel can be through reversible or irreversible bonding and forms complete microchannel, described microchannel internal surface can carry out surface treatment as required, and the size of described microchannel can be 1nm～5cm1; Described microchannel can be straight shape passage, spirally coiled passage, rectangular channel, spirality channel etc.; Described microchannel can be single passage, can be also many parallel channels.
In step 3) in, the motivating force that described liquid flows in the microchannel provides by electric field or by the syringe pump that is connected to channel outlet, gravity etc.; Described probe can be not only nucleic acid, can also be micromolecular compound, polypeptide, protein, antigen, polysaccharide, part, medicine, acceptor, cell or tissue etc.
The invention has the advantages that: in the situation that reagent consumption is few, can be simply and effectively at the microchannel of sealing processing probe array, and the density of the speed setting probe that can switch by feed flow, and reduction is to the dependence of expensive instrument, cut down finished cost low, accelerate manufacturing speed.
Description of drawings
Fig. 1 is the schematic diagram of processing probe array on micro-fluidic chip of the present invention.
Fig. 2 is the schematic diagram of many parallel channels being processed simultaneously probe array of the present invention.
Fig. 3 is the schematic diagram of processing probe array on the micro-fluidic chip with helical channel of the present invention.
Fig. 4 is the schematic diagram of processing probe array on the micro-fluidic chip of employing of the present invention capillary inlet.
Fig. 5 is that employing syringe of the present invention is as the probe array machining sketch chart of motivating force.
Fig. 6 is fixing probe array fluorescence signal intensity of the present invention.In Fig. 6, X-coordinate is concentration and probe concentration (μ m), and ordinate zou is fluorescence signal intensity.
Embodiment
The present invention is further illustrated in connection with accompanying drawing for following examples.
Referring to Fig. 1, the microchannel 3 of chip 1 is coil-like, and its entrance 2 is processed into the prong shape, is inserted in liquid storage small test tube A.Liquid storage small test tube A is spaced mineral oil 5 and probe solution 6 is housed respectively.The outlet of microchannel is connected with a horizontal liquid storage tank 4, utilizes the segregation drive liquid-flow, and 3 interiorly realize that mineral oil 7 carries probe drop 8 forma fluens in the microchannel.After flowing to the predetermined position, stop flowing and the fixation reaction of carrying out probe to complete the processing of probe array.
Fig. 6 is the fixing fluorescence signal intensity of 30min gained of nucleic acid probe of different concns, probe contains 20 bases, 3 ' is marked with FITC, and 5 ' is marked with amino, is fixed in passage by the chemical reaction with the free aldehyde generation that is modified at channel inner surface.
Embodiment 2
Referring to Fig. 2, the microchannel 3 of chip 1 is one or more parallel channels, and every passage all is processed with an entrance and exit, processes a plurality of parallel probe arrays and is used for a plurality of parallel analyzing biochips.In Fig. 2, other mark is identical with Fig. 1.
Referring to Fig. 3, microchannel 3 is used for the processing probe array twist.In Fig. 3, other mark is identical with Fig. 1.
Embodiment 4
Referring to Fig. 4, its microchannel entrance is replaced by a bit of quartz capillary, and working method is: aim at the position of microchannel in chip sides, utilize drill bit to get out an aperture, fix as feeder connection with glue after inserting a bit of quartz capillary.In Fig. 4, other mark is identical with Fig. 1.
Referring to Fig. 5, its outlet is connected with a syringe 4, and the negative pressure that syringe produces provides motivating force, drives liquid and flows in the microchannel.In Fig. 5, other mark is identical with Fig. 1.
Claims (11)
1. preparation method who is used for the microfluidic chip probe array of analyzing biochips is characterized in that comprising the following steps:
1) solution is incorporated in the microchannel;
Liquid storage tank or the small test tube that 2) will contain mineral oil and probe solution are spaced as feed flow, by changing the feed flow of microchannel entrance, realize in the microchannel " mineral oil-probe solution-mineral oil-probe solution-mineral oil-... " sequence of droplets, its mineral oil in fluid is as current-carrying, and probe solution is as the drop that flows in current-carrying;
3) moving behind the predetermined position in probe drop sequence flows, stop flowing, probe meeting in drop is spontaneous or be fixed to the microchannel internal surface by reaction or absorption under light, electricity, the extraneous initiation of magnetic, after fixation reaction is completed, discharge mineral oil and probe drop, then clean to complete whole step with damping fluid.
2. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 1, it is characterized in that in step 1) in, described solution is incorporated in the microchannel, that the microchannel entrance is processed into the prong shape, or connect kapillary as entrance, be inserted in liquid storage tank or small test tube solution is incorporated in the microchannel by entrance.
3. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 1, is characterized in that in step 1) in, described microchannel is glass microchannel, quartzy microchannel, silicon micro-channel or superpolymer microchannel.
4. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 1, is characterized in that in step 1) in, described microchannel is to form complete microchannel through reversible or irreversible bonding.
5. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 1, is characterized in that in step 1) in, described microchannel internal surface carries out surface treatment.
6. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 1, is characterized in that in step 1) in, described microchannel is of a size of 1nm～5cm.
7. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 1, is characterized in that in step 1) in, described microchannel is straight shape passage, spirally coiled passage, rectangular channel, spirality channel.
8. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 1, is characterized in that in step 1) in, described microchannel is single passage or many parallel channels.
9. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 1, is characterized in that in step 3) in, the motivating force that described drop flows in the microchannel is provided by gravity or the syringe that is connected to channel outlet.
10. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 1, is characterized in that in step 3) in, described probe is medicine, cell or tissue.
11. the preparation method of a kind of microfluidic chip probe array for analyzing biochips as claimed in claim 10 is characterized in that described medicine is nucleic acid, polypeptide, protein, antigen, polysaccharide, part, acceptor.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010193890 CN101864360B (en) | 2010-06-01 | 2010-06-01 | Method for preparing microfluidic chip probe array for use in biochip analysis |
| PCT/CN2011/000918 WO2011150675A1 (en) | 2010-06-01 | 2011-05-31 | Biochip comprising multiple microchannels |
| US13/553,832 US20120289429A1 (en) | 2010-06-01 | 2012-07-20 | Biochip comprising multiple microchannels |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010193890 CN101864360B (en) | 2010-06-01 | 2010-06-01 | Method for preparing microfluidic chip probe array for use in biochip analysis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101864360A CN101864360A (en) | 2010-10-20 |
| CN101864360B true CN101864360B (en) | 2013-06-19 |
Family
ID=42956287
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010193890 Active CN101864360B (en) | 2010-06-01 | 2010-06-01 | Method for preparing microfluidic chip probe array for use in biochip analysis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101864360B (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI436622B (en) | 2009-12-28 | 2014-05-01 | Mediatek Singapore Pte Ltd | Transmitter and method for transmitter, method in which subcarrier pairs are paired, a computer-readable medium storing instructions |
| WO2011150675A1 (en) * | 2010-06-01 | 2011-12-08 | 厦门大学 | Biochip comprising multiple microchannels |
| WO2013037284A1 (en) * | 2011-09-15 | 2013-03-21 | The Chinese University Of Hong Kong | Microfluidic platform and method for controlling the same |
| CA2984045C (en) * | 2015-04-30 | 2024-01-09 | European Molecular Biology Laboratory | Microfluidic droplet detection and sorting |
| CN104897654B (en) * | 2015-06-02 | 2018-06-01 | 北京纳迅科技股份有限公司 | A kind of microflow controlled biochip detection device and preparation method |
| CN108535239B (en) * | 2018-03-28 | 2021-05-25 | 上海艾瑞德生物科技有限公司 | Micro-fluidic chip and detection system based on micro-droplets |
| CN108587898B (en) * | 2018-05-07 | 2022-04-29 | 东莞东阳光医疗智能器件研发有限公司 | Preparation device and preparation method of digital PCR microdroplet |
| CN111175483B (en) * | 2020-02-19 | 2023-04-11 | 杭州济扶科技有限公司 | High-flux drug screening device and method based on micro-fluidic droplets |
