Culture container

Abstract

To provide a petri dish with alignment marks that can more accurately match a specimen image regardless of rattle between a container body and a lid. [Solution] The petri dish 1 with alignment marks comprises a container body 2 in which a culture medium for culturing microorganisms is provided, a lid body 3 that is detachably attached to the container body 2, and a mark mounting member 6 that is provided on the outside of the container body 2 and has at least two alignment marks 7a, 7b.

Description

[Technical Field]

[0001] The present invention relates to a culture vessel used for culturing microorganisms in a system for detecting microorganisms (bacteria), such as a colony counter. [Background technology]

[0002] Traditionally, culture methods have been used for testing food microbial limits and sterility tests for herbal medicines. Culture methods involve filling a 90 mm diameter Petri dish (also called a petri dish) with a medium, smearing or pouring a sample of microorganisms onto the dish, and culturing the medium at a certain temperature for a certain period of time to grow colonies (collections of microorganisms) that can be seen with the naked eye. The number of colonies is then counted to test the degree of microbial growth over time.

[0003] A method for counting the number of colonies has been proposed in which multiple specimen images taken at different times in a time series are compared, and colonies are extracted and counted based on changes in the specimen images (see Patent Documents 1 and 2 below).

[0004] That is, the invention described in Patent Document 1 compares projection images of the culture medium acquired at predetermined time intervals, extracts the change position where the color changes and the state of color change at that change position, and determines whether the corresponding change position is a colony based on the state of color change.

[0005] The invention described in Patent Document 2 detects actual colonies by determining the difference in corresponding pixels between an initial first image of a growth medium inoculated with microorganisms and a second image of the medium after a selected incubation time interval.

[0006] In both inventions, it was important to accurately match (align) the two specimen images to be compared.

[0007] In this context, a method has been proposed for matching sample images by superimposing the sample images so as to align the center position of the dish and rotating the sample image based on a rotation angle corresponding to the misalignment of a marker attached to the edge of the dish (see Patent Document 3 below).

[0008] The invention described in Patent Document 3 matches sample images by rotating one of the sample images based on the center position of a petri dish, which has alignment markers attached to its edge in advance, so that the markers on the sample image overlap each other. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] Japanese Patent Application Laid-Open No. 2011-92116 [Patent Document 2] Special Publication No. 10-510706 [Patent Document 3] Japanese Patent Publication No. 2022-154207 Summary of the Invention [Problem to be solved by the invention]

[0010] However, the invention described in Patent Document 3 relates to an analytical device that assists human environmental monitoring by detecting microbial colonies from sample images of petri dishes obtained by culturing microorganisms collected in the monitored environment.

[0011] In other words, when capturing an image of a sample in a petri dish, the container body is covered with a lid to prevent the intrusion of foreign matter into the container body where the culture medium is placed. Petri dishes usually have some play between the container body and the lid. Therefore, it is difficult for an analytical device to accurately align the alignment markers provided on the edge of the petri dish.

[0012] Furthermore, in the case of a colony counter that counts the number of smaller colonies, if the alignment marker is misaligned by just 0.1 mm, colonies smaller than 0.1 mm cannot be detected (counted).

[0013] The present invention has been made in consideration of the above, and its purpose is to provide a culture vessel that can more accurately match sample images without being affected by play between the vessel body and the lid. [Means for solving the problem]

[0014] In order to achieve the above object, an aspect of the present invention is characterized by comprising a container body in which a culture medium for culturing microorganisms is provided, a lid body that is detachably attached to the container body, and a mark mounting member that is provided on the outside of the container body and has at least two alignment marks. [Effects of the Invention]

[0015] According to the present invention, it is possible to easily improve the accuracy of matching sample images using alignment marks, thereby providing a culture vessel that can more accurately match sample images regardless of play between the vessel body and the lid. [Brief explanation of the drawings]

