CN107991486A - A kind of liquid-phase chip detection system and method based on quantum dot - Google Patents
A kind of liquid-phase chip detection system and method based on quantum dot Download PDFInfo
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- CN107991486A CN107991486A CN201711146684.6A CN201711146684A CN107991486A CN 107991486 A CN107991486 A CN 107991486A CN 201711146684 A CN201711146684 A CN 201711146684A CN 107991486 A CN107991486 A CN 107991486A
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- G—PHYSICS
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- 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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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Abstract
The invention discloses a kind of liquid-phase chip detection system and method based on quantum dot, the liquid-phase chip detection system includes liquid-phase chip, sheath stream forms module, fluorescence excitation light source, optical signalling detection device and circuit module；Wherein, the liquid-phase chip is the quantum dot coding fluorescence microballoon by chemical modification；The sheath stream, which forms module, includes microchannel, is wrapped liquid-phase chip by sheath fluid, and forms laminar condition in the flow chamber, and microchannel is passed sequentially through with single file streamlined flow, and is detected by optical signalling detection device, obtains fluorescence signal；The circuit module is microprocessor and signal processing circuit.System of the present invention has that structure is simple and easy to implement, and fluoroscopic examination precision is high, avoid because transmitting light it is overlapping caused by interference to testing result, have the characteristics that accuracy height, high sensitivity and cost are low, there is extensive market application foreground.
Description
Technical field
The present invention relates to biomolecular structure detection field, more particularly to a kind of liquid-phase chip detection system based on quantum dot
System and method.
Background technology
Liquid-phase chip technology is using microballoon as reaction carriers, in the liquid-phase reaction system close to biosystem internal environment
Carry out the detection of protein, the large biological molecule such as nucleic acid, can one-time detection many index, complete the purpose of Multiple detection.
Commercialized liquid-phase chip detection system is using the xMAP technologies of Luminex companies of the U.S. as core currently on the market,
The polystyrene microsphere that diameter is about 5.6um is mixed by a certain percentage it is red with infrared two kinds of different organic fluorescent dyes,
And the coding fluorescence and reporter fluorescence of microballoon are detected at the same time using two beam laser, red laser is used to identify microballoon kind
Class, green laser is used for the power of examining report fluorescence signal, so that it is determined that the detectable substance type and quantity being combined, it is fixed to complete
Property and quantitative analysis.But need to be detected using two-beam source in the above method, its complex steps, efficiency is low.
It is mainly multiple rows of continually by wide microchannel by microballoon in existing liquid-phase chip detection system, use
More CCD or CMOS or light spectrum image-forming intake different wave length fluorescence, by sample of high-resolution image card to image at
Reason and image processing software analyze view data.
There are following defect in above-mentioned technical proposal：
1st, wide microchannel makes microballoon is multiple rows of to continue through detection zone, transmitting light have it is overlapping, it is easy to testing result
Interfere, greatly reduce sensitivity and the precision of detection.
2nd, the view data detected by imaging devices such as high-resolution acquisition card acquisition CMOS or CCD, and pass through meter
Calculation machine carries out processing analysis, its space structure is complicated, requires computer hardware height, and algorithm process is complicated, and detection efficiency is low, into
This height.
Therefore, the prior art has yet to be improved and developed.
The content of the invention
The problem of for present technology, mesh of the invention are to provide a kind of liquid-phase chip detection system based on quantum dot
And method, it is intended to overlapping transmitting light can influence the accuracy of testing result when solving to pass through detection zone using multiple rows of microballoon,
And the technical problem for causing hardware facility requirement height, algorithm complicated difficult to realize is detected using CMOS or CCD imaging devices.
To reach above-mentioned purpose, technical scheme is as follows：
A kind of liquid-phase chip detection system based on quantum dot, wherein, including liquid-phase chip, sheath stream formation module, fluorescence
Excitation source, optical signalling detection device and circuit module；Wherein, the liquid-phase chip is the quantum dot by chemical modification
Coding fluorescence microballoon, for being coupled sample to be tested and its reporter fluorescence molecule；The sheath stream, which forms module, includes flow chamber, described
Microchannel is provided with flow chamber, be additionally provided with for inject the sheath stream pipeline of sheath fluid (sheath fluid is formed in flow chamber) with
And for injecting the sample inlet of liquid-phase chip, the sheath fluid and liquid-phase chip form laminar condition in the flow chamber,
Microchannel is passed sequentially through with single file streamlined flow, and is detected by optical signalling detection device, obtains fluorescence signal；
The circuit module includes microprocessor and signal processing circuit.
