CN101699271A - Real-time fluorescence quantitative PCR excitation detection system - Google Patents
Real-time fluorescence quantitative PCR excitation detection system Download PDFInfo
- Publication number
- CN101699271A CN101699271A CN200910218601A CN200910218601A CN101699271A CN 101699271 A CN101699271 A CN 101699271A CN 200910218601 A CN200910218601 A CN 200910218601A CN 200910218601 A CN200910218601 A CN 200910218601A CN 101699271 A CN101699271 A CN 101699271A
- Authority
- CN
- China
- Prior art keywords
- optical filter
- wavelength
- tunable
- filter
- quantitative pcr
- 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.)
- Pending
Links
Images
Abstract
The invention provides a quantitative PCR detection system which is based on a tunable filter and can realize the full wavelength fluorescence excitation detection, and the quantitative PCR detection system comprises a fluorescence excitation module consisting of a light source, a light gathering device and a first adjustable filter, a fluorescence detection module consisting of a second adjustable filter, an optical detector and a photoelectric conversion device, a control unit consisting of a wavelength tuning device and a main control computer, and a heat circulation system consisting of a reaction test tube, a metal warm block and a constant temperature hot cover. The light gathering is carried out on excited light, then the wavelength section is carried out by the tunable filter, the excited fluorescence passes through the tunable filter, the components of the excited light and other interference signals are filtered, and then the optical detector and the photoelectric conversion device are used for delivering to the main control computer for carrying out data analysis.
Description
Technical field
The present invention relates to a kind of fluorescence quantitative PCR detection system, especially a kind of quantitative PCR detection system of realizing long fluorescence excitation of all-wave and detection (acousto-optic tunable, liquid crystal tunable) based on tunable optical filter.
Background technology
Real-time fluorescence quantitative PCR detection system principle of work is to make reactant carry out temperature cycles between specific denaturation temperature, renaturation temperature and elongating temperature, target DNA is carried out millions of times amplification; Simultaneously by the exciting light (exciting) of use different wave length or exciting light (singly exciting) the irradiation test tube of single wavelength more, when the reagent in the test tube is inspired fluorescence, collects fluorescence intensity signals by optical sensor and be sent to computing machine and carry out real-time data analysis.
At present the real-time fluorescence quantitative PCR detection system of main flow (comprise monochromaticly detect, double-colored detection and polychrome detection mode) is main excites with realization and detects by adopting different wave length optical filter (optical filter wheel) or grating to carry out color separation.
A kind of simply to utilize optical filter (optical filter wheel) to carry out fluorescence excitation detection mode groundwork process as follows: by the luminous fluorescence excitation that carries out of halogen tungsten lamp, the fluorescence that inspires arrives CCD by optical filter and detects, and logical filtering wheel carries out the differentiation of multiple fluorescent dye.For being focused into directional light, the light scioptics of LED emission utilize two to focus on less than the light scioptics of certain value to the dichronic mirror reflection wavelength and to excite on the test tube and utilize optical filter to carry out color separation or the monochromatic mode that detects more accurately again; The fluorescence scioptics that inspire are radiated on the dichroic mirror, see through the light of wavelength greater than certain value, are filtered into monochromatic light by optical filter again and are detected by photomultiplier.What the common ground of these two kinds of excitation-detection modes was same utilization is the filtration characteristic of optical filter to certain wavelength light, in case the centre wavelength of optical filter is fixed, the types of agents that detect etc. just need to change optical filter with regard to relative fixed if need the light of other wavelength of application to carry out excitation-detection.
Utilize grating to carry out the main of beam split color separation detection and utilize the mono-colour laser conduct to excite, realize that by 520～660nm grating the color separation simultaneously of the long fluorescence of all-wave utilizes the detection of CCD shooting carrying out fluorescence imaging simultaneously.This method is compared with the detection mode of traditional filter and photomultiplier and be need not to change hardware device when advantage is to upgrade fluorescence chemical.But this mode exist the preparing grating difficulty big, exist secondary lobe influence accuracy of detection, utilization factor defective greatly such as low.
Summary of the invention
Defective or deficiency at prior art exists the objective of the invention is to, and provide a kind of and realize the fluorescence quantitative PCR detection system that the long fluorescence excitation of all-wave detects based on tunable optical filter.This system can realize the multi-wavelength excitation detection and not have machinery switching or mechanical action, can realize exciting and detecting of any wavelength easily by wavelength tuning equipment.
In order to realize above-mentioned task, the present invention takes following technical solution:
A kind of quantitative PCR system based on tunable optical filter realization long fluorescence excitation of all-wave and detection is characterized in that this system comprises:
The fluorescence excitation module of forming by light source, beam condensing unit and first adjustable optical filter;
The fluoroscopic examination module of forming by second adjustable optical filter, photodetector and photoelectric conversion device;
The control module of forming by wavelength tuning equipment and main control computer; Wherein, wavelength tuning equipment is used to regulate the filtration wavelength of the first tunable optical filter and the second tunable optical filter;
Cover the heat circulating system of forming by reaction tube, metal temperature piece and constant temperature heat;
Heat circulating system is communicated with first adjustable optical filter by incident optical, is communicated with second adjustable optical filter by outgoing optical fiber;
Main control computer is connected with light source, photoelectric conversion device and wavelength tuning equipment respectively;
The exciting light that light source sends, beam condensing unit enters first adjustable optical filter after by optically focused, the wavelength of first adjustable optical filter is undertaken tuning by wavelength tuning equipment, arriving by incident optical then places the reaction tube of metal temperature piece to carry out fluorescence excitation, the fluorescence that inspires by outgoing optical fiber through the second tunable optical filter after after elimination exciting light composition and other undesired signals, transfer to photodetector and carry out opto-electronic conversion, light signal is converted to electric signal transfers to the main control computer line data analysis of going forward side by side by photoelectric conversion device.
The filtration wavelength of the described first tunable optical filter and the second tunable optical filter, the wavelength tuning range by wavelength tuning equipment is the wavelength from visible light to infrared region 400nm～800nm, and need not to change optical filter.To satisfy requirement of experiment.
The of the present invention first tunable optical filter and the second tunable optical filter are selected tuning filter able to programme.As acousto-optic tunable filter or liquid crystal tunable optical filter, realize that any wavelength excites and detects.By adjust frequency that acousto-optic tunable filter adds radio frequency can be from complex light (as white light) the specific light that sees through a certain wavelength, reach filter effect.This method tuning range is wide, and the optical filtering spectral line is narrow.Another kind of liquid crystal adjustable optical filter utilizes the electrooptical effect of liquid crystal, realizes wavelength tuning by the alive change of liquid crystal filter institute, can realize that continuous spectrum is tuning in the short time.
The quantitative PCR detection system that detects based on the long fluorescence excitation of tunable optical filter realization all-wave of the present invention, tuned light monochromaticity is good, the narrowest 1 nanometer that reaches of spectral line.
The present invention compares with existing real-time fluorescence quantitative PCR detection system, has the following advantages:
(1) alternative traditional filter wheel and diffraction grating, and the amount of having is light, small and exquisite, to vibrating insensitive advantage, whole color separation detection system does not have movable member, is easier to maintenance and operation.
When (2) tunable optical filter carries out wavelength tuning, need not change optical filter, total system stability is high; Add rf frequency by change and carry out the acousto-optic tunable filter wavelength tuning or change impressed voltage realizing that liquid crystal tunable optical filter wavelength tuning can realize that wavelength regulates continuously, wideer to reagent range of choice in the experimentation.
(3) utilize electronic method to carry out wavelength tuning, tuned speed is fast, can reach the microsecond magnitude; Tuning range is very wide, can cover visible light to infrared region; The optical filtering spectral line is narrow, and I reaches 1nm.
(4) can realize the monochromatic detection of monochromatic excitation easily, can realize that also polychrome excites the polychrome detection and need not add more optical modules.
Description of drawings
Fig. 1 is a quantitative PCR system architecture synoptic diagram of realizing the long fluorescence excitation detection of all-wave based on tunable optical filter of the present invention.
The present invention is described in further detail below in conjunction with accompanying drawing.
Embodiment
With reference to figure, a kind of quantitative PCR system based on tunable optical filter realization long fluorescence excitation of all-wave and detection of the present invention comprises:
The fluorescence excitation module of forming by light source 1, beam condensing unit 2 and adjustable optical filter I (label 3);
The fluoroscopic examination module of forming by adjustable optical filter II (label 5), photodetector 7 and photoelectric conversion device 8;
The control module of forming by wavelength tuning equipment 9 and main control computer 10; Wherein wavelength tuning equipment 9 is used to regulate the filtration wavelength of tunable optical filter I and tunable optical filter II;
By reaction tube 12, metal temperature piece 14 and constant temperature heat lid 13 heat circulating systems of forming 11;
By entering tunable optical filter I behind the optically focused, the wavelength of tunable optical filter I is undertaken tuning the exciting light that light source 1 sends by wavelength tuning equipment 9 by beam condensing unit 2, arrives by incident optical 4 then to place the reaction tube 12 of warm piece 11 to carry out fluorescence excitation.
The fluorescence that inspires by outgoing optical fiber 6 through elimination exciting light composition behind the tunable optical filter II and other undesired signals after, transfer to photodetector 7 and carry out opto-electronic conversion, light signal is converted to electric signal transfers to the main control computer 10 line data analysis of going forward side by side by photoelectric conversion device 8.The filtration wavelength of tunable optical filter (I, II) can be regulated so that it satisfies requirement of experiment by wavelength tuning equipment (9).
Adjustable optical filter I, adjustable optical filter II and the 9 supporting settings of wavelength tuning equipment, adjustable optical filter I, adjustable optical filter II can select tuning filter able to programme for use, for example, acousto-optic tunable filter or liquid crystal adjustable optical filter.Acousto-optic tunable filter changes the wavelength that it passes through by the radiofrequency signal that change is carried on the optical filter; Liquid crystal filter has then utilized the electrooptical effect of liquid crystal, realizes the continuous wavelength adjusting by changing impressed voltage.
Quantitative PCR system based on tunable optical filter realization long fluorescence excitation of all-wave and detection of the present invention is selected the incident excitation wavelength by tunable optical filter, can realize monochromatic excitation and multi-wavelength continuous agitation.By optionally only elimination exciting light and other interference of tunable optical filter wavelength tuning, make testing result more accurate in the testing process by fluorescence.The tunable optical filter that adopts need not to change optical filter does not have machinery operation or movable member, the higher and easy operating of system stability yet.
Claims (4)
1. realize the quantitative PCR system of long fluorescence excitation of all-wave and detection it is characterized in that this system comprises based on tunable optical filter for one kind:
The fluorescence excitation module of forming by light source (1), beam condensing unit (2) and first adjustable optical filter (3);
The fluoroscopic examination module of forming by second adjustable optical filter (5), photodetector (7) and photoelectric conversion device (8);
The control module of forming by wavelength tuning equipment (9) and main control computer (10); Wherein, wavelength tuning equipment (9) is used to regulate the filtration wavelength of the first tunable optical filter (3) and the second tunable optical filter (5);
Cover the heat circulating system (11) that (13) are formed by reaction tube (12), metal temperature piece (14) and constant temperature heat;
Heat circulating system (11) is communicated with first adjustable optical filter (3) by incident optical (4), is communicated with second adjustable optical filter (5) by outgoing optical fiber (6);
Main control computer (10) is connected with light source (1), photoelectric conversion device (8) and wavelength tuning equipment (9) respectively;
The exciting light that light source (1) sends, by entering first adjustable optical filter (3) behind beam condensing unit (2) optically focused, the wavelength of first adjustable optical filter (3) is undertaken tuning by wavelength tuning equipment (9), arriving by incident optical (4) then places the reaction tube (12) of metal temperature piece (11) to carry out fluorescence excitation, the fluorescence that inspires by outgoing optical fiber (6) through the second tunable optical filter (5) back elimination exciting light composition and other undesired signals after, transfer to photodetector (7) and carry out opto-electronic conversion, light signal is converted to electric signal transfers to main control computer (10) and carry out data analysis by photoelectric conversion device (8).
2. the quantitative PCR system that realizes the long fluorescence excitation detection of all-wave based on tunable optical filter as claimed in claim 1, it is characterized in that, the filtration wavelength of the described first tunable optical filter (3) and the second tunable optical filter (5), wavelength tuning range by wavelength tuning equipment (9) is the wavelength from visible light to infrared region 400nm～800nm, and need not to change optical filter.
3. the quantitative PCR system that detects based on the long fluorescence excitation of tunable optical filter realization all-wave as claimed in claim 1 is characterized in that the described first tunable optical filter (3) and the second tunable optical filter (5) are selected tuning filter able to programme.
4. as claimed in claim 3ly realize it is characterized in that the quantitative PCR system of the long fluorescence excitation detection of all-wave that described tuning filter able to programme is based on the optical filter of acousto-optic tunable or based on the wavelength tuning optical filter of liquid crystal based on tunable optical filter.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910218601A CN101699271A (en) | 2009-10-27 | 2009-10-27 | Real-time fluorescence quantitative PCR excitation detection system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910218601A CN101699271A (en) | 2009-10-27 | 2009-10-27 | Real-time fluorescence quantitative PCR excitation detection system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101699271A true CN101699271A (en) | 2010-04-28 |
Family
ID=42147734
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910218601A Pending CN101699271A (en) | 2009-10-27 | 2009-10-27 | Real-time fluorescence quantitative PCR excitation detection system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101699271A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102706849A (en) * | 2012-06-15 | 2012-10-03 | 杭州安杰思医学科技有限公司 | Ectopic multi-wavelength detection-based fluorescence quantitative detection system |
| CN105543073A (en) * | 2016-01-08 | 2016-05-04 | 西安交通大学 | Integrated digital nucleic acid amplification detecting system |
| CN105606581A (en) * | 2016-03-21 | 2016-05-25 | 中国工程物理研究院激光聚变研究中心 | Super-continuum spectrum light source based multicolor fluorescence imaging system |
| CN105866090A (en) * | 2016-06-03 | 2016-08-17 | 苏州百源基因技术有限公司 | Ultraviolet visible light fluorescence detection system |
| CN106929388A (en) * | 2015-12-31 | 2017-07-07 | 苏州百源基因技术有限公司 | A kind of real-time fluorescence quantitative PCR instrument |
| CN109342380A (en) * | 2018-09-28 | 2019-02-15 | 杨帆 | A kind of biological detection system |
| CN109957496A (en) * | 2019-04-17 | 2019-07-02 | 苏州雅睿生物技术有限公司 | Portable PC R real-time fluorescence detection system |
| WO2020142902A1 (en) * | 2019-01-08 | 2020-07-16 | 京东方科技集团股份有限公司 | Fluid detection panels and fluid detection device |
-
2009
- 2009-10-27 CN CN200910218601A patent/CN101699271A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102706849A (en) * | 2012-06-15 | 2012-10-03 | 杭州安杰思医学科技有限公司 | Ectopic multi-wavelength detection-based fluorescence quantitative detection system |
| CN106929388A (en) * | 2015-12-31 | 2017-07-07 | 苏州百源基因技术有限公司 | A kind of real-time fluorescence quantitative PCR instrument |
| CN105543073A (en) * | 2016-01-08 | 2016-05-04 | 西安交通大学 | Integrated digital nucleic acid amplification detecting system |