| CN112570052A (en) * | 2020-12-07 | 2021-03-30 | 厦门大学 | Micro-fluidic device containing probe array and micro-fluidic chip with side channels |
| CN112619720A (en) * | 2020-12-07 | 2021-04-09 | 厦门大学 | Micro-fluidic chip containing probe array and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1279639A2 (en) * | 2001-07-24 | 2003-01-29 | Dalsa Semiconductor Inc. | Micro-fluidic devices |
| CN1645137A (en) * | 2005-01-27 | 2005-07-27 | 复旦大学 | Assistant hot-pressed packing method for polymethylmethacrylate microflow controlled chip solvent |
| CN1880473A (en) * | 2005-06-13 | 2006-12-20 | 中国科学院电子学研究所 | Preprocessed chip of porous silica microfluid sample |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4721227B2 (en) * | 2006-05-22 | 2011-07-13 | アイダエンジニアリング株式会社 | Microchannel chip and manufacturing method thereof |
-
2010
- 2010-06-01 CN CN 201010193890 patent/CN101864360B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1279639A2 (en) * | 2001-07-24 | 2003-01-29 | Dalsa Semiconductor Inc. | Micro-fluidic devices |
| CN1645137A (en) * | 2005-01-27 | 2005-07-27 | 复旦大学 | Assistant hot-pressed packing method for polymethylmethacrylate microflow controlled chip solvent |
| CN1880473A (en) * | 2005-06-13 | 2006-12-20 | 中国科学院电子学研究所 | Preprocessed chip of porous silica microfluid sample |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101864360A (en) | 2010-10-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101864360B (en) | Method for preparing microfluidic chip probe array for use in biochip analysis | |
| US11667907B2 (en) | Method and apparatus for encoding cellular spatial position information | |
| US12103004B2 (en) | Method and apparatus for processing tissue and other samples encoding cellular spatial position information | |
| JP2023071738A (en) | System and method for isolating and analyzing cells | |
| JP6316369B2 (en) | Microfluidic device | |
| US7470540B2 (en) | Method and device for the integrated synthesis and analysis of analytes on a support | |
| CA2636855C (en) | Microfluidic devices and methods of use in the formation and control of nanoreactors | |
| US10065403B2 (en) | Microfluidic assay assemblies and methods of manufacture | |
| US11701656B2 (en) | Multi-droplet capture | |
| CN103191792B (en) | Microfluidic chip for microspheric multi-element biological detection | |
| CN112827517B (en) | Use method and device of micro-fluidic chip | |
| CN104031832B (en) | A kind of nucleic acid sequencing micro-fluidic chip | |
| CN104777316B (en) | A kind of preparation method of Western blotting paper chip based on quantum dot | |
| CN101851680B (en) | Method for biochip high-throughput hybridization | |
| WO2011150675A1 (en) | Biochip comprising multiple microchannels | |
| CN101886126B (en) | Preparation method of capillary probe array used for analyzing biochips | |
| US20040053327A1 (en) | Method and device for analysing chemical or biological samples | |
| KR100960670B1 (en) | Lab-on-a-chip using capillaries and manufacturing method thereof | |
| US20070087353A1 (en) | Microarray biochemical reaction device | |
| US20050118730A1 (en) | Microchannel array component, microchannel array for recovering biomolecules, and method for recovering biomolecules | |
| CN106694066A (en) | Low energy consumption micro-fluidic chip for microspherical biological detection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160617 Address after: 610041, No. 23, No. 2, No. 10, 88, five road, hi tech Zone, Chengdu, Sichuan Patentee after: Chengdu Atena Biological Technology Co. Ltd. Address before: Xiamen City, Fujian Province, 361005 South Siming Road No. 422 Patentee before: Xiamen University |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20211104 Address after: 610000 No. 3, floor 11, building 8, No. 399, west section of Fucheng Avenue, Chengdu hi tech Zone, China (Sichuan) pilot Free Trade Zone, Chengdu, Sichuan Patentee after: Chengdu Atena Biological Technology Co.,Ltd. Patentee after: Jiujiang Research Institute of Xiamen University Address before: 610041 No. 10, floor 23, building 2, No. 88, Jitai fifth road, high tech Zone, Chengdu, Sichuan Patentee before: Chengdu Atena Biological Technology Co.,Ltd. |
|
| TR01 | Transfer of patent right |