CN103087907A - Relative calibration system for verification and correction of biological PCR (polymerase chain reaction) real-time fluorescent detection system - Google Patents
Relative calibration system for verification and correction of biological PCR (polymerase chain reaction) real-time fluorescent detection system Download PDFInfo
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- CN103087907A CN103087907A CN2012105642320A CN201210564232A CN103087907A CN 103087907 A CN103087907 A CN 103087907A CN 2012105642320 A CN2012105642320 A CN 2012105642320A CN 201210564232 A CN201210564232 A CN 201210564232A CN 103087907 A CN103087907 A CN 103087907A
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Abstract
The invention relates to the field of biological and medical detection, and particularly relates to a relative calibration system for verification and correction of a biological PCR (polymerase chain reaction) real-time fluorescent detection system. The relative calibration system is characterized in that a power supply is connected with a signal processing circuit part through a lead wire group internally comprising a signal wire and a current lead wire; and a light insulation part is internally provided with a light source part and a detection part with a light filter lens group, and the light insulation part is provided with a first small hole and a second small hole. The relative calibration system can accurately calibrate and correct the PCR real-time fluorescent detection system; and in addition, the relative calibration system also can be used for the calibration and the correction of other fluorescent detection systems by replacing a light filter lens of a detection part of the light filter lens group and a light source in the light source part.
Description
Technical field
The present invention relates to a kind of PCR(polymerase chain reaction) the metering comparison device of fluorescence spectrum detection system, be mainly used in the calibration of biological PCR fluorescence detecting system, belong to biology and medical science detection field.
Background technology
Along with micro-total analysis system (Miniaturized Total Analysis Systems at the beginning of the nineties in last century, μ-TAS) concept is proposed first, in recent years, different kind organism analysis on the micro-total analysis system conceptual foundation and chemical analysis chip (comprising: the research fast development micro-fluidic biological pcr chip), the micro-total analysis system technology has become the most important researchdevelopment direction of innovation formula analytical and testing instrument and technology, deeply is subjected to the concern of countries in the world.
(μ-TAS) is the analysis experiment lab system of realizing the integrated and structure micromation of the function of whole analytic processes on the basis of microminiaturization to so-called micro-total analysis system.Generally, it is the function in whole laboratory, be many processes and the step of biochemical analysis, comprise that sampling, dilution, reagent adding, reaction, separation, detection etc. are integrated on as far as possible little service platform, the chip that present this micromation service platform is 100 square millimeters of left and right (structure miniature size to tens square millimeters of development).μ-TAS has the outstanding features such as detection speed is fast, sample dosage is few, flux is high, is widely used in the research of different kind organism chemical analysis chip.In μ-TAS, chip volume is very little, and its sample size is only micro updating, and the detection cell volume is very little, and completes in level in second more than analyzing and testing, therefore require very high to sensitivity and the response speed of its detection system.Detection method is the key factor that determines the final detection limit of μ-TAS, is one of core technology of μ-TAS.In the μ that grew up in recent years-TAS detection method, the fluorescence spectrum detection method becomes most popular detection technique in μ-TAS research field gradually because its selectivity is good, the trace qualitative and quantitative analysis is highly sensitive and the characteristics such as nondestructive detection.
The biological PCR fluorescence spectrum detection method of biochemical analysis chip, actual is that fluorescent substance in the microchannel on chip is carried out spectral detection.There are some different from the fluorescence spectrum detection technique in chemical analysis field: at first, small and the signal of the amount of detected object sends a little little, thereby the detected fluorescence spectrum weak output signal that causes, need use the microchannel fluorescence spectrum detecting device of high detection sensitivity; Secondly, also there is no the reference material in a kind of picture chemical analysis field at present, can be used for calibrating biological PCR fluorescence microchannel spectrum detection device.
Summary of the invention
The invention provides a kind of verification system for the optimization of PCR real-time fluorescence detection system.