[0016] [Figure 1] 1 is a schematic diagram showing an example of the configuration of a colony counter that counts the number of microbial colonies from a sample image of a petri dish with alignment marks according to one embodiment of the present invention. FIG. [Figure 2] 1A and 1B show a schematic configuration of a petri dish with alignment marks according to one embodiment, in which FIG. 1A is a plan view and FIG. 1B is a front view. [Figure 3] 1A and 1B show a schematic configuration of a petri dish with alignment marks according to one embodiment, where FIG. 1A is a plan view of a lid, and FIG. 1B is a plan view of a container body. [Figure 4]10 is a flowchart shown to explain a matching process of a specimen image of a petri dish with an alignment mark according to one embodiment. [Figure 5] 10A and 10B are diagrams shown to explain the matching process of a specimen image of a petri dish with alignment marks according to one embodiment. [Figure 6] 10A and 10B show a schematic configuration of a Petri dish with alignment marks according to another embodiment of the present invention, in which FIG. 10A is a plan view and FIG. 10B is a front view. [Figure 7] This shows the general configuration of a petri dish with alignment marks according to another embodiment, where (a) is a plan view of the lid, (b) is a plan view of the container body, and (c) is a plan view of the alignment mark mounting member. DETAILED DESCRIPTION OF THE INVENTION

[0017] Hereinafter, a petri dish with an alignment mark mechanism for a colony counter (a petri dish with alignment marks) to which a culture vessel according to an embodiment of the present invention is applied will be described with reference to the drawings. Note that in this embodiment, the drawings are intended to show a schematic outline of the invention and may differ from the actual product.

[0018] One embodiment Figure 1 shows a system for detecting microorganisms (bacteria) from a sample image of a petri dish 1 with alignment marks as a culture container in one embodiment of the present invention, and here shows a colony counter 10 as an example that counts the number of colonies, which are clusters of microorganisms.

[0019] As shown in Figure 1, the colony counter 10 is composed of a stage glass 12, a photographing camera 13, an upper LED circle light 14, a lower LED circle light 15, a monitor 16, a control unit 17, a memory unit 18, and an operation unit 19.

[0020] The petri dish 1 with alignment marks for photographing a specimen image is set automatically or manually on the stage glass 12. The petri dish 1 with alignment marks is set on the stage glass 12 with the lid 3 attached.

[0021] The photographing camera 13 photographs a sample image (digital image) of the Petri dish 1 with alignment marks set on the stage glass 12. In the colony counter 10 according to this embodiment, the photographing camera 13 reads the sample image before culture and the sample image after culture (or during culture).

[0022] The upper LED circle light 14 illuminates the Petri dish 1 with alignment marks set on the stage glass 12 from above in a circular pattern with a predetermined LED light.

[0023] The lower LED circle light 15 illuminates the Petri dish 1 with alignment marks set on the stage glass 12 from below in a circular pattern with a predetermined LED light.

[0024] The monitor 16 displays an image of the specimen in the Petri dish 1 with alignment marks, which is photographed by the photographing camera 13.

[0025] The control unit 17 controls each part of the colony counter 10, and each part, such as the photographing camera 13, the upper LED circle light 14, the lower LED circle light 15, the monitor 16, the memory unit 18, and the operation unit 19, are connected to it.

[0026] The control unit 17 also includes, for example, an arithmetic processing unit (not shown), and has the function of performing software processing such as matching processing of specimen images and counting processing for counting the number of colonies, as will be described in detail later.

[0027] The storage unit 18 is a storage that stores programs (software) for arithmetic processing, specimen images before and after culture, and the like.

[0028] The operation unit 19 is operated by an operator (such as a laboratory technician) when counting the number of colonies, which are aggregations of microorganisms grown by culture.

[0029] Here, the Petri dish 1 with alignment marks used in the colony counter 10 will be specifically described.

[0030] Fig. 2(a) is a plan view showing the schematic configuration of a petri dish 1 with alignment marks according to one embodiment, Fig. 2(b) is a front view of the same, Fig. 3(a) is a plan view of the lid 3 of the petri dish 1 with alignment marks according to one embodiment, and Fig. 3(b) is a plan view of the container body 2 of the same.

