CN107179303B - Droplet fluorescence detection method, device, system, storage medium and computer equipment - Google Patents
Droplet fluorescence detection method, device, system, storage medium and computer equipment Download PDFInfo
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Abstract
The present invention relates to a kind of droplet fluorescence detection method, device, system, storage medium and computer equipments, droplet fluorescence detection method includes: to obtain the fluorescence fluorescence waveform signal corresponding with scattering light and scattering waveform signal that droplet issues, and fluorescence and scattering light are the optical signal collected under identical frequency acquisition and identical acquisition initial time；Extract the peak position of effective wave band in scattering waveform signal；Photoluminescence peak is obtained according to peak value of the peak position from extracting corresponding peak position in fluorescence waveform signal；The photoluminescence peak of extraction is stored as detection data of the droplet in fluorescence waveform signal corresponding to corresponding peak position.Peak value can be targetedly extracted at corresponding position in fluorescence waveform signal by the peak position according to scattering waveform signal and obtain photoluminescence peak, it does not need to carry out analysis extraction to all wave bands, the data amount of analysis to fluorescence waveform signal can be reduced, to improve fluorescence detection efficiency.
Description
Technical field
The present invention relates to survey control technology field, more particularly to a kind of droplet fluorescence detection method, device, system,
Storage medium and computer equipment.
Background technique
Flow cytometer, digital pcr (Polymerase Chain Reaction polymerase chain reaction) system etc. are high-precision
Close instrument is used to reliably detect the droplet (or microballoon) crossed from runner decision queue.Currently, the common methods of detection droplet
There are two types of method, one is impedance methods, i.e., add electrode in runner two sides, according to droplet by when the impedance variations that generate
To determine the quantity and size of droplet；Another is fluorescence detection.The working principle of fluorescence detection is: with digital pcr
For system, specific for hydrolysis after the probe amplification in the droplet of Water-In-Oil structure issues special under the exciting light of certain wavelength
Fluorescence in wavelength range, such as FAM dyestuff peak absorbance 495nm, peak emission wavelength 520nm；VIC dyestuff peak absorbance
Wavelength is 538nm, peak emission wavelength 554nm；Fluorescence detection has the droplet of fluorescent material, acquisition using light source activation
The corresponding waveform signal of fluorescence when droplet passes through runner simultaneously extracts the data such as peak value, pulsewidth and is stored, to analyze determination
Droplet is negative or positive.
When droplet size is smaller, and in impedance method measurement to impedance variations without obvious contribution when, can only often select
Fluorescence detection.Traditional fluorescence detection is usually directly to carry out peak extraction according to the corresponding waveform signal of the fluorescence of acquisition
And store, it needs to extract whole section of waveform signal, data amount of analysis is big, and detection efficiency is low.
Summary of the invention
Based on this, it is necessary to for the low problem of traditional fluorescence detection detection efficiency, provide a kind of raising detection effect
Droplet fluorescence detection method, device, system, storage medium and the computer equipment of rate.
A kind of droplet fluorescence detection method, comprising:
The fluorescence fluorescence waveform signal corresponding with scattering light and scattering waveform signal that droplet issues are obtained, it is described
Fluorescence and scattering light are the optical signal collected under identical frequency acquisition and identical acquisition initial time；
Extract the peak position of effective wave band in the scattering waveform signal；
It is obtained according to peak value of the peak position from extracting the corresponding peak position in the fluorescence waveform signal
Photoluminescence peak；
The photoluminescence peak of extraction is stored as droplet corresponding to corresponding peak position in the fluorescence waveform signal
Detection data.
A kind of droplet fluorescence detection device, comprising:
Waveform signal obtains module, the fluorescence waveform signal corresponding with scattering light of the fluorescence for obtaining droplet sending
With scattering waveform signal, collected under the fluorescence and the scattering identical frequency acquisition of light and identical acquisition initial time
Optical signal；
Peak position extraction module, for extracting the peak position of effective wave band in the scattering waveform signal；
Photoluminescence peak extraction module, for being extracted from the fluorescence waveform signal according to the peak position described in correspondence
Peak value at peak position obtains photoluminescence peak；
Detection data memory module exists for the photoluminescence peak of extraction to be stored as droplet corresponding to corresponding peak position
Detection data in the fluorescence waveform signal.
Above-mentioned droplet fluorescence detection method and device, by corresponding with scattering light in the fluorescence for obtaining droplet sending
After fluorescence waveform signal and scattering waveform signal, the peak position of effective wave band in scattering waveform signal is extracted, then
Photoluminescence peak is obtained according to peak value of the peak position from extracting corresponding peak position in fluorescence waveform signal, finally by extraction
Photoluminescence peak is stored as detection data of the droplet in fluorescence waveform signal corresponding to corresponding peak position.By using scattering
The peak extraction of waveform signal secondary fluorescence waveform signal, fluorescence and scattering light are that identical frequency acquisition and identical acquisition originate
Collected under time, thus fluorescence waveform signal with scattering waveform signal it is synchronous, the peak position of fluorescence waveform signal and
The peak position for scattering waveform signal is identical, thus can be targetedly glimmering according to the peak position of scattering waveform signal
Peak value is extracted in waveform signal at corresponding position and obtains photoluminescence peak, does not need to carry out analysis extraction to all wave bands, it can
Reduce the data amount of analysis to fluorescence waveform signal, to improve fluorescence detection efficiency.
