JPH0787997A - Recognition of the area inhibiting microorganism to grow there and device therefor - Google Patents

Abstract

PURPOSE:To enable the recognition of the areas in which the growth of the microorganism is inhibited by extracting the boundaries, even when they are photographed in spots in lowered cell body concentrations. CONSTITUTION:In a method for recognizing the areas in which the growth of the microorganism is inhibited by utilizing the difference in optical properties between the growth area and the inhibition area, the areas for recognition are photographed by the camera 23 and the images are mutually treated with the image processor 21 to extract the boundaries between the areas where the growth is inhibited and those where the microorganism has grown and these areas are recognized based on the boundaries.

Description

Detailed Description of the Invention

[0001]

[Industrial field of the present invention] When measuring the efficacy of antibiotics and the like using an agar medium, the bacteria or spores (hereinafter referred to as "bacteria") cannot be killed or grown due to the influence of antibiotics or the like. And an apparatus for recognizing a region that has been grown and a region where bacteria have grown without being influenced by antibiotics, and more specifically, it recognizes the region based on the boundary extracted by image processing. is there.

[0002]

2. Description of the Related Art In order to confirm the quality and efficacy of antibiotics, a cylinder is erected (or pierced) on an agar medium to dispense the antibiotics, and the bacteria growth-inhibiting area where bacteria or the like cannot die or grow ( The determination of the titer by forming a blocking circle) and measuring the diameter of the blocking circle is under the “Japanese Antibiotic Drug Standard”. For example, in the cylindrical plate method in this titer test, as shown in FIG. 8, a base layer agar medium as a nutrient layer is dispensed into a Petri dish 1 having an inner diameter of about 90 mm, and the agar is spread evenly to form a base layer agar plate. Three
To form. Then, an appropriately prepared seed layer agar medium is dispensed onto the base layer agar plate 3 and spread evenly on the surface of the base layer agar plate 3 to form a seed layer agar plate 5. Then, four cylinders 7 are placed on a Petri dish flat plate so that each adjacent one has an interval of 90 ° with respect to the center, and the cylinders 7 are filled with a diluting solution 9 of chemicals having different concentrations. And
After culturing for a predetermined time while keeping it in a certain temperature range,
The extent to which the diluting liquid 9 of the chemical that has filled the cylinder 7 has prevented the growth of bacteria and the like is observed.

A region where bacteria or the like cannot be killed or grown due to the influence of antibiotics or the like forms an inhibition circle which is concentric with the cylinder 7 and has different turbidity. Since a predetermined relationship is established between the titer of the diluting liquid 9 in the cylinder 7 and the diameter of the blocking circle, the titer of the diluting liquid 9 can be obtained by measuring the diameter of the blocking circle. In measuring the diameter of the blocking circle, conventionally, in addition to visual measurement using a caliper, a method of photographing the blocking circle and measuring it by image processing has been adopted. Then, in the case of this image processing, it is general that simple binarization processing is performed. That is, by illuminating a Petri dish flat plate with dark field transmissive illumination, photographing it with a black and white CCD camera, and simply binarizing the luminance value of each pixel of the photographed image, there is no influence of antibiotics or the like. It was determined whether the area was where the bacteria or the like grew, or where the bacteria or the like could not be killed or grown due to the diffusion of the antibiotics or the like.

[0004]

However, under conditions such as a low concentration of bacteria in the seed layer agar medium, the grown bacteria have the same optical properties as those of the region that was killed or could not grow. Areas remained, and these areas were photographed as spots. For this reason, in these regions, only the simple binarization process of binarizing the luminance value of each pixel causes the binarized values to be scattered irregularly, and the boundaries of the regions can be extracted. In the end, the measurement had to be done visually by using a caliper. The present invention has been made in view of the above situation, and even when a region is photographed in spots under a condition in which the concentration of bacteria is low, it is possible to distinguish the region by extracting its boundary. It is an object of the present invention to provide a method of recognizing a growth inhibition area and an apparatus thereof, thereby improving recognition ability in image processing.

[0005]

A method for recognizing a growth inhibition region of a bacterium or the like according to the present invention for achieving the above object is a region in which a bacterium or spore cannot be killed or grown, and a bacterium or spore has grown. A method of recognizing a growth inhibition region of fungi or spores from a captured image of this region by utilizing the difference in optical properties from the region, by capturing the region of recognition target and correlating this captured image. It is characterized in that the boundaries between the regions where the bacteria or spores cannot be killed or grown and the regions where the bacteria or spores have grown are extracted, and these regions are recognized based on these boundaries. Further, the recognition method is that the pixel information of the captured image is subjected to differential absolute value processing, the differential absolute value processed pixel information is binarized, the binarized pixel information is expanded, and the expanded It is preferable that the processed pixel information is subjected to contraction processing, and the region is measured based on a boundary extracted by the contracted pixel information. The recognition device for the growth inhibition region of bacteria and the like according to the present invention is a photographing device, image differential absolute value processing means for performing differential absolute value processing on pixel information of a photographed image photographed by this photographing device, and differential absolute value. Image binarization processing means for binarizing information, image expansion processing means for expanding binary information, image contraction processing means for contracting expansion information, and a boundary between regions obtained from the contraction information. And an image measuring unit for measuring the dimensions thereof.

