CN109060734A - The zearalenone visualization fast screening system and method for fluorescence imaging - Google Patents
The zearalenone visualization fast screening system and method for fluorescence imaging Download PDFInfo
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- CN109060734A CN109060734A CN201810628805.9A CN201810628805A CN109060734A CN 109060734 A CN109060734 A CN 109060734A CN 201810628805 A CN201810628805 A CN 201810628805A CN 109060734 A CN109060734 A CN 109060734A
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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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
Abstract
The invention belongs to technical field of food detection, and in particular to the zearalenone visualization fast screening system and method for fluorescence imaging.Screening system of the present invention includes luminous intensity regulation module, camera bellows and computer；The camera bellows successively includes EMCCD camera, the ultraviolet annular light source mould group of timing controllable multiple-wavelength LED, specimen cup from top to bottom, and specimen cup is nearby placed with photodetector；The computer is electrically connected with EMCCD camera, luminous intensity regulation module respectively；The luminous intensity regulation module is electrically connected with light source module group and photodetector respectively.Using above-mentioned screening system rapid screening can be carried out to zearalenone, specifically: pretreatment sample, acquisition fluorescent color image, high effective liquid chromatography for measuring zearalenone content, establishes simultaneously optimized mathematical model, measurement zearalenone content at instrument testing.Light source module group of the invention is gathered at equal intervals to be arranged, and each sequence LED of independent control increases the uniform and stable property of illumination, improves detection speed and precision of prediction.
Description
Technical field
The invention belongs to technical field of food detection, and in particular to the zearalenone visualization of fluorescence imaging is quickly sieved
Look into system and method.
Background technique
It is counted according to FAO (Food and Agriculture Organization of the United Nation) (FAO), pollution of the grain and oil crop in the whole world average 25% by mycotoxin.By
The influence of the factors such as weather and storage, China are to be polluted one of the countries with the most serious ..., grain security face by mycotoxin in the world
Face severe challenge.Mycotoxin is mycetogenetic secondary metabolite, mainly includes aflatoxin, Gibberella zeae alkene
Ketone, deoxynivalenol, ochratoxin, fumonisin etc..Mycotoxin has virulent property and carcinogenicity, can be dirty
Edible and feeding agricultural product, especially corn, wheat, rice, barley, millet and the oat of all kinds etc. grain is contaminated to make
Object seriously endangers people, animal health.
The seriously polluted threat China grain security of mycotoxin, and traditional grain security management system shortage is effective pre-
Anti- property technological means and quick detection means, such as Gibberella zeae alkene in GB 5009.209-2016 national food safety standard food
The measurement of ketone carries out Gibberella zeae using liquid chromatography, fluorimetry or solid-phase extraction column purification liquid chromatography-mass spectrometry
The measurement of ketenes.Required the device is complicated, complex steps, detection cycle are long, is difficult to realize field quick detection, is unable to satisfy food
The quickly requirement of real-time detection in circulation and process.The rapid detection method of zearalenone such as uses ELISA, glue
The methods of body gold test paper, immune affinity column improve detection speed and detection accuracy, these detection methods are required to acetonitrile or methanol
Processing, complex steps and the time-consumings such as extraction, the purification of equal organic solvents.
The quick knowledge of grain contamination degree is realized using normal grain materials and by the optical signal difference of fungal contamination material
Not, having patent to obtain fluorescence spectrum using high resolution spectrometer can effectively identify that aspergillus flavus pollutes；Related patents disclose one
The identification screening installation and method for separating of kind vomitoxin, aflatoxin detects and sorting unit, one kind are based on FT-NIR skill
The rapid detection method of aflatoxin content in the brown rice of art can be examined quickly by fluorescent differences or near infrared spectrum characterization
Mycotoxin pollution is surveyed, but grain material mycotoxin is unevenly distributed and (is distributed in inside sometimes), mycotoxin and grain
Component combine mode to ultraviolet excitation generate fluorescence capability have differences, cause existing fluorescence detection method False Rate compared with
It is high；The mode of near infrared spectrum point measurement there are problems that taking a part for the whole to Mycotoxins in Cereal assessment of levels, cause to comment
The result stability of valence is poor.The visualization Evaluation on distribution that mycotoxin in grain is realized according to green analytical technology, is being examined
There must be qualitative leap in terms of surveying speed and accuracy.
Summary of the invention
In view of the deficiencies of the prior art, the present invention provides a kind of zearalenone visualization rapid screening of fluorescence imaging
System and method overcomes the problems, such as that conventional Physico-chemical tests speed is slow, chemical reagent is disagreeableness to environment, mycotoxin is overcome to be distributed
Unevenness causes to measure the high problem of False Rate, overcomes the problems, such as that the adaptability of spectral detection point measurement and stability are poor, significantly mentions
The detection speed of zearalenone and the reliability of detection in high grain.
