CN109001150A - A kind of method and apparatus for realizing that bacterium colony is counted based on near-infrared image technology - Google Patents
A kind of method and apparatus for realizing that bacterium colony is counted based on near-infrared image technology Download PDFInfo
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Abstract
The present invention discloses a kind of method and apparatus for realizing that bacterium colony is counted based on near-infrared image technology, is related to calculating, calculate, counting technology field.The method and system that a set of realization bacterium colony counts is constructed using near-infrared image technology, the Colony hybridization under near-infrared light source is obtained by near infrared camera, identification and statistics are carried out to the bacterium colony obtained in image using specific Computer Image Processing and bacterium colony analysis of accounts method, and provide bacterium colony count results.The present invention has the advantages that be influenced that small, Colony hybridization is clear, counting accuracy is high, fireballing by extraneous illumination condition.
Description
Technical field
The invention belongs to the calculating in information technology, reckoning, counting technology fields, in particular to a kind of to be based on near-infrared figure
As technology realizes the method and apparatus that bacterium colony counts.
Background technique
Bacterium colony count be agricultural, food, medical and health analysis in progress quality testing a basic and important job.
It is an important quality index for evaluating the contaminated degree of water that bacterium colony in environment protection field, water, which counts,；In field of food safety,
Total plate count detection in food is the important means to ensure food safety；In medicine and hygiene fields, bacterium colony counts even more must not
The element task that can lack.Ordinary nutrient agar pour plate method (abbreviation colony counting method) is generallyd use to bacterium colony both at home and abroad
It is counted.This method has certain subjectivity by artificial observation counting by artificial counting, time-consuming, low efficiency
Property, cause error big, reproducibility is low.
In recent years, some researchers are it is also proposed that the method for quickly detecting total plate count based on image processing techniques, but
It is that acquired image substantially obtains under visible light.The Colony hybridization obtained under visible light is illuminated by the light, position, vision etc.
The influence of many factors, especially culture dish be easy to it is reflective so that shooting Colony hybridization it is unintelligible, bacterium colony identification mistake
Difference is larger.Develop it is a kind of influenced by external condition small, Colony hybridization is clear, the high method for counting colonies of bacterium colony counting accuracy and
Device is for accurately obtaining bacterium colony statistical result, the efficiency for improving bacterium colony counting researcher, the labor of reduction researcher
Fatigue resistance has great importance.
Summary of the invention
The present invention provides a kind of method and apparatus for realizing that bacterium colony is counted based on near-infrared image technology, the technology to be solved
Problem is: how to obtain clearly Colony hybridization and improves the accuracy and processing speed of bacterium colony counting.
To solve the above-mentioned problems, the present invention provides following technical scheme:
A method of realizing that bacterium colony counts based on near-infrared image technology comprising the steps of:
1) near-infrared light source of circular ring shape 2 is fixed on pedestal 5, culture dish 6 is placed in the circle ring center of near-infrared light source 2
5 plane of pedestal on；The surface of pedestal 5 uses weak reflectorized material, and color is arranged according to the color of bacterium colony, and the principle of setting is
The two color difference is big.
2) camera lens of near infrared camera 1 is mounted in the top center hole of camera bellows 7, makes the camera lens, close of near infrared camera 1
Infrared light supply 2 and culture dish 6 are coaxial on plumb line, and near infrared camera 1 is upper, and near-infrared light source 2 and culture dish 6 are under.
3) focal length for adjusting near infrared camera 1 makes then to clap in the visual field of near infrared camera 1 comprising complete culture dish 6
Take the photograph the photo of culture dish 6.
4) image of the culture dish 6 taken near infrared camera 1 is transferred by computer 4, and completes to scheme by software
As processing and bacterium colony analysis of accounts, and export bacterium colony count results.
