CN109556738A - Analog measurement method, measure data fitting method and chemical luminescence detector - Google Patents
Analog measurement method, measure data fitting method and chemical luminescence detector Download PDFInfo
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Abstract
The present invention relates to a kind of analog measurement method, data fitting method and chemi-luminescent detectors, this method comprises: the direct current signal that the incident photon in acquisition optical signal exports after being enhanced by photomultiplier tube, being amplified；The direct current signal of photomultiplier tube output is converted into voltage signal by dc amplifier, and the voltage signal is exported into filtered voltage signal by filter circuit；The first voltage value in default start time point and the second voltage value in default end time point are obtained, the difference of the second voltage value and the first voltage value is calculated by analog-digital converter, and export ADC value；The first simulation count value of optical signals is calculated according to the ADC value.It by the above method, can guarantee to obtain when counting the optical signal of varying strength good linear, reduce counting error, keep chemiluminescent analyzer more reliable to the measurement of optical signal.
Description
Technical field
The present invention relates to the field of medical instrument technology, more particularly to a kind of analog measurement method, measure data fitting side
Method and chemical luminescence detector.
Background technique
Chemiluminescence immune assay (chemiluminescence immunoassay, CLIA), be will have it is highly sensitive
The product that the chemical luminescent detecting technology of degree is combined with the immune response of high specific.The technology is mainly for detection of analysis
The content of the micro substances such as various antigens, antibody, hormone, marker.Full-automatic chemiluminescence immunoassay analysis meter is exactly with this base
Plinth exploitation forms antigen-antibody mixture by the way that particular agent is added to testing liquid, under the effect of external excitation reagent
Optical signal is generated, optical signal is measured by photomultiplier tube, by measurement result Fitting Analysis, to detect
The content of test substance in liquid.
Currently, up to nearly hundred kinds of the detection project of chemiluminescent analyzer, cover hormone, cancer etc., or even can be right
Some diseases are directly made a definite diagnosis, and play an important role in clinic.With population constantly increase and people's health consciousness plus
By force, immune detection quantity steeply rises, especially liver function, thyroid gland class physical examination project, the analysis to chemiluminescent analyzer
Speed, reliability propose tightened up requirement.
Traditional Single photon counting method can be limited to the range of linearity of photomultiplier tube, when the count value of optical signal in 1s
When more than 1M (1*10^6), counting error is more than 1%, and when the count value of optical signal in 1s is more than 5M, error will increase
Greatly to 10%, wherein the count value of optical signal indicates the relative light unit (relative light unit, RLU) of optical signal.
In clinical application, hepatitis B surface antigen (HBsAg), human chorionic gonadotrophin (HCG) positive sample luminous intensity
It can exceed that 5M, if counting error is big, can not make accurately quantitative or qualitatively judge.
Summary of the invention
Based on this, it is necessary to which for traditional Single photon counting method, when light signal strength is higher, counting error is bigger
Problem provides a kind of analog measurement method, measure data fitting method and chemiluminescence that can effectively improve counting accuracy
Analyzer.
A kind of analog measurement method, for carrying out simulation counting to optical signal, which comprises
The direct current signal that exports after obtaining the incident photon in optical signal and being enhanced by photomultiplier tube, amplified；
The direct current signal of photomultiplier tube output is converted into voltage signal by dc amplifier, and will be described
Voltage signal exports filtered voltage signal by filter circuit；
The first voltage value in default start time point and the second voltage value in default end time point are obtained, is passed through
Analog-digital converter calculates the difference of the second voltage value and the first voltage value, and exports ADC value；
The simulation count value of optical signals is calculated according to the ADC value.
The simulation count value and ADC value are in a linear relationship in one of the embodiments,.
The simulation count value is the linear function of ADC value in one of the embodiments,.
In capacitor, the analog-digital converter is being preset for the voltage signal accumulation in one of the embodiments,
Time point beginning reads the first voltage value on the capacitor, reads second on the capacitor in default end time point
Voltage value, to calculate the difference of the second voltage value and the first voltage value.