The liquid-phase chip detection system based on quantum dot, wherein, the optical signalling detection device swashs including fluorescence
Light emitting source, condenser, spectroscope, the first optical filter, the second optical filter, photocell and photomultiplier；The fluorescence excitation
Light source excites the liquid-phase chip in microchannel by condenser, quantum dot coding fluorescence microballoon and sample to be tested is produced respectively
Fluorescence signal；The spectroscope is in 45 ° of settings, for fluorescence signal to be refracted in the first optical filter and the second optical filter；Institute
State the coding fluorescence signal that photocell receives quantum dot coding fluorescence microballoon by the first optical filter；The photomultiplier passes through
Second optical filter receives the reporter fluorescent signal of sample to be tested.
The liquid-phase chip detection system based on quantum dot, wherein, in the liquid-phase chip, quantum dot coding fluorescence is micro-
A diameter of 0.2~20um of ball.
The liquid-phase chip detection system based on quantum dot, wherein, in the liquid-phase chip, quantum dot coding fluorescence is micro-
The launch wavelength of ball is 480~685nm.
The liquid-phase chip detection system based on quantum dot, wherein, the microchannel is single microchannel, its width
For 0.2-3mm.
The liquid-phase chip detection system based on quantum dot, wherein, the microchannel is made of quartz material.
The liquid-phase chip detection system based on quantum dot, wherein, the fluorescence excitation light source is laser.
A kind of liquid-phase chip detection method based on quantum dot, wherein, comprise the following steps：
A, formed in sheath stream in module：By sheath stream pipeline add sheath fluid, and by sample inlet add coupling have it is to be measured
The liquid-phase chip of sample and its reporter fluorescence molecule, makes the sheath fluid that sheath fluid is formed and liquid-phase chip forming layer in flow chamber
Stream mode, passes through optical signalling detection device successively with single file streamlined flow；
B, in optical signalling detection device：Liquid-phase chip is excited by fluorescence excitation light source, makes quantum dot coding fluorescence
Microballoon produces coding fluorescence signal, sample to be tested is produced reporter fluorescent signal；And coding fluorescence signal and reporter fluorescence are believed
Number be converted to electric signal；
C, in circuit module：Signal processing circuit by amplifying electric signal, and through analog-to-digital conversion circuit send to
Handled in microprocessor, obtain the species and concentration value of sample to be tested.
Beneficial effect：The present invention uses single beam laser while the report of excitation quantum point coding microball fluorescence and molecules of interest
Fluorescence, the complexity of optical system substantially reduced；Sheath liquid syringe (is used to add sheath fluid, and sheath is formed in flow chamber
Fluid) and sample injector (liquid-phase chip for being used to add coupling sample to be tested) by certain speed ratio cause liquid-phase chip and sheath
Fluid is single by laser detection region successively with laminar condition in flow chamber, and speed is per second up to 0.5ul, per second by several
Thousand arrive a microballoons up to ten thousand so that testing result is more accurate, and reporter fluorescence is not easy overlapping；And the present invention only passes through microcontroller
Device processing fluorescence signal can obtain data, reduce workload, avoid the computer hardware requirement of follow-up complexity, effectively reduce setting
Standby cost, has high practicality and application prospect.
Brief description of the drawings
Fig. 1 is the structure diagram of the liquid-phase chip detection system of the present invention based on quantum dot.
Embodiment
To make the objects, technical solutions and advantages of the present invention clearer, clear and definite, develop simultaneously embodiment pair referring to the drawings
The present invention is further described.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and do not have to
It is of the invention in limiting.
