CN108918444A - The detection method of Escherichia coli in cow's milk based on ultraviolet-visible light spectral technology - Google Patents
The detection method of Escherichia coli in cow's milk based on ultraviolet-visible light spectral technology Download PDFInfo
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- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
- G01N21/3151—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths using two sources of radiation of different wavelengths
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
- G01N2021/3155—Measuring in two spectral ranges, e.g. UV and visible
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Abstract
The present invention provides a kind of under the complex background environment of similar cow's milk, can effectively solve the detection spectrum of microorganism is influenced phenomena such as generating characteristic wavelength red shift and obvious noise by unfavorable factor, causes spectral signature to be difficult to pick up, the problems such as quantitative detection result accuracy is low.Activation and secondary culture are carried out to Escherichia coli bacterium powder first, configure bacteria suspension to be measured；Ultraviolet specrophotometer obtains the ultraviolet-visible spectrum of the bacteria suspension to be measured of different incubation times；Noise jamming is eliminated using standard normal correction method and S-G convolution exponential smoothing, spectral signature Red Shift Phenomena is explained from the angle of phenyl ring conjugated structure formation mechenism, characteristic wave bands range is determined with this, the pickup for being carried out characteristic wavelength in characteristic wave bands using successive projection algorithm effectively screens characteristic wavelength according to the rule of microorganism " diauxic growth "；The relationship model that characteristic wavelength and microbial count are established using Partial Least Squares carries out quantitative analysis to the sum of Escherichia coli in cow's milk.
Description
Technical field
The present invention relates to ultraviolet-visible spectrum detection technique more particularly to it is a kind of in cow's milk Escherichia coli sum
Detection method.
Background technique
Escherichia coli are the highest food-borne pathogens of recall rate in cow's milk, are internationally recognized monitoring of hygiene indicator bacterias,
Therefore the specificity of more sensitive capture Escherichia coli, realize that efficiently quickly detection has safely guarantee China's dairy products for it
Significance.Currently, the detection and counting of Escherichia coli rely primarily on conventional bacteriology cultivation in cow's milk：MTF method
And colony counting method, but such method time and effort consuming, operation is strongly professional, and is unable to satisfy quick, non-destructive testing demand.
Uv-visible absorption spectra is usually used in the analysis and research of the protein and other containing conjugated system,
By the spectral signature of conjugated structure can be finally inversed by bacterial micro-organism at be grouped as, the information such as concentration.This technology has nothing
Need to add reaction reagent, it is not damaged, easy to operate the advantages that, be widely used in microorganism quantitative detection, at present state
It is inside and outside to have there is multidigit scholar to realize using this technology pair.But spectral signature and its sum in relation to Escherichia coli in cow's milk are determined
Amount detection research also belongs to blank at home.
The main reason is that cow's milk ingredient is complicated, and the large biological molecules particle such as ox protein of milk, fat
Particle diameter distribution unevenness can generate the noise jammings caused by factors such as light scattering, can also the detection bands of Escherichia coli carry out characteristic light
Spectrum red shift, characteristic wavelength be difficult to pick up, quantitative detection result inaccuracy etc. influences.
Summary of the invention
Brief overview of the invention is given below, in order to provide the basic reason in terms of some of the invention
Solution.It should be appreciated that this summary is not an exhaustive overview of the invention.He is not intended to determine key of the invention
Or pith, nor is it intended to limit the scope of the present invention.Its purpose only provides certain concepts in simplified form, with
This is as the preamble in greater detail discussed later.
In consideration of it, the present invention provides a kind of detection method for Escherichia coli sum in cow's milk, it is existing to solve
Since the large biological molecules such as protein, fat generate detection knot caused by the factors such as light scattering in milk elements complexity and cow's milk
The problem of fruit inaccuracy.
