CN109596582A - Bioluminescence rapid detection method - Google Patents
Bioluminescence rapid detection method Download PDFInfo
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- CN109596582A CN109596582A CN201811409480.1A CN201811409480A CN109596582A CN 109596582 A CN109596582 A CN 109596582A CN 201811409480 A CN201811409480 A CN 201811409480A CN 109596582 A CN109596582 A CN 109596582A
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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
- G01N21/6489—Photoluminescence of semiconductors
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Abstract
The present invention discloses a kind of bioluminescence rapid detection method, and single photocurrent integral measurement is the following steps are included: LED is closed, carried out the conversion of first time adc data, circuit itself the drift voltage V under output LED closed state after integrating capacitor settingOFF Offset；Integrating amplification circuit is successively carried out after resetting and periodically integrating, carries out second of adc data conversion, export the integral voltage V under LED closed stateOFF Integrate；LED is opened, the conversion of third time adc data is carried out, exports circuit itself the drift voltage V under LED open stateON Offset；Integrating amplification circuit is successively carried out after resetting and periodically integrating, carries out the 4th adc data conversion, export the integral voltage V under LED open stateON Integrate；Calculate the effective voltage V of single photocurrent integral measurementVALLD；Then VVALLD=(VON Integrate‑VON Offset)‑(VOFF Integrate‑VOFF Offset).The problem of carrying out single photocurrent integral measurement by the detection method, efficiently solving the drift of circuit voltage and environment trimmed book back noise influence week fluorescent measurement accuracy, improves signal-to-noise ratio.
Description
Technical field
The present invention relates to the technical fields of biological reagent detection, it is more particularly related to which a kind of bioluminescence is fast
Fast detection method.
Background technique
Quick-type biological indicator detection device, mainly for detection of the degree that biological indicator is killed, for assessing
The effect of sterilizing.Biological indicator after sterilizing is placed in culture medium and is cultivated, if biological indicator is not killed completely
It goes out, generates enzymatic reaction after the gemma recovery in biological indicator, strain can mass propagation after after a period of time.Using spy
Fixed fluorescent dye dyes bacterial cell, after bacterium mass propagation, will carry a large amount of fluorescent dyes.
Bioluminescence detection device receives two parts with detection by light source activation and forms.Excitation light source is used to be contaminated with fluorescence
The exciting light for expecting corresponding wavelength, is made of LED light, the voltage-controlled constant-current control circuit with digital-to-analogue conversion is reconfigured, for making
Excitation light source generates constant electric current lightening LED lamp, so that light intensity is kept constant.Detection receive generally use photodiode into
Row photoelectric conversion, and electric current is realized to the conversion of voltage using TIA transreactance amplifier, it is unnecessary to filter out using voltage buffer
High frequency, low frequency signal, it is optional to carry out secondary voltage amplification, after adc data converter is converted into digital signal, by
Microprocessor carries out data operation and processing.
Existing Photoelectric Detection or photoelectric sense technology are carried after bacterial reproduction by the light source activation of specific wavelength
A large amount of fluorescent dye will generate the fluorescence of another wavelength after being excited, accurate to survey by fluorescence detection, fluorescence interpretation
Determine the breeding situation of biological inoculum, so that it may the quality of accurate evaluation sterilization effect.Specifically, the detection of biological reagent is all logical
It crosses fluorescence and carries out interpretation, detection is all using photodiode (PD), and rear end is designed using transimpedance amplifier (TIA).
Traditional Transimpedance amplifier, with very high dynamic characteristic and real-time, extraction and Time-Frequency Analysis for live signal,
Commonly used in various photoelectric sensings, bio-sensing etc., by high dynamic signal amplification and real-time Time-Frequency Analysis, and then to letter
Number feature extracts, and carries out the measurement of component content etc. or the analysis of biological characteristic.However such high dynamic and reality
Shi Xing is not suitable for the quick detection of biological indicator, and in contrast, the detection time of biological indicator is long, usually with small
When discuss, and dynamic detection is not of practical significance in real time, on the contrary, by for a long time, steadily fluorescence measurement, by glimmering
Light data, which carries out statistics interpretation, just can accurately reflect the effect of biological reagent sterilizing.In addition, transreactance amplifier circuit, passes through adjusting
The multiple of mutual resistance amplification, the intensity of Lai Tigao signal amplification, but it is also exaggerated noise signal simultaneously, weaker fluorescence is believed
Number detection, noise is relatively low, it may appear that noise floods the case where measuring signal.Therefore, the existing photoelectricity based on Current amplifier
Detection mode detects weaker fluorescence signal, and noise is relatively low, sensitivity declines, and dynamic characteristic height can not but solve glimmering
It is dry not can avoid the uncertain variation of light intensity and external environment in measurement process for the problems such as light luminous intensity is weak, light intensity continuity
It disturbs, and biological reagent time of measuring is not long, high to requirement of real-time.
