CN207646217U - Biomolecule fluorescence detector metrological standard unit - Google Patents
Biomolecule fluorescence detector metrological standard unit Download PDFInfo
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- CN207646217U CN207646217U CN201720760504.2U CN201720760504U CN207646217U CN 207646217 U CN207646217 U CN 207646217U CN 201720760504 U CN201720760504 U CN 201720760504U CN 207646217 U CN207646217 U CN 207646217U
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Abstract
The utility model discloses a kind of biomolecule fluorescence detector metrological standard units, including computer, data transmission unit, microcontroller, light source module group.Light source module group designs corresponding shape according to different calibration instruments；The corner location of light source module group, center, four sides centre position be respectively arranged with luminescence unit.The calibration of linear and repeated meter characteristic index may be implemented in utility model device.The calibration of multi-wavelength condition lower linear in situ and repeatability may be implemented in the metrological standard unit of the utility model, is not necessarily to movable metering standard when changing wavelength, luminous intensity, ensure that the reliability of repeatability index metering.
Description
Technical field
The utility model is related to biomolecule fluorescence detector technical fields, are examined more particularly to a kind of biomolecule optics
Survey the metrological standard unit of device.
Background technology
With the development of life science and the progress of biotechnology, bio-measurement is more and more widely used to be examined in clinic
Test, food security, bio-safety, drug detection etc. fields, be related to the qualitative and quantitatives work such as nucleic acid, albumen, cell, microorganism
Make, wherein the emphasis for having become biometric and developing is measured to nucleic acid and the accurate of this kind of large biological molecule of protein.
The analysis means of nucleic acid and protein are varied, including chromatography, spectroscopic methodology, mass spectrography etc., wherein being based on light
The instrument and method for learning detection have obtained significant progress in recent years.For example, the common fluorescence real-time quantitative PCR of foranalysis of nucleic acids,
The common fluorescence ELISA of protein analysis and chemiluminescence immune assay etc. are all based on the means of optical detection.For
Ensure based on these optical detection means nucleic acid, protein analysis result it is accurate comparable, it is necessary to biomolecule optics is examined
It surveys device and carries out measurement and calibration, wherein linearly becoming the core meter characteristic index of this kind of detector with repeatability.
To realize that the linear and repeated measurement and calibration of detector, the standard sources that developing can trace to the source become pass therein
Key.It has been reported early period and is calibrated using LED light source, tritium light source etc., but its luminous intensity is traced to the source as difficult point, not
In the case of capable of realizing that Equipnal light intensity is traced to the source, often it is converted into and is traceable in the physical parameters such as absorbance, transmittance,
It can not directly reflect the linear of fluorescence detector.
Therefore, it is necessary to develop a kind of stabilization, can trace to the source, multi-parameter, applied widely biomolecule light
The metrological standard unit of detector is learned, realizes the calibration linearly with repeated meter characteristic index.
Utility model content
The purpose of this utility model is to provide a kind of metrological standard unit of biomolecule fluorescence detector.
A kind of biomolecule fluorescence detector metrological standard unit, including computer, data transmission unit, microcontroller, light
Source module.
Biomolecule fluorescence detector metrological standard unit described in the utility model, wherein the light source module group according to
Different calibration instrument and design corresponding shape；The calibration instrument includes real-time fluorescence quantitative PCR plate and ELISA Plate.
Biomolecule fluorescence detector metrological standard unit described in the utility model, wherein the four of the light source module group
Angle Position, center, four sides centre position be respectively arranged with luminescence unit.
Biomolecule fluorescence detector metrological standard unit described in the utility model, wherein the luminescence unit bottom
Equipped with miniature patch LED；The optical filter of specific wavelength is housed on miniature patch LED；Polymer thin is covered on optical filter
Film.
Biomolecule fluorescence detector metrological standard unit described in the utility model, wherein the miniature patch LED roots
It is different according to the wavelength of instrument to be calibrated, select the LED of different wave length as primary light source；The optical filter after filtering for obtaining
The narrow band light of bandwidth 10nm or so；The thin polymer film plays protection and fixed function.
Furthermore, the useful improvement of biomolecule fluorescence detector metrological standard unit described in the utility model
For that according to actual needs, can be placed around with not in the centre position on the corner location of light source module group, center, four sides
The luminescence unit of co-wavelength.
Biomolecule fluorescence detector metrological standard unit described in the utility model, wherein the data transmission unit
Transmission medium between computer and microcontroller is connected by wired or wireless mode；When using wired connection mode,
Using serial communication mode by being electrically connected between winding displacement and microcontroller；When using radio connection when, computer with connect
It is connected by electrical property of flat cable in a manner of serial communication between receipts unit；
It is electrically connected between microcontroller and transmitter unit；Pass through radio frequency or infrared mode between transmitter unit and receiving unit
Establish wireless data link；
It is connected by wired or wireless way between the data input pin and data transmission unit of computer；The data pass
It is electrically connected between defeated unit and microcontroller；It is electrically connected between the light source module group and microcontroller.
Biomolecule fluorescence detector metrological standard unit described in the utility model, wherein the light source module group is used for
The pulsed light for providing varying strength, the continuous light of different wave length or different duty is realized to fluorescence microplate reader, chemiluminescence point
The calibration of analyzer, fluorescence real-time quantitative PCR instrument optical metrology characteristic；The computer is for providing the information such as frequency, duty ratio
Human-computer interaction interface；Host computer procedure is run on computer, the interface of human-computer interaction is provided, for serial communication ginseng to be arranged
The information such as number, wavelength, pulse frequency, duty ratio；The microcontroller is passing through choosing for receiving the arrange parameter that computer is sent
The mode of the pulse frequency and duty ratio of different I/O ports, change I/O port, driving light source module group is taken to send out different wave length, difference
The optical signal of frequency, different duty.
The biomolecule fluorescence detector metrological standard unit of the utility model compared with prior art, protrudes effect and exists
In：
(1) the utility model is a kind of biomolecule optics stabilization, can trace to the source, multi-parameter, applied widely
The calibration of linear and repeated meter characteristic index may be implemented in the metrological standard unit of detector.
(2) tracing to the source for absolute light intensity is converted into tracing to the source for time by the utility model, can accurately, directly reflect light extraction
Learn the linear of detector.
(3) school of multi-wavelength condition lower linear in situ and repeatability may be implemented in the metrological standard unit of the utility model
Standard is not necessarily to movable metering standard when changing wavelength, luminous intensity, ensure that the reliability of repeatability index metering.
(4) metrological standard unit of the utility model can be communicated wirelessly with host computer, and standard set-up is using
It does not influence instrument in the process and closes the operations such as door, can ensure the state complete one when state and the detection when instrument metering
It causes, ensure that the reliability of metric results.
The biomolecule fluorescence detector measurement criteria of explanation and specific embodiment to the utility model below in conjunction with the accompanying drawings
Device is described further.
Description of the drawings
Fig. 1 is 96 hole elisa Plates optics module schematic diagrames；
Fig. 2 is optics module luminescence unit composition schematic diagram；
Fig. 3 is the optics module schematic diagram comprising different wave length luminescence unit.
Specific implementation mode
Embodiment 1
Biomolecule fluorescence detector metrological standard unit, including computer, data transmission unit, microcontroller, light source die
Group.Transmission medium of the data transmission unit between computer and microcontroller, is connected by wired or wireless mode；Work as use
When wired connection mode, using serial communication mode by being electrically connected between winding displacement and microcontroller；When using wireless connection side
When formula, connected by electrical property of flat cable in a manner of serial communication between computer and receiving unit；
It is electrically connected between microcontroller and transmitter unit；Pass through radio frequency or infrared mode between transmitter unit and receiving unit
Establish wireless data link；
It is connected by wired or wireless way between the data input pin and data transmission unit of computer；The data pass
It is electrically connected between defeated unit and microcontroller；It is electrically connected between the tunable light source and microcontroller.
The pulsed light that light source module group is used to provide varying strength, the continuous light of different wave length or different duty is realized to glimmering
The calibration of light microplate reader, chemiluminescent analyzer, fluorescence real-time quantitative PCR instrument optical metrology characteristic；
Computer is used to provide the human-computer interaction interface of the information such as frequency, duty ratio；Host computer procedure is run on computer,
The interface that human-computer interaction is provided, for the information such as serial communication parameter, wavelength, pulse frequency, duty ratio to be arranged；
Microcontroller is for receiving the arrange parameter that computer is sent, in the arteries and veins by choosing different I/O ports, changing I/O port
The mode of frequency and duty ratio is rushed, driving light source module group sends out the optical signal of different wave length, different frequency, different duty.
Light source module group designs corresponding shape according to different calibration instruments；Wherein, calibration instrument includes real-time fluorescence
Quantitative PCR plate and ELISA Plate etc..
The corner location of light source module group, center, four sides centre position be respectively arranged with luminescence unit 2.It is real-time with 96 holes
For quantitative fluorescent PCR plate, ELISA Plate, optics module schematic diagram as shown in Figure 1,96 hole elisa Plates exemplary position A1,
Optics module is housed on the position A6, A12, D1, D6, D12, H1, H6, H12.
As shown in Fig. 2, 2 bottom of luminescence unit is equipped with miniature patch LED5；Specific wavelength is housed on miniature patch LED5
Optical filter 4；Thin polymer film 3 is covered on optical filter 4.Miniature patch LED5 is different according to the wavelength of instrument to be calibrated,
Select the LED of different wave length as primary light source；Optical filter 4 after filtering for obtaining the narrow band light of bandwidth 10nm or so；Polymerization
Object film 3 plays protection and fixed function.
It, according to actual needs, can be in the interposition of the corner location of light source module group, center, four sides in conjunction with shown in Fig. 3
It sets and is placed around the luminescence unit (6,7,8,9) with different wave length, realize same plate to different wave length optics detector
Calibration.
The application method of biomolecule fluorescence detector metrological standard unit：
Wavelength, frequency and duty cycle parameters are sent by computer, the adjustment to luminous intensity and frequency is realized, passes through inspection
The waveform of fixed qualification oscillograph detection single-chip I/O mouth, obtains the time that can be traced to the source and frequency parameter, thus by luminous intensity
The problem of tracing to the source is converted into tracing to the source for time and frequency.
When carrying out linear gauging, the light source module group of corresponding wavelength is selected according to the Detection wavelength of instrument to be calibrated first,
Light source module group is put into instrument to be calibrated；Then duty ratio is arranged 100%, it, will be to be calibrated by changing flicker frequency
The luminous signal value of instrument is adjusted near the maximum value of the range of linearity, be then respectively set duty ratio be 100%, 80%,
60%, 40%, 20% and 0, the luminous signal value of difference register instrument；By duty ratio and corresponding luminous signal value into line
Property fitting, and calculate the linearity error under each signal value according to formula (1)；
In formula：
--- slope average value；
N --- the number of different duty setting；
Ii --- the signal strength of instrument under different duty；
Di --- duty ratio；
In formula：
Δi--- linearity error；
Ki --- the slope under a certain duty ratio；
When carrying out repeatability calibration, on the basis of above-mentioned parameter optimizes, duty ratio is set as 20%, then with waiting for school
Accurate instrument calculates the relative standard deviation of signal according to formula (3) to the luminous intensity replication 20 times under the duty ratio
Characterization as repeatability；
In formula：
xi--- ith measured value；
--- the average value of n times measured value；
N--- pendulous frequencies, herein n=20.
Embodiment 2
Biomolecule fluorescence detector metrological standard unit, including computer, data transmission unit, microcontroller, light source die
Group.Transmission medium of the data transmission unit between computer and microcontroller, in host computer side, by control software gui interface
The wavelength of input, period, duty cycle information are sent into transmitter unit by way of serial communication, by AX5043 radio frequency chips with
433MHz carrier waves carry out wireless radio frequency transmission；At metrology and measurement end, radiofrequency signal is equally received by AX5043, and therefrom demodulate
Go out to control 51 microcontrollers that information is sent into STC, drives different I/O ports to light therewith according to wavelength information by the microcontroller
Connected light source module group realizes the selection function of wavelength, while generating PWM modulation according to the period of setting and duty cycle information
Signal controls the brightness of light source module group.Light source module group by a 520nm LED and centre wavelength 520nm, bandwidth 10nm
Interferometric filter composition, the light of the 520nm sent out is used to simulate the fluorescence signal of FAM.
When carrying out linear gauging, metrology and measurement end is put into quantitative PCR apparatus, duty ratio is set as 100% first,
By change the period, luminous intensity is adjusted to 300000, be then respectively set duty ratio be 100%, 80%, 60%, 40%,
20% and 0, measure luminous signal value respectively, under each duty ratio duplicate measurements be averaged for 3 times, value is respectively 300758,
239001,181147,128591,69845,5642, linearly dependent coefficient 0.999 reflects that instrument is linearly good.
When carrying out repeatability calibration, under the conditions of above-mentioned parameter, duty ratio is set as 20%, then with instrument to be calibrated
Device is to the luminous intensity replication 20 times under the duty ratio, and it is 68197 to measure luminous intensity average value, and relative standard deviation is only
It is 0.93%, it was demonstrated that Instrument measuring repeatability is good.
For prominent the beneficial effects of the utility model, following comparative example experiment has also been carried out,
Comparative example 1
The linear and repeated of instrument is calibrated using standard items.By FAM fluoresceins standard items boric acid salt buffer
Liquid is diluted, and constantly adjusts concentration, and 40 μ L FAM fluorescein standard solution are then added in the hole of PCR amplification plate, is measured
Its fluorescence intensity level makes its fluorescence intensity close to 300000, then by this solution according to 1：2 gradient dilution step by step, until most
The fluorescence intensity of low concentration standard items is 5000 or so or near detection limit.Each standard solution that dilution is obtained is each
Analysis three times, measures its fluorescence intensity, is averaged, then linearly returned to the concentration of standard items and its fluorescence signal intensity
Return, linearly dependent coefficient 0.993.The standard solution for being 70000 or so to fluorescence signal intensity continuously measures 20 times, calculates
The relative standard deviation of fluorescence signal intensity is 2.1%.
The mode that by the way of standards calibration and the metering device of the utility model embodiment is calibrated is compared, for same
The linear and repeated measurement criteria dress being all not so good as using the utility model embodiment when one instrument is calibrated using calibration object
The calibration result set.
This is primarily due to the molecular fluorescence in solution and receives energy between interaction, molecule and solvent between molecule
Amount exchange etc., makes its fluorescence signal intensity deviate with theoretical value, while also causing the repeatability of signal measurement bad.Cause
This, using the calibrating mode of fluorescein standard items, it is impossible to the performance of true reflection instrument.
Embodiment described above is only that preferred embodiments of the present invention are described, not to this practicality
Novel range is defined, and under the premise of not departing from the spirit of the design of the utility model, those of ordinary skill in the art are to this
The various modifications and improvement that the technical solution of utility model is made should all fall into the protection of the utility model claims book determination
In range.
Claims (6)
1. a kind of biomolecule fluorescence detector metrological standard unit, it is characterised in that：Including computer, data transmission unit,
Microcontroller, light source module group；The corner location of the light source module group, center, four sides centre position be respectively arranged with luminescence unit
(2)；Luminescence unit (2) bottom is equipped with miniature patch LED (5)；The filter of specific wavelength is housed on miniature patch LED (5)
Mating plate (4)；Thin polymer film (3) is covered on optical filter (4).
2. biomolecule fluorescence detector metrological standard unit according to claim 1, it is characterised in that：The light source die
Group designs corresponding shape according to different calibration instruments；The calibration instrument includes real-time fluorescence quantitative PCR plate and enzyme mark
Plate.
3. biomolecule fluorescence detector metrological standard unit according to claim 2, it is characterised in that：The miniature patch
Piece LED (5) is different according to the wavelength of instrument to be calibrated, selects the LED of different wave length as primary light source；The optical filter (4)
Narrow band light for obtaining bandwidth 10nm or so after optical filtering；The thin polymer film (3) plays protection and fixed function.
4. biomolecule fluorescence detector metrological standard unit according to claim 2, it is characterised in that：According to practical need
It wants, the luminous list with different wave length can be placed around in the centre position on the corner location of light source module group, center, four sides
Member.
5. according to claim 1-4 any one of them biomolecule fluorescence detector metrological standard units, it is characterised in that：Institute
Transmission medium of the data transmission unit between computer and microcontroller is stated, is connected by wired or wireless mode；Work as use
When wired connection mode, using serial communication mode by being electrically connected between winding displacement and microcontroller；When using wireless connection side
When formula, connected by electrical property of flat cable in a manner of serial communication between computer and receiving unit；
It is electrically connected between microcontroller and transmitter unit；It is established by radio frequency or infrared mode between transmitter unit and receiving unit
Wireless data link；
It is connected by wired or wireless way between the data input pin and data transmission unit of computer；The data transmission list
It is electrically connected between member and microcontroller；It is electrically connected between the light source module group and microcontroller.
6. biomolecule fluorescence detector metrological standard unit according to claim 5, it is characterised in that：The light source die
The pulsed light of continuous light or different duty of the group for providing varying strength, different wave length is realized to fluorescence microplate reader, chemistry
The calibration of luminescence analyzer, fluorescence real-time quantitative PCR instrument optical metrology characteristic；The computer is for providing frequency, duty ratio
Etc. information human-computer interaction interface；Host computer procedure is run on computer, and the interface of human-computer interaction is provided, it is logical for serial ports to be arranged
Interrogate the information such as parameter, wavelength, pulse frequency, duty ratio；The microcontroller is for receiving the arrange parameter that computer is sent, logical
Cross choose different I/O ports, the pulse frequency and duty ratio that change I/O port mode, driving light source module group sends out different wave length, no
The optical signal of same frequency, different duty.
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| CN201720760504.2U CN207646217U (en) | 2017-06-27 | 2017-06-27 | Biomolecule fluorescence detector metrological standard unit |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107129931A (en) * | 2017-06-27 | 2017-09-05 | 中国计量科学研究院 | Biomolecule fluorescence detector metrological standard unit and application method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107129931A (en) * | 2017-06-27 | 2017-09-05 | 中国计量科学研究院 | Biomolecule fluorescence detector metrological standard unit and application method |
| CN107129931B (en) * | 2017-06-27 | 2023-04-18 | 中国计量科学研究院 | Biomolecule optical detector metering standard device and use method |
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