CN1595144A - Micro channel array type biological chip by utilizing photon particle coding and method of use thereof - Google Patents
Micro channel array type biological chip by utilizing photon particle coding and method of use thereof Download PDFInfo
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- CN1595144A CN1595144A CN 200410041365 CN200410041365A CN1595144A CN 1595144 A CN1595144 A CN 1595144A CN 200410041365 CN200410041365 CN 200410041365 CN 200410041365 A CN200410041365 A CN 200410041365A CN 1595144 A CN1595144 A CN 1595144A
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Abstract
It is micro-path array biological chip by use of photon particle code, which is a high-flux detection chip for multiple proteins and genes in the biology sample. The photon crystal micro ball 3 of the chip is located with probe molecule and distributes on the designed order in the micro path 1 with sample-in mouth 2 and sample-out 4, wherein, the sample-out mouth has a smaller radium than that of sample-in mouth and blocks the photon particle in the micro path 1. When in use, it separately connects multiple-position sample valve 5 with sample-in mouth 2 and sample-out mouth 4 through micro peristaltic pump 6 and T shaped pipe 7 to form a seal cross-breed reaction path.
Description
Technical field
The present invention be a kind of be used for carrying out multiple proteins and genetic test in the biological sample fast, convenient, cheap high throughput testing chip.The detection, legal medical expert that can be widely used in biological sample in clinical detection, drug screening, the environment identified, customs's agricultural product are imported and exported inspection and quarantine, gene order and functional analysis and military detection of going up various fatal or pathogenic microorganismss in the bacteriological warfare.
Background technology
Detect range protein and dna sequence dna relevant in the biological sample fast, high-throughout in scientific research and various fields such as clinical detection, drug screening and environment measuring, can provide for prevention, diagnosis, treatment and the life science of disease and instruct and reference with human various diseases, health and important vital movement.Biochip is a kind of high-throughout Measurement for Biotechnique that grew up in recent years, and the difference according to the biological sample of its detection can be divided into genetic chip, protein-chip and cell chip etc.The prior biological chip mainly is based on probe molecule is fixed on the solid phase carriers such as glass substrate, utilizes the positional information of probe molecule to encode, hybridizes with fluorescently-labeled target molecule then.After the hybridization, determine the kind of probe according to the positional information at probe place, the existence of determining molecules of interest according to the signal of target molecule fluorescence whether.This method is to utilize positional information that probe molecule is encoded, and by the xy coordinate kind of biomolecule is discerned.Therefore, probe molecule must have good positioning system accurately each probe molecule to be fixed on the position that designs at substrate surface fixedly the time.The micro-machined method of general employing.This method has good designability and can be used for preparing high-density biochip.But the cost of this method is than higher.In order to reduce the cost of biochip, the research of the simple and convenient preparation method of relevant biochip has been subjected to very big attention.Inkjet printing, serial of methods such as fiber array are developed.Except the manufacturing cost of chip, the sensitivity and the detection speed that how to improve detection in the use of biochip also are key issues.Having only these problems to obtain good solution biochip just might be used widely.
The photonic crystal particulate is a kind of particulate with special optical characteristic and nanostructured.It is made of two or more materials with different optical constant.These materials can be to form concentrically ringed arrangement architecture or layered arrangement in the photonic crystal particulate.Its cycle arrives between thousands of nanometers at tens nanometer.Because cycle that material is arranged in the photonic crystal particulate and optical wavelength are in same yardstick, photonic crystal particulate emblem reveals some distinctive optical characteristics.Such as, the photonic crystal particulate has the light rejection zone.Can be reflected with the consistent light of frequency of light rejection zone when light incides on the photonic crystal particulate, the light of other frequencies then can pass through photonic crystal.When utilizing reflectance spectrum that the photonic crystal particulate is detected, can observe a very strong reflection peak consistent with the light rejection zone.Because the position of the light rejection zone of photonic crystal particulate can be controlled by the optical constant and the periodic structure of composition material, so the control of the reflection peak position of photonic crystal particulate can realize by the structural design to photonic crystal.