The analog-digital converter carries out voltage acquisition in one of the embodiments, and the time of analog-to-digital conversion is less than
The voltage of the capacitor saves the time.
A kind of measure data fitting method is carried out for the measurement data to analog measurement method and Single photon counting method
Fitting, which comprises
Obtain the count value that Single photon counting method counts the incident photon of optical signal；
The count value is extended linearly when the count value is greater than the first preset threshold, is obtained to not sharing the same light
The simulation count value of signal strength；
Corresponding ADC value under different light signal strengths, the analogue measurement side are measured by analog measurement method
The light signal strength of method measurement and the light signal strength of Single photon counting method measurement correspond；
Data fitting is carried out by least square method according to the ADC value and the simulation count value, is counted
According to matched curve.
The Single photon counting method includes: in one of the embodiments,
The pulse current that exports after obtaining the incident photon in optical signal and being enhanced by photomultiplier tube, amplified；
The pulse signal that the pulse current exports after preamplifier carries out voltage amplification is obtained, and to described
Pulse signal obtains the gentle pulse signal of waveform after being filtered；
The gentle pulse signal of the waveform is sent to microprocessor count after shaping pulse, scaling down processing.
The simulation count value and ADC value are in a linear relationship in one of the embodiments,.
The simulation count value is the linear function of ADC value in one of the embodiments,.
A kind of chemical luminescence detector, including measuring chamber, the measuring chamber uses above-mentioned measure data fitting method, right
The measurement data of analog measurement method and Single photon counting method is fitted analysis.
Above-mentioned analog measurement method, measure data fitting method and chemical luminescence detector, by analogy aerotriangulation by light
Signal is converted to voltage signal, and exports ADC value according to the voltage difference in preset time period by analog-digital converter,
The first simulation count value of optical signals is calculated according to the ADC value；Also, when the count value of optical signal is greater than first in advance
If when threshold value, the second simulation count value that the ADC value of analog measurement method measurement is measured with Single photon counting method is led to
It crosses least square method and carries out data fitting, obtain data matched curve.It can guarantee to count the optical signal of varying strength
When, obtains good linear, reduces counting error, keeps chemiluminescent analyzer more reliable to the measurement of optical signal.
Detailed description of the invention
Fig. 1 is the flow diagram of analog measurement method in one embodiment；
Fig. 2 is the flow diagram of Single photon counting method in one embodiment；
Fig. 3 is the flow diagram of measure data fitting method in one embodiment；
Fig. 4 is the flow diagram of Single photon counting method in one embodiment；
Fig. 5 is the flow diagram of analog measurement method in one embodiment.
Fig. 6 is analog measurement method and counting method ideal Value Data matched curve figure in one embodiment.
Specific embodiment
To facilitate the understanding of the present invention, a more comprehensive description of the invention is given in the following sections with reference to the relevant attached drawings.In attached drawing
Give presently preferred embodiments of the present invention.But the invention can be realized in many different forms, however it is not limited to herein
Described embodiment.On the contrary, purpose of providing these embodiments is keeps the understanding to the disclosure more saturating
It is thorough comprehensive.
Unless otherwise defined, all technical and scientific terms used herein and belong to technical field of the invention
The normally understood meaning of technical staff it is identical.Term as used herein in the specification of the present invention is intended merely to retouch
State the purpose of specific embodiment, it is not intended that the limitation present invention.It is appreciated that term " first " used in the present invention,
" second " etc. can be used to describe various elements herein, but these elements should not be limited by these terms.These terms are only used to
It distinguishes the first element from the other element.
A kind of analog measurement method presented below, by carrying out analogue measurement to optical signal to obtain the simulation of optical signal
Count value, as shown in fig.1, this approach includes the following steps S110~S130.
S110: obtain optical signal in incident photon enhanced by photomultiplier tube, amplify after export direct current letter
Number.