[0006]

According to the recognition method of the present invention, basically, the photographed images of the region to be recognized are subjected to the correlation processing so that the region in which the bacterium or spore could not be killed or grown and the bacterium or spore grew. Boundaries with the regions are extracted, the dimensions of these regions are measured based on the boundaries, and the regions can be recognized. More specifically, when the captured image is subjected to the image differential absolute value processing, the luminance value of the pixel in the region where the bacteria or the like could not be killed or grown becomes a uniform small value, and this pixel information is binarized into the image. As a result of the processing, only a large value portion is subjected to extraction processing to represent a speckled area. Next, the binarized information is expanded, whereby adjacent spot areas are combined to obtain expanded information that can be regarded as an area in which bacteria and the like are extracted. In this expansion information, the boundary with the area that could not be killed or grown is also expanded, so image contraction processing obtains contraction information that returns the boundary to the state before expansion processing, and as a result, the boundary of the area It will be extracted and the dimensions will be measured based on this boundary and the area will be recognized. Further, in the device for recognizing the growth inhibition area of bacteria, etc., photographing, image differential absolute value processing,
The boundary is extracted by the image binarization process, the image expansion process, and the image contraction process, and the region is recognized by the image measuring means based on the boundary.

[0007]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A preferred embodiment of a method for recognizing a growth inhibition region for bacteria and the like and an apparatus therefor according to the present invention will be described in detail below with reference to the drawings. FIG. 1 is a block diagram showing the outline of the configuration of the recognition apparatus of the present invention. A photographing device (black and white CCD camera or the like) 23 is connected to the image preprocessing means 21, and the photographing device 23
Uses the Petri dish 25 arranged below as a subject. The petri dish 25 has, for example, an inner diameter of 90 mm, on which a base layer agar plate and a seed layer agar plate are formed. The agar plate has an outer diameter of 8.0 mm, an inner diameter of 6.0 mm, and a height of 10.0 m.
A cylinder of m is erected, and a diluted antibiotic is dispensed in the cylinder (see FIG. 8). Then, the agar plate is cultured in a state of being kept in a constant temperature range, and is in a state in which an inhibition circle concentric with the cylinder is formed. An illumination device 27 is provided below the petri dish 25, and the illumination device 27 illuminates the petri dish 25 with dark field transmission illumination.
Therefore, the image taken from above by the imaging device 23 is
Areas where bacteria and the like cannot be killed or grown will appear uniformly dark.

The image preprocessing means 21 comprises an image differential absolute value processing means 29, an image binarization processing means 31, an image expansion processing means 33 and an image contraction processing means 35.
3 is connected to the image differential absolute value processing means 29. The image differential absolute value processing means 29 performs differential absolute value processing on the information (luminance value) for each pixel of the photographed image by Laplacian conversion in the horizontal and vertical directions to obtain a differential absolute value image. An image binarization processing unit 31 is connected to the image differential absolute value processing unit 29, and the image binarization processing unit 31 outputs the differential absolute value information obtained by the image differential absolute value processing unit 29 to 2
By performing the binarization process, only the pixels having a large luminance value are extracted to obtain the spot-like region. Image binarization processing means 3
An image expansion processing unit 33 is connected to the image processing unit 1, and the image expansion processing unit 33 performs an expansion process on the binarized information and combines adjacent spot areas to obtain an expanded image. An image contraction processing unit 35 is connected to the image expansion processing unit 33. The image contraction processing unit 35 performs contraction processing on the expansion information, and a contracted image that restores the boundary unnecessarily expanded by the image expansion processing unit 33 to the state before expansion. I'm supposed to get it.

The image preprocessing means 21 has an image measuring means 37.
Are connected, and the image measuring means 37 can measure the dimension based on the boundary of the finally obtained area. As the image preprocessing unit 21, for example, one having a horizontal pixel number of 512, a vertical pixel number of 480, and a luminance value of 8 bits / pixel is used. The image capturing device 23, the image differential absolute value processing means 29, the image binarization processing means 31, the image expansion processing means 33, the image contraction processing means 35, and the image measuring means 37 constitute a recognition device 39 according to the present embodiment. There is.

The area recognition procedure of the recognition device 39 thus configured will be described with reference to FIGS. 2 is a flowchart showing the procedure of the recognition method, FIG. 3 is a diagram showing the brightness value of a part of the captured image, FIG. 4 is a diagram showing the brightness value of a part of the differential absolute value image, and FIG. 5 is a part of the binarized image. FIG. 6 is a diagram showing the luminance value of FIG.
FIG. 7 is a diagram showing the luminance value of a part of the contracted image. The blocking circle formed on the agar plate is black and white under dark field transmitted illumination.
When photographed by a CD camera (41 in FIG. 2), a luminance value is digitally output with a precision of 8 bits / pixel in the photographed image. Then, the area in which the bacteria and the like are not killed or cannot be grown looks dark. Therefore, as shown in FIG. 3, the luminance value of the pixel in this portion has a small numerical value.