The present invention provides a kind of zearalenone visualization fast screening system of fluorescence imaging, specifically, of the invention
It is carried out by following scheme:
A kind of zearalenone visualization fast screening system of fluorescence imaging, including luminous intensity regulate and control module, camera bellows
With computer；The camera bellows from top to bottom successively include EMCCD camera, the ultraviolet annular light source mould group of timing controllable multiple-wavelength LED,
Specimen cup；The EMCCD camera is placed in the top of camera bellows, and the camera lens of EMCCD camera can protrude into the inside of camera bellows；The timing can
The control ultraviolet annular light source mould group of multi-wavelength LED is fixed on the middle and lower part of camera bellows by lifting device；The specimen cup is placed in camera bellows
Bottom, specimen cup is nearby placed with photodetector；
Luminous intensity regulation module is located at outside camera bellows, and respectively with the ultraviolet annular light source mould of timing controllable multiple-wavelength LED
Group and photodetector electrical connection, are equipped with acquisition control processing software in the computer, the computer respectively with EMCCD
Camera, luminous intensity regulation module electrical connection；
The ultraviolet annular light source mould group of timing controllable multiple-wavelength LED be circular ring shape, including multiple sequence LED light source groups and
Show wide indicator lamp groups；Multiple sequence LED light source groups are successively equidistantly covered with entire annulus, and positioned at the inner ring of annulus and outside
Center between circle；The sequence LED light source group includes the First ray LED equidistantly arranged, the second sequence LED and
Three sequence LED；It is described show wide indicator lamp groups include 4 or 6 it is circumferentially evenly arranged show wide indicator light, and close to annulus
Inner ring setting；
It includes constant pressure and flow controller, the light for individually adjusting First ray LED illumination intensity that luminous intensity, which regulates and controls module,
Coupling adjuster a, the optocoupler adjuster b for individually adjusting the second sequence LED illumination intensity and for individually adjust third sequence
The optocoupler adjuster c of LED illumination intensity；It is connected in parallel between the optocoupler adjuster a, optocoupler adjuster b and optocoupler adjuster c
Optocoupler adjuster group is formed, the optocoupler adjuster group and constant pressure and flow controller are connected in series.
Preferably, the outer diameter of the First ray LED, the second sequence LED and third sequence LED are 1.6mm；First sequence
The excitation wavelength for arranging LED is 236nm, and the excitation wavelength of the second sequence LED is 274nm, and the excitation wavelength of third sequence LED is
365nm；
Preferably, the center of the specimen cup is placed in EMCCD camera focusing center line and timing controllable multiple-wavelength LED is ultraviolet
On the line of the center line of annular light source mould group；The ultraviolet annular light source mould group of the timing controllable multiple-wavelength LED and specimen cup
Vertical range is 30-45cm；The photodetector is placed in the uniform light of the ultraviolet annular light source mould group of timing controllable multiple-wavelength LED
According in area；
Preferably, the response wave band of the photodetector is 200-700nm；
Preferably, the EMCCD camera uses the EMCCD chip of resolution ratio 512*512 Pixel Dimensions, in EMCCD camera
Sensor devices using EX2 coating technique make, using FPGA carry out EMCCD camera time mark；The computer uses
USB2.0 or 3.0 computer interfaces；
Preferably, the acquisition control processing software realizes the acquisition control processing using Visual Studio programming
Software development environment be 7 Ultimate of computer system Windows (64) and 2010 Ultimate of Visual Studio, exploitation
The executable file of the acquisition control processing software, which may be mounted at, to be met in the computer or industrial personal computer of minimalist configuration.
The present invention also provides a kind of zearalenone of fluorescence imaging visualize quick screening method, especially by with
Lower step is realized:
S1. grain sample is pre-processed:
Batch obtains the grain sample that sickle-like bacteria natural pollution contains zearalenone, and grain is made through pulverizer crushing
Powdered sample；Food Powder sample is divided into multiple sample sample；Multiple sample sample is laid in the center of specimen cup, and will
Specimen cup is placed in the image acquisition areas right-angled intersection point of EMCCD camera；
S2. instrument testing:
By the clarity, saturation degree, lightness of sample specimens fluorescent color image, best fluorescent color image condition is determined
When luminous intensity, the standard value of as three sequence LED light source mould groups；The LED light of three sequences is obtained by photodetector
The luminous intensity of source mould group feeds back to computer, if luminous intensity is not standard value, regulates and controls module increasing by luminous intensity and adds deduct
The luminous intensity of low current intensity, three sequence LED light source mould groups of adjust automatically becomes standard value；Then to EMCCD camera
Acquisition parameter is standardized, and is set as the Optimizing Mode of zearalenone, including the time for exposure, area-of-interest size,
Field angle, linkage trigger signal, object distance；
S3. fluorescent color image is acquired:
Different wave length is issued by the ultraviolet annular light source mould group of acquisition control processing software control sequential controllable multiple-wavelength LED
Exciting light；All First ray LED, all second sequence LED, all third sequence LED are successively lighted, after lighting every time,