The relative Calibration system that is used for the calibrating of biological PCR real-time fluorescence detection system and proofreaies and correct is characterized in that: power supply be connected with signal treatment circuit and is contained signal wire by inside and be connected the connection of wire group with current lead; Be provided with probe portion and the Lights section with the optical filtering group every the light partial interior, be provided with the first aperture and second orifice every the light part;
The exciting light of the 470nm that PCR fluorescence detecting system the Lights section sends enters the probe portion with the optical filtering group in light part by the first aperture, can obtain an electrical signal this moment, be transmitted in power supply signal treatment circuit part by the signal wire in the wire group, obtain the value of electrical signal; Simultaneously, by the control of power supply and signal treatment circuit part, electric current is transmitted to the Lights section by the current lead in the wire group, and this moment, the light of the 530nm that the Lights section sends penetrated through second orifice.
What the calibrating of fluorescence detecting system was adopted at present is all that excitation wavelength is in the 350nm left and right, emission wavelength is at the Quinine Sulphate Di HC standardized solution of 450nm left and right, and the excitation wavelength that is used for the fluorescing system that PCR detects in real time is in the 470nm left and right, emission wavelength is in the 530nm left and right, the proposition of novelty of the present invention a kind of detection method of physics, can accurately examine and determine and proofread and correct the PCR real-time fluorescence detection system.In addition, by changing with light source in the optical filtering of the probe portion of optical filtering group and the Lights section, the present invention is not limited to the calibrating of PCR real-time fluorescence detection system, can be used for equally calibrating and the correction of other fluorescence detecting system.
Description of drawings
Fig. 1 is system schematic of the present invention.
Fig. 2 is system applies mode one schematic diagram of the present invention.
Fig. 3 is system applies mode two schematic diagram of the present invention.
Embodiment
As shown in Figure 1, the present invention mainly by 1, power supply and signal treatment circuit part, 2, wire group (signal wire and current lead are contained in inside), 3, every the light part, 4, with the probe portion of optical filtering group, 5, the Lights section, 6, the first aperture, 7, second orifice.The verification system mode of operation is as follows: the exciting light of 470nm that PCR fluorescence detecting system the Lights section sends left and right enters the probe portion 4 with the optical filtering group in light part 3 by 6 first apertures, can obtain an electrical signal this moment, be transmitted in power supply signal treatment circuit part 1 by the signal wire in wire group 2, at this moment can obtain the value of electrical signal; Simultaneously, by the control of power supply and signal treatment circuit part 1, electric current is transmitted to the Lights section 5 by the wire in wire group 2, and the light about the 530nm that this moment, the Lights section sent penetrates this verification system through 7 second orifices.
Below in conjunction with Fig. 2-Fig. 3, concrete case study on implementation of the present invention is described.
Scheme 1: describe in conjunction with Fig. 2
This programme mainly by 8, Re Gai, 9, the heating/refrigerating unit, 10, test tube, 11, the first fiber optic conduction system, 12, the second fiber optic conduction system, 13, the excitation light source part, the verification system that the PCR real-time fluorescence detection system that 14 fluoroscopic examinations partly consist of and the present invention propose is carried out.The verification system that the present invention proposes in Fig. 2 by 1, power supply and signal circuit treating part, 2 wire groups, 3 form every light part (inside is contained with the probe portion 4 of optical filtering group and the Lights section 5).