[0031] This Petri dish 1 with alignment marks is, for example, a Petri dish made of transparent plastic with a diameter of approximately 90 mm, and is equipped with a container body 2, a lid body 3, and a mark mounting member 6 having at least two alignment marks 7a, 7b.

[0032] As shown in Figure 2(b), the container body 2 has a culture medium for cultivating microorganisms provided in the inner bottom portion, and a scratch prevention member (clog) 5 is provided on the bottom surface along the outer edge portion to prevent scratches on the bottom surface when placed on a stage glass 12, etc.

[0033] As shown in FIG. 3(b), for example, the container body 2 is provided with a mark attachment member 6 corresponding to at least a part of the outer periphery.

[0034] 3(a), the lid 3 is provided to prevent foreign matter (such as bacteria) from getting into the culture medium, and has some play and is provided so as to be detachable from the container body 2. The lid 3 is preferably not simply configured to cover the container body 2, but is preferably configured so as to be fixed by, for example, rotation.

[0035] The lid body 3 is provided with anti-slip members 4 along its outer edge to prevent the lid body 3 from sliding sideways when stacked.

[0036] The mark mounting member 6 is made of transparent plastic and is preferably integrally molded into the lower end portion of the container body 2 (close to the culture medium, approximately 4-5 mm from the bottom surface), as shown in Figure 2(b).

[0037] The mark mounting member 6 is a plate-like member having an approximately U-shape, as shown in Figures 2(a) and 3(b), for example, and has alignment marks 7a and 7b consisting of painted markings or through holes attached near the outer corners.

[0038] In addition, the mark attachment member 6 can be used to attach barcodes or write characters, particularly to areas that do not affect the capture of the sample image, and further has an information area (display area) 8 where identification information such as a sample number can be added as needed.

[0039] In this embodiment, the recognition areas of the alignment marks 7a and 7b can be set on the outer side of the container body 2 of the Petri dish 1 with alignment marks, which serves as the sample image. This makes it possible to avoid interference with recognition of the alignment marks 7a and 7b due to slight rattle of the lid 3 relative to the container body 2 or changes over time due to food residue or foreign matter other than bacteria (microorganisms) inside the container body 2, thereby enabling more accurate image matching.

[0040] Next, with reference to FIGS. 4 and 5, a processing flow (sequence) when the colony counter 10 counts the number of colonies from the specimen image on the Petri dish 1 with alignment marks will be described.

[0041] That is, when matching the alignment marks 7a and 7b of the specimen image before and after cultivation in the petri dish 1 with the alignment marks and counting the number of colonies that have grown over time, the operator first takes a photograph of the specimen image before cultivation (before the change over time) in the petri dish 1 with the alignment marks in the colony counter 10 (step S01 in Figure 4).

[0042] At this time, the operator fills the container body 2 of the Petri dish 1 with a culture medium, and smears or pours a microorganism sample onto the culture medium. The Petri dish 1 is then set on the stage glass 12 of the colony counter 10 with the lid 3 attached. Note that the setting of the Petri dish 1 with alignment marks onto the stage glass 12 can also be automated by a robot arm (not shown) or the like.

[0043] In this state, when the operator operates the operating unit 19, the control unit 17 turns on the upper LED circle light 14 and the lower LED circle light 15, and the photographing camera 13 captures an image SP of the specimen before cultivation in the petri dish 1 with alignment marks (Figure 5(a)).

[0044] The image SP of the specimen before culture on the Petri dish 1 with alignment marks, photographed by the photographing camera 13, is taken into the storage unit 18 via the control unit 17.

[0045] Thereafter, the Petri dish 1 with the alignment marks is placed in an incubator (not shown) and cultured under predetermined conditions (constant temperature and period). After a preselected incubation time, the Petri dish 1 with the alignment marks is moved from the incubator onto the stage glass 12 of the colony counter 10. This allows the operator to capture a specimen image AP of the Petri dish 1 with the alignment marks after culture (after changes over time) in the colony counter 10 (step S02 in FIG. 4).