A kind of storage medium, is stored with computer program, realizes when the computer program of storage is executed by processor above-mentioned
The step of droplet fluorescence detection method.
A kind of computer equipment can be run on a memory and on a processor including memory, processor and storage
The step of computer program, the processor realizes above-mentioned droplet fluorescence detection method when executing the computer program.
Above-mentioned storage medium and computer equipment can equally reduce to fluorescence, it can be achieved that above-mentioned droplet fluorescence detection method
The data amount of analysis of waveform signal, to improve fluorescence detection efficiency.
A kind of droplet fluorescence detecting system, including scattering optical acquisition device, fluorescent collecting device and data processing equipment, institute
It states scattering optical acquisition device and the fluorescent collecting device is located at the runner two sides flowed through for droplet, and be all connected with described
Data processing equipment；
The scattering optical acquisition device and the fluorescent collecting device are in identical frequency acquisition and identical acquisition initial time
The scattering light and fluorescence of the lower acquisition droplet respectively obtain scattering waveform signal and fluorescence waveform signal and are sent to described
Data processing equipment；
The data processing equipment extracts the peak position of effective wave band in the scattering waveform signal, according to the peak
Peak value of the value position from extracting the corresponding peak position in the fluorescence waveform signal obtains photoluminescence peak, and by extraction
Photoluminescence peak is stored as detection data of the droplet in the fluorescence waveform signal corresponding to corresponding peak position.
Above-mentioned droplet fluorescence detecting system acquires droplet by using scattering optical acquisition device and fluorescent collecting device respectively
Scattering light and fluorescence obtain scattering waveform signal and fluorescence waveform signal and be sent to data processing equipment, data processing dress
The peak position for extracting effective wave band in scattering waveform signal is set, is then extracted from fluorescence waveform signal according to peak position
Peak value at corresponding peak position obtains photoluminescence peak, and finally the photoluminescence peak of extraction is stored as corresponding to corresponding peak position
Detection data of the droplet in fluorescence waveform signal.By using the peak value of scattering waveform signal secondary fluorescence waveform signal
It extracts, fluorescence and scattering light is collect under identical frequency acquisition and identical acquisition initial time, therefore fluorescence waveform signal
Synchronous with scattering waveform signal, the peak position of fluorescence waveform signal is identical as the scattering peak position of waveform signal, from
And peak can targetedly be extracted in fluorescence waveform signal according to the peak position of scattering waveform signal at corresponding position
Value obtains photoluminescence peak, does not need to carry out analysis extraction to all wave bands, can reduce the data amount of analysis to fluorescence waveform signal,
To improve fluorescence detection efficiency.
Detailed description of the invention
Fig. 1 is the flow chart of droplet fluorescence detection method in an embodiment；
Fig. 2 is the schematic diagram of scattering waveform signal and fluorescence waveform signal in the case of the first；
Fig. 3 is the schematic diagram that waveform signal and fluorescence waveform signal are scattered under second situation；
Fig. 4 is the schematic diagram of scattering waveform signal and fluorescence waveform signal in the case of the third；
Fig. 5 scatters the schematic diagram of waveform signal and fluorescence waveform signal in the case of being the 4th kind；
Fig. 6 is the flow chart of droplet fluorescence detection method in another embodiment；
Fig. 7 is the function structure chart of droplet fluorescence detection device in an embodiment；
Fig. 8 is the structure chart of droplet fluorescence detecting system in an embodiment；
Fig. 9 is the operating position schematic diagram of droplet fluorescence detecting system in an embodiment.
Specific embodiment
With reference to Fig. 1, droplet fluorescence detection method in an embodiment includes the following steps.
S110: the fluorescence fluorescence waveform signal corresponding with scattering light and scattering waveform letter that droplet issues are obtained
Number.
Wherein, fluorescence and scattering light are the optical signal collected under identical frequency acquisition and identical acquisition initial time.
Droplet carries fluorescent dye, and the droplet crossed using decision queue in light source activation runner inspires fluorescence and production
Raw scattering light.The fluorescence of the corresponding one or more droplets of fluorescence waveform signal, scattering waveform signal are corresponding one or more micro-
The scattering light of drop.Due to fluorescence and scatter the frequency acquisition of light and to acquire initial time identical, it is ensured that sample-synchronous, then fluorescence
Fluorescence waveform signal corresponding with scattering light is synchronous with scattering waveform signal.Specifically, it by fluorescent collecting device and can dissipate
It penetrates optical acquisition device and acquires fluorescence and scattering light respectively under identical frequency acquisition and identical acquisition initial time, and pass through photoelectricity
The available fluorescence waveform signal of the conversion of signal and scattering waveform signal.