Summary of the invention
Technical matters: it is low to the purpose of this invention is to provide a kind of cost, highly sensitive, the micro channel array formula biochip that utilizes photon particulate coding that hybridization speed is fast, easy to use.
Technical scheme: photonic crystal micro channel array formula biochip of the present invention can be prepared into one dimension or two-dimensional array structure.Injection port is positioned at an end of passage, and outlet is positioned at the other end of passage.In channel outlet by the blocking mechanism that constitutes with the microflute of sand core or micro etch or filter paper etc.In the microchannel, begin to be arranged with in order photonic crystal particulate with different colours from blocking mechanism.Be fixed with different probes on the photon crystal micro-ball of each color.When the hybridization buffer that contains biological sample or elution buffer flowed in passage, protein, the nucleic acid molecules that only can hold in the biological sample passed through, and photon crystal micro-ball then is blocked within the microchannel.
Chip of the present invention structurally is made up of microchannel, injection port, photon crystal micro-ball, outlet; Be fixed with probe molecule on the photon crystal micro-ball, the photonic crystal particulate by the series arrangement that sets in the microchannel, two of microchannel is respectively injection port and outlet, wherein the outlet inner end diameter is less than the injection port internal diameter, its size design can make biomolecule pass through, and the photonic crystal carrier is blocked in the microchannel, avoid probe fixed thereon thereupon outside the flow pass.The photon crystal micro-ball that is fixed with probe molecule is the photonic crystal particulate with different colours, fixes a kind of probe on the photonic crystal particulate of each color.
During use the multidigit sampling valve is communicated with injection port and outlet respectively by micro-creep pump, T type pipe, constitute the hybridization reaction passage of a sealing, tested biomolecule moves in circles mobile in passage under the driving of micro-creep pump in the microchannel, till hybridization reaction fully.In hybridization reaction multidigit sampling valve, micro-creep pump being in series with outlet with injection port is connected, and in wash-out T type pipe is connected with outlet, and peristaltic pump is connected with outlet indirectly, makes damping fluid flow into waste liquid pool by outlet.
Photonic crystal coding micro channel array formula biochip of the present invention is that biological sample circulates in the microchannel, hybridizes with the bioprobe molecule that is fixed on the photonic crystal under the effect of the driving of micro-creep pump.Inject the biological sample that need detect at the microchannel entrance point earlier before the hybridization, when injecting biological sample, the endpiece sealing is in order to avoid the test sample flow pass.Hybridization is carried out in regulating the warm bath of temperature, and temperature is arranged on the suitableeest hybridization reaction temperature; Remove the micro-creep pump after the hybridization, the endpiece of passage adds a vacuum draw pump, injects elution buffer at entrance point, at endpiece elution buffer is extracted out then.Wash-out is afterwards with microchannel reading of data under the laser confocal scanning instrument.Whether be combined in existence that having or not of fluorescent marker on the probe molecule declare target molecule according to hybridization then,
The present invention is with the carrier of photonic crystal particulate as probe molecule, by the arrangement of photonic crystal particulate in the microchannel of kapillary or etching prepared biochip.This chip utilizes the rejection zone of photon crystal micro-ball and positional information that probe molecule is carried out hybrid coding, and target molecule enters the microchannel and flows under the effect that adds driving force, so that accelerate hybridization reaction speed.The chip that utilizes this technology to prepare has characteristics such as cost is low, highly sensitive, and hybridization speed is fast, easy to use.
We are by preparing biochip having the arrangement of rejection zone photonic crystal particulate in the microchannel of not sharing the same light in the present invention.This method has been compared preparation with existing method simple, and selection is wide, and it is big to be utilized to the information encoded amount, tests advantages such as simple.
We utilize photonic crystal particulate and micro-fluid chip technology to reduce the manufacturing cost of biochip at above these problems of array biochip in the present invention, improve sensitivity and detection speed.
Beneficial effect: the present invention has realized having fixed photonic crystal arrangement in order in the microchannel of probe molecule, utilize photon crystal micro-ball itself color and and positional information probe is encoded, with the kind of sign probe, utilize existence that the recognition reaction of probe molecule comes the recognition objective molecule whether simultaneously.Characteristics of the present invention are: the hybrid coding of (1) photonic crystal color and positional information, can utilize the brilliant microsphere of photon of different colours to carry out permutation and combination encodes to probe molecule, coding information quantity is big, and it is convenient to read sign indicating number, even with the naked eye just can read sign indicating number.(2) hybridization speed is fast, because hybridization reaction is limited in the microchannel, compares with the dull and stereotyped biochip of two dimension, can adopt modes such as concussion, fluid drives to accelerate hybridization reaction speed when hybridization, shortens detection time.(3) susceptibility improves: because the fixing surface of oligonucleotides increases, this device can improve the sensitivity of detection, thereby can the interior low-abundance biomolecule of test sample.(4) since hybridization reaction be enclosed in the microchannel and carry out, can with device logotype such as The pretreatment, improve the robotization that detects.
Description of drawings
Fig. 1 is the structural representation of photonic crystal of the present invention.Wherein Fig. 1 a is the photon crystal structure synoptic diagram of spherical shape; Fig. 1 b is the photon crystal structure synoptic diagram of microplate shape; Fig. 1 c is complex layered photon crystal structure synoptic diagram.
Fig. 2 is that the present invention utilizes in the micro channel array formula biochip of photon microballoon coding the synoptic diagram of 1-D photon crystal microchannel chip detecting system.
Fig. 3 is the schematic flow sheet of chip manufacturing of the present invention and use.
Among the above figure microchannel 1, injection port 2, photon crystal micro-ball 3, outlet 4, multidigit sampling valve 5, micro-creep pump 6, T type pipe 7, sample introduction bottle 8, damping fluid bottle 9, waste liquid bottle 10 are arranged.
Embodiment
The micro channel array formula biochip of photon crystal micro-ball coding of the present invention is made up of microchannel 1, injection port 2, photon crystal micro-ball 3, outlet 4, be fixed with probe molecule on the photon crystal micro-ball 3, photonic crystal particulate 3 by the series arrangement that sets in microchannel 1, two of microchannel 1 is respectively injection port 2 and outlet 4, wherein outlet 4 inner end diameter are less than the injection port internal diameter, its size design can make biomolecule pass through, and photonic crystal carrier 3 is blocked in the microchannel 1, avoid probe fixed thereon thereupon outside the flow pass.The photon crystal micro-ball 3 that is fixed with probe molecule is the photonic crystal particulates with different colours, fixes a kind of probe on the photonic crystal particulate of each color.
Concrete preparation flow is as follows:
The preparation of photonic crystal particulate: dual mode is adopted in the preparation of photonic crystal particulate among the present invention.Utilize synthetic good SiO
2Or macromolecule dispersed nano particulate forms orderly micron ball or micron membranes by self assembly.Perhaps the photonic crystal particulate clad surface of utilization with multilayer order membrane structure itself has the high molecular polymer of functional group, as polystyrene etc.
The probe mark of photonic crystal: utilize silicone agent that photonic crystal is carried out finishing, will design and synthesize good probe molecule with bifunctional reagent again and be fixed on the photonic crystal particulate.
The preparation method of microchannel: adopt die or laser lithography method; on PDMS, PMMA or glass sheet, make stair-stepping microchannel as cover plate, adopt methods such as bonding, chemical bonding, thermal bonding, plasma bonding or telegraph key close to make cover plate same substrate bonding then.
The arrangement of photonic crystal particulate in the microchannel: the photonic crystal particulate is arranged with dual mode among the present invention in the microchannel.To be connected with the sampling valve of micro-creep pipe and the microchannel of hybridization reaction before the sample introduction earlier splits.The negative pressure of utilizing syringe pump to form at the outlet end sucks photon crystal micro-ball in the microchannel, the photon crystal micro-ball of the color of the same race of each suction fixed qty, after the suction, the photon crystal micro-ball of different colours aligns in the microchannel by the order in when suction several times.Another mode is that funnel is connected with the passage injection port, inject in the microchannel in order the photonic crystal carrier that is fixed with the different colours of different probe molecule respectively, stopped at endpiece photonic crystal carrier, the photonic crystal particulate of different colours is arranged in the microchannel in order.So far, finished the preparation of the micro channel array formula biochip that utilizes photon particulate coding.
During use multidigit sampling valve 5 is communicated with injection port 2 and outlet 4 respectively by micro-creep pump 6, T type pipe 7, constitute the hybridization reaction passage of a sealing, tested biomolecule in passage in microchannel 1, move in circles under the driving of micro-creep pump 6 mobile, till hybridization reaction is fully.In hybridization reaction, multidigit sampling valve 5, micro-creep pump 6 be in series with outlet 4 with injection port 2 and be connected, in wash-out, T type pipe 7 is connected with outlet 4, peristaltic pump is connected with outlet 4 indirectly, makes damping fluid pass through outlet 4 and flow into waste liquid pool 10.
The pretreatment and fluorescence labeling: detected biological sample needed to extract earlier the target molecule that band detects before hybridization reaction.The nucleic acid samples that extracts also needs can carry out fluorescence labeling in amplification through the multiplex PCR amplification; When the target molecule that band detects is protein,, also need enrichment if content is low.
The hybridization reaction of prehybridization and test sample and wash-out: the injection port of prehybridization damping fluid in the microchannel that configures injected in the microchannel, then with injection port and outlet micro-creep pump be connected on the sampling valve, utilizing the micro-creep pump to drive the prehybridization damping fluid back and forth flows in the microchannel, after treating hybridization fully, the micro-creep pump separates with the microchannel, and damping fluid is reserved from the microchannel.Hybridization reaction operation and prehybridization reacting phase are together, hybridization solution is transferred in the sample introduction bottle, open the switch of micro-creep pump, it is pumped in the microchannel, regulate after the flow velocity, T type pipe is communicated with the micro-creep pump by the multidigit sampling valve, it is mobile that hybridization solution is moved in circles in passage, after hybridization fully, T type pipe is communicated with waste liquid pool, make hybridization solution flow into waste liquid pool, from the buffering liquid pool, elution buffer is pumped in the microchannel then by injection port, again T type pipe is communicated with the micro-creep pump again by the multidigit sampling valve, by effect elution buffer back and forth mobile wash-out in passage of micro-creep pump, after wash-out is finished, elution buffer is poured the liquid pool that abandons one's profession, different is that hybridization reaction carries out in can regulating the temperature water-bath, in the time of hybridization reaction, temperature is set in best hybridization reaction temperature.
Claims (4)
1, a kind of microchannel configuration biochip that utilizes photonic crystal particulate coding is characterized in that this chip is made up of microchannel (1), injection port (2), photonic crystal particulate (3), outlet (4); Photon crystal micro-ball is fixed with probe molecule on (3), photonic crystal particulate (3) by the series arrangement that sets in microchannel (1), two of microchannel (1) is respectively injection port (2) and outlet (4), wherein outlet (4) inner end diameter is less than the injection port internal diameter, its size design can make biomolecule pass through, and photonic crystal carrier (3) is blocked in the microchannel (1), avoid probe fixed thereon thereupon outside the flow pass.
2, the microchannel configuration biochip that utilizes the photon crystal micro-ball coding according to claim 1, the photon crystal micro-ball (3) that it is characterized in that being fixed with probe molecule is the photonic crystal particulate with different colours, fixes a kind of probe on the photonic crystal particulate of each color.
3, a kind of using method of utilizing the microchannel configuration biochip of photonic crystal particulate coding as claimed in claim 1, when it is characterized in that using multidigit sampling valve (5) is communicated with injection port (2) and outlet (4) respectively by micro-creep pump (6), T type pipe (7), constitute the hybridization reaction passage of a sealing, tested biomolecule in passage in microchannel (1), move in circles under the driving of micro-creep pump (6) mobile, till hybridization reaction is fully.
