CN106694066A - Low energy consumption micro-fluidic chip for microspherical biological detection - Google Patents
Low energy consumption micro-fluidic chip for microspherical biological detection Download PDFInfo
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- CN106694066A CN106694066A CN201611196504.0A CN201611196504A CN106694066A CN 106694066 A CN106694066 A CN 106694066A CN 201611196504 A CN201611196504 A CN 201611196504A CN 106694066 A CN106694066 A CN 106694066A
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- Prior art keywords
- microballoon
- micro
- chip
- fluidic chip
- coding
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Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
Abstract
The invention discloses a low energy consumption micro-fluidic chip for microspherical biological detection. The micro-fluidic chip can be used for detecting biological macromolecules such as proteins and nucleic acids and has a wide application prospect in the fields of clinical test, inspection and quarantine, environmental monitoring, drug screening, microbiological assay and nucleic acid and protein function analysis and the like. The micro-fluidic chip is composed of a cover piece located on the upper half portion and a substrate located on the lower half portion, wherein a micro-channel network is arranged in the substrate, a solid phase carrier as a probe molecule in biological detection with coded microspheres is arranged in the microchannel, and an inlet (1), an outlet (2), a long channel (3), a reaction pond (4), a group of auxiliary pipelines (5) and a screening pipeline (6) are arranged at each of two ends of the micro-channel. In a using process of the chip, sampling and reaction of a sample and detection of a reaction result are performed on the chip, so that the chip has the advantages of being high in integration level, convenient to use, small in sample consumption, high in detecting sensitivity and the like.
Description
Technical field
This patent refers to micro-fluidic chip of microballoon biological detection of a kind of low power consuming and preparation method thereof.It can be with
For detecting the large biological molecules such as protein, nucleic acid, clinical detection, inspection and quarantine, environmental monitoring, medicine are can be widely applied for
The field such as screening, microbial identification and nucleic acid and protein-function assays.
Background technology
Microfluidic analysis chip refer to by microelectronics, micro-processing technology square centimeter size solid-phase media surface structure
The micro-analysis system built, with realize in tissue and cell DNA, protein and other biological component it is quick, efficient, sensitive
Treatment with analysis.It is a kind of analytical technology that early 1990s mid-term grows up in analytical chemistry field, to divide
Analysis chemistry and analytical biochemistry based on, using microelectronic processing technique, processed on microchip micron-sized container,
The micro-structural network such as pump, valve, pipeline, integrated micro-total analysis system is carried out by this process of the preparation of sample, reaction and detection
System.The size of chip in several square centimeters or so, from silicon chip, glass, silicon rubber, plastic or other material as substrate and lid
Piece, microchannel is processed by methods such as etching, photoetching or dies, and the stream in passage is driven using modes such as electricity, pressure, gravity
Body, is finally detected using chemiluminescence, electrochemistry, fluorescence detector etc..The chip is mainly characterized by it on device
The resulting structure (including its passage, reative cell and other functional parts) for accommodating fluid is micron order chi at least in a dimension
Degree.Compared with the experimental provision of macro-size, the micron scale construction of microfluidic analysis chip can significantly increase fluid environment
The ratio of area volume.This change causes a series of its property of the decision relevant with body surface in microfluidic system
Peculiar effect, such as laminar flow effect, surface tension and capillary effect, rapid thermal conduction effect and diffusion effect etc..These effects
The analytical performance of microfluid analysis chip should be made to be significantly improved, including can reduce the volume of analytical equipment,
The more integrated automation of equipment, can significantly improve analysis efficiency and be remarkably decreased sample and reagent consumption.