Sample specimens are excited, while EMCCD camera acquires corresponding sequence fluorescent color image；To three sequences of acquisition
Fluorescent color image extracts R, G and B color component respectively；
S4. the content of high effective liquid chromatography for measuring zearalenone:
Multiple sample sample by fluorescent color Image Acquisition, measures Food Powder using high performance liquid chromatography respectively
The content of zearalenone in sample, measurement result is as reference value；
S5. founding mathematical models:
To R, G and B color component and high performance liquid chromatography measurement result of the multiple sample sample of acquisition, it is averaged respectively
It is worth the color component and high performance liquid chromatography measured value as Food Powder sample；
Using in stepwise multiple linear regression, local weighted linear regression, weighted least-squares method, Partial Least Squares
One kind or combinations thereof mode establishes the number between R, G and B color component of Food Powder sample and high performance liquid chromatography measured value
Model is learned, specifically:
CZEN=a1×R1+b1×G1+c1×B1+a2×R2+b2×G2+c2×B2+a3×R3+b3×G3+c3×B3+d；
C in formulaZENFor image space site f(x,y)The content of zearalenone, a in lower grain1、a2、a3Value is respectively to scheme
Image space site f(x,y)Lower R component regression coefficient, R1、R2、R3The red color component value respectively extracted in fluorescent color image, b1、
b2、b3Value is respectively image space site f(x,y)Lower G Quantile Regression coefficient, G1、G2、G3It is green to be extracted in fluorescent color image
Colouring component value, c1、c2、c3Value is image space site f(x,y)Lower B component regression coefficient, B1、B2、B3To be mentioned in fluorescent color image
The blue color component value taken, d are the correction value of mathematical regression model；
S6. optimized mathematical model:
The performance for the zearalenone mathematical model that evaluation is established is verified with independent sample collection and is advanced optimized；
S7. zearalenone content in grain sample to be measured is measured:
Grain sample to be measured is extracted into R, G and B color of three sequence fluorescent color images after the processing of step S2, S3
Component, the zearalenone mathematical model established using step S6 optimization, the corn for calculating each space site of color image are red
Mould ketenes content, with zearalenone content at each space site in pseudo color image expression grain sample.
Preferably, in step S2, the time for exposure is 20ms, area-of-interest size diameter is 8cm, field angle is
24 °, linkage trigger signal be 1-1-1, object distance 55cm.
Preferably, it in step S5, is modeled when the mathematical model is established using point sample, tile when test sample batch sample
Spatial distribution prediction.
Preferably, in step S7, the pseudo color image expression indicates grain by 64 color linear transitions of green to red setting
The pollution level of zearalenone in sample.
Compared with prior art, beneficial effects of the present invention embody as follows:
(1) present invention discloses the zearalenone visualization fast screening system and method for a kind of fluorescence imaging, this hair
The bright fluorescent color image that Grain Powder sample is obtained using multi-wavelength ultraviolet LED as excitaton source, detection process do not need to try using chemistry
Agent overcomes the problems, such as that conventional Physico-chemical tests speed is slow, chemical reagent is disagreeableness to environment；In addition more importantly the method
It is more convenient to detect speed, use cost and maintenance cost are far below high performance liquid chromatography and solid-phase extraction column purification liquid phase color
Spectrum-mass spectrography.
(2) present invention overcomes the problems, such as that mycotoxin is unevenly distributed the adaptability that caused point measures and stability is poor, existing
Have the mycotoxin using online colour-selection technique detection grain or oil crops, be unevenly distributed there are surface because of mycotoxin pollution and
The inside and outside root problem being distributed, the present invention excites the crushing sample of grain using annular LED ultraviolet source mould group, in grain
The spatial distribution visualization of zearalenone content is accurately comprehensively expressed, and testing result is precisely reliable.
(3) the ultraviolet annular light source mould group of timing controllable multiple-wavelength LED in the present invention is gathered at equal intervals using annular and is arranged,
And regulate and control the intensity of illumination of each sequence LED of module independent control by luminous intensity, the uniformity and stability of illumination are increased, with
Standardized ultraviolet LED excitation guarantees the precision of prediction of zearalenone mathematical model.
(4) present invention passes through the intensity of illumination of each sequence LED of photodetector feedback control, eliminates since light source uses the longevity
Light intensity variation, improves the stability of fluorescent color image original signal caused by life or external circuit disturb.
Detailed description of the invention
Fig. 1 shows the zearalenone visualization fast screening system structural schematic diagram of fluorescence imaging；
Fig. 2 shows the ultraviolet annular light source mould group arrangement schematic diagrames of timing controllable multiple-wavelength LED；
In figure, the ultraviolet annular light source mould group of 1- timing controllable multiple-wavelength LED, 2-EMCCD camera, 3- luminous intensity regulate and control mould
Block, 4- camera bellows, 5- computer, 6- photodetector, 7- fluorescent color Image Acquisition Special sample cup, 8- First ray LED, 9-
Second sequence LED, 10- third sequence LED, 11- shows wide indicator light.