[0046] In other words, when the operator operates the operation unit 19, the control unit 17 turns on the upper LED circle light 14 and the lower LED circle light 15, and the photographing camera 13 captures an image AP of the specimen after cultivation in the petri dish 1 with alignment marks (Figure 5(b)).

[0047] The specimen image AP after culture on the Petri dish 1 with alignment marks, photographed by the photographing camera 13, is taken into the storage unit 18 via the control unit 17.

[0048] Here, the position and orientation (angle) of the specimen images SP and AP before and after cultivation change each time depending on the state of the Petri dish 1 with alignment marks set on the stage glass 12. In other words, because the Petri dish 1 with alignment marks is circular, it is difficult to set it in the same position and with the same orientation with good reproducibility every time, regardless of whether it is done manually or by a robot arm.

[0049] Therefore, image matching is performed to correct the deviation in position and orientation of the specimen images SP and AP before and after culture.

[0050] That is, in the colony counter 10, a matching process is performed between the pre-culture specimen image SP and the post-culture specimen image AP (step S03 in FIG. 4). In this matching process, the post-culture specimen image AP is aligned with the pre-culture specimen image SP using alignment marks 7a and 7b, as shown in FIG. 5(c), for example.

[0051] In this embodiment, for example, the control unit 17 operates in accordance with a calculation processing program stored in the memory unit 18 to align one alignment mark 7a of the post-culture specimen image AP with one alignment mark 7a of the pre-culture specimen image SP. Then, using the one alignment mark 7a as the starting point, a rotation angle is calculated for aligning the other alignment mark 7b of the post-culture specimen image AP with the other alignment mark 7b of the pre-culture specimen image SP.

[0052] If the minimum diameter of the colonies counted by the colony counter 10 is about 0.1 mm, the post-culture specimen image AP and the pre-culture specimen image SP must be matched with an accuracy of 0.1 mm or less.

[0053] After this, on the screen of the monitor 16, the pre-culture specimen image SP and the post-culture specimen image AP are brought into a state in which both alignment marks 7a are superimposed, and from that state, the post-culture specimen image AP is rotated according to the determined rotation angle, for example as shown in Figure 5(d), so that both alignment marks 7b are superimposed.

[0054] The images before and after culture, which have been matched in this way, are then subjected to a predetermined inter-image calculation process (step S04 in FIG. 4) and a residual image extraction process (step S05 in FIG. 4) by the control unit 17. This allows for the cutting out of unnecessary portions of the image, as shown in FIG. 5(e), and the extraction of residual images of portions that have changed over time, as shown in FIG. 5(f).

[0055] Next, the residual image of the portion that has changed over time is subjected to binarization processing (step S06 in FIG. 4) and noise removal processing (step S07 in FIG. 4) by the control unit 17. As a result, as the noise component is reduced by eliminating the misalignment between the specimen images SP and AP, even minute residual images can be faithfully detected as colonies CC without being removed as noise.

[0056] Finally, the control unit 17 counts the number of remaining images of the portion detected as the colony CC (step S08 in FIG. 4), thereby enabling the number of colonies to be counted more accurately.

[0057] As described above, according to this embodiment, it is possible to easily improve the accuracy of matching the sample images SP and AP using the alignment marks 7a and 7b, and it is possible to more accurately match the sample images SP and AP regardless of any play between the container body 2 and the lid body 3.

[0058] That is, the Petri dish 1 with alignment marks is provided with a mark attachment member 6 on which at least alignment marks 7a, 7b are provided in correspondence with a part of the outer periphery of the container body 2.

[0059] As a result, the alignment marks 7a and 7b are less susceptible to the rattle of the lid body 3 with the container body 2 or changes over time, and in the case of the colony counter 10, the misalignment between the specimen images SP and AP in the matching images of the specimen image SP before culture and the specimen image AP after culture can be almost eliminated.