Summary of the invention
Shortcoming present in view of the above technology, the present invention provide a kind of bioluminescence rapid detection method, effectively
Ground solves the problems, such as that the drift of circuit voltage and environment trimmed book back noise influence week fluorescent measurement accuracy, improves noise
Than the precision and stability of, duplicate measurements.
In order to realize these purposes and other advantages according to the present invention, the invention is realized by the following technical scheme:
The present invention provides a kind of bioluminescence rapid detection method, carries out single photocurrent integral measurement, including following
Step:
It is closed, after integrating capacitor setting in LED, carries out the conversion of first time adc data, export the electricity under LED closed state
Road itself drift voltage, is denoted as VOFF Offset；
Integrating amplification circuit is successively carried out to reset and timing integrates, the time of integration arrives, and carries out second of adc data and turns
It changes, exports the integral voltage under LED closed state, be denoted as VOFF Integrate；
LED is opened, the conversion of third time adc data is carried out, exports circuit itself drift voltage under LED open state, note
For VON Offset；
Integrating amplification circuit is successively carried out to reset and timing integrates, the time of integration arrives, and carries out the 4th adc data and turns
It changes, exports the integral voltage under LED open state, be denoted as VON Integrate；
The effective voltage for calculating single photocurrent integral measurement, is denoted as VVALLD；Then,
VVALLD=(VON Integrate-VON Offset)-(VOFF Integrate-VOFF Offset)。
Preferably, the conversion of first time adc data is carried out, circuit itself drift voltage under LED closed state, packet are exported
Include following steps:
Controller control trigger triggering by optical signal change into current signal photodiode and integrating amplification circuit it
Between electric current input switch open；
The capacitance integral of controller control timer clocked flip integrating amplification circuit opens guarantor after closing the switch reset again
It holds；
Adc data conversion circuit carries out the conversion of first time adc data, exports circuit itself drift under LED closed state
Voltage.
Preferably, integrating amplification circuit is successively carried out to reset and timing integrates, the time of integration arrives, and carries out second
Adc data conversion, exports the integral voltage under LED closed state, comprising the following steps:
The capacitance integral of controller control timer clocked flip integrating amplification circuit closes the switch reset；
After capacitance integral closes the switch reset, controller control timer clocked flip electric current input switch closure,
Capacitance integral switch, which is opened, to be kept, and integrating amplification circuit is in current integration state；
The time of integration arrives, and controller control timer clocked flip electric current input switch disconnects；
Adc data conversion circuit carries out second of adc data conversion, exports the integral voltage under LED closed state.
Preferably, the closed reduction time of capacitance integral switch is much smaller than the opening retention time；
The closed reduction time range of capacitance integral switch is between 5uS-10uS；
Single photocurrent integral measurement time range is in 50mS-100mS.
Preferably, the step of third time adc data is converted repeats the step of first time adc data is converted；4th time
The step of adc data is converted repeats the step of second of adc data is converted.
Preferably, integrating capacitor is arranged, comprising the following steps:
Its corresponding capacitor is accessed into integrating amplification circuit by one closed in several capacitance selections switch；
Or, being combined access integral amplification by the several capacitors that corresponded to closed in several capacitance selections switch
Circuit.
Preferably, between two single photocurrent integral measurements, integrating capacitor setting further includes integrating capacitor size
The step of adjusting:
Controller generates control sequential control capacitance selection by the timing of control timer and the triggering of trigger and opens
The switching of pass is combined access integrating amplification circuit to adjust its correspondence capacitor.
Preferably, LED is closed and is opened, the reset of integrating amplification circuit and whether carries out adc data conversion, is control
Device processed generates control sequential by the timing of control timer and the triggering of trigger and triggers corresponding selection switch realization.
The present invention is include at least the following beneficial effects:
Bioluminescence rapid detection method provided by the invention, when carrying out single photocurrent integral measurement,
It in the case where LED is closed and opens two states, is measured twice respectively, the circuit under output LED closed state is certainly
Body drift voltage and circuit itself drift voltage and integral voltage under integral voltage, LED opening state, then to carry out single photosensitive
Current integration measures the calculating of effective voltage, efficiently solves the drift of circuit voltage and the influence of environment trimmed book back noise is micro-
The problem of hypofluorescence measurement accuracy, improves the precision and stability of signal-to-noise ratio, duplicate measurements.
Further advantage, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Detailed description of the invention
Fig. 1 is the flow chart that bioluminescence rapid detection method carries out single photocurrent integral measurement；
Fig. 2 is the schematic diagram that bioluminescence rapid detection method carries out single photocurrent integral measurement；
Fig. 3 is the detection system detected using bioluminescence rapid detection method and its communication example.
Specific embodiment
Present invention will be described in further detail below with reference to the accompanying drawings, to enable those skilled in the art referring to specification text
Word can be implemented accordingly.
It should be appreciated that such as " having ", "comprising" and " comprising " term used herein are not precluded one or more
The presence or addition of a other elements or combinations thereof.
Fig. 1 is the flow chart that bioluminescence rapid detection method carries out single photocurrent integral measurement；Fig. 2 is that biology is glimmering
The schematic diagram of light rapid detection method progress single photocurrent integral measurement；It is quick that Fig. 3 gives a kind of application bioluminescence
The detection system and its communication example that detection method is detected, detection method of the invention includes but is not limited to that can be applied to
Detection system shown in Fig. 3.
As depicted in figs. 1 and 2, present embodiment provides a kind of bioluminescence rapid detection method, carries out the photosensitive electricity of single
Flow integral measurement, comprising the following steps:
S10 is closed, after integrating capacitor setting in LED, is carried out the conversion of first time adc data, export under LED closed state
Circuit itself drift voltage, be denoted as VOFF Offset；
S20 successively carries out integrating amplification circuit to reset and timing integrates, and the time of integration arrives, and carries out second of adc data
Conversion exports the integral voltage under LED closed state, is denoted as VOFF Integrate；
S30 opens LED, carries out the conversion of third time adc data, exports circuit itself the drift electricity under LED open state
Pressure, is denoted as VON Offset；
S40 successively carries out integrating amplification circuit to reset and timing integrates, and the time of integration arrives, and carries out the 4th adc data
Conversion exports the integral voltage under LED open state, is denoted as VON Integrate；
S50 calculates the effective voltage of single photocurrent integral measurement, is denoted as VVALLD；Then,
VVALLD=(VON Integrate-VON Offset)-(VOFF Integrate-VOFF Offset)。
In above embodiment, the integral voltage under LED closed state includes detecting environment light integral voltage and circuit certainly
Body drift voltage, detection environment light integral voltage is background ambient light noise signal；Integral voltage packet under LED open state
Effective voltage and circuit itself drift voltage are included, effective voltage is the background signal measured.Therefore, it closes and opens in LED
It under two states, is measured twice respectively, exports circuit itself drift voltage and integral voltage, LED under LED closed state
Circuit itself drift voltage and integral voltage under opening state, then carry out the meter of single photocurrent integral measurement effective voltage
The problem of calculating, efficiently solving the drift of circuit voltage and background ambient light noise influence week fluorescent measurement accuracy, mentions
High signal-to-noise ratio, duplicate measurements precision and stability.
It should be noted that the integrated capacitor of integrating capacitor size is set during single photocurrent integral measurement
It sets and is remained unchanged always after determining, to guarantee the identical time of integration.
As the preferred of above embodiment, the conversion of first time adc data is carried out, exports the circuit under LED closed state
Itself drift voltage, comprising the following steps:
Controller control trigger triggering by optical signal change into current signal photodiode and integrating amplification circuit it
Between electric current input switch open；
The capacitance integral of controller control timer clocked flip integrating amplification circuit opens guarantor after closing the switch reset again
It holds；
Adc data conversion circuit carries out the conversion of first time adc data, exports circuit itself drift under LED closed state
Voltage.
The embodiment can obtain itself drift voltage of adc data converter in the circuit under LED closed state.
As the preferred of above embodiment, integrating amplification circuit is successively carried out to reset and timing integrates, the time of integration
It arrives, carries out second of adc data conversion, export the integral voltage under LED closed state, comprising the following steps:
The capacitance integral of controller control timer clocked flip integrating amplification circuit closes the switch reset；
After capacitance integral closes the switch reset, controller control timer clocked flip electric current input switch closure,
Capacitance integral switch, which is opened, to be kept, and integrating amplification circuit is in current integration state；
The time of integration arrives, and controller control timer clocked flip electric current input switch disconnects；
Adc data conversion circuit carries out second of adc data conversion, exports the integral voltage under LED closed state.
The embodiment can obtain the integral voltage under LED closed state, i.e. background ambient light noise signal.
As the preferred of above embodiment, the closed reduction time of capacitance integral switch is much smaller than the opening retention time；
The closed reduction time range of capacitance integral switch is between 5uS-10uS；Single photocurrent integral measurement time range exists
50mS-100mS。
Therefore during (3-5 hours) Bacteria Culture for a long time, the single photocurrent integral measurement time can be ignored,
Effectively improve measurement repeatable accuracy and stability.
As the preferred of above embodiment, the step of third time adc data is converted, the conversion of first time adc data is repeated
The step of；The step of 4th adc data conversion, repeat the step of second of adc data is converted.
As the preferred of above embodiment, integrating capacitor setting, comprising the following steps:
Its corresponding capacitor is accessed into integrating amplification circuit by one closed in several capacitance selections switch；Or,
Access integrating amplification circuit is combined by the several capacitors that corresponded to closed in several capacitance selections switch.More into one
Step, between two single photocurrent integral measurements, integrating capacitor setting further includes the steps that the big minor adjustment of integrating capacitor: control
Device processed by the timing of control timer and the triggering of trigger generate control sequential control the switching of capacitance selection switch with
It adjusts its correspondence capacitor and is combined access integrating amplification circuit.
By the embodiment it is found that the integration capacitor value of either single photocurrent integral measurement is still repeatedly photosensitive
Integration capacitor value between current integration measurement, the control sequential that can be generated by controller, clocked flip capacitance selection
Switch is to be selected and be adjusted.
As the preferred of above embodiment, LED close with unlatching, integrating amplification circuit reset and whether carry out
Adc data conversion is that controller is corresponded to by the timing of control timer and the triggering generation control sequential triggering of trigger
Selection switch realize.
By the embodiment it is found that controller is by adjusting control sequential, can be realized more with a variety of switches of clocked flip
The switching of kind state, is convenient for accurate, stable, prolonged biological reagent fluorescence detection.
Below by taking Fig. 3 as an example, it is specifically described the process that detection method of the invention carries out single photocurrent integral measurement.
Integrating amplification circuit has multiple groups capacitor model, and every group of capacitor model is provided with concatenated capacitance selection switch S2 and electricity
Hold, the capacitor's capacity of multiple groups capacitor model can it is identical also cannot be different, such as Fig. 3 gives 3 groups of capacitor models and amounts to 3
The example of capacitor, the capacitance of this 3 capacitors can be preferably 10pF, 30pF, 60pF.The timing that controller passes through control timer
And the triggering of trigger generates control sequential, the switching of clocked flip control capacitance selection switch S2 makes it correspond to capacitor progress
Combination access integrating amplification circuit completes the selection setting of integrating capacitor.When repeating photocurrent integral measurement, survey twice
Between amount, also the different integrating capacitor of adjustable control sequential clocked flip selects setting to controller, to realize integral electricity
The adjusting for holding size improves the size of effective voltage further to adjust the time of integration.
After integrating capacitor setting, when the timing of controller control timer and the triggering of trigger generate a control
Sequence, triggering selection switch S4 are closed, then LED 3 is closed.At this point, optical signal is changed into electric current letter by the triggering of controller control trigger
Number photodiode 1 and integrating amplification circuit 2 between electric current input switch S3 open；Controller control timer timing is touched
Holding is opened again after sending out the capacitance integral switch S1 closed reduction of integrating amplification circuit；Adc data conversion circuit carries out for the first time
Adc data conversion, exports circuit itself the drift voltage V under 3 closed state of LEDOFF Offset。
The capacitance integral switch S1 closed reduction of controller control timer clocked flip integrating amplification circuit；Capacitance integral
After switch S1 of short duration closed reduction, controller control timer clocked flip electric current input switch S3 closure, Power Capacity
Switch-dividing S1, which is opened, to be kept, and integrating amplification circuit is in current integration state；The time of integration arrives, controller control timer timing
Trigger current input switch S3 is disconnected；Adc data conversion circuit carries out second of adc data conversion, exports 3 closed state of LED
Under integral voltage VOFF Integrate。
The timing of controller control timer and the triggering of trigger generate a control sequential, triggering selection switch S4
It opens, then LED 3 is opened.The step of being repeated in the conversion of first time adc data and second of adc data conversion, obtains respectively
Circuit itself drift voltage V under 3 open state of LEDON OffsetAnd integral voltage, it is denoted as VON Integrate。
Calculate the effective voltage V of single photocurrent integral measurementVALLD；Then,
VVALLD=(VON Integrate-VON Offset)-(VOFF Integrate-VOFF Offset)。
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With.It can be applied to various suitable the field of the invention completely.It for those skilled in the art can be easily
Realize other modification.Therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details and legend shown and described herein.
Claims (8)
1. a kind of bioluminescence rapid detection method, which is characterized in that carry out single photocurrent integral measurement, including following step
It is rapid:
It is closed, after integrating capacitor setting in LED, carries out the conversion of first time adc data, the circuit under output LED closed state is certainly
Body drift voltage, is denoted as VOFF Offset；
Integrating amplification circuit is successively carried out to reset and timing integrates, the time of integration arrives, and carries out second of adc data conversion, defeated
Integral voltage under LED closed state out, is denoted as VOFF Integrate；
LED is opened, the conversion of third time adc data is carried out, circuit itself drift voltage under LED open state is exported, is denoted as VON Offset；
Integrating amplification circuit is successively carried out to reset and timing integrates, the time of integration arrives, and carries out the 4th adc data conversion, defeated
Integral voltage under LED open state out, is denoted as VON Integrate；
The effective voltage for calculating single photocurrent integral measurement, is denoted as VVALLD；Then,
VVALLD=(VON Integrate-VON Offset)-(VOFF Integrate-VOFF Offset)。
2. bioluminescence rapid detection method as described in claim 1, which is characterized in that the conversion of first time adc data is carried out,
Export circuit itself drift voltage under LED closed state, comprising the following steps:
The triggering of controller control trigger changes into optical signal between the photodiode of current signal and integrating amplification circuit
Electric current input switch is opened；
The capacitance integral of controller control timer clocked flip integrating amplification circuit opens holding after closing the switch reset again；
Adc data conversion circuit carries out the conversion of first time adc data, exports circuit itself drift voltage under LED closed state.
3. bioluminescence rapid detection method as described in claim 1, which is characterized in that successively carried out to integrating amplification circuit
It resetting and timing integrates, the time of integration arrives, and carries out second of adc data conversion, the integral voltage under LED closed state is exported,
The following steps are included:
The capacitance integral of controller control timer clocked flip integrating amplification circuit closes the switch reset；
After capacitance integral closes the switch reset, controller control timer clocked flip electric current input switch closure, capacitor
Integral restrictor, which is opened, to be kept, and integrating amplification circuit is in current integration state；
The time of integration arrives, and controller control timer clocked flip electric current input switch disconnects；
Adc data conversion circuit carries out second of adc data conversion, exports the integral voltage under LED closed state.
4. bioluminescence rapid detection method as claimed in claim 2 or claim 3, which is characterized in that the closure of capacitance integral switch
Resetting time is much smaller than the opening retention time；
The closed reduction time range of capacitance integral switch is between 5uS-10uS；
Single photocurrent integral measurement time range is in 50mS-100mS.
5. bioluminescence rapid detection method as described in claim 1, which is characterized in that the step of third time adc data conversion
Suddenly, the step of repeating the conversion of first time adc data；The step of 4th adc data conversion, repeats second of adc data conversion
The step of.
6. bioluminescence rapid detection method as described in claim 1, which is characterized in that integrating capacitor setting, including it is following
Step:
Its corresponding capacitor is accessed into integrating amplification circuit by one closed in several capacitance selections switch；
Or, being combined access integral amplification electricity by the several capacitors that corresponded to closed in several capacitance selections switch
Road.
7. bioluminescence rapid detection method as described in claim 1 or 6, which is characterized in that two single photocurrent products
Divide between measurement, integrating capacitor setting further includes the steps that the big minor adjustment of integrating capacitor:
Controller generates control sequential by the timing of control timer and the triggering of trigger and controls capacitance selection switch
Switching is combined access integrating amplification circuit to adjust its correspondence capacitor.
8. bioluminescence rapid detection method as described in claim 1, which is characterized in that LED is closed and opened, integrates amplification
The reset of circuit and whether adc data conversion is carried out, is that controller passes through the timing of control timer and the touching of trigger
The control sequential that hair generates triggers corresponding selection switch and realizes.
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