Specific operation process:
to put into test tube 10 every light part 3, the exciting light that excitation light source part 13 is sent receives every light part 3 by 11 first accredited systems of fiber optic conduction system, optical signal can be converted into electrical signal, electrical signal is transmitted to power supply and signal circuit treating part 1 by wire group 2, just can know the value of accurate electrical signal this moment, this verification system is put into the same PCR real-time fluorescence detection system of different PCR real-time fluorescence detection systems or different times, by the size of the value of electrical signals that obtains and the quality that stability can judge PCR real-time fluorescence detection system the Lights section, the electrical signal that obtains is large and stablely illustrate that the PCR real-time fluorescence detection system has the Lights section preferably.in addition, electric current is regulated in power supply in the verification system that proposes by the present invention and signal circuit treating part 1, can send an optical signal every light part 3, optical signal is transmitted to fluoroscopic examination part 14 by 12 second fiber optic conduction systems, this moment, the PCR real-time fluorescence detection system can obtain an electrical signal, by regulating the verification system power supply and signal circuit treating part 2 makes electric current constant, to go out light intensity constant for verification system, this verification system is put into the same PCR real-time fluorescence detection system of different PCR real-time fluorescence detection systems or different times, the size of the value of electrical signals that obtains by the PCR real-time fluorescence detection system just can be known the quality of the fluoroscopic examination part of PCR real-time fluorescence detection system, its fluoroscopic examination part of the explanation that under same current, electrical signal is large better.Can also examine and determine the height of the minimum detectability of PCR real-time fluorescence detection system with the verification system of the present invention's proposition, by constantly turning power supply and signal circuit treating part 1 down, the going out light intensity and can constantly weaken thereupon of verification system is until till fluoroscopic examination part 14 can't detect.Just can know the height of its minimum detectability by the same PCR real-time fluorescence detection system of verification system being put into different PCR real-time fluorescence detection systems or different times, the PCR real-time fluorescence detection system minimum detectability that when fluoroscopic examination part 14 can't detect, corresponding current is low is higher.
Scheme 2: the use of this verification system (after miniature) in the biochip with microchannel fluorescence detection function device is described in conjunction with Fig. 3.
this programme is mainly by 15, the point import and export layer of detecting unit, 16, the semiconductor light responsive detectors, 17 first combined filters, 18 first optical waveguide tops (not necessarily shape shown), 19 first multilayer film (to the high reflection of light wave in conduit, the outer light wave of conduit being intercepted fully), 20, chip microchannel, 21, the chip microchannel cover plate, 22, excite the electric import and export microchannel of unit, 23 second combined filters, 24, the luminescence unit of excitation light source, 25, the chip microchannel slide glass, 26, the second multilayer film, 27, the second optical waveguide top, 28 first reserved locations, 29, the second reserved location (as the case may be, can be in chip same check point increases integrated another set of little fluorescence detecting system, to improve biological faint light spectrum signal detection sensitivity) biochip with microchannel fluorescence detection function device that forms and by power supply and signal circuit treating part 1, wire group 2, containing probe portion 4 and the Lights section 5 with the optical filtering group every light part 3(inside) verification system that forms carries out.
Specific implementation method:
the light that the luminescence unit 24 of excitation light source sends is through 23 second combined filters, the second multilayer film 26, have enter behind the second optical waveguide top 27 of optical coupled and focusing effect the verification system that is placed in chip microchannel 20 every light part 3 and received, optical signal can be converted into electrical signal, electrical signal is transmitted to power supply and signal circuit treating part 1 by wire group 2, just can know the value of accurate electrical signal this moment, this verification system is put into the different biochips with microchannel fluorescence detection function device or the same biochip with microchannel fluorescence detection function device of different times, by the size of the value of electrical signals that obtains and the quality that stability can judge the Lights section in the biochip with microchannel fluorescence detection function device, large and that biochip with microchannel fluorescence detection function device corresponding to stable explanation of the electrical signal that obtains has the Lights section preferably.in addition, regulate electric current by the power supply in verification system and signal circuit treating part 1, can send an optical signal every light part 3, optical signal is by having the first optical waveguide top 18 and the first rear arrival semiconductor light of combined filters 17 responsive detectors 16 of optical coupled and focusing effect, this moment, biological chips detection system obtained an electrical signal, by regulating the verification system power supply and signal circuit treating part 1 makes electric current constant, to go out light intensity constant for verification system, this verification system is put into the different biochips with microchannel fluorescence detection function device or the same biochip with microchannel fluorescence detection function device of different times, the size of the value of electrical signals that obtains by the biochip with microchannel fluorescence detection function device just can be known the quality of the fluoroscopic examination part in the biochip with microchannel fluorescence detection function device, its fluoroscopic examination part of the explanation that under same current, electrical signal is large better.Can also examine and determine the height of the minimum detectability of the biochip with microchannel fluorescence detection function device with the verification system of the present invention's proposition, by constantly turning power supply and signal circuit treating part 1 down, the going out light intensity and can constantly weaken thereupon of verification system is until till semiconductor light responsive detectors 16 can't detect.Just can know the height of its minimum detectability by the same biochip with microchannel fluorescence detection function device of verification system being put into the different biochips with microchannel fluorescence detection function device or different times, the biochip minimum detectability with microchannel fluorescence detection function device that when semiconductor light responsive detectors 16 can't detect, corresponding current is low is higher.