| CN105543073B (en) * | 2016-01-08 | 2019-09-20 | 西安交通大学 | A kind of integrated digital nucleic acid amplification detection system |
| CN105606581A (en) * | 2016-03-21 | 2016-05-25 | 中国工程物理研究院激光聚变研究中心 | Super-continuum spectrum light source based multicolor fluorescence imaging system |
| CN105606581B (en) * | 2016-03-21 | 2019-02-26 | 中国工程物理研究院激光聚变研究中心 | A kind of multicolor fluorescence imaging system based on super continuum source |
| CN105866090A (en) * | 2016-06-03 | 2016-08-17 | 苏州百源基因技术有限公司 | Ultraviolet visible light fluorescence detection system |
| CN109342380A (en) * | 2018-09-28 | 2019-02-15 | 杨帆 | A kind of biological detection system |
| WO2020142902A1 (en) * | 2019-01-08 | 2020-07-16 | 京东方科技集团股份有限公司 | Fluid detection panels and fluid detection device |
| US11255790B2 (en) | 2019-01-08 | 2022-02-22 | Boe Technology Group Co., Ltd. | Fluid detection panel with filter structure and fluid detection device with filter structure |
| CN109957496A (en) * | 2019-04-17 | 2019-07-02 | 苏州雅睿生物技术有限公司 | Portable PC R real-time fluorescence detection system |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101699271A (en) | Real-time fluorescence quantitative PCR excitation detection system | |
| US11366052B2 (en) | Multi-modal fluorescence imaging flow cytometry system | |
| US20230349812A1 (en) | Fluorescence Imaging Flow Cytometry With Enhanced Image Resolution | |
| CN104388307B (en) | A kind of drop style product fluorescence detecting system and method | |
| US8120002B2 (en) | Multi-color biosensor for detecting luminescence sites on a substrate having a refractive optical element for adjusting and focusing at least two incident irradiation beams of different wavelengths | |
| US20020109100A1 (en) | Light detection device | |
| US20110278471A1 (en) | Fluorescence detecting device and fluorescence detecting method | |
| WO2010084719A1 (en) | Fluorescence detecting device and fluorescence detecting method | |
| JP6501714B2 (en) | Optical survey device | |
| US20120085933A1 (en) | Fluorescence detection device and fluorescence detection method | |
| US20090218518A1 (en) | Apparatus for carrying out real-time pcr reactions | |
| GB2404013A (en) | Measuring fluorescence lifetime | |
| US20160230213A1 (en) | Non-motorized optical multiplexing for the simultaneous detection of dna target amplicons in a polymerase chain reaction solution | |
| WO2022218329A1 (en) | Fluorescence quantitative pcr processing method and system | |
| JPWO2010079731A1 (en) | Fluorescence detection apparatus and fluorescence detection method | |
| CN106153575A (en) | A kind of acousto-optic modulation double light path double detector type near infrared spectrometer and method of testing | |
| CN105092836A (en) | Up-converting phosphor immunochromatography detection apparatus and detection method | |
| WO2007081692A2 (en) | Method and apparatus for dense spectrum unmixing and image reconstruction of a sample | |
| CN209784193U (en) | Equipment capable of measuring Raman spectrum of substance under strong fluorescence background | |
| JP2005091701A (en) | Fluorescence microscope and exciting light source control method thereof | |
| US20130105708A1 (en) | Narrow band fluorophore exciter | |
| JP2009145044A (en) | Fluorescence detector | |
| EP3922978A1 (en) | Optical device for detecting fluorescence emission | |
| TWI470202B (en) | Biochemical measurement system | |
| US20230221591A1 (en) | Systems and methods for controlling molecules |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Open date: 20100428 |