As shown in Figure 1, the present invention provides a kind of liquid-phase chip detection system based on quantum dot, it include liquid-phase chip,
Sheath stream forms module, fluorescence excitation light source, optical signalling detection device and circuit module.Liquid-phase chip detection of the present invention
System is by technology of quantum dots, using single beam laser while excitation quantum point coding fluorescence microballoon and sample to be tested (purpose minute
Son), the two is produced fluorescence signal, and located by circuit module (being microprocessor and signal processing circuit in the present invention)
Reason, is converted to electric signal, so as to quickly detect the type and quantity of sample to be tested by fluorescence signal.
The improvement of liquid-phase chip detection system of the present invention has two：First, using single microchannel 103, with laminar flow shape
Form has sample to be tested by coupling one by one successively and its reports the liquid-phase chip of molecule, reduces the interference to testing result, has
Effect improves sensitivity and the precision of detection；Second, using microprocessor and signal processing circuit as circuit module to acquisition
Fluorescent signal data is handled, and structure is simple and easy to implement for it, effectively reduces equipment cost.
Specifically, liquid-phase chip of the present invention is the quantum dot coding fluorescence microballoon by chemical modification, for embedding
Sample to be tested.More specifically, a diameter of 0.2~20um of the quantum dot coding fluorescence microballoon, its launch wavelength for 480~
685nm.Quantum dot is narrow due to having the characteristics that fluorescent stabilization, exciting light spectrum width, emission spectrum, and the present invention is using in microballoon
Middle embedded quantum dots, are made quantum dot coding fluorescence microballoon, only need to use a branch of light source excite at the same time microballoon coding fluorescence and
Reporter fluorescence, you can detect the fluorescence signal of quantum dot coding fluorescence microballoon and sample to be tested.
In the present embodiment, the sheath stream, which forms module, includes flow chamber 10, the microchannel being connected with flow chamber 10
103.The microchannel 103 is made of quartz material.The sample to be tested that the flow chamber 10 is used to focus in sheath fluid is (even
It is associated in liquid-phase chip), sheath stream pipeline 101 for injecting sheath fluid is additionally provided with the flow chamber 10 and for injecting liquid
The sample inlet 102 of phase chip (coupling has sample to be tested and its reporter fluorescence molecule).Liquid-phase chip is clamped or surrounded to sheath fluid,
It is tapered to microsphere diameter size on the direction of flowing, and laminar condition is formed in the flow chamber 10, with list
Row streamlined flow passes sequentially through microchannel 103, and is detected by optical signalling detection device, obtains fluorescence signal.
Sheath fluid and sample to be tested are injected using propulsion device for injector in the embodiment of the present invention, in addition it can also pass through
Pneumatic shuttle replaces propulsion device for injector, and the propulsion effect of pneumatic shuttle is relatively stable, can reach and uniformly produce sheath fluid
Effect.Preferably, the embodiment of the present invention can control each valve switch and two syringes to push away by fluid path control device
Into the motor movement speed of device and time, 20um or so sheath fluid is produced according to fluid focus principle and (sheath is formed in flow chamber
Fluid) parcel sample flow (coupling has the liquid-phase chip of sample to be tested), sheath fluid and sample flow are in stable laminar flow at this time
State, streamlined flow is sequentially transmitted microchannel to the sample to be tested of modified quantum dot coding fluorescence microballoon capture in single file
It is detected in 103.
In the present embodiment, the microchannel 103 is single microchannel 103, its width is 0.2-3mm, in same a period of time
The liquid-phase chip number that interior each cross-sectional area unit passes through is 1, so it is possible to prevente effectively from overlapping, the increasing of fluorescence signal
Add the accuracy of detection.
Further, optical signalling detection device of the present invention includes two detection contents：Quantum point coding microball is glimmering
The fluorescence signal detection of light and the fluorescence signal of sample to be tested detect.Specifically, including fluorescence excitation light source, condenser 21, point
Light microscopic 22, the first optical filter 24, the second optical filter 23, photocell 26 and photomultiplier 25.The fluorescence excitation light source leads to
The liquid-phase chip in the excitation microchannel 103 of condenser 21 is crossed, quantum dot coding fluorescence microballoon and sample to be tested is produced respectively
Fluorescence signal；The spectroscope 22 is in 45 ° of settings, for fluorescence signal to be refracted to the first optical filter 24 and the second optical filter 23
In；The photocell 26 receives the fluorescence signal of quantum dot coding fluorescence microballoon by the first optical filter 24；The photomultiplier transit
Pipe 25 receives the fluorescence signal of sample to be tested by the second optical filter 23.