Claims (1)
1. the relative Calibration system that is used for the calibrating of biological PCR real-time fluorescence detection system and proofreaies and correct is characterized in that: power supply be connected with signal treatment circuit and is contained signal wire by inside and be connected the connection of wire group with current lead; Be provided with probe portion and the Lights section with the optical filtering group every the light partial interior, be provided with the first aperture and second orifice every the light part;
The 470nm exciting light that PCR fluorescence detecting system the Lights section sends enters the probe portion with the optical filtering group in light part by the first aperture, can obtain an electrical signal this moment, be transmitted in power supply signal treatment circuit part by the signal wire in the wire group, obtain the value of electrical signal; Simultaneously, by the control of power supply and signal treatment circuit part, electric current is transmitted to the Lights section by the current lead in the wire group, and this moment, the light of the 530nm that the Lights section sends penetrated through second orifice.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI500450B (en) * | 2014-08-22 | 2015-09-21 | ||
| CN111269971A (en) * | 2020-02-17 | 2020-06-12 | 上海科源电子科技有限公司 | Method for automatically calibrating polymerase chain reaction fluorescent signal |
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| CN1643128A (en) * | 2002-03-26 | 2005-07-20 | 来德行 | Novel integrated circuit chip for bioassays |
| CN102106720A (en) * | 2009-12-23 | 2011-06-29 | 麦克内尔-Ppc股份有限公司 | Device and method for detecting plaque in the oral cavity |
| CN102768203A (en) * | 2012-07-03 | 2012-11-07 | 北京工业大学 | Space-oriented minitype cylindrical microfluidic PCR (polymerase chain reaction) real-time fluoroscopic detection system |
| CN102782477A (en) * | 2010-03-02 | 2012-11-14 | 皇家飞利浦电子股份有限公司 | Optical examinations with controlled input light |
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- 2012-12-21 CN CN201210564232.0A patent/CN103087907B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1643128A (en) * | 2002-03-26 | 2005-07-20 | 来德行 | Novel integrated circuit chip for bioassays |
| CN102106720A (en) * | 2009-12-23 | 2011-06-29 | 麦克内尔-Ppc股份有限公司 | Device and method for detecting plaque in the oral cavity |
| CN102782477A (en) * | 2010-03-02 | 2012-11-14 | 皇家飞利浦电子股份有限公司 | Optical examinations with controlled input light |
| CN102768203A (en) * | 2012-07-03 | 2012-11-07 | 北京工业大学 | Space-oriented minitype cylindrical microfluidic PCR (polymerase chain reaction) real-time fluoroscopic detection system |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI500450B (en) * | 2014-08-22 | 2015-09-21 | ||
| CN111269971A (en) * | 2020-02-17 | 2020-06-12 | 上海科源电子科技有限公司 | Method for automatically calibrating polymerase chain reaction fluorescent signal |
| CN111269971B (en) * | 2020-02-17 | 2023-06-20 | 上海科源电子科技有限公司 | Method for automatically calibrating fluorescent signal of polymerase chain reaction |
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