CN105928911A - Calibration method of fluorescence detection instrument - Google Patents
Calibration method of fluorescence detection instrument Download PDFInfo
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- CN105928911A CN105928911A CN201610220080.0A CN201610220080A CN105928911A CN 105928911 A CN105928911 A CN 105928911A CN 201610220080 A CN201610220080 A CN 201610220080A CN 105928911 A CN105928911 A CN 105928911A
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- light source
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- 238000001917 fluorescence detection Methods 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title abstract description 10
- 238000001514 detection method Methods 0.000 claims abstract description 42
- 239000000758 substrate Substances 0.000 claims description 12
- 230000003287 optical effect Effects 0.000 claims description 5
- 238000005259 measurement Methods 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 238000003908 quality control method Methods 0.000 abstract description 3
- 239000000463 material Substances 0.000 description 15
- 238000010586 diagram Methods 0.000 description 5
- 238000005286 illumination Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 2
- 241000876446 Lanthanotidae Species 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000005281 excited state Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 239000012925 reference material Substances 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
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/06—Illumination; Optics
- G01N2201/061—Sources
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Optics & Photonics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a calibration method of a fluorescence detection instrument. The method is used for calibration of fluorescence detection instrument before leaving the factory. The light intensity detection of the fluorescence detection instrument and calibration of signal detection module parameters are carried out into two steps independently. The light intensity detection uses a light intensity measurement instrument; and the calibration of the signal detection module parameters uses a standard light source as a detection light source. The invention adopts the existing standard equipment after measurement determination for the calibration, has standard traceability and uncertainty, and is suitable for mass and accurate measurement. At the same time, the method of the invention can also be used in quality control of instruments to detect light source attenuation after long time operation; if attenuation exists, the light source can be replaced. The invention improves the consistency of fluorescence detection instrument in mass production.
Description
Technical field
The present invention relates to the fluorescence detection equipment in biologic medical field, particularly relate to the calibration steps of a kind of fluorescence detection equipment.
Background technology
Fluorescent material characteristic is: when fluorescent material irradiates through the incident illumination of certain wavelength, absorb luminous energy laggard enter excited state, and immediately de excitation send out and send be different from incident illumination the emergent light of wavelength；And once stopping incident illumination, luminescence phenomenon disappears the most immediately.The emergent light with this character is just referred to as fluorescence.
As shown in Figure 1, fluorescence detection equipment (luminoscope) includes light source 1, optical filter 3, signal detection module 4 and mirror group, its operation principle is by irradiating on the fluorescent substance with the light (referred to as exciting light) of the first wavelength, fluorescent material launches the light (referred to as launching light) of the second wavelength, by detecting the light intensity of the second wavelength, indirectly obtain the content of the material of fluorescent material labelling.
At present, the calibration of luminoscope is as default standard with constant concentration fluorescent material, is put on fluorescent instrument measurement.If having multiple stage instrument to calibrate just to calibrate as standard with this fixing fluorescence content material.The problem of do so is that the fluorescence intensity of constant concentration fluorescent material can not be fixed.Because fluorescent material can decay after after fluorescent characteristic is irradiation or placing for a long time, the light launched will weaken, and will go wrong, because standard substance fluorescence intensity has changed and for if being used for calibrating multiple stage instrument.Or the instrument of calibration today, will calibrate by same fluorescence tomorrow again, and value all can change.
This scheme cannot ensure the concordance of machine when big measuring appratus produces, and can only ensure one probably.
Summary of the invention
The main technical problem to be solved in the present invention is to provide the calibration steps of a kind of fluorescence detection equipment, and it can improve the concordance of fluorescence detection equipment in volume production.
For solving above-mentioned technical problem, the present invention provides the calibration steps of a kind of fluorescence detection equipment, the steps include:
A kind of calibration steps of fluorescence detection equipment, the intensity of light source of fluorescence detection equipment is detected, the most independently carrying out with signal detection module parametric calibration, intensity of light source detection uses light intensity measuring instrument measured light intensity, and signal detection module parametric calibration uses standard light source as detection light source.
Described intensity of light source detecting step is: light intensity measuring instrument is placed on detection position to examine and determine the luminous intensity of light source；By adjusting light source power supply circuit parameter, light source luminescent intensity being adjusted to preset value, the light source of the most all fluorescence detection equipment just can be calibrated to same standard.
Described signal detection module parametric calibration step is: is placed on detection position by the standard light source that can send the light that wavelength is l2, turns off the light source of fluorescence detection equipment, adjusts the parameter of signal detection module, signal detection module measurement to value be adjusted to preset value.
Described light intensity measuring instrument includes substrate, is provided with sensor devices, light intensity indicating circuit and power supply on substrate, and three's circuit couples；Sensor devices receives light source and irradiates, and converts optical signals to the signal of telecommunication, inputs light intensity indicating circuit, and when light signal strength reaches preset value, light intensity indicating circuit is pointed out；Power supply is that light intensity indicating circuit is powered.
Described standard light source includes substrate, is provided with luminescent device, drive circuit and power supply on substrate, and three's circuit couples, and power supply is drive circuitry, and drive circuit driven for emitting lights device is luminous.
The invention has the beneficial effects as follows: the calibration steps of a kind of fluorescence detection equipment, the intensity of light source of fluorescence detection equipment is detected and signal detection module parametric calibration is the most independently carried out, intensity of light source detection uses light intensity measuring instrument measured light intensity, and signal detection module parametric calibration uses standard light source as detection light source；The present invention uses the existing standard device determined through metering to calibrate, and has standard trackability and definitiveness, can be suitable for high-volume and accurately measure；The inventive method may further be used to do the Quality Control of instrument simultaneously, and whether detection light source can decay by long operational time, if light source decay can change light source, the present invention improves the concordance of fluorescence detection equipment in volume production.
Accompanying drawing explanation
Fig. 1 is calibration (test) schematic diagram of existing fluorescence detection equipment；
Fig. 2 is the schematic diagram of the step one of an embodiment of the present invention；
Fig. 3 is the schematic diagram of the step 2 of an embodiment of the present invention；
Fig. 4 is standard light source electrical block diagram of the present invention；
Fig. 5 is light intensity measuring instrument electrical block diagram of the present invention.
Detailed description of the invention
As it is shown in figure 1, be a kind of calibrating principle (light path) figure of fluorescence detection equipment instrument 5.Actual test there is fluorescent material in test strips 2, is irradiated in test strips 2 with the light source 1 of l1, excites the fluorescence in test strips 2, by the filtration of optical filter 3, the fluorescence of l2 can only be allowed to enter into signal detection module 4.Prior art is with being as default standard (calibration) with constant concentration fluorescent material, is put on fluorescent instrument 5 measurement.The fluorescence that fluorescent material sends can be decayed (fluorescent material can change over time or the change of illumination and change), has uncertainty with fluorescent material as reference material, difference after the meeting on calibration machine performance out.Certainly, prior art also has the luminoscope light path of other kind, but essence is all with l1(wavelength 1) light source activation, detect l2(wavelength 2) light intensity, the most no longer enumerate.
The present invention, a kind of calibration steps of fluorescence detection equipment, by light source 1 intensity detection of fluorescence detection equipment, the most independently carry out with signal detection module 4 parametric calibration, light source 1 intensity detection uses light intensity measuring instrument 6 measured light intensity, and signal detection module 4 parametric calibration uses standard light source 7 as detection light source.
With reference to shown in Fig. 2-5, the calibration steps of fluorescence detection equipment of the present invention, by measuring light source 1, and the mode with standard light source 7 demarcation signal detection module 4 with light intensity measuring instrument 6, it is achieved calibration.
Combine accompanying drawing below by detailed description of the invention the present invention is described in further detail.
Refer to shown in Fig. 1,2,3, the present invention uses two kinds of equipment to calibrate luminoscope detecting instrument 5.
(1) it being placed on test strips 2(as shown in Figure 1 with miniature light intensity measuring instrument 6) position i.e. monitors position and examines and determine light source 1 luminous intensity.By adjusting light source 1 power supply circuits parameter, light source 1 luminous intensity is adjusted to preset value.The light source 1 of the most all luminoscope detecting instruments 5 just can be calibrated to same standard.
(2) it is placed on test strips 2(as shown in Figure 1 by the standard light source 7 of the light that can send wavelength l2) position, turn off the light source 1 of fluorescence detection equipment 5.Adjusting the parameter of signal detection module 4, the value that signal detection module 4 is measured is adjusted to preset value.
Adjusted by this two pacings amount, just all fluorescence detection equipment performance parameters can be adjusted consistent.
It is adjusted to parameter preset when this preset value of preset value dispatches from the factory with reference to existing luminoscope detecting instrument.
As shown in Figure 4, standard light source 7 includes substrate, is provided with luminescent device 9, drive circuit 10 and power supply 8 on substrate, and three's circuit couples, and power supply 8 is powered for drive circuit 10, and drive circuit 10 driven for emitting lights device 9 is luminous.
During enforcement, luminescent device 9 can be LED light source.
During enforcement, the outside sheathed housing of standard light source 7, corresponding luminescent device 9 position is arranged windows so that luminescent device 9 exposes, and forms test card pattern.
As shown in Figure 5, light intensity measuring instrument 6 includes substrate, substrate is provided with sensor devices 12, light intensity indicating circuit 13 and power supply 11, three's circuit couples, sensor devices 12 receives light source 1 and irradiates, and convert optical signals to the signal of telecommunication, and input light intensity indicating circuit 13, when light signal strength reaches preset value, light intensity indicating circuit 13 is pointed out.Power supply 11 is powered for light intensity indicating circuit 13.
The key point of the present invention is can be by existing method to luminoscope Registration.Registration process the Registration process of two pacing amounts can not be changed into what tradition one step surveying carried out calibrating.First light intensity measuring instrument scaling light source is used, then with standard light source calibration signal detection module.With existing standard accurate calibration luminoscope.
The present invention be exactly use will not in time with method and the material of illumination variation, realize the calibration of fluorescence analyser.
The present invention uses the existing standard device determined through metering to calibrate, and has standard trackability and definitiveness, can be suitable for high-volume and accurately measure；The inventive method may further be used to do the Quality Control of instrument simultaneously, and whether detection light source can decay by long operational time, if light source decay can change light source.
Above content is to combine specific embodiment further description made for the present invention, it is impossible to assert the present invention be embodied as be confined to these explanations.For general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, it is also possible to make some simple deduction or replace, all should be considered as belonging to protection scope of the present invention.
Claims (5)
1. the calibration steps of a fluorescence detection equipment, it is characterized in that: by light source (1) intensity detection of fluorescence detection equipment, the most independently carry out with signal detection module (4) parametric calibration, light source (1) intensity detection uses light intensity measuring instrument (6) measured light intensity, and signal detection module (4) parametric calibration uses standard light source (7) as detection light source.
2. the calibration steps of fluorescence detection equipment as claimed in claim 1, it is characterised in that described light source (1) intensity detection step is: light intensity measuring instrument is placed on detection position to examine and determine the luminous intensity of light source (1)；By adjusting light source (1) power supply circuits parameter, light source (1) luminous intensity being adjusted to preset value, the light source (1) of the most all fluorescence detection equipment just can be calibrated to same standard.
3. the calibration steps of fluorescence detection equipment as claimed in claim 1, it is characterized in that, described signal detection module (4) parametric calibration step is: be placed on detection position by the standard light source (7) that can send the light that wavelength is l2, turn off the light source (1) of fluorescence detection equipment (5), adjusting the parameter of signal detection module (4), the value that signal detection module (4) is measured to is adjusted to preset value.
4. the calibration steps of fluorescence detection equipment as claimed in claim 1 or 2, it is characterised in that described light intensity measuring instrument (6) includes substrate, is provided with sensor devices (12), light intensity indicating circuit (13) and power supply (11) on substrate, and three's circuit couples；Sensor devices (12) receives light source (1) and irradiates, and converts optical signals to the signal of telecommunication, and input light intensity indicating circuit (13), when light signal strength reaches preset value, light intensity indicating circuit (13) is pointed out；Power supply (11) is light intensity indicating circuit (13) power supply.
5. the calibration steps of the fluorescence detection equipment as described in claim 1 or 3, it is characterized in that, described standard light source (7) includes substrate, substrate is provided with luminescent device (9), drive circuit (10) and power supply (8), three's circuit couples, power supply (8) is drive circuit (10) power supply, and drive circuit (10) driven for emitting lights device (9) is luminous.