In the above-described embodiments, fluorescence excitation light source selects laser 20, such as semiconductor laser 20, it is big in volume
Small, price, the type of cooling and preheating time are dominant.It is very fast additionally, due to sheath fluid velocity ratio, quantum dot coding fluorescence microballoon
There is direct relation with the strong and weak intensity with illuminated time and exciting light of fluorescence signal being excited in sample to be tested, therefore treat
Sample needs enough intensities of illumination to produce the fluorescence signal of sufficient intensity.Semiconductor laser as a kind of coherent source,
The illumination of single wavelength, high intensity and stabilization is capable of providing, ensures focusing illumination to the quantum dot microsphere being coupled with determinand,
Therefore the present invention uses excitation apparatus of the semiconductor laser as fluorescence signal.
In addition, heretofore described circuit module is microcontroller and signal processing circuit, including photocell 26 and light
Electric multiplier tube 25, by the way that the fluorescence signal detected is converted to electric signal, so as to calculate the species and concentration of sample to be tested.
It realizes that process is as follows：Semiconductor laser 20 produces laser, and the sheath fluid and liquid-phase chip in flow chamber 10 form laminar flow
State (individual layer state), and flow forward is single successively focuses on and excite by condenser 21 by single microchannel 103, laser
Liquid-phase chip in single microchannel 103 produces fluorescence signal, then is divided into by the spectroscope 22 that 45 ° of angles of inclination are set
Two-way fluorescence signal：One is the coding fluorescence signal of quantum dot coding fluorescence microballoon, is launched by first optical filter 24
Handled into photocell 26；Secondly be the reporter fluorescent signal of sample to be tested, by second optical filter 23 launch to
Handled in photomultiplier 25 (PMT), optical signalling (i.e. fluorescence signal) is converted into current signal (about uA grades), and
And current signal passes through current-voltage conversion process, the size by the intensity-conversion of two-way fluorescence signal into corresponding magnitude of voltage is defeated
Enter into analog-to-digital conversion circuit, realize the qualitatively and quantitatively detection to sample to be tested, detect sample to be tested species and
Concentration value.
In addition, the present invention also provides a kind of liquid-phase chip detection method based on quantum dot, it comprises the following steps：
A, formed in sheath stream in module：By sheath stream pipeline add sheath fluid, and by sample inlet add coupling have it is to be measured
The liquid-phase chip of sample and its reporter fluorescence molecule, makes the sheath fluid that sheath fluid is formed and liquid-phase chip forming layer in flow chamber
Stream mode, passes through optical signalling detection device successively with single file streamlined flow.
B, in optical signalling detection device：Liquid-phase chip is excited by fluorescence excitation light source, makes quantum dot coding fluorescence
Microballoon produces coding fluorescence signal, sample to be tested is produced reporter fluorescent signal；And coding fluorescence signal and reporter fluorescence are believed
Number be converted to electric signal.
C, in circuit module：Signal processing circuit by amplifying electric signal, and through analog-to-digital conversion circuit send to
Handled in microprocessor, obtain the species and concentration value of sample to be tested.
The operating process of liquid-phase chip detection method of the present invention based on quantum dot and the liquid-phase chip detection system
The detecting step of system is identical, and details are not described herein.
In conclusion a kind of liquid-phase chip detection system and method based on quantum dot of the present invention, it utilizes quantum
Point exciting light spectrum width, the excellent fluorescent characteristic such as emission spectrum is narrow, fluorescence intensity is strong, stability is good, make sample flow in sheath fluid band
Dynamic lower single file, which is stablized, quickly move through optical signalling detection device, and using single beam laser excitation quantum point coding fluorescence microballoon and
The fluorescence signal of sample to be tested, wherein photocell collect the coding fluorescence signal of quantum dot coding fluorescence microballoon, photomultiplier
The reporter fluorescent signal of sample to be tested is collected, and then realizes the qualitative and quantitative detection to sample to be tested.This hair system knot
Structure simply easily realizes that it has the characteristics that high accuracy, high sensitivity and cost are low, available for tumor-marker analyte detection, allergy
Former examination, infectious diseases detection etc..