The present invention provides a kind of for the inspection for Escherichia coli sum in cow's milk based on ultraviolet-visible light spectral technology
Survey method, integral experiment process include：Escherichia coli bacterium powder is subjected to activation and secondary culture, obtaining has the of growth activity
Three generations's coli strain；In an aseptic environment by bacterial strain culture transferring into sterile full-cream fresh cow milk, continuous culture respectively after packing；
The cultured cow's milk that carries disease germs is put into cuvette, is inhaled using the ultraviolet-visible that ultraviolet specrophotometer obtains its 200 ~ 400nm
Receive spectrum；At the same time, the cow's milk that carries disease germs of 12 groups of incubation times is subjected to plate count, as scaling experiment, and obtains cow's milk
The true value of the quantity of middle Escherichia coli；The uv-visible absorption spectra of Escherichia coli imports at data in the cow's milk that will acquire
It manages software and carries out reasonable spectroscopic data processing, and using uv-visible absorption spectra to the conjugated structure in organic compound
Have the characteristics that higher sensibility, explains that spectral signature wave band red shift is existing from the angle of phenyl ring conjugated structure formation mechenism
As determining characteristic wave bands range with this, establishing regressive prediction model, pass through absorbance value and the total true value phase of the cow's milk that carries disease germs
Closing property r and root-mean-square error value RMSE carrys out the stability and predictive ability of valuation prediction models.
Further, the Escherichia coli bacterium powder, which is activated with subculture step, includes：The activation of strain：It is living according to strain
Change explanation, in an aseptic environment, take appropriate bacterium powder in redissolving in liquid, after mixing, bacterium solution streak inoculation is entered into the training of LB solid
It supports in base, is inverted in 37 DEG C of constant incubators and cultivates 18h；The separation of thallus：Culture dish after removal activation；The single bacterium of picking
It falls in LB nutrient broth medium, is placed in 37 DEG C of constant incubators and cultivates 18h；The purifying of thallus：By obtained bacterium solution into
Row centrifugation-deionized water washing-suction abandons supernatant liquor-centrifugation etc. and repeats above step three times, to ensure thallus not because high speed is revolved
Then destroyed, the speed of centrifuge is set as 3000r/min, and setting time 5min, Escherichia coli bacterium derived above
Suspension.
Further, the preparation step of the milk sample to be measured that carries disease germs includes：The full-cream cow's milk of offering of 300mL is poured into taper
In bottle, and take sterile fresh cow milk 25mL in test tube, number 0, as a control group：200 μ L Escherichia coli are drawn with pipettor
Bacteria suspension is in cow's milk, and mixing is sub-packed in 11 test tubes after shaking up, every 25mL, respectively number 1,2,3 ... 11, for experiment
Group；The sample of experimental group is put into constant incubator, respectively under 37 DEG C of constant temperatures culture 1.5,3,4,6,8,10,12,
14、16、18、20h。
Further, the obtaining step of the ultraviolet-visible spectrum of Escherichia coli includes in the cow's milk：Using ultraviolet spectrometry
Photometer carries out the acquisition of spectroscopic data, according to the priori spectral information of milk elements, spectral detection range is chosen to be 200 ~
400nm, while in order to avoid omitting valuable characteristic information, the sampling interval is set as 0.5nm, using deionized water as ginseng
Than：5mL is pipetted in cuvette from the cultured cow's milk that carries disease germs, and the sample duplicate measurements of each time point three times, and takes it
Mean value is as measurement result.The experiment of 4 independent completions has been carried out herein, has obtained 144 groups of original spectral data altogether, by taking
Value and to the preliminary screening of effective spectrum, obtains can be used for further totally 36 groups of pretreated spectroscopic data, every group of data include
201 wavelength absorbance values.
Further, include the step of Escherichia coli Ultraviolet-visible Spectroscopic Characteristics waveband selection in the cow's milk：In cow's milk
Continuously there is feature wave crest in 300 ~ 400nm wave band in the uv-visible absorption spectra of Escherichia coli, and peak intensity increases over time
And characteristic wave bands red shift, basic reason be there is phenyl ring conjugated structure in three kinds of amino acid moleculars that Escherichia coli contain, and
Benzene ring substitution group presence-COOH and-NH in amino acid molecular2Inhomogeneity contraposition replaces, substituent group internal structure is complicated and deposits
With the active-OH of parental materials ,-NH2Equal substituent groups transmit electronic effect by conjugated structure and cause conjugated double bond number
Situations such as mesh increases collective effect, promotes conjugation intensity to increase, the increase of feature peak intensity, absorption band red shift, therefore selects
Select characteristic wave bands of 300 ~ 400nm wave band as spectrum.