When this image is processed by the image differential absolute value processing means 29 (43 in FIG. 2), the obtained differential absolute value image has a uniform small luminance value of the pixel in that portion. . However, since the concentration of bacteria etc. is low, in a region where bacteria etc. grow in spots, brightly visible regions are distributed in spots, so the differential absolute value image of that part is
As shown in FIG. 4, pixels having large values surround each spot. Next, the differential absolute value image is subjected to extraction processing by the image binarization processing means 31, for example, with a fixed threshold value set to 50 (45 in FIG. 2). The binarized image thus obtained (see FIG. 5) represents a spot-like region.

[0012] Between the spotted areas, the cells were not extracted because the fungus or the like did not exist at the beginning, but there was a possibility that the area could grow if the fungus or the like existed. Can be judged. Therefore, the image expansion processing means 3
By performing expansion processing once in the vicinity of 8 according to 3 (see FIG.
(47), the adjacent spot areas are combined and regarded as an area in which bacteria and the like are extracted, and an expanded image (see FIG. 6) is obtained. In this expansion processing, the boundary with the area that has not been killed or cannot be grown is also expanded. Therefore, the image contraction unit 35 performs the contraction processing once in 8 neighborhoods (49 in FIG. 2) and expands the boundary. A contracted image (see FIG. 7) returned to the previous state is obtained.
As a result, the boundary L of the area (see FIG. 7) is extracted. The area where the boundary L is extracted is the image measuring means 37.
Measures the dimension based on the boundary L (5 in FIG. 2).
1) The area will be recognized.

[0013]

As described in detail above, according to the method for recognizing the growth inhibition region of bacteria and the like according to the present invention, the photographed image is image-processed so that the bacteria or spores cannot be killed or grown. The boundaries between the regions and the regions where the fungi or spores have grown are extracted, and the dimensions of these regions are measured based on these boundaries, so when the regions are photographed in spots under conditions where the concentration of bacteria is low. Also in the above, the area can be discriminated, and as a result, the recognition ability in the image processing can be remarkably improved. According to the apparatus for recognizing the growth inhibition region of bacteria or the like according to the present invention, the boundary between the region in which the bacterium or spore cannot be killed or grown and the region in which the bacterium or spore has grown can be extracted by each image processing means. It is possible to recognize the area by measuring the size of the area based on this boundary.

[Brief description of drawings]

FIG. 1 is a block diagram showing a schematic configuration of a recognition device of the present invention.

FIG. 2 is a flowchart showing a procedure of a recognition method.

FIG. 3 is a diagram illustrating a luminance value of a part of a captured image.

FIG. 4 is a diagram showing a luminance value of a part of a differential absolute value image.

FIG. 5 is a diagram showing a luminance value of a part of a binarized image.

FIG. 6 is a diagram showing a luminance value of a part of an expanded image.

FIG. 7 is a diagram illustrating a luminance value of a part of a contracted image.

FIG. 8 is a side view illustrating a cylindrical flat plate method in a titer test.

[Explanation of symbols]

 21 image pre-processing means 23 photographing apparatus 29 image differential absolute value processing means 31 image binarization processing means 33 image expansion processing means 35 image contraction processing means 37 image measuring means 39 recognition device for a growth inhibition area of bacteria etc.

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Claims (3)

[Claims] 1. Utilizing the difference in optical properties between a region in which a bacterium or spore cannot be killed or grown and a region in which the bacterium or spore has grown, a growth-inhibited region of the bacterium or spore is identified by a photographed image of the region. A method of recognizing, wherein the region of the recognition target is photographed, and the photographed image is subjected to correlation processing so that the region in which the bacterium or spore cannot die or grow and the region in which the bacterium or spore grows A method for recognizing a growth inhibition region for bacteria and the like, which comprises extracting boundaries and recognizing these areas based on the boundaries. 2. The pixel information of the captured image is subjected to differential absolute value processing, the differential absolute value processed pixel information is binarized, and the binarized pixel information is expanded, and the expanded The method for recognizing a growth inhibition region for bacteria and the like according to claim 1, wherein the processed pixel information is contracted, and the region is measured based on a boundary extracted by the contracted pixel information. 3. A photographing device, image differential absolute value processing means for performing differential absolute value processing on pixel information of a photographed image photographed by the photographing device, and image binarization for binarizing the differential absolute value information. Processing means, image expansion processing means for expanding the binarized information, image contraction processing means for contracting the expansion information, and image measurement for measuring the dimensions based on the boundary of the region obtained from the contraction information. A device for recognizing a growth inhibition region for bacteria and the like, which comprises: