CN201553741U - Multiwavelength fluorescence detection device of quantitative PCR - Google Patents
Multiwavelength fluorescence detection device of quantitative PCR Download PDFInfo
- Publication number
- CN201553741U CN201553741U CN2009202000602U CN200920200060U CN201553741U CN 201553741 U CN201553741 U CN 201553741U CN 2009202000602 U CN2009202000602 U CN 2009202000602U CN 200920200060 U CN200920200060 U CN 200920200060U CN 201553741 U CN201553741 U CN 201553741U
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- 238000003753 real-time PCR Methods 0.000 title claims abstract description 20
- 238000001917 fluorescence detection Methods 0.000 title claims abstract description 17
- 230000003287 optical effect Effects 0.000 claims abstract description 16
- 239000013307 optical fiber Substances 0.000 claims abstract description 15
- 239000000835 fiber Substances 0.000 claims description 16
- 230000004913 activation Effects 0.000 claims description 12
- 230000003595 spectral effect Effects 0.000 claims description 9
- 238000012546 transfer Methods 0.000 claims description 3
- 229910052736 halogen Inorganic materials 0.000 claims description 2
- 150000002367 halogens Chemical class 0.000 claims description 2
- 230000005693 optoelectronics Effects 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 20
- 230000001360 synchronised effect Effects 0.000 abstract description 5
- 238000004458 analytical method Methods 0.000 abstract description 3
- 238000012360 testing method Methods 0.000 description 10
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- 238000000137 annealing Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
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- 239000000654 additive Substances 0.000 description 1
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- 230000009286 beneficial effect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
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- 230000004069 differentiation Effects 0.000 description 1
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- 238000002474 experimental method Methods 0.000 description 1
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- 230000003252 repetitive effect Effects 0.000 description 1
- 239000012780 transparent material Substances 0.000 description 1
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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The utility model discloses a multiwavelength fluorescence detection device of a quantitative PCR, which comprises n fluorescent wavelength detection channels which are spaced to be a modularized standard hole site, a polyhedral reflecting mirror and an optical sensor, wherein the polyhedral reflecting mirror is provided with n working surfaces, n=2-96, each fluorescent wavelength detection channel is provided with a light source exciting optical fiber path and a fluorescent receiving optical fiber path, and a light source, a first lens, a first light filter and a first focusing mirror are arranged on the light source exciting optical fiber path along the transmitting direction of fluorescence in turn; a second lens, a second light filter, a second focusing mirror and a plane reflecting mirror are arranged on the fluorescent receiving optical fiber path along the transmitting direction of fluorescence in turn; fluorescence of n fluorescent wavelength detection channels is gathered on the polyhedral reflecting mirror of the optical sensor and is arranged on the reflecting light path of the plane reflecting mirror. The multiwavelength fluorescence detection device can achieve real-time and synchronous detection of multiwavelength fluorescence, greatly improves detection efficiency, does not generate exciting light crosstalk when in detection, and has accurate quantitative PCR analysis results.
Description
Technical field
The utility model relates to a kind of multi-wavelength fluorescence detection device that is used for quantitative PCR.
Background technology
The quantitative PCR detection system principle of work is the instrument that makes reactant automated cycle between specified denaturation temperature, annealing temperature and elongating temperature, and the temperature cycle by sex change, annealing and extension can increase millions of times with target DNA at short notice.
Quantitative PCR detection system is by using the excitation light irradiation test tube of different wave length, when the reagent in the test tube is inspired specific wavelength fluorescence, use optical pickocff, collect fluorescence intensity signals and in time be sent to computer as PMT (photomultiplier), PD (photodiode), APD (avalanche photodide) etc. and carry out real time data demonstration and analysis.
Along with modern round pcr development, need in same pipe or homogeneous experiment, realize the detection of multiple dye fluorescence sometimes, to adapt to the needs that reagent additive (object of reference, reagent integrated level increase) is on the increase.
The multi-wavelength scanning work pattern of main flow quantitative PCR detection system (Real time PCR, real-time fluorescence quantitative PCR) is at present: adopt exciting light optical filter wheel and fluorescent optical filter wheel.It is characterized in that: use stepper-motor, select the exciting light spectral filter and the fluorescent optical filter that need with mechanical drive mode one by one.Under this pattern, the repetitive positioning accuracy of optical filter wheel is limited by running gear, and its reliability is owing to motion reduces.Realize that multi-wavelength detects, must switch filter set successively, control exciting light successively, have only a kind of luminous light work of wavelength at every turn, its total detection time is quite long.With 4 passages is example, and the scanning of standard 96 orifice plates generally all needs more than the 20s.Because the reagent enzymic activity is subjected to the remarkably influenced of time and temperature, so the prolongation of time will cause reagent fluorescence decay and degraded inevitably, and influence the detected result of quantitative PCR.
At studies show that of the full skirtboard of single PCR test tube and 96 holes, at interval when position, 1 standard module hole (96 * 0.2ml test tube usefulness, i.e. excitation beam spacing 18mm), crosstalking substantially below 0.1% between the exciting light detects to use at PCR and can ignore substantially.If exciting light is an adjacent holes, exciting light can be transmitted to by the test tube of transparent material on the adjacent holes, forms background and causes that mistake excites, and it influences at more than 10% of typical fluorescence signal intensity, just can not meet the demands.
Summary of the invention
At the defective that existing multi-wavelength detection technique exists, the purpose of this utility model provides a kind of multi-wavelength fluorescence detection device that can realize the quantitative PCR of real-time synchronous detection.
The multi-wavelength fluorescence detection device of quantitative PCR of the present utility model, comprise that n space is the wavelength of fluorescence sense channel of position, a module standard hole, polyhedron reflective mirror and optical pickocff, the polyhedron reflective mirror has n working face, n=2～96, each wavelength of fluorescence sense channel has a light source activation fiber path and a fluorescence receives fiber path, be provided with light source at the light source activation fiber path successively along the light source activation direction, first lens, first spectral filter and first condensing lens, receive fiber path at fluorescence and be provided with second lens successively along the fluorescence transfer direction, second spectral filter, second condensing lens and plane mirror, the polyhedron reflective mirror that the fluorescence of n fluorescence detection channel is converged to optical pickocff is positioned on the reflected light path of plane mirror.
On the light source stated can be photodiode, laser diode or halogen lamp.Photo-sensor can be photomultiplier, photodiode, avalanche optoelectronic pipe or charge coupled device.First condensing lens and second condensing lens all can adopt prism.
It can be two independently optical fiber that light source activation optical fiber in the utility model and fluorescence receive optical fiber, also can be Y shape bifurcation fiber.
The beneficial effects of the utility model are:
1) can realize the real-time synchronous detection of multi-wavelength fluorescence, detection efficiency improves greatly.
2) position, a module standard hole, wavelength of fluorescence sense channel space does not produce exciting light and crosstalks during detection.
3) shorten detection total time to greatest extent, helped overcoming the influence of material in the reagent sample (enzyme, primer, confidential reference items product etc.) decay degraded, obtained quantitative PCR analysis result accurately.
4) optical fiber of Cai Yonging does not have relative movement, does not have optical fiber problem fatigue lifetime, is convenient to the system integration and proofing unit miniaturization.
5) by the whole move mode of proofing unit, can realize multi-wavelength fluoroscopic examination to any hole.
Description of drawings
Fig. 1 is the utility model device synoptic diagram.
Embodiment
Further specify the utility model below in conjunction with accompanying drawing.
With reference to Fig. 1, the multi-wavelength fluorescence detection device of quantitative PCR of the present utility model, comprise that n space is the wavelength of fluorescence sense channel of position, a module standard hole, polyhedron reflective mirror 11 and optical pickocff 12, in the illustrated example, n=2, polyhedron reflective mirror 11 has 2 working faces, each wavelength of fluorescence sense channel has a light source activation fiber path 1 and a fluorescence receives fiber path 2, this example, and light source activation optical fiber and fluorescence receive optical fiber and adopt Y shape bifurcation fiber.Be provided with light source 3 at the light source activation fiber path successively along the light source activation direction, first lens 4, first spectral filter 5 and first condensing lens 6, receive fiber path at fluorescence and be provided with second lens 7, second spectral filter 8, second condensing lens 9 and plane mirror 10 successively along the fluorescence transfer direction, polyhedron reflective mirror 11 is positioned on the reflected light path of plane mirror 10, and the fluorescence of 2 fluorescence detection channel is converged on the photoelectric surface of optical pickocff 12.
In illustrated example when work,, 2 light sources are lighted simultaneously, and light source obtains the exciting light of required wavelength separately through first collimated and exciting light spectral filter, again after first condensing lens focuses on along fiber-optic illuminated to test tube 1 and test tube 3 bottoms or the top excite the reagent sample.Reagent sample fluorescence enters fluorescence respectively and receives fiber path in 2 test tubes, the fluorescence that obtains required wavelength through second collimated and second fluorescent optical filter shines 2 working faces of plane mirror, converge on the photoelectric surface of optical pickocff 12 by the fluorescence of plane mirror, realize 2 real-time synchronous detection of wavelength of fluorescence sense channel with 2 fluorescence detection channel.The moment among the figure, test tube 2 is not detected, but by the whole move mode of proofing unit, can realize multi-wavelength fluorescence synchronous detection any hole of standard module (96 * 0.2ml hole test tube plate), need not machinery switches, and used optical fiber and outage have also been eliminated the integrity problem that the fine exercise fatigue of light causes.