In one embodiment, the process flow of analog measurement method is as shown in figure 5, firstly, by the incidence in optical signal
Photon exports pulse current after photomultiplier tube 501 is enhanced, amplified.Wherein, the selection of photomultiplier tube 501 is answered
Meet the requirement of reagent system.Optionally, it in the luminescence system applied by one embodiment, is excited after marker is added
Substrate generates optical signal, and the optical signal peak wavelength generated is 420nm, and with micro stray light.Therefore, in order to
Highest quantum efficiency (from the photoelectron number of photocathode surface launching and the ratio of incident light subnumber) is obtained at 420nm, and can be use up
Possibly it is collected into the optical signal that luminous determinand generates, 501 chosen spectrum response wave length scope of photomultiplier tube is 300~
650nm, peak wavelength 420nm, linear counting range are 0-400M type per second.
S120: the direct current signal of photomultiplier tube output is converted into voltage signal by dc amplifier, and will
The voltage signal exports filtered voltage signal by filter circuit.
Specifically, voltage is converted to by the direct current signal that dc amplifier 502 exports photomultiplier tube 501
Signal.Wherein, due to the electric current very little that photomultiplier tube 501 generates, down to uA rank, therefore it is required that the biased electrical of amplifier
Stream, offset voltage are very small.In addition, temperature will be slow rising over time, therefore lack of proper care inside measuring system
Voltage temperature drift is also an important type selecting index.Therefore the biasing of dc amplifier 502 can be chosen in one embodiment
Electric current is 0.2pA, and offset voltage 5uV, offset voltage temperature drift is 0.26uV/ DEG C to meet the needs of measuring system.
Further, the voltage signal is exported into filtered electricity after low-pass filter 503 carries out low-pass filtering
Press signal.Optionally, filter circuit can be selected as RC low pass circuit in one embodiment, and the RC low pass circuit
Cutoff frequency should be set to be slightly greater than the maximum frequency of optical signal.
S130: obtaining the first voltage value in default start time point and the second voltage value in default end time point,
The difference of the second voltage value and the first voltage value is calculated by analog-digital converter, and exports ADC value.
Specifically, in capacitor, analog-digital converter 504 is read in default start time point for the voltage signal accumulation
The first voltage value on the capacitor reads the second voltage value on the capacitor in default end time point, to calculate
The difference of second voltage value and the first voltage value is stated, and ADC value is exported according to voltage difference.
In one embodiment, analog-digital converter 504 can measure the voltage in the capacitor, to count
Calculate the count value of optical signal in the set time.For improving measurement accuracy, analog-digital converter 504 carries out voltage acquisition and mould
The voltage that the time of number conversion is less than the capacitor is saved the time, i.e., the acquisition of analog-digital converter 504 conversion time is less than capacitor
The charging time of device.
S140: the first simulation count value of optical signals is calculated according to the ADC value.
Specifically, the first simulation count value is to be calculated simultaneously by voltage strength of the analog measurement method to optical signal
The ADC value can be exported to microprocessor and carry out that the first simulation meter is calculated by the ideal count value simulated
Numerical value.The first simulation count value and ADC value are in a linear relationship, and optionally, the first simulation count value is mould
The linear function of number conversion value.
Above-mentioned analog measurement method converts optical signals to voltage signal by analogy aerotriangulation, and passes through analog-to-digital conversion
Device exports ADC value according to the voltage difference in preset time period, calculates the of optical signals according to the ADC value
One simulation count value.It can guarantee to obtain when counting the optical signal of varying strength good linear, reduce counting and miss
Difference keeps chemiluminescent analyzer more reliable to the measurement of optical signal.
A kind of data fitting method presented below, combine Single photon counting method and analog measurement method to optical signal into
Row counts, and can guarantee to obtain when counting the optical signal of varying strength good linear, reduces counting error.Ginseng
It reads shown in Fig. 3, this approach includes the following steps S310~S340.
S310: the count value that Single photon counting method counts the incident photon of optical signal is obtained.
Specifically, light source is detected by photomultiplier tube, when light emitting source is very faint, in photomultiplier tube
Temporal resolution in almost without state existing for more than two photoelectrons, referred to as monochromatic light subdomains, at this time using single
Photon counting method can be stablized, accurately count to optical signal.
As shown in Fig. 2, Single photon counting method includes the following steps S210~S230.
S210: obtain optical signal in incident photon enhanced by photomultiplier tube, amplify after export pulse electricity
Stream.
S220: the pulse signal that the pulse current exports after preamplifier carries out voltage amplification is obtained, and right
The pulse signal obtains the gentle pulse signal of waveform after being filtered.
S230: the gentle pulse signal of the waveform is sent to microprocessor meter after shaping pulse, scaling down processing
Number.
In one embodiment, the realization process of Single photon counting method is as shown in figure 4, by the incident photon in optical signal
Pulse current is exported after photomultiplier tube 401 is enhanced, amplified.The processing of the process and above-mentioned analog measurement method
Process is identical, is not repeated herein.
Further, pulse current photomultiplier tube 401 exported carries out voltage amplification by preamplifier 402
Output pulse signal afterwards.Wherein, it in pre-amplification circuit, needs a low noise, Low-bias Current, low maladjustment voltage, put
Big multiple is high, the biggish amplifier 402 of bandwidth, more faint (uA grades) to cope with input signal, and frequency is higher, bandwidth compared with
Big feature.Therefore, the current noise of selection amplifier 402 is in one embodimentBias current is 5uA,
Offset voltage is 6mV, bandwidth 725MHz, Open loop gain cofficient 80dB.In the present embodiment, amplifying circuit is put using exchange
UA grades of pulse signals are converted to mV grades of pulse signals by large model.
Further, by the amplified pulse signal, output waveform is gentle after discriminator 403 filters out dark noise
Pulse signal, and by the gentle pulse signal of the waveform after shaping pulse, frequency divider 404 divide output to micro- place
Device 405 is managed to count.Wherein, discriminator 403 is made of the comparator and linear voltage regulator that switching speed is nanosecond, frequency divider
404 effects are to reduce pulse signal frequency, are promoted and are counted linearly, reduce the requirement of subsequent counter channel frequency.Microprocessor 405
By the pulse number in the pulse signal after calculation processing to obtain the incident photon quantity in optical signal.
S320: the count value is extended linearly when the count value is greater than the first preset threshold, is obtained to not
Second with light signal strength simulates count value.
In one embodiment, the first preset threshold can be big according to counting deviation required during actual test
It is small to be set, for example, first preset threshold can be set as 1M.The count value and light signal strength of Single photon counting method
In a linear relationship, the second simulation count value is that the reason for the Single photon counting method that linear analogue goes out is carried out by Single photon counting method
Want to be worth.
S330: corresponding ADC value under different light signal strengths is measured by analog measurement method, the simulation is surveyed
The light signal strength of amount method measurement and the light signal strength of Single photon counting method measurement correspond.
Specifically, it is influenced due to the count accuracy of Single photon counting method by the time response of photomultiplier tube, when adjacent
When time interval between two pulses is too small, photomultiplier tube can not be identified as two effective impulses.Work as light signal strength
When excessively high, the counting error of Single photon counting method can also be increased with it, and therefore, when light signal strength is high, simulation can be used and survey
The measurement of amount method carries out modified result to Single photon counting method.It is measured using analog measurement method with Single photon counting method
With measuring under light signal strength, the ADC value under different light signal strengths is obtained.
S340: data are carried out by least square method and are intended according to the ADC value and the second simulation count value
It closes, obtains data matched curve.