4, the using method of utilizing the microchannel configuration biochip of photonic crystal particulate coding according to claim 4, it is characterized in that in hybridization reaction multidigit sampling valve (5), micro-creep pump (6) be in series with outlet (4) with injection port (2) and be connected, in wash-out, T type pipe (7) is connected with outlet (4), peristaltic pump is connected with outlet (4) indirectly, makes damping fluid pass through outlet (4) and flow into waste liquid pool (10).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CNB2004100413655A CN1295347C (en) | 2004-07-13 | 2004-07-13 | Micro channel array type biological chip by utilizing photon particle coding and method of use thereof |
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| CNB2004100413655A CN1295347C (en) | 2004-07-13 | 2004-07-13 | Micro channel array type biological chip by utilizing photon particle coding and method of use thereof |
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| CN1595144A true CN1595144A (en) | 2005-03-16 |
| CN1295347C CN1295347C (en) | 2007-01-17 |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101221168B (en) * | 2008-01-08 | 2011-08-10 | 东南大学 | Microfluidic chip based on microsphere biological detection |
| CN101762571B (en) * | 2009-12-30 | 2013-07-10 | 东南大学 | Flow-type gas sensor array and preparation method thereof |
| CN103267749A (en) * | 2013-04-27 | 2013-08-28 | 东南大学 | Photon encoding microsphere-based multielement biomarker assay kit-type chip |
| CN105158310A (en) * | 2015-09-21 | 2015-12-16 | 东南大学 | Microfluidic detection chip based on micropore electrode and application thereof |
| CN110304599A (en) * | 2019-07-04 | 2019-10-08 | 广西农喜作物科学有限公司 | A kind of big flow high-precision filling apparatus |
| CN112378889A (en) * | 2020-10-12 | 2021-02-19 | 南京师范大学 | Photonic crystal microsphere capillary column for detecting ochratoxin A and preparation method and application thereof |
| CN113484283A (en) * | 2021-05-19 | 2021-10-08 | 南京医科大学第二附属医院 | Detection system and method for simultaneously measuring multiple biological information |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5736332A (en) * | 1995-11-30 | 1998-04-07 | Mandecki; Wlodek | Method of determining the sequence of nucleic acids employing solid-phase particles carrying transponders |
| CZ301174B6 (en) * | 1999-04-16 | 2009-11-25 | Biocartis Sa | Encoded micro-carrier and method for preparing and reading thereof, method of detecting analyte and chemical library |
| WO2002033419A1 (en) * | 2000-10-19 | 2002-04-25 | Tibotec Bvba | Method and device for the manipulation of microcarriers for an identification purpose |
-
2004
- 2004-07-13 CN CNB2004100413655A patent/CN1295347C/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101221168B (en) * | 2008-01-08 | 2011-08-10 | 东南大学 | Microfluidic chip based on microsphere biological detection |
| CN101762571B (en) * | 2009-12-30 | 2013-07-10 | 东南大学 | Flow-type gas sensor array and preparation method thereof |
| CN103267749A (en) * | 2013-04-27 | 2013-08-28 | 东南大学 | Photon encoding microsphere-based multielement biomarker assay kit-type chip |
| CN103267749B (en) * | 2013-04-27 | 2015-07-29 | 东南大学 | A kind of multielement biomarker assay kit-type chip based on photon coding microball |
| CN105158310A (en) * | 2015-09-21 | 2015-12-16 | 东南大学 | Microfluidic detection chip based on micropore electrode and application thereof |
| CN110304599A (en) * | 2019-07-04 | 2019-10-08 | 广西农喜作物科学有限公司 | A kind of big flow high-precision filling apparatus |
| CN110304599B (en) * | 2019-07-04 | 2021-02-26 | 广西农喜作物科学有限公司 | Large-traffic high accuracy filling device |
| CN112378889A (en) * | 2020-10-12 | 2021-02-19 | 南京师范大学 | Photonic crystal microsphere capillary column for detecting ochratoxin A and preparation method and application thereof |
| CN113484283A (en) * | 2021-05-19 | 2021-10-08 | 南京医科大学第二附属医院 | Detection system and method for simultaneously measuring multiple biological information |
| CN113484283B (en) * | 2021-05-19 | 2024-02-06 | 南京医科大学第二附属医院 | Detection system and method for simultaneously measuring multiple biological information |
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| CN1295347C (en) | 2007-01-17 |
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