[0060] Therefore, it is possible to reduce noise components in the matching image, and even when extremely fine noise components are removed, it is possible to prevent minute colonies from being mistakenly removed, thereby enabling more accurate counting of the number of colonies that change over time.

[0061] In the above-described embodiment, the alignment mark-equipped petri dish 1 is described as having a mark mounting member 6 with alignment marks 7a, 7b integrally molded with the container body 2, but of course this is not limited to this, and other embodiments will be described below.

[0062] Other embodiments Fig. 6(a) is a plan view showing the schematic configuration of a Petri dish 1 with alignment marks according to another embodiment of the present invention, and Fig. 6(b) is a front view of the same. Fig. 7(a) is a plan view of the lid 3 of the Petri dish 1 with alignment marks, disassembled, Fig. 7(b) is a plan view of the container body 2, and Fig. 7(c) is a plan view of the mark attachment member 60.

[0063] Here, the same or similar reference numerals are used to designate the same parts as those in the Petri dish 1 with alignment marks according to the above-described embodiment, and detailed explanations will be omitted.

[0064] The petri dish 1 with alignment marks according to this other embodiment comprises a container body 2, a lid body 3, and a mark mounting member 60 having at least two alignment marks 7a, 7b, as shown in Figures 6 and 7, for example.

[0065] As shown in Figure 7(c), the mark mounting member 60 is configured such that a mating portion 62 for mating with the container body 2 is opened in the approximate center of a rectangular plate-like member 61 made of transparent plastic.

[0066] That is, the Petri dish 1 with alignment marks according to this other embodiment is configured such that the mark attachment member 60 is detachably fixed to the lower end portion of the container body 2 by fitting.

[0067] In such a configuration, as shown in Figure 6(a), for example, a protrusion 9 is provided on the outer periphery of the container body 2, and when mating, the protrusion 9 of the container body 2 is inserted into a notch 63 provided in the mating portion 62 of the mark mounting member 60, thereby allowing the two to be positioned and fixed without any misalignment.

[0068] In the case of the Petri dish 1 with alignment marks according to this other embodiment, the colony counter 10 can more accurately count the number of colonies that changes over time, in a manner similar to that of the above-described embodiment.

[0069] The above describes aspects of the present invention by illustrating several embodiments, but each embodiment is merely an example, and the scope of the invention described in the claims can be modified in various ways without departing from the gist of the invention. [Explanation of symbols]

[0070] 1... Petri dish (culture vessel) with alignment marks 2...Container body 3...Lid 4...Skid prevention member 5...Scratch prevention material 6,60...Mark mounting member 7a, 7b...Alignment marks 8...Information area (display area) 9...Convex part 10...Colony Counter 12...Stage glass 13...Camera 16...Monitor 17...Control unit 18...Storage section 61... Plate-shaped member 62...Matching part 63...Notch CC…Colony AP: Image of specimen after cultivation SP: Image of specimen before culture

Claims

1. a container body in which a culture medium for culturing microorganisms is provided; a lid body that is detachably attached to the container body; a mark attachment member provided on the exterior of the container body and having at least two alignment marks; A culture vessel comprising:

2. 2. The culture vessel according to claim 1, wherein the mark attachment member is integrally molded with the vessel body.

3. 2. The culture vessel according to claim 1, wherein the mark attachment member is made of a plate-like member into which the vessel body is fitted.

4. The container body is provided with a protrusion, 4. The culture vessel according to claim 3, wherein the mark attachment member is provided with a recess into which the protrusion is engaged.

5. 2. The incubation vessel according to claim 1, wherein the mark attachment member further comprises a display area to which identification information can be added.

6. 2. The culture vessel according to claim 1, wherein the vessel body and the lid are made of plastic.

7. 2. The culture vessel according to claim 1, wherein the lid is provided with a lateral slip prevention member along the outer edge of the surface thereof to prevent lateral slippage during stacking.

8. 2. The culture vessel according to claim 1, wherein the vessel body is provided with a scratch prevention member along the outer edge of the bottom surface thereof to prevent scratches on the bottom surface.

Images