CN101718698A - Laser-induced fluorescence analyzer with PCR-CE coupled microfluidic chip - Google Patents
Laser-induced fluorescence analyzer with PCR-CE coupled microfluidic chip Download PDFInfo
- Publication number
- CN101718698A CN101718698A CN200910272874A CN200910272874A CN101718698A CN 101718698 A CN101718698 A CN 101718698A CN 200910272874 A CN200910272874 A CN 200910272874A CN 200910272874 A CN200910272874 A CN 200910272874A CN 101718698 A CN101718698 A CN 101718698A
- Authority
- CN
- China
- Prior art keywords
- pcr
- laser
- unit
- induced fluorescence
- chip
- 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.)
- Granted
Links
- 238000001499 laser induced fluorescence spectroscopy Methods 0.000 title claims abstract description 34
- 238000012545 processing Methods 0.000 claims abstract description 16
- 238000000926 separation method Methods 0.000 claims abstract description 11
- 238000006243 chemical reaction Methods 0.000 claims description 18
- 230000003321 amplification Effects 0.000 claims description 12
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 12
- 238000013519 translation Methods 0.000 claims description 12
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 230000003287 optical effect Effects 0.000 claims description 5
- 230000008878 coupling Effects 0.000 claims description 4
- 238000010168 coupling process Methods 0.000 claims description 4
- 238000005859 coupling reaction Methods 0.000 claims description 4
- 229910052697 platinum Inorganic materials 0.000 claims description 4
- 229910052751 metal Inorganic materials 0.000 claims description 2
- 239000002184 metal Substances 0.000 claims description 2
- 238000012946 outsourcing Methods 0.000 claims description 2
- 230000005622 photoelectricity Effects 0.000 claims description 2
- 201000010099 disease Diseases 0.000 abstract description 14
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 14
- 238000004458 analytical method Methods 0.000 abstract description 12
- 238000001514 detection method Methods 0.000 abstract description 12
- 238000003745 diagnosis Methods 0.000 abstract description 7
- 108090000623 proteins and genes Proteins 0.000 abstract description 6
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 238000000034 method Methods 0.000 description 18
- 239000000523 sample Substances 0.000 description 18
- 108020004414 DNA Proteins 0.000 description 16
- 230000008569 process Effects 0.000 description 11
- 238000005251 capillar electrophoresis Methods 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 8
- 238000012408 PCR amplification Methods 0.000 description 5
- 238000001962 electrophoresis Methods 0.000 description 5
- 238000011161 development Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000002795 fluorescence method Methods 0.000 description 2
- 230000004907 flux Effects 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000004065 semiconductor Substances 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 238000004159 blood analysis Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 238000001818 capillary gel electrophoresis Methods 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005323 electroforming Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000005357 flat glass Substances 0.000 description 1
- 235000021393 food security Nutrition 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 229920000620 organic polymer Polymers 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 238000001259 photo etching Methods 0.000 description 1
- 229920003229 poly(methyl methacrylate) Polymers 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 239000004926 polymethyl methacrylate Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 238000009666 routine test Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000001039 wet etching Methods 0.000 description 1
Images
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a laser-induced fluorescence analyzer with a PCR-CE coupled microfluidic chip, relating to a biomedical detection analyzer. In the structure of the laser-induced fluorescence analyzer, a machine shell (00) is internally provided with a PCR unit (10), a CE unit (20), a laser-induced fluorescence detection unit (30) and a signal collecting and processing unit (40); after mutually connecting with t, the PCR unit (10) and the CE unit (20) are connected with each other firstly and then connected with the laser-induced fluorescence detection unit (30); and the laser-induced fluorescence detection unit (30) is connected with the signal collecting and processing unit (40). The invention integrates PCR, electrophoretic separation, and laser-induced fluorescence detection into a whole, has the advantages of small size, simple structure, easy operation, low cost, stable performance, high analysis and detection sensitivity, and short disease diagnosis period, and is suitable for disease diagnosis of genes related to diseases.
Description
Technical field
The present invention relates to a kind of biomedical detection instrument, relate in particular to a kind of PCR-CE (PCR-Capillary Electrophoresis) coupling micro-fluidic chip laser-induced fluorescence analyzer.
Background technology
The micro-total analysis unit is a brand-new field that begins to rise the beginning of the nineties in last century.The micro-total analysis unit relates to a lot of ambits, as analytical chemistry, physics, biology, medical science, pharmacy, micro-processing technology, electronics, computing machine etc., by integrated, the microminiaturization of sample preparation, separation and detection being realized the basic function of Routine Test Lab, thereby realize low cost, high flux, intellectuality, portable target.
Lab-on-chip technology is with whole breadboard function, having only on several square centimeters the microchip from process integration such as application of sample, reaction, separation, detections, at present in chemosynthesis, cellular incubation, medicament high flux screening, medical diagnosis on disease, food security, aspects such as blood analysis and environment measuring make important progress.The development of micro-fluidic chip technology makes that round pcr is achieved on the chip.Along with the development of micro-processing technology, microfluid the channel dimensions of process entered into nanoscale from micron order, make the research object of microflow control technique enter into molecular level from cell grade.Microflow control technique has become current research forward position and focus.
The detection method of micro-fluidic chip mainly contains: laser inductive fluorescence method, electrochemical process, chemoluminescence method, mass spectroscopy etc.Wherein, laser inductive fluorescence method has highly sensitive, and principle is simple, is integrated in easily on the micro-fluidic chip, becomes the main at present detection method that adopts.The optical unit of laser-Induced Fluorescence Detection mainly contains two kinds: a kind of is that the exciting light light path is vertical mutually with the fluorescence light path; Another kind is focusing altogether, and it is with microcobjective unpolarized light beam to be focused on the micro-fluidic chip raceway groove, and fluorescence excitation incides on the photomultiplier by same object lens and detects.The subject matter of the laser-Induced Fluorescence Detection technology of Chu Xianing is that the volume ratio of detecting device is bigger in the market, costs an arm and a leg, and complex structure is difficult to realize the microminiaturization of analytic unit.
Capillary electrophoresis technique has become requisite means in the analysis field since coming out.What use always in the Capillary Electrophoresis is gel electrophoresis, and amount of samples is bigger, and during operating cost, the perfusion gel is difficult, and analysis time is long, the applied voltage height.Micro flow control chip capillary electrophoresis adopts the low polymkeric substance sieving media of viscosity to serve as electrophoretic medium, and it is easier to operate.Microcurrent controlled capillary tube electrophoresis has only several centimeter length owing to separate raceway groove, and disengaging time dwindles greatly, only needs a few minutes, even tens seconds.Microcurrent controlled capillary tube electrophoresis, the agents useful for same consumption is few, and separation voltage is low, and sensitivity is higher.
At present, when carrying out the disease gene diagnosis, all be that sample extraction is come out, on the pcr amplification instrument, increase, carry out electrophoresis detection then.Detection method has: conventional capillary gel electrophoresis and microcurrent controlled capillary tube gel electrophoresis.Conventional pcr amplification instrument is because volume is big, and it is slow with cooling rate to heat up, and makes the amplification procedure length that expends time in.And the micro-fluidic chip technology makes in the enterprising performing PCR amplification of micro-fluidic chip, and proliferation time is short, and it is fast with cooling rate to heat up, and has shortened the amplification required time greatly.And the sample that amplification is come out also will spend certain sample preparation time before carrying out electrophoresis detection, makes that the whole medical diagnosis on disease cycle is longer.
Summary of the invention
Purpose of the present invention is exactly to overcome long, problem such as amount of samples is big, labor workload is big, cost is high and the cycle is long of the processing sample time that exists in existing disease gene diagnosis sample extraction, PCR sample amplification, electrophoretic separation, the whole process of laser-Induced Fluorescence Detection, and a kind of PCR-CE coupling micro-fluidic chip laser-induced fluorescence analyzer is provided.
The object of the present invention is achieved like this:
In casing, be provided with PCR unit, CE unit, laser-Induced Fluorescence Detection unit and signals collecting and processing unit;
The PCR unit be connected with the laser-Induced Fluorescence Detection unit again after the CE unit interconnects, the laser-Induced Fluorescence Detection unit is connected with processing unit with signals collecting.
Principle of work of the present invention is:
By the micro-fluidic chip technology, the PCR process and the Capillary Electrophoresis Separation Process of disease DNA sample integrate, and analysis result carries out fluorescence signal by the laser-Induced Fluorescence Detection unit and detects, and carries out the demonstration of analysis result on computers.The concentration of disease DNA sample is very low, and the PCR process is to realize that by well heater 3 different warm areas carry out amplification procedure to disease DNA, to reach the level that can detect.Realize the separation of each component in the disease DNA sample after DNA cloning is intact through Capillary Electrophoresis, analyze by result, thereby disease is diagnosed separation.
The present invention has the following advantages and good effect:
1, integrated PCR, electrophoretic separation, laser-Induced Fluorescence Detection be in one, and volume is little, simple in structure, easy operating, cost are low;
2, stable performance, analyzing and testing is highly sensitive, and the medical diagnosis on disease cycle is short.
3, be applicable to the gene clinical diagnosis of diseases related.
Description of drawings
Fig. 1 is a structured flowchart of the present invention;
Fig. 2 is structural representation of the present invention (a main pseudosection);
Fig. 3 is a light path principle figure of the present invention.
Among the figure:
The 00-casing,
The 01-support;
The 10-PCR unit,
The 11-PCR well heater, the 12-chip set, the three-dimensional mobile platform of 13-XYZ,
The integrated PCR-CE chip of 14-;
The 20-CE unit,
The 21-electrode holder, 22-controllable high-voltage out-put supply,
23-spiral translation device;
30-laser-Induced Fluorescence Detection unit,
31-semiconductor solid state laser, the 32-plane mirror, the 33-half-reflecting half mirror,
The 34-object lens, the 35-convex lens, the 36-pin hole,
The 37-optical filter, the 38-photomultiplier;
40-signals collecting and processing unit
41-signals collecting, filtering, amplification, analog to digital conversion circuit, the 42-computing machine.
Abbreviation:
1, PCR-PCR (Polymerase Chain Reaction) is a kind of Protocols in Molecular Biology, is used for amplifying specific dna fragmentation. The special dna replication dna that can regard in vitro as. The basic principle of round pcr is similar to the natural reproduction process of DNA, and its specificity depends on the Oligonucleolide primers with the complementation of target sequence two ends. PCR is made of sex change-annealing-extension three basic reactions steps: the 1. sex change of template DNA: template DNA is after being heated to 93 ℃ of left and right sides certain hours, template DNA double-stranded DNA double-stranded or that form through pcr amplification is dissociated, make it to become strand, so that it is combined with primer, for the lower whorl reaction is prepared; 2. the annealing (renaturation) of template DNA and primer: template DNA is after heat denatured becomes strand, and temperature is down to about 55 ℃, the complementary series pairing combination of primer and template DNA strand; 3. the extension of primer: dna profiling-primer bond is under the effect of TaqDNA polymerase, take dNTP as reaction raw materials, target sequence is template, press base complementrity pairing and semi-conservative replication principle, synthetic new and the semi-conservative replication chain complementation of template DNA chain, repetitive cycling sex change--fire--three processes of extending just can obtain more " semi-conservative replication chain ", and this new chain can become again the template of circulation next time. Whenever finish a circulation and need 2～4 minutes, just can genes of interest amplification be expanded amplify millions of times in 2～3 hours.
2, CE-Capillary Electrophoresis (Capillary electrophoresis) is a kind of separate analytical technique. Have fast, efficient, high sensitivity, easily quantitatively, the advantages such as favorable reproducibility and automation, be widely used in the compartment analysis research of little molecule, small ion, polypeptide and protein. It demonstrates again huge potentiality aspect separate nucleic acid. The basic device of Capillary Electrophoresis is one and is full of the capillary of electrophoretic buffer and enters capillary from an end capillaceous by " pressure " or " electromigration " with two bottle micro-examples that link to each other with the capillary two ends. During electrophoresis, two electrodes that are connected with high voltage source soak respectively in the buffer solution of people's capillary two ends bottle. Sample court and self electrically charged opposite polarity electrode direction swimming. Each component because of its molecular size, the difference migration rate difference of the character such as electrically charged number, isoelectric point, move to successively near the photodetector of capillary output, detect, record absorbance, and on screen take transit time as abscissa, absorbance is that ordinate is dynamically recorded each component intuitively with the form of absworption peak.
Embodiment
Describe in detail below in conjunction with drawings and Examples:
One, overall
As Fig. 1, the present invention is provided with PCR unit 10, CE unit 20, laser-Induced Fluorescence Detection unit 30 and signals collecting and processing unit 40 in casing 00;
Two, functional block
0, casing 00
As Fig. 2, in casing 00, be provided with support 01, use for fixing other functional block.
1, the PCR unit 10
As Fig. 2, PCR unit 10 comprises PCR well heater 11, chip set 12, the three-dimensional mobile platform 13 of XYZ and integrated PCR-CE chip 14;
The support 01 at middle part is provided with the three-dimensional mobile platform 13 of XYZ in the casing 00, is connected with chip set 12 at the left of the three-dimensional mobile platform 13 of XYZ, is respectively arranged with integrated PCR-CE chip 14 and PCR well heater 11 at the upper and lower of chip set 12;
Described PCR well heater 11 is a kind of electric heaters of being made up of 1～3 heat block (heating material commonly used);
Described chip set 12 is a kind of for the metal frame that connects and support usefulness;
The three-dimensional mobile platform 13 of described XYZ is a kind of outsourcing pieces commonly used;
Described integrated PCR-CE chip 14 is a kind of micro-fluidic chips of design voluntarily, partly is made up of PCR part and CE; PCR partly is a PCR reaction tank or a PCR reaction channel; The CE part is made up of sample feeding passage and sample separation passage, is right-angled intersection or double T and links to each other; The sample feeding passage of the reaction tank of PCR part or reaction channel and CE part is interconnected.
This integrated PCR-CE chip 14 is to utilize classical photoetching development and wet etching method to produce micron-sized passage on glass, utilize drill bit to get out liquid storage tank in relevant position on glass, again with the flat glass of an other same material at high temperature bonding obtain; This integrated PCR-CE chip 14 also can be by the electroformed mould general laws organic polymer polymethylmethacrylate heating and pressurizing on the electroforming mould to be prepared from.
2, the CE unit 20
As Fig. 2, CE unit 20 comprises electrode holder 21, controllable high-voltage out-put supply 22 and spiral translation device 23;
Described electrode holder 21 is pcb boards of a kind of 2-4 of including platinum electrode;
Described controllable high-voltage out-put supply 22 is a kind of 0～6000V adjustable high voltage power supply;
Described spiral translation device 23 has the listing product;
3, the laser-Induced Fluorescence Detection unit 30
As Fig. 2,3, laser-Induced Fluorescence Detection unit 30 comprises solid state laser 31, plane mirror 32, half-reflecting half mirror 33, object lens 34, convex lens 35, pin hole 36, optical filter 37 and photomultiplier 38;
Its light path is: the laser that solid state laser 31 produces enters half-reflecting half mirror 33 after plane mirror 32 reflections; Its reflected light promptly produces fluorescence through object lens 34 behind integrated PCR-CE chip 14, fluorescence is gone into photoelectricity multiplier tube 38 by object lens 34, half-reflecting half mirror 33, convex lens 35, pin hole 36 and optical filter 37 are laggard again successively, detects fluorescence signal (simulating signal).
4, signals collecting and processing unit 40
As Fig. 2, signals collecting and processing unit 40 comprise preceding latter linked signals collecting, filtering, amplification, analog to digital conversion circuit 41 and computing machine 42;
Signals collecting, filtering, amplification, analog to digital conversion circuit 41 are a kind of circuit commonly used, realize collection, filtering, amplification and the analog to digital conversion of fluorescence signal, obtain fluorescence signal (digital signal);
Three, the operation steps of this analysis instrument is as follows:
The integrated PCR-CE chip 14 that 1. will add sample solution and buffering solution is positioned on the chip set 12, adjustable screw translation device 23 moves down electrode holder 21, thereby the electrode on the electrode holder 21 is inserted respectively in the liquid storage tank of integrated PCR-CE chip 14 correspondences; Use the three-dimensional mobile platform 13 of XYZ to adjust the position of integrated PCR-CE chip 14, make the sense channel of integrated PCR-CE chip 14 and laser-Induced Fluorescence Detection light path at grade.
2. open semiconductor solid state laser 31, regulate the three-dimensional mobile platform 13 of XYZ again, the focus point of laser beam is detected on the check point of raceway groove at integrated PCR-CE chip 14, build the lid of shell 00.
3. open PCR well heater 11, treat to open photomultiplier 38 after pcr amplification finishes, opening signal collection, filtering, amplification, analog to digital conversion circuit 41 are opened controllable high-voltage out-put supply 22, regulate the operating voltage of photomultiplier 38, select suitable detection sensitivity.
4. regulate controllable high-voltage out-put supply 22, on integrated PCR-CE chip 14, carry out sample introduction, the detachment process of sample simultaneously synchronously; In this process, the demonstration of the detection of fluorescence signal, the collection of data and processing, fluorescence pattern all can be carried out simultaneously.
5. after treating that the sample analysis process finishes, integrated PCR-CE chip 14 is taken out, clean up with the liquid storage tank and the raceway groove of deionized water with integrated PCR-CE chip 14.
Claims (5)
1.PCR-CE coupling micro-fluidic chip laser-induced fluorescence analyzer is characterized in that:
In casing (00), be provided with PCR unit (10), CE unit (20), laser-Induced Fluorescence Detection unit (30) and signals collecting and processing unit (40);
PCR unit (10) be connected with laser-Induced Fluorescence Detection unit (30) again after CE unit (20) interconnect, laser-Induced Fluorescence Detection unit (30) are connected with processing unit (40) with signals collecting.
2. by the described laser-induced fluorescence analyzer of claim 1, it is characterized in that:
Described PCR unit (10) comprises PCR well heater (11), chip set (12), the three-dimensional mobile platform (13) of XYZ and integrated PCR-CE chip (14);
The support (01) at middle part is provided with the three-dimensional mobile platform (13) of XYZ in the casing (00), left at the three-dimensional mobile platform of XYZ (13) is connected with chip set (12), is respectively arranged with integrated PCR-CE chip (14) and PCR well heater (11) at the upper and lower of chip set (12);
Described PCR well heater (11) is a kind of electric heater of being made up of 1～3 heat block;
Described chip set (12) is a kind of for the metal frame that connects and support usefulness;
The three-dimensional mobile platform of described XYZ (13) is a kind of outsourcing piece commonly used;
Described integrated PCR-CE chip (14) is a kind of micro-fluidic chip, partly is made up of PCR part and CE; PCR partly is a PCR reaction tank or a PCR reaction channel; The CE part is made up of sample feeding passage and sample separation passage, is right-angled intersection or double T and links to each other; The sample feeding passage of the reaction tank of PCR part or reaction channel and CE part is interconnected.
3. by the described laser-induced fluorescence analyzer of claim 1, it is characterized in that:
Described CE unit (20) comprises electrode holder (21), controllable high-voltage out-put supply (22) and spiral translation device (23);
Described electrode holder (21) is the pcb board of a kind of 2-4 of including platinum electrode;
Described controllable high-voltage out-put supply (22) is a kind of 0～6000V adjustable high voltage power supply;
Described spiral translation device (23) has the listing product;
Spiral translation device (23) is fixed on the chip set (12), and electrode holder (21) links to each other with spiral translation device (23), by adjustable screw translation device (23) electrode holder (21) is moved up and down; Electrode holder (21) is connected with controllable high-voltage out-put supply (22) by lead, powers to platinum electrode.
