CN107817235A - A kind of real time fluorescent quantitative augmentation detection instrument - Google Patents
A kind of real time fluorescent quantitative augmentation detection instrument Download PDFInfo
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- CN107817235A CN107817235A CN201711310372.4A CN201711310372A CN107817235A CN 107817235 A CN107817235 A CN 107817235A CN 201711310372 A CN201711310372 A CN 201711310372A CN 107817235 A CN107817235 A CN 107817235A
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- 238000001514 detection method Methods 0.000 title claims abstract description 26
- 230000003416 augmentation Effects 0.000 title claims abstract description 20
- 230000003287 optical effect Effects 0.000 claims abstract description 25
- 238000010438 heat treatment Methods 0.000 claims abstract description 20
- 230000003321 amplification Effects 0.000 claims abstract description 17
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 17
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 15
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 15
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 15
- 239000013307 optical fiber Substances 0.000 claims abstract description 10
- 239000004065 semiconductor Substances 0.000 claims abstract description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 14
- NJPPVKZQTLUDBO-UHFFFAOYSA-N novaluron Chemical compound C1=C(Cl)C(OC(F)(F)C(OC(F)(F)F)F)=CC=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F NJPPVKZQTLUDBO-UHFFFAOYSA-N 0.000 claims description 10
- 229910052736 halogen Inorganic materials 0.000 claims description 6
- 150000002367 halogens Chemical class 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 4
- 238000003491 array Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 2
- 229910052724 xenon Inorganic materials 0.000 claims description 2
- FHNFHKCVQCLJFQ-UHFFFAOYSA-N xenon atom Chemical compound [Xe] FHNFHKCVQCLJFQ-UHFFFAOYSA-N 0.000 claims description 2
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 230000007306 turnover Effects 0.000 abstract description 2
- 238000010276 construction Methods 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000000835 fiber Substances 0.000 description 3
- 239000004606 Fillers/Extenders Substances 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000005611 electricity Effects 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 230000009471 action Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000005357 flat glass Substances 0.000 description 1
- 230000017525 heat dissipation Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Optics & Photonics (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The present invention relates to a kind of real time fluorescent quantitative augmentation detection instrument, comprising casing, controller, is arranged on the turnover case lid in casing upper end, the nucleic acid amplification module being separately positioned in casing and fluoroscopic examination module；The nucleic acid amplification module include be sequentially placed and be bonded to each other from top to bottom 96 hole heating plates, semiconductor chilling plate, heat conducting pipe, radiator, blower fan；The fluoroscopic examination module includes frame, the UV IR filters that are separately positioned in frame and focus lamp, is horizontally set on frame upper end and is arranged on that the optical filter, two beam one end that circumferentially direction is uniformly placed on rotating disk and wavelength is different are arranged on above rotating disk and the other end is connected with 96 hole heating plates and extends to the incident optical beam and the output optical fiber beam, the light source and fluorescence detector that are separately positioned on frame lower end of each hole wall respectively by stepper motor driven rotating disk, multi-disc；The present invention can not only improve detection sensitivity, reduce manufacturing cost, and simple in construction, easy to use.
Description
Technical field
The present invention relates to PCR augmentation detections field, refers in particular to a kind of real time fluorescent quantitative augmentation detection instrument.
Background technology
Nucleic acid amplification fluorescent detecting system is the reagent being irradiated to exciting light in sample cell, is then received out of sample cell
The fluorescence that reagent moment inspires, to realize sample is qualitative and quantitatively detects system.
At present, existing nucleic acid amplification fluorescent detecting system, which is substantially, is irradiated to exciting light at the top of sample cell, sample
After reagent excites fluorescence in pipe, then the fluorescence excited through returning to reception at the top of sample cell, such as prior art 201710286911.9
In disclose a kind of real-time fluorescence quantitative PCR gene magnification detector, in use, have the disadvantages that：1st, caused by light source
For light after semi-transparency type filter and optical filter, the spectrum for forming particular color passes through the sheet glass in through hole to enter in reagent wells,
Default fluorescent material is absorbed into the light in reagent wells in reagent wells, the light that fluorescent material is sent by through hole through optical filter and
Semi-transparency type filter transmission and reflection is into fluorescence collector, because the fluorescence excited will connect again after the scattering at the top of reagent wells
Receive, light intensity has greater loss, causes the sensitivity of detection to decline；2nd, pan is engaged with connecting hole so that fluoroscopic examination module
Gene magnification module can be surrounded to overturn, be easy to fluoroscopic examination module to flip over so that it is very laborious during replacing reagent, and turn over
Turning number excessively easily causes the reduction of fluoroscopic examination module precision；3rd, light caused by light source is after semi-transparency type filter, then passes through
The spectrum that optical filter forms particular color enters in reagent wells, and device for revolving and driving rotates, and the optical filter of different colours is right successively
Accurate reagent wells circumferentially, because each passage is required for setting semi-transparency type filter and optical filter, considerably increases and be manufactured into
This.
The content of the invention
A kind of real time fluorescent quantitative augmentation detection instrument is provided the invention aims to overcome the deficiencies in the prior art, no
It is only capable of putting forward detection sensitivity, reduces manufacturing cost, and be simple in structure and convenient in use.
To reach above-mentioned purpose, the technical solution adopted by the present invention is：A kind of real time fluorescent quantitative augmentation detection instrument, comprising
Casing, controller, the case lid that is arranged on casing upper end and can flip over, it is separately positioned on what is be connected with controller line in casing
Nucleic acid amplification module and fluoroscopic examination module；The nucleic acid amplification module include be placed in below case lid pedestal, from top to bottom according to
The secondary 96 hole heating plates for being arranged on pedestal and being bonded to each other, semiconductor chilling plate, heat conducting pipe, radiator, blower fan；It is described
Fluoroscopic examination module includes frame, the UV-I R filters being separately positioned in frame and focus lamp, is horizontally set on frame upper end
And the optical filtering that circumferentially direction is uniformly placed on rotating disk and wavelength is different is arranged on by stepper motor driven rotating disk, multi-disc
Piece, two beam one end are arranged on above rotating disk and the other end is connected with 96 hole heating plates and extends to entering for each hole wall respectively
Penetrate fibre bundle and the output optical fiber beam, the light source and fluorescence detector that are separately positioned on frame lower end；The light that the light source is sent according to
It is secondary by entering 96 hole heating plates after UV-IR filters, wherein a tablet filter, incident optical beam, what tested reagent excited
Fluorescence enters fluorescence detector after passing sequentially through the output optical fiber beam, wherein another tablet filter, focus lamp.
Preferably, the frame includes lower cover, is arranged on the upper shell cover of lower cover upper end, is arranged on lower cover lower end
Support；The UV-IR filters and focus lamp are arranged on lower cover, and rotating disk is arranged between upper shell cover and lower cover, and two beams enter
The one end and stepper motor for penetrating fibre bundle and the output optical fiber beam are separately positioned on upper shell cover, and light source and fluorescence detector are set respectively
Put on support；The upper and lower ends of the rotating disk are respectively arranged with and upper shell cover and lower casing in the circumference that multiple optical filters are formed
The annular retaining ring that lid gap is placed.
Preferably, the semiconductor chilling plate is provided with multi-disc, and arranged in arrays.
Preferably, horizontal positioned heating cover plate is provided with the case lid above pedestal.
Preferably, it is in three-back-shaped sealing ring to be additionally provided with a circle at the edge of heating cover plate on the case lid.
Preferably, it is additionally provided with the casing and gives the switch of nucleic acid amplification module and fluoroscopic examination module for power supply electricity respectively
Source.
Preferably, the both sides of the casing are respectively arranged with radiator window.
Preferably, the light source is one kind in LED, xenon lamp, Halogen lamp LED or LASER Light Source.
Preferably, the fluorescence detector is one kind in CCD-detector, photodiode or PMT photomultipliers.
Due to the utilization of above-mentioned technical proposal, the present invention has following advantages compared with prior art：
Real time fluorescent quantitative augmentation detection instrument of the present invention at hole wall position by that directly can realize exciting for fluorescence
And reception, the phenomenons of fluorescent scattering and light intensity decays is removed, improves the detection sensitivity of fluorescence quantitative gene extender；And core
Acid amplification module is set directly at below case lid, and sample cell can directly be changed by opening case lid, and use is more convenient；And multi-disc is not
The optical filter of co-wavelength is arranged on rotating disk simultaneously, it is only necessary to is according to optical filter corresponding to fluorescent dye rotation rotating disk selection
Can, it is not only simple in structure, purposes is wider, and can reduce manufacturing cost.
Brief description of the drawings
Technical solution of the present invention is described further below in conjunction with the accompanying drawings：
Accompanying drawing 1 is the structural representation of real time fluorescent quantitative augmentation detection instrument of the present invention；
Accompanying drawing 2 removes the structural representation after casing and case lid for the present invention；
Accompanying drawing 3 is the structural representation of fluoroscopic examination module in the present invention；
Accompanying drawing 4 is the sectional view of fluoroscopic examination module in the present invention；
Accompanying drawing 5 is the sectional view that amplifying nucleic acid of the present invention expands module；
Accompanying drawing 6 is the light path schematic diagram in single hole in 96 hole heating plates in the present invention.
Wherein：1st, casing；11st, radiator window；2nd, case lid；21st, cover plate is heated；22nd, sealing ring；3rd, nucleic acid amplification module；31、
Pedestal；32nd, 96 hole heating plate；33rd, semiconductor chilling plate；34th, heat conducting pipe；35th, radiator；36th, blower fan；4th, fluoroscopic examination
Module；40th, frame；401st, lower cover；402nd, support；403rd, upper shell cover；41st, incident optical beam；42nd, the output optical fiber beam；43rd, turn
Disk；431st, annular retaining ring；44th, optical filter；45th, Halogen lamp LED；46th, CCD-detector；47th, stepper motor；48th, UV-I R filters；
49th, focus lamp；5th, Switching Power Supply；6th, sample cell.
Embodiment
Below in conjunction with the accompanying drawings and specific embodiment the present invention is described in further detail.
Accompanying drawing 1-6 is real time fluorescent quantitative augmentation detection instrument of the present invention, comprising casing 1, controller, is arranged on case
The upper end of body 1 and can flip over case lid 2, be separately positioned on the nucleic acid amplification module 3 that is connected with controller line in casing 1 and glimmering
Light detection module 4；The nucleic acid amplification module 3 includes being placed in the pedestal 31 of the lower section of case lid 2, is successively set on pedestal from top to bottom
On 31 and be bonded to each other 96 hole heating plates 32, semiconductor chilling plate 33, heat conducting pipe 34, radiator 35, blower fan 36；It is described
Fluoroscopic examination module 4 include be made up of upper shell cover 403, lower cover 401, support 402 frame 40, be separately positioned on lower cover
UV-I R filters 48 and focus lamp 49 on 401, it is horizontally set between upper shell cover 403 and lower cover 401 and by stepping electricity
The rotating disk 43 of the driving of machine 47, multi-disc are arranged on the optical filter 44, two that circumferentially direction is uniformly placed on rotating disk 43 and wavelength is different
Shu Yiduan is arranged on upper shell cover 403 and the other end is connected with 96 hole heating plates 32 and extends to entering for each hole wall respectively
The Halogen lamp LED 45 and CCD-detector 46 for penetrate fibre bundle 41 and the output optical fiber beam 42, being separately positioned on support 402；The halogen
The light that lamp 45 is sent enters the heating of 96 holes after passing sequentially through UV-IR filters 48, wherein a tablet filter, incident optical beam 41
Plate 32, the fluorescence that reagent excites in detected sample QC 6 pass sequentially through the output optical fiber beam 42, wherein another tablet filter, focus lamp
Enter CCD-detector 46 after 49.
Further, the upper and lower ends of the rotating disk 43 are respectively arranged with and upper shell cover in the circumference that multiple optical filters are formed
403 and the gap of lower cover 401 place annular retaining ring 431, play every light action, the interference of light CCD for preventing Halogen lamp LED 45 from sending
Detector 46 receives fluorescence, causes measurement inaccurate.
Further, the semiconductor chilling plate 33 is provided with multi-disc, and arranged in arrays, with per it is a piece of be a region,
Realize multi-region temperature control technique, can effectively improve the uniformity of accuracy of temperature control and temperature, and can gradient set, purposes is wider.
Further, horizontal positioned heating cover plate 21 is provided with the case lid 2 in the top of pedestal 31 so that temperature is more
Add uniformly.
Further, it is in three-back-shaped sealing ring 22 to be additionally provided with a circle at the edge of heating cover plate 21 on the case lid 2, is prevented
Only reagent volatilizees in heating, and amplification efficiency reduces.
Further, it is additionally provided with the casing 1 and is opened respectively to what nucleic acid amplification module 3 and fluoroscopic examination module 4 were powered
Powered-down source 5, avoid powering off suddenly in the course of the work, it is necessary to detect again, not only influence testing result, also greatly increases inspection
Survey the time.
Further, the both sides of the casing 1 are respectively arranged with radiator window 11, play heat dissipation ventilation effect, strengthen radiating effect
Fruit.
Due to the utilization of above-mentioned technical proposal, the present invention has following advantages compared with prior art：
Real time fluorescent quantitative augmentation detection instrument of the present invention at hole wall position by that directly can realize exciting for fluorescence
And reception, the phenomenons of fluorescent scattering and light intensity decays is removed, improves the detection sensitivity of fluorescence quantitative gene extender；And core
Acid amplification module is set directly at below case lid, and sample cell can directly be changed by opening case lid, and use is more convenient；And multi-disc is not
The optical filter of co-wavelength is arranged on rotating disk simultaneously, it is only necessary to is according to optical filter corresponding to fluorescent dye rotation rotating disk selection
Can, it is not only simple in structure, purposes is wider, and can reduce manufacturing cost.
It the above is only the concrete application example of the present invention, protection scope of the present invention be not limited in any way.All uses
Equivalent transformation or equivalent replacement and the technical scheme formed, all fall within rights protection scope of the present invention.
Claims (9)
- A kind of 1. real time fluorescent quantitative augmentation detection instrument, it is characterised in that：Comprising casing, controller, be arranged on casing upper end and The case lid that can flip over, it is separately positioned on the nucleic acid amplification module and fluoroscopic examination module being connected in casing with controller line； The nucleic acid amplification module includes the pedestal being placed in below case lid, be successively set on pedestal and be bonded to each other from top to bottom nine 16 hole heating plates, semiconductor chilling plate, heat conducting pipe, radiator, blower fan；The fluoroscopic examination module includes frame, set respectively The UV-I R filters in frame and focus lamp are put, is horizontally set on frame upper end and by stepper motor driven rotating disk, more Piece is arranged on the optical filter that circumferentially direction is uniformly placed on rotating disk and wavelength is different, two beam one end are arranged on rotating disk top and another One end is connected with 96 hole heating plates and extends to the incident optical beam of each hole wall and the output optical fiber beam, set respectively respectively Light source and fluorescence detector in frame lower end；The light that the light source is sent passes sequentially through UV-I R filters, wherein a piece of optical filtering Enter 96 hole heating plates after piece, incident optical beam, the fluorescence that tested reagent excites passes sequentially through the output optical fiber beam, wherein another Enter fluorescence detector after one tablet filter, focus lamp.
- 2. real time fluorescent quantitative augmentation detection instrument according to claim 1, it is characterised in that：The frame includes lower casing The support cover, be arranged on the upper shell cover of lower cover upper end, being arranged on lower cover lower end；The UV-I R filters and focus lamp are set On lower cover, rotating disk is arranged between upper shell cover and lower cover, one end and the step of two beam incident optical beams and the output optical fiber beam Stepper motor is separately positioned on upper shell cover, and light source and fluorescence detector are separately positioned on support；The upper and lower ends of the rotating disk The annular retaining ring with upper shell cover and the placement of lower cover gap is respectively arranged with the circumference that multiple optical filters are formed.
- 3. real time fluorescent quantitative augmentation detection instrument according to claim 1 or 2, it is characterised in that：The semiconductor refrigerating Piece is provided with multi-disc, and arranged in arrays.
- 4. real time fluorescent quantitative augmentation detection instrument according to claim 3, it is characterised in that：In pedestal on the case lid Top is provided with horizontal positioned heating cover plate.
- 5. real time fluorescent quantitative augmentation detection instrument according to claim 4, it is characterised in that：In heating cover on the case lid It is in three-back-shaped sealing ring that the edge of plate, which is additionally provided with a circle,.
- 6. real time fluorescent quantitative augmentation detection instrument according to claim 1, it is characterised in that：It is additionally provided with the casing Respectively to nucleic acid amplification module and the Switching Power Supply of fluoroscopic examination module for power supply.
- 7. real time fluorescent quantitative augmentation detection instrument according to claim 1, it is characterised in that：The both sides difference of the casing It is provided with radiator window.
- 8. real time fluorescent quantitative augmentation detection instrument according to claim 1, it is characterised in that：The light source is LED, xenon One kind in gas lamp, Halogen lamp LED or LASER Light Source.
- 9. real time fluorescent quantitative augmentation detection instrument according to claim 1, it is characterised in that：The fluorescence detector is One kind in CCD-detector, photodiode or PMT photomultipliers.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711310372.4A CN107817235A (en) | 2017-12-11 | 2017-12-11 | A kind of real time fluorescent quantitative augmentation detection instrument |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711310372.4A CN107817235A (en) | 2017-12-11 | 2017-12-11 | A kind of real time fluorescent quantitative augmentation detection instrument |
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| Publication Number | Publication Date |
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| CN107817235A true CN107817235A (en) | 2018-03-20 |
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| CN201711310372.4A Pending CN107817235A (en) | 2017-12-11 | 2017-12-11 | A kind of real time fluorescent quantitative augmentation detection instrument |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109971643A (en) * | 2019-05-08 | 2019-07-05 | 中国人民解放军军事科学院军事医学研究院 | Portable constant temperature augmentation detection instrument |
| CN110161007A (en) * | 2019-06-03 | 2019-08-23 | 杭州安誉科技有限公司 | A kind of optical detection apparatus for the synchronous detection of multi-wavelength fluorescence |
| CN110218647A (en) * | 2019-06-27 | 2019-09-10 | 杭州安塔生物科技有限公司 | A kind of PCR reaction unit |
| CN110656046A (en) * | 2019-11-01 | 2020-01-07 | 中国人民解放军军事科学院军事医学研究院 | Detection device and PCR instrument |
| CN110699249A (en) * | 2019-11-01 | 2020-01-17 | 宁波胤瑞生物医学仪器有限责任公司 | Nucleic acid amplification instrument |
| CN111949055A (en) * | 2020-08-03 | 2020-11-17 | 中国科学院长春光学精密机械与物理研究所 | Independent temperature control system and method for microfluidic culture chip |
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|---|---|---|---|---|
| CN109971643A (en) * | 2019-05-08 | 2019-07-05 | 中国人民解放军军事科学院军事医学研究院 | Portable constant temperature augmentation detection instrument |
| CN110161007A (en) * | 2019-06-03 | 2019-08-23 | 杭州安誉科技有限公司 | A kind of optical detection apparatus for the synchronous detection of multi-wavelength fluorescence |
| CN110218647A (en) * | 2019-06-27 | 2019-09-10 | 杭州安塔生物科技有限公司 | A kind of PCR reaction unit |
| CN110656046A (en) * | 2019-11-01 | 2020-01-07 | 中国人民解放军军事科学院军事医学研究院 | Detection device and PCR instrument |
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| CN110699249B (en) * | 2019-11-01 | 2021-08-06 | 宁波胤瑞生物医学仪器有限责任公司 | Nucleic acid amplification instrument |
| CN111949055A (en) * | 2020-08-03 | 2020-11-17 | 中国科学院长春光学精密机械与物理研究所 | Independent temperature control system and method for microfluidic culture chip |
| CN111949055B (en) * | 2020-08-03 | 2021-10-26 | 中国科学院长春光学精密机械与物理研究所 | Independent temperature control system and method for microfluidic culture chip |
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