Further, Escherichia coli ultraviolet-visible spectrum data prediction step includes in the cow's milk：Due to causing light
It is ingredient complicated in cow's milk that spectrum signature, which is difficult to the main reason for extracting, and fat, protein and other partial size point
Light scattering phenomenon caused by cloth is uneven, therefore selection can effectively correct the standard normal of the scattering phenomenon for the cow's milk spectrum that carries disease germs
Variate calibration method（SNV）, eliminate or weaken the noise jamming in spectrum；And it further uses the smooth convolution method of S-G and eliminates baseline
The noises such as drift, inclination, to improve the robustness and predictive ability of model；For the selection of characteristic wave bands in previous step,
Successive projection method is used to the ultraviolet-visible spectrum data information of 300 ~ 400nm（SPA）Carry out picking up for further feature wavelength
It taking, the absorbance value of the characteristic wavelength picked up, the change curve of absorbance meets the growth curve of bacterium at any time, and
Occur " diauxic growth " this regularity variation at 3h and be also fully consistent with the distinctive growth rhythm of bacterium, shows SPA
The extracted characteristic wavelength of algorithm can be used to characterization Escherichia coli Growth feature, further demonstrate SNV-SG-SPA pairs
The accuracy that Pretreated spectra and characteristic wavelength pick up.
For the detection of microbial count under the similar relative complex background of ox composition of milk, examined using spectrum of the invention
Survey method and data processing method can fully consider that complex environment is dry to microorganism detection spectrum bring spectral noise
Disturb, feature red shift the problems such as, so as to obtain more accurate quantitative spectrometric testing result.
Total detection method for microorganism under the complex background environment based on ultraviolet-visible light spectral technology of the invention
And the preprocess method for spectroscopic data under complex background environment, using SNV-SG method combine to original spectral data into
Row calibration one by one, noise smoothing, SPA algorithm further extracts the characteristic wavelength in pretreated spectrum, and utilizes
The growth rhythm of microorganism distinctive " diauxic growth " is further preferred to characteristic wavelength, can effectively reduce synteny shadow
It rings, improves modeling conditions, improve the predictive ability and precision of established model.
Control group of the present invention using colony counting method as experimental result, and utilization can represent Escherichia coli Growth trend
Characteristic wavelength and the true value of Escherichia coli sum that measures of colony counting method, it is total to construct Escherichia coli in cow's milk
PLS prediction model.
Detailed description of the invention
The present invention can be by reference to better understanding, wherein all attached below in association with description given by attached drawing
The same or similar appended drawing reference has been used in figure to indicate same or similar component.The attached drawing is together with following detailed
Illustrate that instrument includes in the present specification and to form a part of this specification, and be used to that the present invention is further illustrated
Preferred embodiment and explain the principle of the present invention and advantage.
In the accompanying drawings：
Fig. 1 be the present invention realize complex background environment under, microorganism detection spectroscopic data processing and regressive prediction model establish
Flow chart；
Fig. 2 is the primary light spectrogram of E. coli detection in cow's milk in the present invention；
Fig. 3 is the characteristic wave bands spectrogram of E. coli detection in cow's milk in the present invention；
Fig. 4 is the pretreated spectrogram of SNV-SG method in E. coli detection in cow's milk in the present invention；
Fig. 5 is SPA algorithm preferred feature wave location drawing in spectrum in E. coli detection in cow's milk in the present invention；
Fig. 6 is characteristic wave strong point Escherichia coli Growth curve in E. coli detection in cow's milk in the present invention；
Fig. 7 is the PLS prediction model that characteristic spectrum and Escherichia coli sum are established in E. coli detection in cow's milk in the present invention
Figure；
It will be appreciated by those skilled in the art that schematically shown for the sake of attached drawing is answered and understood just for the sake of letter, without
For limiting technical solution of the present invention, to help to improve the understanding to the embodiment of the present invention.