4. by the described laser-induced fluorescence analyzer of claim 1, it is characterized in that:
Described laser-Induced Fluorescence Detection unit (30) comprises solid state laser (31), plane mirror (32), half-reflecting half mirror (33), object lens (34), convex lens (35), pin hole (36), optical filter (37) and photomultiplier (38);
Its light path is: the laser that solid state laser (31) produces enters half-reflecting half mirror (33) after plane mirror (32) reflection; Its reflected light promptly produces fluorescence through object lens (34) behind integrated PCR-CE chip (14), fluorescence is gone into photoelectricity multiplier tube (38) by object lens (34), half-reflecting half mirror (33), convex lens (35), pin hole (36) and optical filter (37) are laggard again successively, detects fluorescence signal.
5. by the described laser-induced fluorescence analyzer of claim 1, it is characterized in that:
Signals collecting and processing unit (40) comprise preceding latter linked signals collecting, filtering, amplification, analog to digital conversion circuit (41) and computing machine (42).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102728741A CN101718698B (en) | 2009-11-20 | 2009-11-20 | Laser-induced fluorescence analyzer with PCR-CE coupled microfluidic chip |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102728741A CN101718698B (en) | 2009-11-20 | 2009-11-20 | Laser-induced fluorescence analyzer with PCR-CE coupled microfluidic chip |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101718698A true CN101718698A (en) | 2010-06-02 |
| CN101718698B CN101718698B (en) | 2011-08-17 |
Family
ID=42433303
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009102728741A Expired - Fee Related CN101718698B (en) | 2009-11-20 | 2009-11-20 | Laser-induced fluorescence analyzer with PCR-CE coupled microfluidic chip |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101718698B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103063646A (en) * | 2013-01-14 | 2013-04-24 | 武汉大学 | Dual-purpose laser-induced fluorescence detecting system |
| CN103194383A (en) * | 2013-04-09 | 2013-07-10 | 湖北民族学院 | Chip level PCR-LVCE integrated system |
| CN104007091A (en) * | 2013-02-26 | 2014-08-27 | 中国科学院天津工业生物技术研究所 | High-throughput detection system for microbe based on droplet microfluidic chip |
| CN106906137A (en) * | 2017-03-19 | 2017-06-30 | 北京化工大学 | A kind of high-throughput nucleic acid analysis device based on micro-fluidic chip |
| CN107338189A (en) * | 2011-05-04 | 2017-11-10 | 卢米耐克斯公司 | Apparatus and method for for integrated sample preparation, reaction and detection |
| CN107807114A (en) * | 2017-09-12 | 2018-03-16 | 华南师范大学 | Gene based on functionalization capillary is quick, quantified system analysis and method |
| CN111610173A (en) * | 2020-05-27 | 2020-09-01 | 中国水利水电科学研究院 | Three-dimensional fluid concentration field calibration device and calibration method |
-
2009
- 2009-11-20 CN CN2009102728741A patent/CN101718698B/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107338189A (en) * | 2011-05-04 | 2017-11-10 | 卢米耐克斯公司 | Apparatus and method for for integrated sample preparation, reaction and detection |
| CN103063646A (en) * | 2013-01-14 | 2013-04-24 | 武汉大学 | Dual-purpose laser-induced fluorescence detecting system |
| CN103063646B (en) * | 2013-01-14 | 2015-04-29 | 武汉大学 | Dual-purpose laser-induced fluorescence detecting system |
| CN104007091A (en) * | 2013-02-26 | 2014-08-27 | 中国科学院天津工业生物技术研究所 | High-throughput detection system for microbe based on droplet microfluidic chip |
| CN103194383A (en) * | 2013-04-09 | 2013-07-10 | 湖北民族学院 | Chip level PCR-LVCE integrated system |
| CN103194383B (en) * | 2013-04-09 | 2014-09-03 | 湖北民族学院 | Chip level PCR-LVCE integrated system |
| CN106906137A (en) * | 2017-03-19 | 2017-06-30 | 北京化工大学 | A kind of high-throughput nucleic acid analysis device based on micro-fluidic chip |
| CN106906137B (en) * | 2017-03-19 | 2019-02-12 | 北京化工大学 | A high-throughput nucleic acid analysis device based on microfluidic chip |
| CN107807114A (en) * | 2017-09-12 | 2018-03-16 | 华南师范大学 | Gene based on functionalization capillary is quick, quantified system analysis and method |
| CN111610173A (en) * | 2020-05-27 | 2020-09-01 | 中国水利水电科学研究院 | Three-dimensional fluid concentration field calibration device and calibration method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101718698B (en) | 2011-08-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101718698B (en) | Laser-induced fluorescence analyzer with PCR-CE coupled microfluidic chip | |
| Gong et al. | New advances in microfluidic flow cytometry | |
| CN201581079U (en) | Polymerase chain reaction-capillary electrophoresis combined micro-fluidic chip laser-induced fluorescence analyzing device | |
| US8053214B2 (en) | Apparatus and method of extracting and optically analyzing an analyte from a fluid-based sample | |
| Liu et al. | Chemiluminescence detection for a microchip capillary electrophoresis system fabricated in poly (dimethylsiloxane) | |
| Haab et al. | Single-molecule detection of DNA separations in microfabricated capillary electrophoresis chips employing focused molecular streams | |
| Sandlin et al. | Microfluidic electrophoresis chip coupled to microdialysis for in vivo monitoring of amino acid neurotransmitters | |
| CN102083533B (en) | Microfluidic chip devices and their use | |
| CN101614655B (en) | Detection method of microfluidic chip based on photoacoustic technique and device thereof | |
| CN201548547U (en) | Flow cell analysis device based on microfluidic chip | |
| JP2002536962A (en) | Integrated portable biological detection system | |
| JP2006504957A (en) | Microfluidic system for analyzing nucleic acids | |
| US20120082985A1 (en) | Sensing And Identifying Biological Samples On Microfluidic Devices | |
| CN101692047A (en) | Microfluidic chip for capillary electrophoresis separation and chemiluminescence detection | |
| Kim et al. | Microfluidic pipette tip for high-purity and high-throughput blood plasma separation from whole blood | |
| WO2012020257A1 (en) | Method and system for analyzing a sample | |
| CN103667012A (en) | Microfluidic PCR (Polymerase Chain Reaction) chip fluorescence fluid detection device based on CCD (Charge Coupled Device) image sensor | |
| CN101943697A (en) | Blood examination method, system and microchip | |
| JP5011313B2 (en) | Proteolytic method and kit using polymethine marker dye | |
| WO2021239010A1 (en) | Cell sorting chip, apparatus, and method based on dielectric deterministic displacement | |
| CN205556678U (en) | A single cell sorting device based on a microfluidic chip | |
| Xu et al. | Electrochemical and optical detectors for capillary and chip separations | |
| EP2049901B1 (en) | Method for analyzing samples including a gas evolving means | |
| CN201517993U (en) | Micro-fluidic chip detecting device based on photoacoustic technique | |
| Nguyen et al. | On-line dual-stage enrichment via magneto-extraction and electrokinetic preconcentration: A new concept and instrumentation for capillary electrophoresis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: GENEINN BIOTECHNOLOGY (NINGBO) CO., LTD. Free format text: FORMER OWNER: NINGBO POCEL MICROFLUIDICS CO., LTD. Effective date: 20110418 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20110418 Address after: West Street in the official Zhejiang city of Ningbo province Zhenhai District 315200 Village No. 777 Applicant after: Geneinn Biotechnology (Ningbo) Co., Ltd. Address before: West Street in the official Zhejiang city of Ningbo province Zhenhai District 315200 Village No. 777 Applicant before: Ningbo Pusai Microfluidic Technology Co., Ltd. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110817 Termination date: 20111120 |