The parameter of exciting light and fluorescent optical filter is selected according to the dyestuff spectral response curve of required detection reagent, with accurate differentiation background and signal, can be any wavelength in the 200-900nm scope.
Claims (5)
1. the multi-wavelength fluorescence detection device of a quantitative PCR, it is characterized in that comprising that n space is the wavelength of fluorescence sense channel of position, a module standard hole, polyhedron reflective mirror (11) and optical pickocff (12), polyhedron reflective mirror (11) has n working face, n=2～96, each wavelength of fluorescence sense channel has a light source activation fiber path (1) and a fluorescence receives fiber path (2), be provided with light source (3) at the light source activation fiber path successively along the light source activation direction, first lens (4), first spectral filter (5) and first condensing lens (6), receive fiber path at fluorescence and be provided with second lens (7) successively along the fluorescence transfer direction, second spectral filter (8), second condensing lens (9) and plane mirror (10), the polyhedron reflective mirror (11) that the fluorescence of n fluorescence detection channel is converged to optical pickocff (12) is positioned on the reflected light path of plane mirror (10).
2. the multi-wavelength fluorescence detection device of quantitative PCR according to claim 1 is characterized in that light source (3) is photodiode, laser diode or halogen lamp.
3. the multi-wavelength fluorescence detection device of quantitative PCR according to claim 1 is characterized in that it is two independently optical fiber that light source activation optical fiber and fluorescence receive optical fiber, or Y shape bifurcation fiber.
4. the multi-wavelength fluorescence detection device of quantitative PCR according to claim 1 is characterized in that photo-sensor (12) is photomultiplier, photodiode, avalanche optoelectronic pipe or charge coupled device.
5. the multi-wavelength fluorescence detection device of quantitative PCR according to claim 1 is characterized in that first condensing lens and second condensing lens are prism.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009202000602U CN201553741U (en) | 2009-11-16 | 2009-11-16 | Multiwavelength fluorescence detection device of quantitative PCR |
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| CN2009202000602U CN201553741U (en) | 2009-11-16 | 2009-11-16 | Multiwavelength fluorescence detection device of quantitative PCR |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102128799A (en) * | 2010-12-21 | 2011-07-20 | 无锡荣兴科技有限公司 | Water quality detection sensor |
| CN102445439A (en) * | 2010-10-09 | 2012-05-09 | 北京网新易尚科技有限公司 | Chemiluminescence detection device |
| CN104677870A (en) * | 2015-02-06 | 2015-06-03 | 余家昌 | Superminiaturization multi-channel real-time fluorescent spectrum detector |
| CN106399075A (en) * | 2016-08-26 | 2017-02-15 | 浙江科技学院 | Reflector based fluorescent quantitative PCR detection system |
| CN107367496A (en) * | 2017-07-27 | 2017-11-21 | 苏州合惠生物科技有限公司 | A kind of fluorescence detection device |
| CN109957496A (en) * | 2019-04-17 | 2019-07-02 | 苏州雅睿生物技术有限公司 | Portable PCR real-time fluorescence detection system |
| WO2021169069A1 (en) * | 2020-02-25 | 2021-09-02 | 杭州博日科技有限公司 | Pcr detection device and method |
-
2009
- 2009-11-16 CN CN2009202000602U patent/CN201553741U/en not_active Expired - Lifetime
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102445439A (en) * | 2010-10-09 | 2012-05-09 | 北京网新易尚科技有限公司 | Chemiluminescence detection device |
| CN102445439B (en) * | 2010-10-09 | 2015-09-02 | 易尚明天科技有限公司 | chemical fluorescence detector |
| CN102128799A (en) * | 2010-12-21 | 2011-07-20 | 无锡荣兴科技有限公司 | Water quality detection sensor |
| CN104677870A (en) * | 2015-02-06 | 2015-06-03 | 余家昌 | Superminiaturization multi-channel real-time fluorescent spectrum detector |
| CN106399075A (en) * | 2016-08-26 | 2017-02-15 | 浙江科技学院 | Reflector based fluorescent quantitative PCR detection system |
| CN107367496A (en) * | 2017-07-27 | 2017-11-21 | 苏州合惠生物科技有限公司 | A kind of fluorescence detection device |
| CN109957496A (en) * | 2019-04-17 | 2019-07-02 | 苏州雅睿生物技术有限公司 | Portable PCR real-time fluorescence detection system |
| WO2021169069A1 (en) * | 2020-02-25 | 2021-09-02 | 杭州博日科技有限公司 | Pcr detection device and method |
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Granted publication date: 20100818 |