A method of realizing that bacterium colony counts based on near-infrared image technology, the image procossing and bacterium colony counting include
Following steps:
1) greyscale image transitions for using thresholding method to get near infrared camera 1 for bianry image, to bianry image into
Row holes filling carries out connected component labeling to the filled image of hole, is removed outside culture dish 6 according to the area of connected region
The noise in portion.
2) using the portion other than 6 inner circle of culture dish in connected component labeling method and area attribute metric function removal image
Point, the bacterium colony within 6 inner circle of reservation culture dish.
3) the original of bacterium colony that be connected with 6 inner circle of culture dish is obtained with operation using closed operation, image subtraction operation and image
Image.Using connected component labeling method, area attribute metric function, nonlinear gray transforming function transformation function, range conversion method, watershed
Algorithm and thresholding method are partitioned into adhesion and overlapping within 6 inner circle of the bacterium colony being connected with 6 inner circle of culture dish and culture dish
Bacterium colony；The nonlinear gray transforming function transformation function is using the imadjust function in MATLAB, the range conversion method
It is using the bwdist function in MATLAB.
4) using connected component labeling method and area attribute metric function to all bacterium colonies inside the culture dish 6 identified
It is counted, and provides statistical result.
A kind of bacterium colony counting device based on near-infrared imaging technology includes near infrared camera 1, near-infrared light source 2, light source
Power controller 3, computer 4, pedestal 5, culture dish 6, camera bellows 7；Near-infrared light source 2 is installed on the pedestal 5, it is close red
Outer light source 2 is circular ring shape；Culture dish 6 is mounted in 5 plane of pedestal of the circle ring center of near-infrared light source 2；The surface of pedestal 5
Using weak reflectorized material, both color is arranged according to the color of bacterium colony, and the principle of setting is color difference is big；It is installed on pedestal 5
Camera bellows 7 constitutes a closed space, culture dish 6 and near-infrared light source 2 in the bottom centre of camera bellows 7；By near infrared camera 1
Camera lens be mounted in the top center hole of camera bellows 7 downward, make the camera lens, near-infrared light source 2 and culture dish 6 of near infrared camera 1
It is coaxial on plumb line；Near infrared camera 1 is upper, and near-infrared light source 2 and culture dish 6 are under；Light source power controller 3 with it is close red
Outer light source 2 connects；Computer 4 is connected by data line or WIFI near infrared camera 1.
A kind of bacterium colony counting device based on near-infrared imaging technology, the photosensitive wavelength range of the near infrared camera 1
Including but not limited near infrared band.
A kind of bacterium colony counting device based on near-infrared imaging technology, the culture dish 6 are disposable plastic culture dish
6；The pedestal 5 uses opaque material.
Bacterium colony Counting software inside self-editing culture dish is installed in the computer 4；Bacterium colony counts inside culture dish
It include to open camera to obtain image, determine the specified image of acquisition image, load, start counting, count and mark in original in software
On figure, empty interface, bacterium colony number shows 7 modules.
Working principle:
A kind of method and apparatus for realizing that bacterium colony is counted based on near-infrared image technology, the course of work includes the following: nearly red
Outer ring light source 2 is fixed on pedestal 5, and culture dish 6 is placed in the upper surface of pedestal 5, the center of circular ring shape near-infrared light source 2；It will
Near infrared camera 1 is mounted on the top of camera bellows 7；The focal length for adjusting camera 1, makes culture dish 6 within the visual field, shoots culture dish 6
Photo；The near-infrared image of the culture dish 6 taken is transferred by computer 4, is based on connected component labeling method, area attribute
The methods of metric function, nonlinear gray transforming function transformation function, range conversion method, watershed algorithm, thresholding method are successively realized: being gone
Fall the noise outside culture dish 6, removes the part in image other than 6 inner circle of culture dish, be partitioned into respectively and 6 inner circle phase of culture dish
Bacterium colony even, the bacterium colony of adhesion and overlapping within 6 inner circle of culture dish, to all bacterium colonies inside the culture dish 6 identified into
Row counts and exports bacterium colony count results.
It has the beneficial effect that:
1) a kind of method for realizing that bacterium colony is counted based on near-infrared image technology of design, is influenced small, bacterium by extraneous illumination condition
The accuracy rate for falling counting is high, average relative error 0.8%；It is 2.5GHz for cpu frequency, inside saves as the computer of 8GB, locates
Managing a width culture dish diameter is 90mm, and the time of image of the clump count between 30 ~ 300 is 7 ~ 10s, improves bacterium colony counting
The working efficiency of researcher.
2) light source acquired using near infrared light as Colony hybridization, is increased the contrast of bacterium colony and background, improved
The acquisition quality of Colony hybridization ensure that the accuracy that bacterium colony counts.
Appended drawing reference:
1. 5. pedestal of near infrared camera 2. near-infrared light source, 3. light source power controller, 4. computer, 6. culture dish 7. is dark
Case
Detailed description of the invention:
Attached drawing 1: bacterium colony counting device composition schematic diagram
Attached drawing 2: method for counting colonies flow chart
Attached drawing 3: the culture dish original image containing bacterium colony
Attached drawing 4: the noise outside removal culture dish
Attached drawing 5: the process of image other than removal culture dish inner circle
Attached drawing 6: the bacterium colony being connected with culture dish inner circle is split
Attached drawing 7: the bacterium colony of adhesion and overlapping is split
Attached drawing 8: indicia framing selects the bacterium colony identified
Attached drawing 9: count results are exported on bacterium colony Counting software interface.
Specific embodiment:
Methods and apparatus of the present invention is described in further detail with reference to the accompanying drawings and detailed description.
It is a kind of based on near-infrared image technology realize bacterium colony count method and apparatus include: near infrared camera 1, circular ring shape
Near-infrared light source 2, light source power controller 3, computer 4, pedestal 5, culture dish 6, camera bellows 7.
A kind of course of work of method and apparatus for realizing bacterium colony counting based on near-infrared image technology is as follows:
A method of realizing that bacterium colony counts based on near-infrared image technology comprising the steps of:
1) as shown in Figure 1, the near-infrared light source 2 of circular ring shape is fixed on pedestal 5, culture dish 6 is placed in near-infrared light source 2
Circle ring center 5 plane of pedestal on；The surface of pedestal 5 uses weak reflectorized material, and color is arranged according to the color of bacterium colony, if
The principle set is that the two color difference is big.Furthermore color (such as black, dark blue, blue, brown, the red, purple of different pedestals 5 are analyzed
Color) influence to bacterium colony statistical result, as a result illustrate, when the color of the color of pedestal 5 and bacterium colony has obvious difference,
This method can identify bacterium colony well, this to identify the bacterium colony under different colours culture medium, illustrate the dress
Setting has good universality with method.
2) camera lens of near infrared camera 1 is mounted in the top center hole of camera bellows 7, makes the camera lens, close of near infrared camera 1
Infrared light supply 2 and culture dish 6 are coaxial on plumb line, and near infrared camera 1 is upper, and near-infrared light source 2 and culture dish 6 are under, such as
Shown in Fig. 1；
3) focal length for adjusting near infrared camera 1 makes in the visual field of near infrared camera 1 comprising complete culture dish 6, then shooting training
The photo of ware 6 is supported, as shown in Figure 3；
4) image of the culture dish 6 taken near infrared camera 1 is transferred by computer 4, and is completed at image by software
Reason and bacterium colony analysis of accounts, and count results are exported, as shown in Figure 9.
A kind of method for realizing that bacterium colony is counted based on near-infrared image technology, as shown in Fig. 2, at the image
Reason and bacterium colony analysis of accounts comprise the steps of:
1) greyscale image transitions for using thresholding method to get near infrared camera 1 is bianry images, as a result such as Fig. 4 a institute
Show.Holes filling is carried out to bianry image, as a result as shown in Figure 4 b.Connected component labeling is carried out to the filled image of hole,
Remove the noise outside culture dish according to the area of connected region, retains the area that connected region area is greater than 360000 pixels
Domain carries out image to Fig. 4 c and negates operation, as shown in figure 4d, and subtract Fig. 4 d with Fig. 4 a, then obtain Fig. 4 e as illustrated in fig. 4 c,
The near-infrared image and noise outside culture dish 6 are eliminated at this time.
2) using the portion other than 6 inner circle of culture dish in connected component labeling method and area attribute metric function removal image
Point, the bacterium colony within 6 inner circle of reservation culture dish: first removing the region other than 6 inner circle of culture dish, carries out connected region to Fig. 4 e
Label, and using area attribute metric function retain the region that connected region area is greater than 50000 pixels, as shown in Figure 5 a.
The noise (Fig. 5 b) inside culture dish 6 is rejected according to the area of connected region, image is carried out to Fig. 5 b and negates operation, and subtracts figure
4d obtains Fig. 5 c；It is greater than the region of 250000 pixels using connected component labeling and Retention area to Fig. 5 c, extracts culture
The inner circle region (Fig. 5 d) of ware 6.To Fig. 5 d and Fig. 3 progress and operation, as a result as depicted in fig. 5e.At this point, only being cultivated in image
Bacterium colony within 6 inner circle of ware.
3) the original of bacterium colony that be connected with 6 inner circle of culture dish is obtained with operation using closed operation, image subtraction operation and image
Image.Using connected component labeling method, area attribute metric function, nonlinear gray transforming function transformation function, range conversion method, watershed
Algorithm and thresholding method are partitioned into adhesion and overlapping within 6 inner circle of the bacterium colony being connected with 6 inner circle of culture dish and culture dish
Bacterium colony.The nonlinear gray transforming function transformation function is using the imadjust function in MATLAB, the range conversion method
It is using the bwdist function in MATLAB.It first extracts the bacterium colony that is connected with 6 inner circle of culture dish: closed operation being carried out to Fig. 5 d first,
Smoothly, as a result as shown in Figure 6 a inner circle is carried out.Subtraction is carried out to Fig. 6 a and Fig. 5 d, acquisition is connected with 6 inner circle of culture dish
The mask image of bacterium colony, as shown in Figure 6 b.Fig. 6 b and Fig. 3 progress are extracted to obtain the bacterium colony that is connected with 6 inner circle of culture dish with operation
Original image, as fig. 6 c.The enhancing of nonlinear gray changing image is carried out to Fig. 6 c, then carries out Threshold segmentation and obtains and train
The bianry image of the connected bacterium colony of 6 inner circle of ware is supported, as shown in fig 6d.To Fig. 5 e progress nonlinear gray changing image enhancing, then into
Row threshold division, obtain with the disjunct bacterium colony of 6 inner circle of culture dish, as shown in Figure 7a.Secondly the bacterium inside culture dish 6 is extracted
It falls: Fig. 7 a is separated adhesion and the bacterium colony to overlap using range conversion method, watershed algorithm etc., as a result such as Fig. 7 b
It is shown.Finally, counted using connected component labeling method and area of the area attribute metric function to connected region in Fig. 7 b,
The bacterium colony in Fig. 5 e is divided into petite (Fig. 7 c) and macrocolony (figure according to whether connected region area is greater than 100 pixels
7d).The gray value of petite in statistical chart 7c carries out two-value to gray level image Fig. 7 d based on gray value setting segmentation threshold
Change, will be separated with macrocolony adhesion or overlapping petite, as a result as shown in figure 7e.
4) using connected component labeling method and area attribute metric function to all bacterium inside the culture dish 6 identified
Capable statistics is dropped into, the bacterium colony in Fig. 6 d, Fig. 7 b and Fig. 7 e is counted, circle is selected and marked and identifies in different colors
Bacterium colony, as shown in Figure 8.Count results are shown in software interface, as a result as shown in Figure 9.
The photosensitive wavelength range of the near infrared camera 1 is including but not limited near infrared band.It is described in the present embodiment
Near infrared camera 1 using PointGrey company of Canada produce model BFLY-120 PGE-13E4M-CS camera,
The sensitive volume of the camera is 300-1100nm；The near-infrared light source 2 is raw using thousand Electro-optical Technology, INC. (US) 62 Martin Road, Concord, Massachusetts 017 of Shanghai side
The circular ring shape near-infrared light source of the model VR180-75-IR of production, the wavelength of the light source are 860nm.
As can be seen from the above embodiments, the present invention devises a kind of bacterium colony counting side based on near-infrared image technology
Method, influenced by extraneous illumination condition it is small, bacterium colony count accuracy rate it is higher, using this method to 30 width bacterium colonies 30 ~ 300 it
Between culture dish 6 in bacterium colony counted, and be compared, as a result illustrate, this method is to bacterium colony with the result manually counted
The average relative error of total statistical result is 0.8%；It is 2.5GHz for cpu frequency, inside saves as the computer of 8GB, processing one
Width culture dish diameter is 90mm, and the time of image of the clump count between 30 ~ 300 is 7 ~ 10s, is the 18 of artificial statistical time
~ 25%, improve the working efficiency that bacterium colony counts researcher.
A kind of bacterium colony counting device based on near-infrared image technology of the invention, as shown in Figure 1, including near infrared camera
1, near-infrared light source 2, light source power controller 3, computer 4, pedestal 5, culture dish 6, camera bellows 7.It is installed on the pedestal 5
Near-infrared light source 2, near-infrared light source 2 are circular ring shape；Culture dish 6 is mounted on to the pedestal 5 of the circle ring center of near-infrared light source 2
In plane；Both the surface of pedestal 5 uses weak reflectorized material, and color is arranged according to the color of bacterium colony, and the principle of setting is color
It is poor big；Camera bellows 7 is installed on pedestal 5, constitutes a closed space, culture dish 6 and near-infrared light source 2 are in the bottom of camera bellows 7
Center；The camera lens of near infrared camera 1 is mounted on downward in the top center hole of camera bellows 7, the camera lens, close of near infrared camera 1 is made
Infrared light supply 2 and culture dish 6 are coaxial on plumb line；Near infrared camera 1 is upper, and near-infrared light source 2 and culture dish 6 are under；Light
Source power controller 3 is connect with near-infrared light source 2；Computer 4 is connected by data line or WIFI near infrared camera 1.It is described
It is a kind of based on near-infrared image technology realize bacterium colony count device, the culture dish 6 be disposable plastic culture dish 6；
The pedestal 5 uses opaque material.
As shown in figure 9, being equipped with bacterium colony Counting software inside self-editing culture dish in the computer 4；In culture dish
It include to open camera to obtain image, determine the specified image of acquisition image, load, start counting, count in portion's bacterium colony Counting software
And mark in original image, empty interface, bacterium colony number shows 7 modules.
As can be seen from the above embodiments, the light source that the present invention uses near infrared light to acquire as Colony hybridization, increases
The contrast of bacterium colony and background improves the acquisition quality of Colony hybridization, ensure that the accuracy that bacterium colony counts.
As described above, the plan content in conjunction with given by attached drawing, embodiment of the method and Installation practice, can derive class
As technical solution.But all contents without departing from the technical program, according to the technical essence of the invention to above embodiments institute
Any simple modification, equivalent variations and the modification made, all of which are still within the scope of the technical scheme of the invention.
Claims (6)
1. a kind of method for realizing that bacterium colony is counted based on near-infrared image technology, which is characterized in that comprise the steps of:
1) near-infrared light source of circular ring shape (2) is fixed on pedestal (5), culture dish (6) is placed in near-infrared light source (2)
On the pedestal (5) of center portion position；The surface of pedestal (5) is weak reflectorized material, and color is arranged according to the color of bacterium colony, the original of setting
It is then that the two color difference is big；
2) camera lens of near infrared camera (1) is mounted in the top center hole of camera bellows (7), make near infrared camera (1) camera lens,
Near-infrared light source (2) and culture dish (6) are coaxial on plumb line, and near infrared camera (1) is in upper, near-infrared light source (2) and culture
Ware (6) is under；
3) comprising complete culture dish (6) in the visual field of near infrared camera (1), then the focal length for adjusting near infrared camera (1), makes
Shoot the photo of culture dish (6)；
4) image that the culture dish (6) taken near infrared camera (1) is transferred by computer (4), passes through self-editing culture
Bacterium colony Counting software completes image procossing and bacterium colony analysis of accounts inside ware, finally exports bacterium colony count results.
2. a kind of method for realizing that bacterium colony is counted based on near-infrared image technology according to claim 1, which is characterized in that
The image procossing and bacterium colony analysis of accounts comprises the steps of:
1) greyscale image transitions for using thresholding method to get near infrared camera (1) is bianry images, to bianry image
Holes filling is carried out, connected component labeling is carried out to the filled image of hole, culture dish is removed according to the area of connected region
(6) external noise；
2) part in image other than culture dish (6) inner circle is removed using connected component labeling method and area attribute metric function,
Bacterium colony within reservation culture dish (6) inner circle；
3) it is obtained being connected with culture dish (6) inner circle the original graph of bacterium colony using closed operation, image subtraction operation and image and operation
Picture；
Using connected component labeling method, area attribute metric function, nonlinear gray transforming function transformation function, range conversion method, watershed
Algorithm and thresholding method be partitioned into adhesion within the bacterium colony being connected with culture dish (6) inner circle and culture dish (6) inner circle and
The bacterium colony of overlapping；
4) using connected component labeling method and area attribute the metric function all bacterium colonies internal to the culture dish (6) identified into
Row statistics, and provide count results.
3. it is a kind of based on near-infrared image technology realize bacterium colony count device, which is characterized in that comprising near infrared camera (1),
Near-infrared light source (2), light source power controller (3), computer (4), pedestal (5), culture dish (6), camera bellows (7)；Described
Near-infrared light source (2) are installed on pedestal (5), near-infrared light source (2) is circular ring shape；Culture dish (6) is mounted on near-infrared light source
(2) in pedestal (5) plane of circle ring center；The surface of pedestal (5) uses weak reflectorized material, and color is according to the color of bacterium colony
Setting, the principle of setting are that the two color difference is big；Camera bellows (7) are installed on pedestal (5), constitute a closed space, culture dish
(6) and near-infrared light source (2) camera bellows (7) bottom centre；The camera lens of near infrared camera (1) is mounted on camera bellows (7) downward
Top center hole in, keep the camera lens, near-infrared light source (2) and culture dish (6) of near infrared camera (1) coaxial on plumb line；
Near infrared camera (1) is upper, and near-infrared light source (2) and culture dish (6) are under；Light source power controller (3) and near-infrared light source
(2) it connects；Computer (4) is connected by data line or WIFI near infrared camera (1).
4. a kind of device for realizing that bacterium colony is counted based on near-infrared image technology according to claim 3, described is close red
The photosensitive wavelength range of outer camera (1) is including but not limited near infrared band.
5. a kind of device for realizing that bacterium colony is counted based on near-infrared image technology according to claim 3, which is characterized in that
The culture dish (6) is disposable plastic culture dish (6)；The pedestal (5) uses opaque material.
6. a kind of device for realizing that bacterium colony is counted based on near-infrared image technology according to claim 3, which is characterized in that
Bacterium colony Counting software inside self-editing culture dish is installed in the computer (4)；Inside culture dish in bacterium colony Counting software
Comprising opening camera acquisition image, determining acquisition image, the specified image of load, starting counting, count and mark in original image, clearly
Empty interface, bacterium colony number show 7 modules.
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