S130: the peak position of effective wave band in scattering waveform signal is extracted.
Scattering waveform signal includes multiple wave bands continuous in time, and effective wave band refers to the wave band for meeting preset condition.
Peak position, which refers to, there is the position of peak value in effective wave band.By being analyzed scattering waveform signal to determine significant wave
Section, to extract the peak position of effective wave band.
S150: fluorescence peak is obtained according to peak value of the peak position from extracting corresponding peak position in fluorescence waveform signal
Value.
Due to scattering waveform signal it is synchronous with fluorescence waveform signal, fluorescence waveform signal occur peak value position and
The position that peak value occurs in scattering waveform signal is identical, to can determine needs according to the peak position in scattering waveform signal
The position of peak value is extracted from fluorescence waveform signal.
S170: the photoluminescence peak of extraction is stored as droplet corresponding to corresponding peak position in fluorescence waveform signal
Detection data.
The corresponding relationship of peak position and droplet can corresponding relationship according to the pre-stored data determine, for example, can be stored in advance
The corresponding relationship between wave band in the serial number and scattering peak waveform signal of each droplet, i.e. which wave band correspond to which serial number
Droplet；By determining which wave band peak position belongs to, the droplet which serial number peak position corresponds to can be obtained, thus by root
Detection data of the photoluminescence peak extracted according to peak position as corresponding droplet in the fluorescence detection channel where photoluminescence peak.
Specifically, the quantity of fluorescence waveform signal can have multiple.That is, synchronization, carries out fluorescent collecting to droplet
Fluorescent collecting device has multiple, and the synchronous fluorescence waveform signal obtained from has multiple.For example, in the runner for thering is droplet to flow through
One scattering optical acquisition device of side setting, the other side is arranged two fluorescent collecting devices, scatters optical acquisition device and two fluorescence
Acquisition device acquires simultaneously, can obtain the synchronous waveform of scattering all the way signal and two-way fluorescence waveform signal.At this point, step
S150 are as follows: the peak value for extracting corresponding position from each fluorescence waveform signal respectively according to peak position obtains each fluorescence waveform
Signal corresponds to the photoluminescence peak of peak position；Step S170 are as follows: it is right that the photoluminescence peak of extraction is stored as corresponding peak position institute
Detection data of the droplet answered in the fluorescence waveform signal where photoluminescence peak.
For example, corresponding droplet is first droplet for peak position X, extracted from first via fluorescence waveform signal
Obtained photoluminescence peak is a1, and the photoluminescence peak extracted from the second road fluorescence waveform signal is b1, then needs to store
Information includes: that detection data of first droplet in first via fluorescence waveform signal is a1, in the second road fluorescence waveform signal
Detection data be b1.
Above-mentioned droplet fluorescence detection method, by obtaining the fluorescence fluorescence wave corresponding with scattering light that droplet issues
After shape signal and scattering waveform signal, the peak position of effective wave band in scattering waveform signal is extracted, then according to peak
Peak value of the value position from extracting corresponding peak position in fluorescence waveform signal obtains photoluminescence peak, finally by the fluorescence peak of extraction
Value is stored as detection data of the droplet in fluorescence waveform signal corresponding to corresponding peak position.By using scattering waveform
The peak extraction of signal secondary fluorescence waveform signal, fluorescence and scattering light are under identical frequency acquisition and identical acquisition initial time
It collects, therefore fluorescence waveform signal is synchronous with scattering waveform signal, the peak position and scattering light of fluorescence waveform signal
The peak position of waveform signal is identical, thus can be targetedly in fluorescence waveform according to the peak position of scattering waveform signal
Peak value is extracted in signal at corresponding position and obtains photoluminescence peak, does not need to carry out analysis extraction to all wave bands, can reduce pair
The data amount of analysis of fluorescence waveform signal, to improve fluorescence detection efficiency.
In addition, above-mentioned droplet fluorescence detection method passes through the corresponding relationship of peak position and droplet, by the fluorescence peak of extraction
Value is stored as corresponding detection data of the droplet in fluorescence waveform signal, a pair of so as to which photoluminescence peak and droplet are carried out one
The phenomenon that answering, specifying the corresponding relationship between photoluminescence peak and droplet, avoid the occurrence of detection data entanglement between different droplets, mentions
The accuracy of high data acquisition.