Laboratory large scale equipment is integrated on operating platform as small as possible, is used to complete different experimentations, and can enter product
The technology of row analysis.It not only reduces the consumption of reagent, and improves speed of experiment, expense reduction, has fully demonstrated and has worked as
Modern laboratory equipment miniaturization, integrated and portability development trend.Present micro-fluidic chip in biochemical analysis and
Environmental analysis aspects is sent out to have and is widely applied and quickly develops.Because micro-fluidic chip is by the mistake of whole sample analysis
Journey, including sampling and the treatment of sample, preconcentration, dilution and mixing, separation, chemical reaction and being fully integrated of signal detection exist
On one piece of small chip, it realizes the reagent and sample consumption that sub-micro rises even nanoliter level compared with traditional analytical equipment,
Being substantially reduced for reaction compartment improves the homogeneity of reaction, and reaction speed is accelerated, while reducing production cost and being easy to take
Band.
Patent content
Technical problem：The purpose of this patent is to provide a kind of micro-fluidic chip of the microballoon biological detection of low power consuming, is used for
Biomolecule detection is carried out by solid phase carrier of microballoon.The coding microball that bioprobe molecule will be fixed with enters to advance by chip
Sample and testing sample reaction, washing and detection, there is provided a kind of chip platform of detection and analysis.
Technical scheme：The purpose of this patent can be achieved through the following technical solutions：
The micro-fluidic chip based on microballoon biological detection of this patent is by the cover plate positioned at the first half and positioned at lower half
Substrate is constituted；Functional microchannel network is provided with substrate or cover plate, is provided with microchannel network with coding
Microballoon as biomolecule detection carrier, in microchannel network, the periphery of reaction tank is screening pipeline, screening pipeline it is outer
Week is subsidiary conduit, and subsidiary conduit connects with outlet, and reaction tank taps into mouth by long-channel.
Using microballoon as carrier, microballoon has coding to biomolecule detection, and being fixed with biomolecule on the surface of microballoon visits
Pin, probe biomolecule used can be the one kind in nucleic acid, protein, polypeptide.
The reaction of microballoon and testing sample is carried out in chip reaction tank；Solution to be measured and lavation buffer solution are entered by import
Enter reaction tank, reacted or washing microballoon with microballoon in reaction tank, reaction terminates microballoon and be transfused in passage to detect and export.
Used as probe molecule carrier, its material can be glass or polystyrene and rubber to microballoon, and the size of microballoon is 50
μm~300 μm between；The host material of chip can be glass, silicon, polymethyl methacrylate dimethyl silicone polymer and gather
One kind in carbonic ester.
Coding microball is barcode encoding, fluorescence-encoded, quantum-dot coding, photonic crystal is encoded, Raman tag is encoded, red
External spectrum coding, shape coding, radio frequency encoding, size coding and it is position encoded in one kind.
Between 200 μm~1200 μm, between 60 μm~550 μm, length exists long-channel width the diameter of reaction tank
Between 10mm~50mm.
Its operation principle is：
A) a reciprocation type syringe pump or peristaltic pump are connected in chip exit, chip outer coding microballoon is connected into its delivery
Buffer solution is by long-channel, sucting reaction pond.Due to sieving the diameter of the size less than coding microball of passage, therefore by pump
Continue to aspirate, microballoon is stayed in reaction tank, and the buffer solution for delivering microballoon is then blotted；
B) now, testing sample solution is being sucked to reaction tank by import, is making testing sample solution fully anti-with microballoon
Should；
C) reaction is finished, and continues through import suction lavation buffer solution；
D) after washing completely, by exporting injection microballoon vehicle buffer, microballoon is made to enter long logical with vehicle buffer
Road, when microballoon passes through long-channel, detects the reaction result of microsphere surface.
Beneficial effect：According to this patent, detection carrier, sample note sample, reaction, washing and the company detected are carried out using chip
Continuousization is operated, and the solid phase carrier with coding microball as biomolecule detection has advantages below：
(1) detection speed is fast：Because hybridization reaction is only carried out in microchannel, hybridization solution is circulated, it is possible to reduce molten
The volatilization of liquid and purpose target molecule improve the speed of reaction to the time of tat probe, shorten detection time；
(2) can realize that multivariate analysis is detected：The microballoon of each sample introduction has coding, therefore can encode multi-element biologic point
Son detection, while the multiple indexs in detecting same sample.Ball type carrier has specific surface area big simultaneously, the spy such as can roll
Point, so the sensitivity of detection reaction is high, sample requirement is few, and reaction speed is fast；
(3) degree of accuracy is high：Microballoon, because channel size is slightly larger than microsphere diameter, is much smaller than when long-channel is entered
Two diameters of microballoon, therefore microballoon single can only pass through, and be detected successively, accuracy is high；
(4) scalability is high：As a result of the form of micro-fluidic chip, in that context it may be convenient to miniflows such as sample pretreatments
Control integrated chip, promotes the miniaturization and automation of analysis system；
(5) this patent is in combination with microsphere supported and micro-fluidic chip biological detection advantage, to the micro- of small size
Ball is simple to operate, with distinct advantage.