It should be appreciated that the application of the present invention is not limited to above-mentioned citing, for those of ordinary skills, can
To be improved or converted according to the above description, all these modifications and variations should all belong to the guarantor of appended claims of the present invention
Protect scope.
Claims (8)
- A kind of 1. liquid-phase chip detection system based on quantum dot, it is characterised in that including liquid-phase chip, sheath stream formed module, Fluorescence excitation light source, optical signalling detection device and circuit module；Wherein,The liquid-phase chip is the quantum dot coding fluorescence microballoon by chemical modification, glimmering for being coupled sample to be tested and its report Optical molecule；The sheath stream, which forms module, includes flow chamber, and microchannel is provided with the flow chamber, is additionally provided with for injecting sheath The sheath stream pipeline of liquid and the sample inlet for injecting liquid-phase chip, the sheath fluid and liquid-phase chip that the sheath fluid is formed exist Laminar condition is formed in the flow chamber, microchannel is passed sequentially through with single file streamlined flow, and by optical signalling detection dress Put and be detected, obtain fluorescence signal；The circuit module includes microprocessor and signal processing circuit.
- 2. the liquid-phase chip detection system based on quantum dot according to claim 1, it is characterised in that the optical signalling inspection Surveying device includes fluorescence excitation light source, condenser, spectroscope, the first optical filter, the second optical filter, photocell and photomultiplier transit Pipe；The fluorescence excitation light source by condenser excite microchannel in liquid-phase chip, make quantum dot coding fluorescence microballoon and Sample to be tested produces fluorescence signal respectively；The spectroscope is in 45 ° of settings, for fluorescence signal to be refracted in the first optical filter and the second optical filter；The photocell receives the coding fluorescence signal of quantum dot coding fluorescence microballoon by the first optical filter；The photomultiplier receives the reporter fluorescent signal of sample to be tested by the second optical filter.
- 3. the liquid-phase chip detection system based on quantum dot according to claim 1, it is characterised in that the liquid-phase chip In, a diameter of 0.2~20um of quantum dot coding fluorescence microballoon.
- 4. the liquid-phase chip detection system based on quantum dot according to claim 3, it is characterised in that the liquid-phase chip In, the launch wavelength of quantum dot coding fluorescence microballoon is 480~685nm.
- 5. the liquid-phase chip detection system based on quantum dot according to claim 1, it is characterised in that the microchannel is Single microchannel, its width are 0.2-3mm.
- 6. the liquid-phase chip detection system based on quantum dot according to claim 1, it is characterised in that the flow chamber uses Quartz material is made.
- 7. the liquid-phase chip detection system based on quantum dot according to claim 2, it is characterised in that the fluorescent exciting Source is laser.
- 8. a kind of liquid-phase chip detection method based on quantum dot, it is characterised in that comprise the following steps：A, formed in sheath stream in module：Sheath fluid is added by sheath stream pipeline, and add coupling by sample inlet to have sample to be tested And its liquid-phase chip of reporter fluorescence molecule, the sheath fluid that sheath fluid is formed is formed laminar flow shape in flow chamber with liquid-phase chip State, passes through optical signalling detection device successively with single file streamlined flow；B, in optical signalling detection device：Liquid-phase chip is excited by fluorescence excitation light source, makes quantum dot coding fluorescence microballoon Coding fluorescence signal is produced, sample to be tested is produced reporter fluorescent signal；And coding fluorescence signal and reporter fluorescent signal are turned It is changed to electric signal；C, in circuit module：Signal processing circuit is sent to micro- place by amplifying electric signal, and through analog-to-digital conversion circuit Handled in reason device, obtain the species and concentration value of sample to be tested.
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| CN114137324A (en) * | 2021-11-19 | 2022-03-04 | 浙江工业大学 | Method for detecting electric field effect of quantum dots |
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