For example, for including two-way fluorescence waveform signal, fluorescence A indicates the first via with reference to Fig. 2, Fig. 3, Fig. 4 and Fig. 5
The corresponding waveform diagram of fluorescence waveform signal, fluorescence B indicate the corresponding waveform diagram of the second road fluorescence waveform signal, and scattering light C is indicated
Scatter the waveform diagram of waveform signal.The case where being likely to occur in droplet detection process includes: scattering waveform signal and two-way
Fluorescence waveform signal is normal, as shown in Figure 2；Due to runner, wherein fluorescence waveform signal is excessively faint to be flooded all the way
Not in noise, such asFig. 3It is shown；When having air, fluid velocity shakiness in runner, waveform signal and fluorescence waveform signal are scattered
It all very disorderly, is clutter, as shown in Figure 4；Droplet, which divides fusion phenomenon, to be occurred, and is caused droplet to be likely to occur and is excessively greater than, is small
In normal droplet, to scatter waveform signal and fluorescence waveform signal is excessive or too small, as shown in Figure 5.Traditional fluorescence inspection
Survey method needs to extract and judge to per each wave band in fluorescence waveform signal all the way, computationally intensive.Meanwhile for figure
Situation shown in 3, at the peak point of first via fluorescence waveform signal, the second road fluorescence waveform signal occurs without obvious wave crest
Second road fluorescence waveform signal is excessively faint and is submerged in noise, at this point, extracting not appearance in the second road fluorescence waveform signal
Value, the data storage for droplet easily occur are omitted；At this point, traditional fluorescence detection will appear the peak value of two fluorescence waveform signals
It is different, it saves in photoluminescence peak array, two photoluminescence peaks under same index location are not necessarily mentioning for the same droplet
It takes as a result, to data entanglement occur.Using above-mentioned droplet fluorescence detection method, calculation amount can be reduced and improve detection efficiency, together
When by photoluminescence peak it is corresponding with droplet, can avoid data misplace and omission.
It in one embodiment, include step S131 to step S135 with reference to Fig. 6, step S130.
S131: the peak value of each wave band and pulsewidth in scattering waveform signal are extracted and judges whether to extract successfully.
The specific extraction operation that peak value and pulsewidth are carried out to wave band can be used conventionally known technology and realize.If extracting
Peak value and pulsewidth, expression are extracted successfully, execute step S133 at this time.
S133: judge the peak value extracted and pulsewidth whether respectively in preset peak ranges and preset pulse duration range
It is interior.
Peak ranges and pulse duration range are respectively used to limit the normal value range of peak value and pulsewidth；If step S133's sentences
Disconnected result be it is yes, indicate the peak value extracted in peak ranges and the pulsewidth of extraction be in pulse duration range, execute step at this time
S135。
Specifically, peak ranges and pulse duration range can be stored in advance based on experience value, can also be real by receiving user
When the numberical range that inputs acquire.For example, in one embodiment, before step S133, further including that range determines step: connecing
Receive the peak ranges inputted and pulse duration range and respectively as preset peak ranges and preset pulse duration range.Specifically, model
It encloses determining step and can be and executed after step S131, can also execute, may be used also after step silo, before step S131
To be executed before step S110.
S135: the wave band of peak value and pulsewidth will be extracted as effective wave band, and extract the peak position of effective wave band.
The peak value of extraction is in peak ranges and the pulsewidth of extraction is in pulse duration range, then it represents that corresponding wave band meets pre-
If condition, using corresponding wave band as effective wave band, to extract peak position.
Whether step S131 to step S135 is by the peak value and Pulse Width Analysis of extraction wave band in corresponding peak value model
Enclose in pulse duration range, so that it is determined that whether corresponding wave band is effective wave band, can the wave band effectively except effective wave band pick
It removes, improves the accuracy that peak position extracts.It is appreciated that in other embodiments, other ginsengs of wave band can also be extracted
Number analyzes whether it is effective wave band.
In one embodiment, with continued reference to Fig. 6, after step S131, if peak value and pulsewidth are extracted unsuccessful, step is executed
Rapid S132.
S132: droplet corresponding to corresponding wave band is recorded as clutter droplet.
Do not extract peak value and pulsewidth, then it represents that extraction is unsuccessful, can regard as corresponding wave band and be flooded by noise, example
In waveform as shown in Figure 4, all wave bands are extracted less than peak value and pulsewidth；At this point it is possible to the wave band pair that will do not extract
The data answered abandon, and droplet corresponding to corresponding wave band is recorded as clutter droplet.
Specifically, after step S133, if the peak value and pulsewidth that extract be not respectively in peak ranges and pulse duration range
It is interior, then follow the steps S134.
S134: droplet corresponding to corresponding wave band is recorded as size exception droplet.
The peak value and pulsewidth extracted be not respectively in peak ranges and pulse duration range, and include the case where having: peak value does not have
Have in peak ranges and pulsewidth not in pulse duration range, peak value not in peak ranges and pulsewidth in pulse duration range,
Peak value is in peak ranges and pulsewidth is not in peak ranges；Wherein, peak value is not less than in peak ranges including peak value
The minimum value and peak value of peak ranges are greater than two kinds of situations of maximum value of peak ranges, and pulsewidth includes arteries and veins not in pulse duration range
Wide minimum value and pulsewidth less than pulse duration range is greater than two kinds of situations of maximum value of pulse duration range.When the peak value and pulsewidth extracted
Not respectively in peak ranges and pulse duration range when, situation shown in Figure 5 indicates that the waveform shape of corresponding wave band is excessive
Or it is too small, so that corresponding droplet is excessive or too small, corresponding droplet is recorded as size exception droplet at this time.
By the way that the corresponding droplet of wave band except effective wave band is carried out the book of final entry, will extract less than peak value and pulsewidth
The corresponding droplet of wave band is recorded as clutter droplet, by the peak value extracted and pulsewidth not respectively in peak ranges and pulse duration range
Droplet corresponding to interior correspondence wave band is recorded as size exception droplet, can check convenient for user to understand details.
In one embodiment, fluorescence waveform signal with scattering waveform signal include in preset duration each moment it is corresponding
Wave data.I.e. a droplet can correspond to the Wave data of the fluorescence at multiple and different moment, the fluorescence of one or more droplets
Wave data form fluorescence waveform signal.
It further include step S120 before step S150 after step S110 with continued reference to Fig. 6.
S120: the Wave data at caching fluorescence waveform signal corresponding each moment.
By caching Wave data, subsequent lookup is facilitated to use.Specifically, scattering light can also be cached after step S120
The Wave data at waveform signal corresponding each moment.
Specifically, step S150 includes step S151 to step S154.
S151: it at the time of acquisition peak position corresponds to, searches where at the time of peak position corresponds in fluorescence waveform signal
Preset time range in wave band.Wherein, as median at the time of preset time range is corresponded to using peak position.
Scattering peak waveform signal is made of the Wave data arranged according to time order and function, and different moments correspond to different positions
It sets, therefore specifically peak position can be corresponded to using the moment.As the preset time model of median at the time of correspondence using peak position
Enclose, be including peak position correspond at the time of including period.
S152: it extracts the peak value of the wave band found and judges whether to extract successfully.If so, thening follow the steps S153；It is no
Then follow the steps S154.
Since fluorescence waveform signal is synchronous with scattering waveform signal, scatter peak position pair in waveform signal
The Wave data of position where at the time of the Wave data answered is corresponding with peak position in fluorescence waveform signal corresponds to same
Detection data of the droplet at same position；Due to during fluorescence and scattering light collection there may be error, by
Searched in fluorescence waveform signal using at the time of peak position as the wave band of the preset time range of median, to the wave band of lookup into
The accuracy for obtaining peak value can be improved in row peak extraction.
S153: using the peak value of extraction as the photoluminescence peak at corresponding peak position.
S154: Wave data corresponding at the time of peak position corresponds to is searched from the Wave data of caching as correspondence
Photoluminescence peak at peak position.
For the extracted wave band for extracting peak position from scattering waveform signal, indicate that corresponding droplet is positive
Normal droplet, two kinds of situations existing for fluorescence waveform signal are: fluorescence waveform signal is normal, can extract peak value, with reference to Fig. 2；It is glimmering
Waveform signal is flooded by noise, is extracted less than peak value, with reference to the second road fluorescence waveform signal flooded in Fig. 3 by noise.If
Peak value is successfully extracted from the wave band of lookup, then using the peak value of extraction as photoluminescence peak, accuracy is high；If extracting less than peak
Value, then search corresponding Wave data as photoluminescence peak from the data of caching；In this way, no matter peak value can be extracted
Ensure to get corresponding photoluminescence peak, the data that can avoid occurring in conventional fluorescent detection method omit phenomenon, and data acquisition
Accuracy is high.
Specifically, the quantity of peak position has multiple.Therefore, each peak position corresponds to respective photoluminescence peak.Please after
Continue and refer to Fig. 6, further includes step S180 and step S190 after step S170.
S180: judge whether all peak positions are corresponding with photoluminescence peak.
If not, then it represents that do not extracted in fluorescence signal there are also peak value, execute step S190 at this time.
S190: it obtains next peak position and obtains new peak position, and return step S150.
Specifically, step S190 return step S151.In the present embodiment, step S150 and step S170 are according to peak value position
The sequencing set successively executes, i.e. step S150 is glimmering according to previous peak position acquisition photoluminescence peak, step S170 storage
After photopeak value is the detection data of corresponding droplet, step S150 is returned again to according to next peak position and obtains photoluminescence peak, directly
Photoluminescence peak is obtained to according to all peak positions.So, it can be ensured that all peak positions correspondence obtains a fluorescence
Peak value avoids missing data.
A kind of storage medium, is stored with computer program, realizes when the computer program of storage is executed by processor above-mentioned
The step of droplet fluorescence detection method.
A kind of computer equipment can be run on a memory and on a processor including memory, processor and storage
The step of computer program, processor realizes above-mentioned droplet fluorescence detection method when executing computer program.
Above-mentioned storage medium and computer equipment can similarly reduce pair due to including above-mentioned droplet fluorescence detection method
The data amount of analysis of fluorescence waveform signal, to improve fluorescence detection efficiency；And the accuracy of data acquisition is high.
With reference to Fig. 7, a kind of droplet fluorescence detection device, including waveform signal obtain module 110, peak position extraction module
130, photoluminescence peak extraction module 150 and detection data memory module 170.
Waveform signal obtains the fluorescence fluorescence waveform letter corresponding with scattering light that module 110 is used to obtain droplet sending
Number and scattering waveform signal, fluorescence and scattering light are the light that collects under identical frequency acquisition and identical acquisition initial time
Signal.
Peak position extraction module 130 is used to extract in scattering waveform signal the effectively peak position of wave band.
Photoluminescence peak extraction module 150 is used for according to peak position from the corresponding peak position of extraction in fluorescence waveform signal
Peak value obtain photoluminescence peak.
Detection data memory module 170 is used to for the photoluminescence peak of extraction to be stored as droplet corresponding to corresponding peak position
Detection data in fluorescence waveform signal.
Above-mentioned droplet fluorescence detection device obtains the fluorescence and dissipate that droplet issues by obtaining module 110 in waveform signal
It penetrates after the corresponding fluorescence waveform signal of light and scattering waveform signal, peak position extraction module 130 extracts scattering light
The peak position of effective wave band in waveform signal, then photoluminescence peak extraction module 150 is believed according to peak position from fluorescence waveform
The peak value at corresponding peak position is extracted in number and obtains photoluminescence peak, and last detection data memory module 170 is by the fluorescence of extraction
Peak value is stored as detection data of the droplet in fluorescence waveform signal corresponding to corresponding peak position.By using scattering light wave
The peak extraction of shape signal secondary fluorescence waveform signal, fluorescence and scattering light are identical frequency acquisition and identical acquisition initial time
Under collect, therefore fluorescence waveform signal with scattering waveform signal it is synchronous, the peak position of fluorescence waveform signal and scattering
The peak position of waveform signal is identical, thus can be targetedly in fluorescence wave according to the peak position of scattering waveform signal
Peak value is extracted in shape signal at corresponding position and obtains photoluminescence peak, does not need to carry out analysis extraction to all wave bands, can reduce
To the data amount of analysis of fluorescence waveform signal, to improve fluorescence detection efficiency.
In addition, passing through the corresponding relationship of peak position and droplet, the photoluminescence peak of extraction is stored as corresponding droplet glimmering
Detection data in waveform signal specifies photoluminescence peak and droplet so as to correspond photoluminescence peak and droplet
Between corresponding relationship, the phenomenon that avoiding the occurrence of detection data entanglement between different droplets, improve the accuracy of data acquisition.
Specifically, peak position extraction module 130 specifically can be used in droplet fluorescence detection method include step S131 extremely
The method of step S135 extracts the peak position of effective wave band in scattering waveform signal, and this will not be repeated here.
Specifically, fluorescence waveform signal and scattering waveform signal include each moment corresponding waveform in preset duration
Data.Above-mentioned droplet fluorescence detection device further includes cache module, obtains fluorescence waveform for obtaining module 110 in waveform signal
After signal and scattering waveform signal, the Wave data at caching fluorescence waveform signal corresponding each moment.Photoluminescence peak extracts
The method that step S151 to step S154 in droplet fluorescence detection method specifically can be used in module 150 obtains glimmering according to peak position
Photopeak value, this will not be repeated here.
Specifically, above-mentioned droplet fluorescence detection device further includes loop detection module (not shown), all for judging
Whether peak position is corresponding with photoluminescence peak, if otherwise obtaining next peak position obtains new peak position, and controls
Photoluminescence peak extraction module 150 executes corresponding function.
With reference to Fig. 8, a kind of droplet fluorescence detecting system, including scattering optical acquisition device 210,220 and of fluorescent collecting device
Data processing equipment 230, scatters optical acquisition device 210 and fluorescent collecting device 220 is located at the runner flowed through for droplet
300 (referring to Fig. 9) two sides, and it is all connected with data processing equipment 230.
Optical acquisition device 210 and fluorescent collecting device 220 are scattered under identical frequency acquisition and identical acquisition initial time
The scattering light and fluorescence for acquiring droplet respectively obtain scattering waveform signal and fluorescence waveform signal and are sent to data processing dress
Set 230.
Data processing equipment 230 extract scattering waveform signal in effectively wave band peak position, according to peak position from
The peak value at corresponding peak position is extracted in fluorescence waveform signal and obtains photoluminescence peak, and the photoluminescence peak of extraction is stored as pair
Answer detection data of the droplet corresponding to peak position in fluorescence waveform signal.
Above-mentioned droplet fluorescence detecting system is adopted respectively by using scattering optical acquisition device 210 and fluorescent collecting device 220
The scattering light and fluorescence for collecting droplet obtain scattering waveform signal and fluorescence waveform signal and are sent to data processing equipment 230,
Data processing equipment 230 extracts in scattering waveform signal the effectively peak position of wave band, then according to peak position from fluorescence
The peak value at corresponding peak position is extracted in waveform signal and obtains photoluminescence peak, finally is stored as corresponding to by the photoluminescence peak of extraction
Detection data of the droplet corresponding to peak position in fluorescence waveform signal.By using scattering waveform signal secondary fluorescence
The peak extraction of waveform signal, fluorescence and scattering light are to collect under identical frequency acquisition and identical acquisition initial time, because
This fluorescence waveform signal is synchronous with scattering waveform signal, the peak position of fluorescence waveform signal and the peak of scattering waveform signal
Be worth position it is identical, thus according to scattering waveform signal peak position can targetedly in fluorescence waveform signal it is corresponding
Peak value is extracted at position and obtains photoluminescence peak, is not needed to carry out analysis extraction to all wave bands, can be reduced to fluorescence waveform signal
Data amount of analysis, to improve fluorescence detection efficiency.
In addition, passing through the corresponding relationship of peak position and droplet, the photoluminescence peak of extraction is stored as corresponding droplet glimmering
Detection data in waveform signal specifies photoluminescence peak and droplet so as to correspond photoluminescence peak and droplet
Between corresponding relationship, the phenomenon that avoiding the occurrence of detection data entanglement between different droplets, improve the accuracy of data acquisition.
In one embodiment, with reference to Fig. 9, the quantity of fluorescent collecting device 220 is multiple, 230 basis of data processing equipment
Peak position corresponds to the peak value from peak position from extraction in the corresponding fluorescence waveform signal of each fluorescent collecting device 220 respectively,
The photoluminescence peak that each fluorescence waveform signal corresponds to peak position is obtained, the photoluminescence peak of extraction is stored as corresponding peak position institute
Detection data of the corresponding droplet in the fluorescence waveform signal where photoluminescence peak.
Specifically, scattering optical acquisition device 210 includes scattering optical channel optical system and the first ADC (Analog-to-
Digital Converter analog-digital converter), the first ADC connection scatters optical channel optical system and data processing equipment 230；
Fluorescent collecting device 220 includes fluorescence channel optical system and the 2nd ADC, the 2nd ADC connection fluorescence channel optical system sum number
According to processing unit 230.The frequency acquisition of first ADC and the 2nd ADC is identical with acquisition initial time.
It scatters the scattering light of optical channel collection optical system droplet and carries out photoelectric conversion and obtain corresponding electric signal transmission
To the first ADC, the first ADC is dissipated according to preset frequency acquisition electric signal corresponding with acquisition initial time collection of scattered light
It penetrates waveform signal and is sent to data processing equipment 230；The fluorescence of fluorescence channel collection optical system droplet simultaneously carries out photoelectricity
It is converted to corresponding electric signal and is sent to the 2nd ADC, the 2nd ADC is acquired according to preset frequency acquisition and acquisition initial time
The corresponding electric signal of fluorescence obtains fluorescence waveform signal and is sent to data processing equipment 230.By to different channel opticals
System is correspondingly arranged respective ADC, carries out classification acquisition, and the accuracy of data processing can be improved.
It is light source with reference to Fig. 9, D, E is light source optical system；A, B is respectively that the light in two fluorescence channel optical systems is visited
Device is surveyed, C is the optical detector scattered in optical channel optical system, and optical detector is for converting optical signals to electric signal；F is scattered
Penetrate the acquisition channel of optical channel optical system；G, H is respectively the acquisition channel of two fluorescence channel optical systems.Light source activation stream
Decision queue in road 300 and cross droplet inspire fluorescence and generate scattering light；Fluorescence is received by fluorescence channel optical system
Collection is received by detector A and detector B respectively；It scatters light to collect by scattering optical channel optical system, by detector for scattered light
C is received, and detector C is placed on to be tested on the spherical surface of the droplet centre of sphere, and θ is its angle, and θ angular dimension can be according to the letter of signal
It makes an uproar than adjusting.
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 protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (9)
1. a kind of droplet fluorescence detection method characterized by comprising
Obtain the fluorescence fluorescence waveform signal corresponding with scattering light and scattering waveform signal that droplet issues, the fluorescence
It is the optical signal collected under identical frequency acquisition and identical acquisition initial time with scattering light；
Extract the peak position of effective wave band in the scattering waveform signal；The quantity of the peak position has multiple；
Fluorescence is obtained according to peak value of the peak position from extracting the corresponding peak position in the fluorescence waveform signal
Peak value；
The photoluminescence peak of extraction is stored as detection of the droplet in the fluorescence waveform signal corresponding to corresponding peak position
Data；
Judge whether all peak positions are corresponding with photoluminescence peak；
If it is not, obtaining next peak position obtains new peak position, and return it is described according to the peak position from described
The step of peak value at the corresponding peak position obtains photoluminescence peak is extracted in fluorescence waveform signal.
2. droplet fluorescence detection method according to claim 1, which is characterized in that described to extract the scattering waveform letter
The peak position of effective wave band in number, comprising:
It extracts the peak value of each wave band and pulsewidth in the scattering waveform signal and judges whether to extract successfully；
If whether extracting the peak value for successfully judging to extract and pulsewidth respectively in preset peak ranges and preset pulsewidth model
In enclosing；
If so, the wave band of peak value and pulsewidth will be extracted as effective wave band, and extract the peak position of effective wave band.
3. droplet fluorescence detection method according to claim 2, which is characterized in that described to extract the scattering waveform letter
The peak value of each wave band and pulsewidth and judge whether after extracting successfully in number, further includes:
If extraction is unsuccessful, droplet corresponding to corresponding wave band is recorded as clutter droplet；
It is described judge the peak value that extracts and pulsewidth whether respectively in preset peak ranges and preset pulse duration range after,
Further include:
If it is not, droplet corresponding to corresponding wave band is then recorded as size exception droplet.
4. droplet fluorescence detection method according to claim 1, which is characterized in that the fluorescence waveform signal and it is described dissipate
Waveform signal is penetrated and includes the corresponding Wave data of each moment in preset duration, it is described to obtain the fluorescence and dissipate that droplet issues
Penetrate after the corresponding fluorescence waveform signal of light and scattering waveform signal, it is described according to the peak position from the fluorescence
It further include caching the fluorescence waveform before peak value in waveform signal at the corresponding peak position of extraction obtains photoluminescence peak
The step of Wave data at signal corresponding each moment；
It is described to be obtained according to peak value of the peak position from extracting the corresponding peak position in the fluorescence waveform signal
Photoluminescence peak, comprising:
At the time of obtaining the peak position and correspond to, searches peak position described in the fluorescence waveform signal and correspond at the time of institute
Preset time range in wave band, as median at the time of the preset time range is corresponded to using the peak position；
It extracts the peak value of the wave band found and judges whether to extract successfully；
If so, using the peak value of extraction as the photoluminescence peak at corresponding peak position；
If it is not, then searching Wave data conduct pair corresponding at the time of the peak position corresponds to from the Wave data of caching
Answer the photoluminescence peak at peak position.
5. a kind of storage medium, is stored with computer program, which is characterized in that when the computer program of storage is executed by processor
It realizes such as the step of any one of claim 1-4 the method.
6. a kind of computer equipment including memory, processor and stores the meter that can be run on a memory and on a processor
Calculation machine program, which is characterized in that the processor is realized when executing the computer program such as any one of claim 1-4
The step of the method.
7. a kind of droplet fluorescence detection device characterized by comprising
Waveform signal obtains module, for obtaining the fluorescence fluorescence waveform signal corresponding with scattering light and dissipate that droplet issues
Waveform signal is penetrated, the fluorescence and scattering light are the light letter collected under identical frequency acquisition and identical acquisition initial time
Number；
Peak position extraction module, for extracting the peak position of effective wave band in the scattering waveform signal；The peak value
The quantity of position has multiple；
Photoluminescence peak extraction module, for extracting the corresponding peak value from the fluorescence waveform signal according to the peak position
Peak value at position obtains photoluminescence peak；
Detection data memory module, for the photoluminescence peak of extraction to be stored as droplet corresponding to corresponding peak position described
Detection data in fluorescence waveform signal；
Loop detection module, for judging whether all peak positions are corresponding with photoluminescence peak, if otherwise obtaining next
Peak position obtains new peak position, and controls the photoluminescence peak extraction module and execute corresponding function.
8. a kind of droplet fluorescence detecting system, which is characterized in that at scattering optical acquisition device, fluorescent collecting device and data
Device is managed, the scattering optical acquisition device and the fluorescent collecting device are located at the runner two sides flowed through for droplet, and
It is all connected with the data processing equipment；
The scattering optical acquisition device and the fluorescent collecting device are adopted under identical frequency acquisition and identical acquisition initial time
The scattering light and fluorescence for collecting the droplet respectively obtain scattering waveform signal and fluorescence waveform signal and are sent to the data
Processing unit；
The data processing equipment extracts the peak position of effective wave band in the scattering waveform signal, the peak position
Quantity has multiple；The data processing equipment extracts the corresponding peak according to the peak position from the fluorescence waveform signal
Peak value at value position obtains photoluminescence peak, and the photoluminescence peak of extraction is stored as droplet corresponding to corresponding peak position and is existed
Detection data in the fluorescence waveform signal, judges whether all peak positions are corresponding with photoluminescence peak, if it is not, obtaining
Next peak position obtains new peak position, and return it is described according to the peak position from the fluorescence waveform signal
Extract the step of peak value at the corresponding peak position obtains photoluminescence peak.
9. droplet fluorescence detecting system according to claim 8, which is characterized in that the quantity of the fluorescent collecting device is
Multiple, the data processing equipment is according to the peak position respectively from the corresponding fluorescence waveform signal of each fluorescent collecting device
It extracts the peak value at the corresponding peak position and obtains the corresponding photoluminescence peak of each fluorescence waveform signal.
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