Specific embodiment
Below in association with attached drawing, the present invention will be further described.
The present invention provides a kind of for the inspection for Escherichia coli sum in cow's milk based on ultraviolet-visible light spectral technology
Survey method, integral experiment process include：Escherichia coli bacterium powder is subjected to activation and secondary culture, obtaining has the of growth activity
Three generations's coli strain；In an aseptic environment by bacterial strain culture transferring into sterile full-cream fresh cow milk, continuous culture respectively after packing；
The cultured cow's milk that carries disease germs is put into cuvette, is inhaled using the ultraviolet-visible that ultraviolet specrophotometer obtains its 200 ~ 400nm
Receive spectrum；At the same time, the cow's milk that carries disease germs of 12 groups of incubation times is subjected to plate count, as scaling experiment, and obtains cow's milk
The true value of the quantity of middle Escherichia coli；The uv-visible absorption spectra of Escherichia coli imports at data in the cow's milk that will acquire
It manages software and carries out reasonable spectroscopic data processing, and using uv-visible absorption spectra to the conjugated structure in organic compound
Have the characteristics that higher sensibility, explains that spectral signature wave band red shift is existing from the angle of phenyl ring conjugated structure formation mechenism
As determining characteristic wave bands range with this, establishing regressive prediction model, pass through absorbance value and the total true value phase of the cow's milk that carries disease germs
Closing property r and root-mean-square error value RMSE carrys out the stability and predictive ability of valuation prediction models.
Ultraviolet-visible spectrum detection side for Escherichia coli sum in cow's milk of the invention is described below with reference to Fig. 1
The detailed process flow of method.
As shown in Figure 1, executing step S110 after process flow starts.
In step s 110, activation and secondary culture carried out to microbial germ powder, and cow's milk mixed configuration bacteria suspension to be measured,
Continuous culture at the appointed time.Then, step S120 is executed.
Wherein culture environment is continuously to cultivate 1.5,3,4,6,8,10,12,14,16,18,20h under sterile, 37 DEG C of constant temperature.
In the step s 120, obtained using ultraviolet specrophotometer the bacteria suspension to be measured of different incubation times it is ultraviolet-can
Light-exposed spectrum.Then, step S130 is executed.
Wherein, the spectral detection range of ultraviolet specrophotometer is chosen to be 200 ~ 400nm, while valuable in order to avoid omitting
The characteristic information of value, sampling interval are set as 0.5nm, using deionized water as reference.
In step s 130, standard normal variable correction method is utilized（SNV）It eliminates or weakens in conjunction with S-G convolution exponential smoothing and make an uproar
Acoustic jamming.Then, step S140 is executed.
Wherein S-G convolution exponential smoothing carries out 3 Integral Processings.
In step S140, spectral signature wave band Red Shift Phenomena is explained from the angle of phenyl ring conjugated structure formation mechenism,
300 ~ 400nm of characteristic wave bands range is determined with this.Then, step S150 is executed.
Wherein, continuously there is characteristic wave in 300 ~ 400nm wave band in the uv-visible absorption spectra of Escherichia coli in cow's milk
Peak, peak intensity increases over time and characteristic wave bands red shift, and basic reason is in three kinds of amino acid moleculars that Escherichia coli contain
There is phenyl ring conjugated structure, and benzene ring substitution group presence-COOH and-NH in amino acid molecular2Inhomogeneity contraposition replaces, replaces
Base internal structure is complicated and exists with the active-OH of parental materials ,-NH2Equal substituent groups transmit electricity by conjugated structure
Sub- effect causes situations such as conjugated double bond number increase collective effect, and conjugation intensity is promoted to increase, and feature peak intensity increases
Greatly, absorption band red shift, therefore select 300 ~ 400nm wave band as the characteristic wave bands of spectrum
In step S150, using successive projection algorithm（SPA）The pickup that characteristic wavelength is carried out in characteristic wave bands, according to micro- life
The growth rhythm of object " diauxic growth " carries out significantly more efficient screening to characteristic wavelength.Then.Execute S160.
In step S160, Partial Least Squares is utilized（PLS）Establish the model between characteristic wavelength and microbial count
Relationship realizes that the sum to microorganism under complex background environment carries out quantitative analysis.
Claims (7)
1. the ultraviolet-visible light spectrum detection method for Escherichia coli sum in cow's milk, which is characterized in that the detection method packet
It includes：
Activation and secondary culture is carried out to microbial germ powder to connect at the appointed time with pure cow's milk mixed configuration bacteria suspension to be measured
Continuous culture；
The ultraviolet-visible spectrum of the bacteria suspension to be measured of different incubation times is obtained using ultraviolet specrophotometer；
Utilize standard normal variable correction method（SNV）Noise jamming is eliminated or weakened in conjunction with S-G convolution exponential smoothing；
Spectral signature wave band Red Shift Phenomena is explained from the angle of phenyl ring conjugated structure formation mechenism, and characteristic wave bands model is determined with this
It encloses；
Using successive projection algorithm（SPA）The pickup that characteristic wavelength is carried out in characteristic wave bands, according to microorganism " diauxic growth "
Growth rhythm significantly more efficient screening is carried out to characteristic wavelength；
Utilize Partial Least Squares（PLS）The relationship model between characteristic wavelength and microbial count is established, realizes and complexity is carried on the back
The sum of microorganism carries out quantitative analysis under scape environment.
2. detection method according to claim 1, which is characterized in that the detection microorganism and complex background environment,
And the step of carrying out activation and secondary culture to microbial germ powder, configuring bacteria suspension to be measured, includes：
The activation of strain：Illustrate according to actication of culture, in an aseptic environment, takes appropriate bacterium powder in redissolving in liquid, after mixing,
Bacterium solution streak inoculation is entered in LB solid medium, is inverted in 37 DEG C of constant incubators and cultivates 18h；The separation of thallus：Removal
Culture dish after activation；Picking single bacterium colony is placed in 37 DEG C of constant incubators in LB nutrient broth medium and cultivates 18h；
The purifying of thallus：Obtained bacterium solution is carried out to be centrifuged the repetition above steps such as-deionized water washing-suction abandoning supernatant liquor-centrifugation
Three times, to ensure that thallus is not destroyed because of high speed rotation, the speed of centrifuge is set as 3000r/min, and when setting
Between 5min, Escherichia coli bacteria suspension derived above.
3. the obtaining step of the ultraviolet-visible spectrum of Escherichia coli includes in cow's milk according to claim 1 or 2：
Spectrum is examined according to the priori spectral information of milk elements using the acquisition that ultraviolet specrophotometer carries out spectroscopic data
It surveys range and is chosen to be 200 ~ 400nm, while in order to avoid omitting valuable characteristic information, the sampling interval is set as 0.5nm, with
Deionized water is as reference：5mL is pipetted in cuvette from the cultured cow's milk that carries disease germs, and the sample of each time point repeats to survey
Amount three times, and takes its mean value as measurement result；
The experiment of 4 independent completions has been carried out herein, 144 groups of original spectral data is obtained altogether, by taking mean value and to effective light
The preliminary screening of spectrum obtains can be used for further totally 36 groups of pretreated spectroscopic data, and every group of data include 201 wavelength extinctions
Angle value.
4. spectroscopic data preprocess method according to claim 3, which is characterized in that described due to causing spectral signature difficult
The main reason for extract is ingredient complicated in cow's milk, and fat, protein and other particle diameter distribution unevenness are drawn
The light scattering phenomenon risen, therefore selection can effectively correct the standard normal variable correction of the scattering phenomenon for the cow's milk spectrum that carries disease germs
Method（SNV）, eliminate or weaken the noise jamming in spectrum；And it further uses the smooth convolution method of S-G and eliminates baseline drift, inclination
Equal noises, to improve the robustness and predictive ability of model.