In one embodiment, using feedback-type, can digital regulated brightness blue-ray LED module as control light source, drive
Streaming current and luminous intensity have good linear relationship.In other embodiments, can be used any one control digit value and
The directly proportional control light source of light source luminescent intensity.Under multiple and different standard sources brightness, while using analog measurement method
Optical signal is measured with Single photon counting method.Theoretically, analog measurement method, Single photon counting method ideal value and
Light-source brightness is linear, therefore the ideal value of the measured value of analog measurement method and Single photon counting method also linear pass
System, and the relational expression of the count value of the measured value and Single photon counting method of analog measurement method are as follows:
Y0=b*X+a；
Wherein, Y0 is the ideal value of Single photon counting method, and the as second simulation count value, X is the survey of analog measurement method
Magnitude, as ADC value, b are straight slope, and a is Linear intercept.
Further, when the count value of Single photon counting method is more than 1M, Single photon counting method ideal value Y0 can be by small
Actual measurement count results when 1M are extended linearly.Analog measurement method and Single photon counting method test result is as follows table
It is shown.
Analog measurement method and Single photon counting method test result contrast table
From test result as can be seen that analog measurement method embodies good line when photon count value reaches 20M
Property, it counts deviation and is respectively less than 1%, and Single photon counting method counts deviation and just compares analogue measurement when photon count value reaches 1M
The counting deviation of method is big, so being no longer applicable in when photon count value reaches 1M using Single photon counting method.Implement herein
In example, it is desirable that count deviation and be respectively less than 1%, then the first preset threshold is set as 1M.In other embodiments, the first default threshold
Depending on the counting deviation that value can be required during actual test.It should be understood that counting, deviation is smaller, and measurement is smart
It spends higher.
As shown in fig. 6, analog measurement method and counting method ideal value are used when the count value RLU of optical signal reaches 1M
Least square method carries out data fitting, obtains analog measurement method and counting method ideal Value Data matched curve figure, simulates in figure
Measurement method A/D value is ADC value.
Analog measurement method and counting method ideal Value Data matched curve figure
It can be obtained by the analog measurement method and counting method ideal Value Data matched curve figure, be measured through analog measurement method
Obtained simulation count value=1147.5 × ADC value -30543784.
As it can be seen that being less than the luminous intensity optical signal of 1M for the count value RLU of optical signal, Single photon counting method pair should be chosen
Optical signal is counted, and linear, stability, signal-to-noise ratio and measurement accuracy show relatively good when optical signal is weaker.Work as light
It when the count value RLU of signal is greater than 1M, is measured by using voltage value of the analog measurement method to optical signal, then by institute
It states voltage value and is converted to simulation count value, it is linear to obtain good measurement.In conclusion being less than 20M for count value RLU
Any luminous intensity optical signal measurement result may be expressed as:
Above-mentioned measure data fitting method, by the way that when the count value of optical signal is greater than the first preset threshold, simulation is surveyed
The second simulation count value that the ADC value of amount method measurement is measured with Single photon counting method is counted by least square method
According to fitting, data matched curve is obtained.Meter is converted by simulation measured value (i.e. ADC value) by data matched curve
Number method count value, for upper computer software display counting as a result, obtaining measurement data.Above-mentioned measure data fitting method can
Guarantee obtains when counting to the optical signal of varying strength good linear, reduces counting error, makes chemiluminescence analysis
Instrument is more reliable to the measurement of optical signal.
In one embodiment, a kind of chemical luminescence detector is also provided, which includes measuring chamber,
The measuring chamber is using the measure data fitting method in the various embodiments described above, to analog measurement method and Single photon counting method
Measurement data be fitted analysis.Good linear, drop can be guaranteed to obtain when counting the optical signal of varying strength
Low counting error, it is more reliable to the measurement of optical signal.
Each technical characteristic of embodiment described above can be combined arbitrarily, for simplicity of description, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, all should be considered as described in this specification.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to guarantor of the invention
Protect range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of analog measurement method, for carrying out simulation counting to optical signal, which comprises
The direct current signal that exports after obtaining the incident photon in optical signal and being enhanced by photomultiplier tube, amplified；
The direct current signal that photomultiplier tube exports is converted into voltage signal by dc amplifier, and the voltage is believed
Number filtered voltage signal is exported by filter circuit；
The first voltage value in default start time point and the second voltage value in default end time point are obtained, is turned by modulus
Parallel operation calculates the difference of the second voltage value and the first voltage value, and exports ADC value；
The first simulation count value of optical signals is calculated according to the ADC value.