Fig. 3 shows LED light source mould group timing control flow chart；
Fig. 4 shows the zearalenone visualization quick screening method Establishing process figure of fluorescence imaging.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical solution in the embodiment of the present invention is explicitly described, it is clear that described embodiment is the present invention
A part of the embodiment, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art are not having
Every other embodiment obtained under the premise of creative work is made, shall fall within the protection scope of the present invention.
Specific embodiments of the present invention will be described in further detail with reference to the accompanying drawings of the specification.
Embodiment 1
As shown in Figure 1, 2, 3, a kind of zearalenone of fluorescence imaging visualizes fast screening system, including luminous intensity
Regulate and control module 3, camera bellows 4 and computer 5；The camera bellows 4 successively includes EMCCD camera 2, timing controllable multiple-wavelength from top to bottom
The ultraviolet annular light source mould group 1 of LED and specimen cup 7；The EMCCD camera 2 is placed in the top of camera bellows 4, the camera lens of EMCCD camera 2
The inside of camera bellows 4 can be protruded into；The ultraviolet annular light source mould group 1 of timing controllable multiple-wavelength LED is fixed in camera bellows 4 by bracket
Lower part, bracket are moved up and down by slideway；The specimen cup 7 is placed in the bottom of camera bellows 4, and specimen cup 7 is nearby placed with photodetection
Device 6, photodetector 6 are placed in the uniform illumination area of the ultraviolet annular light source mould group 1 of timing controllable multiple-wavelength LED；The timing
The ultraviolet annular light source mould group 1 of controllable multiple-wavelength LED and the vertical range of specimen cup 7 are 30-45cm；
When grain sample fluorescence Image Acquisition, sample is placed in sample cup 7, and the center of specimen cup 7 is placed in EMCCD camera 2
On the line of focusing center line and ultraviolet 1 center line of annular light source mould group of timing controllable multiple-wavelength LED, and adopted in 4 image of camera bellows
Ji Qu identifies central point；
Luminous intensity regulation module 3 is located at outside camera bellows 4, and respectively with the ultraviolet annular light source of timing controllable multiple-wavelength LED
Mould group 1 and photodetector 6 are electrically connected, and acquisition control processing software is equipped in the computer 5, and the computer 5 is distinguished
It is electrically connected with EMCCD camera 2, luminous intensity regulation module 3；
The ultraviolet annular light source mould group 1 of timing controllable multiple-wavelength LED is circular ring shape, including multiple sequence LED light source groups
With show wide indicator lamp groups；Multiple sequence LED light source groups are successively equidistantly covered with entire annulus, and be located at the inner ring of annulus with
Center between outer ring；The sequence LED light source group includes the First ray LED 8 equidistantly arranged, the second sequence LED
9 and third sequence LED 10；It is described show wide indicator lamp groups include 4 or 6 it is circumferentially evenly arranged show wide indicator light 11, and
Inner ring close to annulus is arranged.
The First ray LED 8, the second sequence LED 9 and third sequence LED 10 outer diameter be 1.6mm；Described
The excitation wavelength of one sequence LED 8 is 236nm, and excitation generates 418nm fluorescence；The excitation wavelength of second sequence LED 9 is
274nm, excitation generate 440nm fluorescence；The excitation wavelength of third sequence LED 10 is 365nm, and excitation generates 450nm fluorescence；Institute
The LED excitation wavelength for stating three kinds of sequences is to optimize determination by a large number of experiments, and the LED of three kinds of sequences all has high directivity,
Light emitting angle is 6-20 °, and the LED light source mould group of each sequence is respectively formed uniform illumination in image acquisition areas；To eliminate environment
The interference of the ambient lights such as light, the ultraviolet annular light source mould group 1 of timing controllable multiple-wavelength LED are installed in camera bellows 4, and when detection closes
Close camera bellows 4；The LED light source mould group of each sequence regulates and controls 3 independent control luminous intensity of module by luminous intensity；
Show that wide indicator light 11 is white light LEDs, shows that wide indicator lamp groups are known as white LED light source mould group；For sample fluorescence
Before Image Acquisition, naked eyes determine that grain sample illumination range and collected region, confirmation grain sample are placed in image acquisition areas；
EMCCD chip of the EMCCD camera 2 using resolution ratio 512*512 Pixel Dimensions, the photoreceptor of EMCCD camera 2
Part is made of EX2 coating technique, improves quantum efficiency response range, and control EMCCD cryogenic temperature reduces the shadow of dark noise
It rings；EMCCD camera 2 carries out time label exact timing using FPGA, and EMCCD camera 2 uses USB2.0/3.0 computer interface
Improve data transfer speed；The camera lens of EMCCD camera 2 configures 400nm long wave pass filter, and exciting light and stray light is avoided to enter
EMCCD camera 2；Selecting EMCCD camera 2 is determined by repetition test, and content of toxins height is indefinite in grain sample, in purple
Fluorescence reaction is different under the excitation of outer light, and the conventional photosensitive sensitivity of CCD is poor, and the fluorescent image noise of acquisition is relatively low, EMCCD phase
Machine 2 can be improved the intensity of zearalenone fluorescent assay signal in grain sample, obtain lower detection limit, present invention choosing
EMCCD peak quantum efficiency is greater than 90%.
It includes constant pressure and flow controller, the light for individually adjusting First ray LED illumination intensity that luminous intensity, which regulates and controls module 3,
Coupling adjuster a, for individually adjusting the optocoupler adjuster b of the second sequence LED illumination intensity, for individually adjusting third sequence
The optocoupler adjuster of LED illumination intensity forms；It is in parallel between the optocoupler adjuster a, optocoupler adjuster b and optocoupler adjuster c
Connection forms optocoupler adjuster group, and the optocoupler adjuster group and constant pressure and flow controller are connected in series；Constant pressure and flow controller
Corresponding sequence LED illumination intensity is adjusted by optocoupler adjuster a, optocoupler adjuster b or optocoupler adjuster c, in addition, constant pressure
Constant-current controller is to show that wide indicator lamp groups (white LED light source mould group) carries out constant pressure power supply；
Luminous intensity regulation module 3 is electrically connected with the ultraviolet annular light source mould group 1 of timing controllable multiple-wavelength LED, can be guaranteed each
Sequence LED light source mould group keeps stable intensity of illumination；Light source can be fluctuated with the variation of power utilization environment, and luminous intensity regulates and controls mould
Block 3 adjusts the intensity of illumination of sequence ultraviolet LED light source group according to the feedback signal of photodetector 6；Photodetector 6 obtains respectively
The luminous intensity of sequence LED light source mould group, compared with luminous intensity when being established with mathematical model, when having differences, by adopting
Collection control processing software, which will be instructed, is transmitted to optocoupler adjuster a, optocoupler adjuster b or optocoupler adjusting via constant pressure and flow controller
Device c, adjusts the size of current of each sequence LED light source mould group of corresponding control, and then automatically adjusts each sequence LED light source mould group
Intensity of illumination guarantees the intensity stabilization of each sequence LED light source mould group；The mathematical model of zearalenone is in light source in grain
It is established on the basis of intensity calibration, to obtain stable, high-precision testing result must adjust in real time.
The response wave band of photodetector 6 is 200-700nm, adaptively regulates and controls to calculate by the acquisition control processing software
Method adjusts the intensity of illumination of LED light source mould group under each sequence, guarantees the stability of each sequence LED light source mould group luminous intensity；It is described
Adaptive regulation algorithm is intensity variation and size of current accounting equation, and sends luminous intensity regulation module for regulating command
3。
The acquisition control processing software being installed in computer 5 is realized using Visual Studio programming, passes through calling
The Software Development Kit that 2 supplier of EMCCD camera provides realizes the information exchange with camera；The acquisition control processing is soft
Part is integrated with every control instruction of EMCCD camera 2 and luminous intensity regulation module 3；The display of the acquisition control processing software
Interface, with real-time display acquisition fluorescent color image, built-in camera parameter setting, luminous intensity regulation and control instruction, image rectification,
Image data extraction, content of toxins computation model, pseudo color image generate and the functions such as data storage and publication；The acquisition control
Processing software processed exploitation environment is 7 Ultimate of computer system Windows (64) and 2010 Ultimate of Visual Studio,
The executable file of the acquisition control processing software of exploitation, which may be mounted at, to be met on the computer or industrial personal computer of minimalist configuration,
With preferable portable and compatibility.
The acquisition control processing software, for feeding back the light intensity of the ultraviolet annular light source mould group 1 of timing controllable multiple-wavelength LED
Regulating command is spent, the ultraviolet annular light source mould group 1 of control sequential controllable multiple-wavelength LED issues the exciting light of different wave length；Acquisition control
The LED 8 of all First ray 236nm is only lighted in processing software control processed, so that EMCCD camera 2 acquires First ray fluorescence coloured silk
Chromatic graph picture；Acquisition control processing software controls the LED 9 for only lighting all second sequence 274nm, so that EMCCD camera 2 acquires
Second sequence fluorescent color image；Acquisition control processing software controls the LED 10 for only lighting all third sequence 365nm, so that
EMCCD camera 2 acquires third sequence fluorescent color image；In addition, acquisition control processing software is also used to each space of grain sample
Under site the calculating of zearalenone content and as the result is shown, storage.
Embodiment 2
As shown in figure 4, a kind of zearalenone visualization quick screening method of fluorescence imaging is using 1 institute of embodiment
What the zearalenone visualization fast screening system for the fluorescence imaging stated was realized, the screening method is especially by following step
It is rapid to realize:
S1. grain sample is pre-processed:
Batch obtains the grain sample that sickle-like bacteria natural pollution contains zearalenone first, and sample is through high speed disintegrator
(revolving speed >=12000r/min) crushes, and Food Powder sample is made；Food Powder sample is divided into four parts of samples using quartering
Sample will wherein three parts of parallel sample samples be laid in specimen cup respectively, the center of specimen cup is placed in the figure of EMCCD camera
As acquisition zone right-angled intersection point；
S2. instrument testing:
The mass parameters such as clarity, saturation degree, lightness by sample specimens fluorescent color image determine that best fluorescence is color
Luminous intensity when color image condition, the standard value of as three sequence LED light source mould groups；Three sequences are obtained by photodetector
The luminous intensity of the LED light source mould group of column feeds back to computer, if luminous intensity is not standard value, regulates and controls module by luminous intensity
Current strength is increasedd or decreased, the luminous intensity of three sequence LED light source mould groups of adjust automatically becomes standard value；Then right
EMCCD camera acquisition parameter is standardized, and is set as the Optimizing Mode of zearalenone, and time for exposure 20ms, sense are emerging
Interesting area size diameter is 8cm, field angle is 24 °, linkage trigger signal is 1-1-1, object distance 55cm.
S3. fluorescent color image is acquired:
Different wave length is issued by the ultraviolet annular light source mould group of acquisition control processing software control sequential controllable multiple-wavelength LED
Exciting light, determine a length of 236nm of the excitation light wave of First ray LED, a length of 274nm of excitation light wave of the second sequence LED, the
The a length of 360nm of excitation light wave of three sequence LED；
It lights all First ray LED to excite sample specimens sample, while EMCCD camera acquisition First ray is glimmering
Light color image；It lights all second sequence LED to excite sample specimens, while the second sequence of EMCCD camera acquisition is glimmering
Light color image；It lights all third sequence LED to excite sample specimens, while EMCCD camera acquisition third sequence is glimmering
Light color image；R, G and B color component are extracted respectively to three sequence fluorescent colors image of acquisition；
S4. the content of high effective liquid chromatography for measuring zearalenone:
Three parts of parallel sample samples by fluorescent color Image Acquisition, measure sample using high performance liquid chromatography respectively
The content of zearalenone in sample, measurement result is as reference value；
S5. founding mathematical models:
Three parts of parallel sample samples of selection, R, G, B color component and high performance liquid chromatography measurement result of acquisition, respectively
It is averaged the color component and high performance liquid chromatography measured value as Food Powder sample；
Using in stepwise multiple linear regression, local weighted linear regression, weighted least-squares method, Partial Least Squares
One kind or combinations thereof establishes the mathematical model between the color component of Food Powder sample and high performance liquid chromatography measured value, tool
Body are as follows:
CZEN=a1×R1+b1×G1+c1×B1+a2×R2+b2×G2+c2×B2+a3×R3+b3×G3+c3×B3+d
C in formulaZENFor image space site f(x,y)The content of zearalenone, a in lower grain1、a2、a3Value is respectively to scheme
Image space site f(x,y)Lower R component regression coefficient, R1、R2、R3The red color component value respectively extracted in fluorescent color image, b1、
b2、b3Value is respectively image space site f(x,y)Lower G Quantile Regression coefficient, G1、G2、G3It is green to be extracted in fluorescent color image
Colouring component value, c1、c2、c3Value is image space site f(x,y)Lower B component regression coefficient, B1、B2、B3To be mentioned in fluorescent color image
The blue color component value taken, d are the correction value of mathematical regression model；
Point sample Accurate Model is used when mathematical model is established, tile when test sample batch sample space forecast of distribution；
S6. optimized mathematical model:
By increasing independent grain sample set, the operation of step S1-S4 is repeated, the color component and efficient liquid of sample are removed
Discrete value in phase chromatographic determination value, the performance for the zearalenone mathematical model that verifying evaluation procedure S5 is established, goes forward side by side one
Walk a in optimized mathematical model1、a2、a3、b1、b2、b3、c1、c2、c3, d etc. numerical value；
S7. zearalenone content in grain sample to be measured is measured:
R, G and B color of three sequence fluorescent color images are extracted after the processing of step S2, S3 to grain sample to be measured
Component, the zearalenone mathematical model established using S6 optimization, calculates the Gibberella zeae alkene in each space site of color image
Ketone content, with zearalenone content at each space site in pseudo color image expression grain sample；
The expression of pseudo color image sets 64 color linear transitions by green (representing 0) to red (representing 64) to indicate Gibberella zeae
The pollution level of ketenes, 0 to 64 color linear correspond to zearalenone content 0-2000ppb, Gibberella zeae alkene in grain
64 (red) are equally used to indicate serious pollution when the content of ketone reaches 2000ppb or more, being similar to traffic lights equally can be straight
See the content and harmfulness of zearalenone in ground display grain.
Fast screening system is visualized using the zearalenone of fluorescence imaging of the invention and method successively demonstrates
Corn, wheat, dregs of beans and rice, traditional detection method detection time is long, and testing cost is high, and traditional detection method is a kind of point survey
The method of amount, haves the defects that Points replacing surfaces, causes False Rate high, and testing result, which only represents to have detected, samples this, the present invention
There is substantial advance between when detecting, in terms of testing cost, detection dimensions and detection accuracy.
The screening technique of the invention of table 1 is compared with the testing result of conventional method
It is found by verification experimental verification, the zearalenone visualization fast screening system of fluorescence imaging of the invention and side
Method is arranged at equal intervals the ultraviolet annular light source mould group uniform illumination of timing controllable multiple-wavelength LED of design, by controlling each sequence
The intensity of illumination of LED has obtained the fluorescent color image of high quality, is controlled by standardized light source, ensure that zearalenone
The applicability and accuracy of mathematical model.
The present invention overcomes the problems, such as that conventional Physico-chemical tests speed is slow, chemical reagent is disagreeableness to environment, overcomes mycotoxin
It is unevenly distributed and causes to measure the high problem of False Rate, overcome the problems, such as that the adaptability of spectral detection point measurement and stability are poor, show
It writes and improves the detection speed of zearalenone and the reliability of detection in grain, compared with prior art, in detectability, inspection
Surveying precision, Visualization etc. present invention has substantive distinguishing features outstanding and significant progress.
Although the embodiments of the invention are described in conjunction with the attached drawings, but those skilled in the art can not depart from this hair
Various modifications and variations are made in the case where bright spirit and scope, such modifications and variations are each fallen within by appended claims
Within limited range.
Claims (10)
1. a kind of zearalenone of fluorescence imaging visualizes fast screening system, which is characterized in that regulate and control including luminous intensity
Module (3), camera bellows (4) and computer (5)；The camera bellows (4) is successively controllably more including EMCCD camera (2), timing from top to bottom
The ultraviolet annular light source mould group (1) of wavelength LED and specimen cup (7)；The EMCCD camera (2) is placed in the top of camera bellows (4), EMCCD
The camera lens of camera (2) can protrude into the inside of camera bellows (4)；The ultraviolet annular light source mould group (1) of timing controllable multiple-wavelength LED passes through
Lifting device is fixed on the middle and lower part of camera bellows (4)；The specimen cup (7) is placed in the bottom of camera bellows (4), and specimen cup (7) is nearby put
There are photodetector (6)；
Luminous intensity regulation module (3) is located at camera bellows (4) outside, and respectively with the ultraviolet annular light source of timing controllable multiple-wavelength LED
Mould group (1) and photodetector (6) are electrically connected, and are equipped with acquisition control processing software, the computer in the computer (5)
(5) it is electrically connected respectively with EMCCD camera (2), luminous intensity regulation module (3)；
The ultraviolet annular light source mould group (1) of timing controllable multiple-wavelength LED be circular ring shape, including multiple sequence LED light source groups and
Show wide indicator lamp groups；Multiple sequence LED light source groups are successively equidistantly covered with entire annulus, and positioned at the inner ring of annulus and outside
Center between circle；The sequence LED light source group includes the First ray LED(8 equidistantly arranged), the second sequence LED
(9) with third sequence LED(10)；It is described to show that wide indicator lamp groups include 4 or 6 and circumferentially evenly arranged show wide indicator light
(11), and close to the inner ring of annulus it is arranged.
2. the zearalenone of fluorescence imaging according to claim 1 visualizes fast screening system, which is characterized in that
The First ray LED(8), the second sequence LED(9) with third sequence LED(10) outer diameter be 1.6mm；First sequence
Arrange LED(8) excitation wavelength be 236nm, the second sequence LED(9) excitation wavelength be 274nm, the third sequence LED
(10) excitation wavelength is 365nm.
3. the zearalenone of fluorescence imaging according to claim 1 visualizes fast screening system, which is characterized in that
Luminous intensity regulation module (3) include constant pressure and flow controller, for individually adjust First ray LED(8) intensity of illumination optocoupler
Adjuster a, for individually adjust the second sequence LED(9) intensity of illumination optocoupler adjuster b and for individually adjust third sequence
LED(10) the optocoupler adjuster c of intensity of illumination；It is in parallel between the optocoupler adjuster a, optocoupler adjuster b and optocoupler adjuster c
Connection forms optocoupler adjuster group, and the optocoupler adjuster group and constant pressure and flow controller are connected in series.
4. the zearalenone of fluorescence imaging according to claim 1 visualizes fast screening system, which is characterized in that
The center of the specimen cup (7) is placed in EMCCD camera (2) focusing center line and the ultraviolet annular light source of timing controllable multiple-wavelength LED
On the line of the center line of mould group (1)；The ultraviolet annular light source mould group (1) of timing controllable multiple-wavelength LED and specimen cup (7)
Vertical range is 30-45cm；The photodetector (6) is placed in the ultraviolet annular light source mould group (1) of timing controllable multiple-wavelength LED
In uniform illumination area；The response wave band of the photodetector (6) is 200-700nm.
5. the zearalenone of fluorescence imaging according to claim 1 visualizes fast screening system, feature exists
In the EMCCD camera (2) uses the EMCCD chip of resolution ratio 512*512 Pixel Dimensions, photosensitive in EMCCD camera (2)
Device is made of EX2 coating technique, is marked using the time that FPGA carries out EMCCD camera (2)；The EMCCD camera (2) is adopted
With USB2.0 or 3.0 computer interfaces.
6. the zearalenone of fluorescence imaging according to claim 1 visualizes fast screening system, feature exists
In the acquisition control processing software is realized using Visual Studio programming；The acquisition control processing software develops ring
Border is 64 7 Ultimates of Windows of computer system and 2010 Ultimate of Visual Studio, and the acquisition control of exploitation is handled
The executable file of software, which may be mounted at, to be met in the computer or industrial personal computer of minimalist configuration.
7. the quick screening method of fast screening system as described in claim 1, which comprises the steps of:
S1. grain sample is pre-processed:
Batch obtains the grain sample that sickle-like bacteria natural pollution contains zearalenone, and Food Powder is made through pulverizer crushing
Sample；Food Powder sample is divided into multiple sample sample；Multiple sample sample is laid in the center of specimen cup, and by sample
Cup is placed in the image acquisition areas right-angled intersection point of EMCCD camera；
S2. instrument testing:
By the clarity, saturation degree, lightness of sample specimens fluorescent color image, when determining best fluorescent color image condition
Luminous intensity, the standard value of as three sequence LED light source mould groups；The LED light source mould of three sequences is obtained by photodetector
The luminous intensity of group feeds back to computer, if luminous intensity is not standard value, regulates and controls module by luminous intensity and increaseds or decreases electricity
The luminous intensity of intensity of flow, three sequence LED light source mould groups of adjust automatically becomes standard value；Then to the acquisition of EMCCD camera
Parameter is standardized, and is set as the Optimizing Mode of zearalenone, including time for exposure, area-of-interest size, visual field
Angle, linkage trigger signal, object distance；
S3. fluorescent color image is acquired:
Swashing for different wave length is issued by the ultraviolet annular light source mould group of acquisition control processing software control sequential controllable multiple-wavelength LED
It shines；All First ray LED, all second sequence LED, all third sequence LED are successively lighted, after lighting every time, to sample
Product sample is excited, while EMCCD camera acquires corresponding sequence fluorescent color image；To three sequence fluorescence of acquisition
Color image extracts R, G and B color component respectively；
S4. the content of high effective liquid chromatography for measuring zearalenone:
Multiple sample sample by fluorescent color Image Acquisition, Food Powder sample is measured using high performance liquid chromatography respectively
The content of middle zearalenone, measurement result is as reference value；
S5. founding mathematical models:
To R, G and B color component and high performance liquid chromatography measurement result of the multiple sample sample of acquisition, it is averaged work respectively
For the color component and high performance liquid chromatography measured value of Food Powder sample；
Using one of stepwise multiple linear regression, local weighted linear regression, weighted least-squares method, Partial Least Squares
Or combinations thereof mode, establish the mathematical modulo between R, G and B color component of Food Powder sample and high performance liquid chromatography measured value
Type, specifically:
CZEN=a1×R1+b1×G1+c1×B1+a2×R2+b2×G2+c2×B2+a3×R3+b3×G3+c3×B3+d；
C in formulaZENFor image space sitef (x,y) The content of zearalenone, a in lower grain1、a2、a3Value is respectively that image is empty
Between sitef (x,y) Lower R component regression coefficient, R1、R2、R3The red color component value respectively extracted in fluorescent color image, b1、b2、b3
Value is respectively image space sitef (x,y) Lower G Quantile Regression coefficient, G1、G2、G3For the green component extracted in fluorescent color image
Value, c1、c2、c3Value is image space sitef (x,y) Lower B component regression coefficient, B1、B2、B3For the indigo plant extracted in fluorescent color image
Colouring component value, d are the correction value of mathematical regression model；
S6. optimized mathematical model:
The performance for the zearalenone mathematical model that evaluation is established is verified with independent sample collection and is advanced optimized；
S7. zearalenone content in grain sample to be measured is measured:
By grain sample to be measured, after the processing of step S2, S3, R, G and B color point of three sequence fluorescent color images is extracted
Amount, the zearalenone mathematical model established using step S6 optimization, calculates the Gibberella zeae in each space site of color image
Ketenes content, with zearalenone content at each space site in pseudo color image expression grain sample.
8. quick screening method according to claim 7, which is characterized in that in step S2, the time for exposure is 20ms,
The area-of-interest size diameter is 8cm, and the field angle is 24 °, and the linkage trigger signal is 1-1-1, the object distance
For 55cm.
9. quick screening method according to claim 7, which is characterized in that in step S5, when the mathematical model is established
It is modeled using point sample, tile when test sample sample batch sample space forecast of distribution.
10. quick screening method according to claim 7, which is characterized in that in step S7, the pseudo color image expression
The pollution level of zearalenone in grain sample is indicated by 64 color linear transitions of green to red setting.
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