5. characteristic spectrum band selection method according to claim 4, which is characterized in that Escherichia coli in the cow's milk
Continuously there is feature wave crest in 300 ~ 400nm wave band in uv-visible absorption spectra, and peak intensity increases over time and characteristic wave bands
Red shift, basic reason are there is phenyl ring conjugated structure, and amino acid molecular in three kinds of amino acid moleculars that Escherichia coli contain
Middle benzene ring substitution group presence-COOH and-NH2Inhomogeneity contraposition replaces, substituent group internal structure is complicated and exists with electrophilic
Replace active-OH ,-NH2Equal substituent groups transmit electronic effect by conjugated structure and cause the feelings such as conjugated double bond number increase
Condition collective effect promotes conjugation intensity to increase, the increase of feature peak intensity, absorption band red shift, therefore selects 300 ~ 400nm
Characteristic wave bands of the wave band as spectrum.
6. characteristic spectrum band selection method according to claim 5, which is characterized in that described to be directed to 300 ~ 400nm's
Ultraviolet-visible spectrum data information uses successive projection method（SPA）The pickup for carrying out further feature wavelength, is picked up
The absorbance value of characteristic wavelength, the change curve of absorbance meets the growth curve of bacterium at any time, and occurs at 3h
" diauxic growth " this regularity variation is also fully consistent with the distinctive growth rhythm of bacterium, and it is extracted to show SPA algorithm
Characteristic wavelength can be used to characterization Escherichia coli Growth feature, further demonstrate SNV-SG-SPA to Pretreated spectra with
And the accuracy that characteristic wavelength picks up.
7. regressive prediction model construction method according to claim 6, which is characterized in that described to utilize Partial Least Squares
（PLS）The relationship model between characteristic wavelength and microbial count is established, realizes the sum to microorganism under complex background environment
Carry out quantitative analysis.
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| CN109668873A (en) * | 2018-12-05 | 2019-04-23 | 邢凤江 | A kind of live vector vaccine device for detecting activity and its application method |
| CN109727280A (en) * | 2019-01-25 | 2019-05-07 | 哈尔滨理工大学 | A kind of high spectrum image abundance estimation method based on orthogonal basis |
| CN110144380A (en) * | 2019-05-06 | 2019-08-20 | 哈尔滨理工大学 | The prediction model of escherichia coli in a kind of dairy products |
| CN110174365A (en) * | 2019-05-27 | 2019-08-27 | 哈尔滨理工大学 | The detection method of content of microorganisms in complicated solution based on ultraviolet-visible spectrum |
| CN111751302A (en) * | 2019-12-30 | 2020-10-09 | 哈尔滨理工大学 | Method for detecting concentration of melamine in whey protein powder |
| CN113340831A (en) * | 2021-05-10 | 2021-09-03 | 哈尔滨理工大学 | Ultraviolet spectral characteristic analysis and quantitative detection method for yeast and escherichia coli in cow's milk |
| CN115747036A (en) * | 2023-01-05 | 2023-03-07 | 中国人民解放军空军特色医学中心 | Method and device for searching and screening laser wavelength capable of being strongly absorbed by flora |
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| CN109668873A (en) * | 2018-12-05 | 2019-04-23 | 邢凤江 | A kind of live vector vaccine device for detecting activity and its application method |
| CN109727280A (en) * | 2019-01-25 | 2019-05-07 | 哈尔滨理工大学 | A kind of high spectrum image abundance estimation method based on orthogonal basis |
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| CN111751302A (en) * | 2019-12-30 | 2020-10-09 | 哈尔滨理工大学 | Method for detecting concentration of melamine in whey protein powder |
| CN113340831A (en) * | 2021-05-10 | 2021-09-03 | 哈尔滨理工大学 | Ultraviolet spectral characteristic analysis and quantitative detection method for yeast and escherichia coli in cow's milk |
| CN115747036A (en) * | 2023-01-05 | 2023-03-07 | 中国人民解放军空军特色医学中心 | Method and device for searching and screening laser wavelength capable of being strongly absorbed by flora |
| CN115747036B (en) * | 2023-01-05 | 2023-05-02 | 中国人民解放军空军特色医学中心 | Method for searching and screening laser wavelength capable of being strongly absorbed by flora |
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