2. analog measurement method according to claim 1, which is characterized in that the first simulation count value and analog-to-digital conversion
It is worth in a linear relationship.
3. analog measurement method according to claim 2, which is characterized in that the first simulation count value is analog-to-digital conversion
The linear function of value.
4. analog measurement method according to claim 1, which is characterized in that the voltage signal is accumulated in capacitor,
The analog-digital converter reads the first voltage value on the capacitor in default start time point, reads in default end time point
The second voltage value on the capacitor is taken, to calculate the difference of the second voltage value and the first voltage value.
5. analog measurement method according to claim 4, which is characterized in that the analog-digital converter carry out voltage acquisition with
And the time of analog-to-digital conversion is less than the voltage savings time of the capacitor.
6. a kind of measure data fitting method, intends for the measurement data to analog measurement method and Single photon counting method
It closes, which comprises
Obtain the count value that Single photon counting method counts the incident photon of optical signal；
The count value is extended linearly when the count value is greater than the first preset threshold, is obtained strong to different optical signals
Second simulation count value of degree；
Corresponding ADC value under different light signal strengths, the analog measurement method measurement are measured by analog measurement method
Light signal strength and Single photon counting method measurement light signal strength correspond；
Data fitting is carried out by least square method according to the ADC value and the second simulation count value, obtains data
Matched curve.
7. measure data fitting method according to claim 6, which is characterized in that the Single photon counting method includes:
The pulse current that exports after obtaining the incident photon in optical signal and being enhanced by photomultiplier tube, amplified；
The pulse signal that the pulse current exports after preamplifier carries out voltage amplification is obtained, and the pulse is believed
The gentle pulse signal of waveform is obtained after number being filtered；
The gentle pulse signal of the waveform is sent to microprocessor count after shaping pulse, scaling down processing.
8. measure data fitting method according to claim 6, which is characterized in that the second simulation count value and modulus
Conversion value is in a linear relationship.
9. measure data fitting method according to claim 8, which is characterized in that the second simulation count value is modulus
The linear function of conversion value.
10. a kind of chemical luminescence detector, which is characterized in that including measuring chamber, the measuring chamber uses claim 6-9 such as to appoint
Measure data fitting method described in one is fitted point the measurement data of analog measurement method and Single photon counting method
Analysis.
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| CN113324938A (en) * | 2021-04-26 | 2021-08-31 | 长春吉大·小天鹅仪器有限公司 | Infrared oil measurement method and oil measurement instrument based on pressure-frequency conversion phase-sensitive detector |
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| CN2409559Y (en) * | 2000-03-06 | 2000-12-06 | 北京医海引航生物技术有限公司 | High linear single photon counter |
| CN1932859A (en) * | 2005-07-21 | 2007-03-21 | 华南师范大学 | Photon counter based on programmable logic |
| CN201229696Y (en) * | 2008-07-24 | 2009-04-29 | 天津港东科技发展股份有限公司 | The single photon counting experimental system |
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| CN2409559Y (en) * | 2000-03-06 | 2000-12-06 | 北京医海引航生物技术有限公司 | High linear single photon counter |
| CN1932859A (en) * | 2005-07-21 | 2007-03-21 | 华南师范大学 | Photon counter based on programmable logic |
| CN201229696Y (en) * | 2008-07-24 | 2009-04-29 | 天津港东科技发展股份有限公司 | The single photon counting experimental system |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113324938A (en) * | 2021-04-26 | 2021-08-31 | 长春吉大·小天鹅仪器有限公司 | Infrared oil measurement method and oil measurement instrument based on pressure-frequency conversion phase-sensitive detector |
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