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| CN201610220080.0A CN105928911A (en) | 2016-04-11 | 2016-04-11 | Calibration method of fluorescence detection instrument |
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| CN201610220080.0A CN105928911A (en) | 2016-04-11 | 2016-04-11 | Calibration method of fluorescence detection instrument |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107064002A (en) * | 2017-03-28 | 2017-08-18 | 中国科学技术大学 | A kind of standard sources of fluorescence analysis |
| CN107271415A (en) * | 2017-06-16 | 2017-10-20 | 中国计量科学研究院 | It is a kind of for standard sample of photo of fluorescent instrument calibration measurement and preparation method thereof |
| CN109668865A (en) * | 2019-01-04 | 2019-04-23 | 上海科源电子科技有限公司 | Fluorescence detector Stability Determination device and measuring method |
| CN110346551A (en) * | 2018-04-04 | 2019-10-18 | 南京东纳生物科技有限公司 | A kind of fluorescence immune chromatography quantitative analysis instrument testing calibration card and its detection method |
| CN110927136A (en) * | 2019-12-25 | 2020-03-27 | 杭州微策生物技术有限公司 | Quality detection device of dry-type immunofluorescence POCT detection instrument and use method thereof |
| CN111157499A (en) * | 2020-01-06 | 2020-05-15 | 深圳市华科瑞科技有限公司 | Method for calibrating fluorescence detection instrument |
| CN111206077A (en) * | 2020-02-12 | 2020-05-29 | 上海科源电子科技有限公司 | Method for calibrating the fluorescence signal of polymerase chain reaction |
| CN111337441A (en) * | 2020-03-12 | 2020-06-26 | 深圳市朗诚科技股份有限公司 | Consistency adjustment method and related equipment |
| CN113933279A (en) * | 2021-11-04 | 2022-01-14 | 珠海丽珠试剂股份有限公司 | Fluorescence analysis assembly detection device and detection method |
| CN114509407A (en) * | 2022-04-19 | 2022-05-17 | 深圳市帝迈生物技术有限公司 | Quality detection component of biometric analyzer and detection method thereof |
| CN119223932A (en) * | 2024-12-03 | 2024-12-31 | 北京林电伟业电子技术有限公司 | A fluorescence microscope measurement calibration method and system |
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107064002A (en) * | 2017-03-28 | 2017-08-18 | 中国科学技术大学 | A kind of standard sources of fluorescence analysis |
| CN107271415A (en) * | 2017-06-16 | 2017-10-20 | 中国计量科学研究院 | It is a kind of for standard sample of photo of fluorescent instrument calibration measurement and preparation method thereof |
| CN107271415B (en) * | 2017-06-16 | 2019-11-29 | 中国计量科学研究院 | A kind of standard sample of photo and preparation method thereof for fluorescent instrument calibration measurement |
| CN110346551A (en) * | 2018-04-04 | 2019-10-18 | 南京东纳生物科技有限公司 | A kind of fluorescence immune chromatography quantitative analysis instrument testing calibration card and its detection method |
| CN109668865A (en) * | 2019-01-04 | 2019-04-23 | 上海科源电子科技有限公司 | Fluorescence detector Stability Determination device and measuring method |
| CN110927136B (en) * | 2019-12-25 | 2021-08-10 | 杭州微策生物技术股份有限公司 | Quality detection device of dry-type immunofluorescence POCT detection instrument and use method thereof |
| CN110927136A (en) * | 2019-12-25 | 2020-03-27 | 杭州微策生物技术有限公司 | Quality detection device of dry-type immunofluorescence POCT detection instrument and use method thereof |
| CN111157499A (en) * | 2020-01-06 | 2020-05-15 | 深圳市华科瑞科技有限公司 | Method for calibrating fluorescence detection instrument |
| CN111157499B (en) * | 2020-01-06 | 2021-05-18 | 深圳市华科瑞科技有限公司 | Method for calibrating fluorescence detection instrument |
| CN111206077A (en) * | 2020-02-12 | 2020-05-29 | 上海科源电子科技有限公司 | Method for calibrating the fluorescence signal of polymerase chain reaction |
| CN111337441A (en) * | 2020-03-12 | 2020-06-26 | 深圳市朗诚科技股份有限公司 | Consistency adjustment method and related equipment |
| CN113933279A (en) * | 2021-11-04 | 2022-01-14 | 珠海丽珠试剂股份有限公司 | Fluorescence analysis assembly detection device and detection method |
| CN113933279B (en) * | 2021-11-04 | 2023-09-15 | 珠海丽珠试剂股份有限公司 | Fluorescence analysis component detection device and detection method |
| CN114509407A (en) * | 2022-04-19 | 2022-05-17 | 深圳市帝迈生物技术有限公司 | Quality detection component of biometric analyzer and detection method thereof |
| CN114509407B (en) * | 2022-04-19 | 2022-10-28 | 深圳市帝迈生物技术有限公司 | Quality detection assembly of biological characteristic analyzer and detection method thereof |
| CN119223932A (en) * | 2024-12-03 | 2024-12-31 | 北京林电伟业电子技术有限公司 | A fluorescence microscope measurement calibration method and system |
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Application publication date: 20160907 |