Brief description of the drawings
Fig. 1 is the top view of this patent chip.
Fig. 2 for this patent chip use when, the structural representation of whole device.
Have in figure above：Import 1, outlet 2, long-channel 3, reaction tank 4, subsidiary conduit 5, screening pipeline 6.
Specific embodiment
This patent is a kind of microfluidic analysis chip based on microballoon, and the biochip is by the cover plate positioned at the first half and position
Substrate in lower half is constituted；Functional microchannel network is provided with substrate or cover plate, is provided with microchannel network
Microballoon with coding as biomolecule detection carrier, in microchannel network, the periphery of reaction tank 4 is screening pipeline 6,
The periphery for sieving pipeline 6 is subsidiary conduit 5, and subsidiary conduit 5 connects with outlet 2, and reaction tank 4 taps into mouth 1 by long-channel 3.
Using microballoon as carrier, microballoon has coding to biomolecule detection, and being fixed with biomolecule on the surface of microballoon visits
Pin, probe biomolecule used can be the one kind in nucleic acid, protein, polypeptide.Carried out in chip reaction tank 4 microballoon with
The reaction of testing sample；Solution to be measured and lavation buffer solution enter reaction tank 4 by import 1, are reacted with microballoon in reaction tank 4
Or washing microballoon, reaction terminates microballoon and is transfused in passage 3 to detect and export., used as probe molecule carrier, its material can for microballoon
To be glass or polystyrene and rubber, the size of microballoon is between 50 μm~300 μm；The host material of chip can be glass
One kind in glass, silicon, polymethyl methacrylate eight, dimethyl silicone polymer and makrolon.Coding microball is that bar code is compiled
Code, fluorescence-encoded, quantum-dot coding, photonic crystal coding, Raman tag coding, infrared spectrum coding, shape coding, radio frequency are compiled
Code, size coding and it is position encoded in one kind.The diameter of reaction tank 4 between 200 μm~1200I μm, the width of long-channel 3
Between 60 μm~550 μm, length is between 10mm~500mm.
As the microballoon of probe molecule carrier, its material can be glass, polystyrene or rubber etc..
Embodiment one：Using PMMA (polymethyl methacrylate) as chip material, glass microsphere is encoded in photonic crystal
Upper fixed probe carries out biological detection, and microsphere diameter is 50 μm.
A. the preparation of substrate：Using laser micromachining methods.Microchannel figure is designed by AutoCAD, CAD diagram shape is turned
Change the recognizable instruction of laser micro-machining system into；Required net is processed on substrate (or cover plate) using laser micro-machining system
Network microchannel, wherein, long-channel width is 60 μm, 20mm long, and a diameter of 250 μm of reaction tank sieves a width of 20 μm of pipeline, passage
Depth be 60 μm.Punching two ends are respectively as injection port and outlet；
B. the preparation of cover plate：Selection is with an equal amount of PMMA slice, thin pieces of substrate as cover plate；The encapsulation of cover plate and substrate:Directly
Connecting glassy state hot key and method carries out the encapsulation of substrate and cover plate；
Probe is fixed on C, microballoon：Clean microballoon is modified through silanization and bifunctional reagent, then by different probes
Molecule is fixed on the glass microsphere of photonic crystal coding；
D. the connection of micro-fluidic chip：Import 1 connects sample cell by pipeline, and outlet 2 is access to compound syringe pump；
E, sample introduction：Coding microball is sucked by long-channel 3 together with its vehicle buffer by syringe pump, subsequently into reaction tank
4, vehicle buffer is finally drained, coding microball is stayed in reaction tank 4；
F, reaction：Solution to be detected is sucked by long-channel 3 by syringe pump, finally enter in reaction tank 4 with coding microball
The probe molecule on surface is reacted；
G, washing：After completion of the reaction, suck lavation buffer solution from entrance 1 by syringe pump is carried out to reaction tank 4 to microballoon
Fully washing：
H, detection：The lavation buffer solution in reaction tank 4 is drained, the delivery for injecting microballoon from outlet 2 using syringe pump is buffered
Liquid, vehicle buffer brings microballoon into long-channel (3), and microballoon passes sequentially through long-channel (3), and is detected successively in long-channel 3
Survey.
Embodiment three：It is fixed on fluorescence-encoded glass microsphere using PDMS (dimethyl silicone polymer) as chip material
Probe carries out biological detection, and microsphere diameter is 200 μm.
The preparation of a, substrate：It is 10 to select the matrix of PDMS and the ratio of curing agent：After 1 (mass ratio) is well mixed, warp
After vacuum suction is to remove bubble, it is cast on the formpiston for processing in advance, carries out the 25min that is heating and curing, wherein, long-channel is wide
It is 250 μm to spend, 50mm long, a diameter of 1200 μm of reaction tank, sieves a width of 50 μm of pipeline, and the depth of passage is 250 μm.It is cured
PDMS is taken off from masterplate afterwards, that is, is made substrate；
The preparation of b, cover plate：It is 5 to select the matrix of PDMS and the ratio of curing agent：After 1 (mass ratio) is well mixed, through true
After sky pumping is to remove bubble, the direct horizontal positioned 20min that is heating and curing is made cover plate；
The encapsulation of c, cover plate and substrate：The substrate and cover plate being cured are fitted after being processed with oxygen plasma again, Ran Houzhi
Connecting overheat bonding method carries out the encapsulation of substrate and cover plate；
Probe is fixed on d, microballoon：Different probe molecules is fixed to the glass microsphere surface of different coding；
The connection of e, micro-fluidic chip：Import 1 connects sample cell by pipeline, and outlet 2 is access to compound micro syringe pump；
F, sample introduction：Coding microball is sucked by long-channel 3 together with its vehicle buffer by syringe pump, subsequently into reaction tank
4, vehicle buffer is finally drained, coding microball is stayed in reaction tank 4；
G, reaction：Solution to be detected is sucked by long-channel 3 by syringe pump, coding microball table in reaction tank 4 is finally entered
The probe molecule in face is reacted；
H, washing：After completion of the reaction, suck lavation buffer solution from entrance 1 by syringe pump is carried out to reaction tank 4 to microballoon
Fully washing；
I, detection：The lavation buffer solution in reaction tank 4 is drained, the delivery for injecting microballoon from outlet 2 using syringe pump is buffered
Liquid, vehicle buffer brings microballoon into long-channel 3, and microballoon passes sequentially through long-channel 3, and is detected successively in long-channel 3.
Claims (6)
1. the micro-fluidic chip of the microballoon biological detection of a kind of low power consuming, it is characterised in that：The biochip is by positioned at the first half
Cover plate and the substrate positioned at lower half constituted：Functional microchannel network is provided with substrate or cover plate, in microchannel
Carrier of the microballoon with coding as biomolecule detection, in microchannel network, the periphery of reaction tank (4) are provided with network
It is screening pipeline (6), the periphery of screening pipeline (6) is that subsidiary conduit (5) connects with outlet (2), and reaction tank (4) is by long-channel
Tap into mouth (1).
2. the micro-fluidic chip of the microballoon biological detection of a kind of low power consuming according to claim 1, it is characterised in that biological
Using microballoon as carrier, microballoon has coding, probe biomolecule, biology used is fixed with the surface of microballoon Molecular Detection
Molecular probe can be the one kind in nucleic acid, protein, polypeptide.
3. the micro-fluidic chip of the microballoon biological detection of a kind of low power consuming according to claim 1 and 2, it is characterised in that
The reaction of microballoon and testing sample is carried out in chip reaction tank (4)；Solution to be measured and lavation buffer solution enter anti-by import (1)
Ying Chi (4), in reaction tank (4)) in microballoon react or washing microballoon, reaction terminate microballoon be transfused to access door (4)) in detection
And export.
4. a kind of micro-fluidic chip of the microballoon biological detection of low power consuming according to claim 2, it is characterised in that microballoon
Used as probe molecule carrier, its material can be glass or polystyrene and rubber, the size of microballoon 50 μm~300 μm it
Between；During the host material of chip can be glass, silicon, polymethyl methacrylate eight, dimethyl silicone polymer and makrolon
One kind.
5. the micro-fluidic chip of the microballoon biological detection of a kind of low power consuming according to claim 1 and 2, it is characterised in that compile
Code microballoon is barcode encoding, fluorescence-encoded, quantum-dot coding, photonic crystal coding, Raman tag coding, infrared spectrum volume
Code, shape coding, radio frequency encoding, size coding and it is position encoded in one kind.
6. a kind of micro-fluidic chip of the microballoon biological detection of low power consuming according to claim 1, it is characterised in that reaction
The diameter in pond (4) between 200 μm~1200 μm, long-channel door (3) width between 60 μm~550 μm, length 10mm~
Between 50mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611196504.0A CN106694066A (en) | 2016-12-22 | 2016-12-22 | Low energy consumption micro-fluidic chip for microspherical biological detection |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611196504.0A CN106694066A (en) | 2016-12-22 | 2016-12-22 | Low energy consumption micro-fluidic chip for microspherical biological detection |
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| CN201611196504.0A Pending CN106694066A (en) | 2016-12-22 | 2016-12-22 | Low energy consumption micro-fluidic chip for microspherical biological detection |
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Citations (6)
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|---|---|---|---|---|
| CN101221168A (en) * | 2008-01-08 | 2008-07-16 | 东南大学 | Microfluidic chip based on microsphere biological detection |
| US20080280378A1 (en) * | 2004-07-16 | 2008-11-13 | Gyros Patent Ab Gyros Ab | Grading of Immune Responses |
| CN104471385A (en) * | 2012-04-19 | 2015-03-25 | 和光纯药工业株式会社 | Methods for real-time sampling of reaction products |
| CN105008895A (en) * | 2012-10-15 | 2015-10-28 | 纳诺赛莱克特生物医药股份有限公司 | Systems, apparatus, and methods for sorting particles |
| WO2016149661A1 (en) * | 2015-03-18 | 2016-09-22 | The Broad Institute, Inc. | Massively parallel on-chip coalescence of microemulsions |
| CN106552683A (en) * | 2016-12-06 | 2017-04-05 | 天津岳达科技有限公司 | Efficient microsphere biological detection chip |
-
2016
- 2016-12-22 CN CN201611196504.0A patent/CN106694066A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080280378A1 (en) * | 2004-07-16 | 2008-11-13 | Gyros Patent Ab Gyros Ab | Grading of Immune Responses |
| CN101221168A (en) * | 2008-01-08 | 2008-07-16 | 东南大学 | Microfluidic chip based on microsphere biological detection |
| CN104471385A (en) * | 2012-04-19 | 2015-03-25 | 和光纯药工业株式会社 | Methods for real-time sampling of reaction products |
| CN105008895A (en) * | 2012-10-15 | 2015-10-28 | 纳诺赛莱克特生物医药股份有限公司 | Systems, apparatus, and methods for sorting particles |
| WO2016149661A1 (en) * | 2015-03-18 | 2016-09-22 | The Broad Institute, Inc. | Massively parallel on-chip coalescence of microemulsions |
| CN106552683A (en) * | 2016-12-06 | 2017-04-05 | 天津岳达科技有限公司 | Efficient microsphere biological detection chip |
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