JP5618810B2 - Fishing fungus device and fishing fungus method - Google Patents

Description

  The present invention relates to a bacterial colony fishing device and a fishing method for collecting bacterial colonies (fishing).

  Bacteria testing is a test that identifies the type of bacteria and examines the susceptibility of bacteria to antibacterial drugs. For example, in a bacterial test in a clinical test, a pathogen is found from clinical materials (specimens) such as stool collected from patients, an identification test that identifies the name of the bacterial species, and how effective an antibacterial agent is for that pathogen Do a drug sensitivity test. The bacteria here include fungi.

  Bacterial testing steps in clinical testing consist of bacterial group classification, isolation culture, pretreatment, identification and drug sensitivity testing. First, the bacterial group classification process will be described. Investigators include smears such as Gram stains (staining tests) and speculums, and comprehensively determine information such as specimen properties and patient condition, so that they are included in the specimen The bacteria to be tested are judged from a plurality of bacteria and classified into roughly groups such as Gram-positive or negative bacteria, cocci or gonococci, and which genera. This group classification is necessary to determine the test conditions (types of antibacterial drugs and pretreatment conditions of specimens to be applied to the test apparatus) to be used in the subsequent test process. Thereafter, in the separation culture process, the specimen is applied on the medium contained in the petri dish, and when it is cultured for about one day, a colony (colon) that is an aggregate of bacteria is formed on the medium.

  Next, a pretreatment process is performed in order to use the formed colony as a test sample. In the pretreatment step, one or a plurality of bacterial colonies to be inspected are collected (fishing) and suspended in saline to prepare a suspension with a predetermined turbidity necessary for the inspection. This predetermined turbidity varies depending on the group of bacteria and the purpose of inspection. The suspension is sometimes called a bacterial solution. The turbidity is proportional to the number of bacteria in the bacterial solution. The turbidity of the suspension is confirmed by measuring with a turbidimeter.

  Finally, in the identification / drug susceptibility testing process, the bacterial solution prepared in the pretreatment process is used as a test sample, for example, Biotech (Biomelieu), BD Phoenix (Becton Dickinson), Microscan WALK AWAY (Siemens), etc. Species identification and drug susceptibility testing are performed on a bacterial test device.

  Japanese Patent Application Laid-Open No. H10-228561 discloses a fishing fungus device that fishes colonies in a pretreatment process of a bacterial test. Patent Document 1 discloses a method for fishing a colony by calculating the position of a colony on a medium from image information of colonies to be fished and position reference means. According to the description of Patent Document 1, it is stated that a target colony can be fished without performing a special image analysis protocol process.

JP 2000-171360 A

  However, in patent document 1, although the image of the colony on a petri dish can fish the target colony by measuring the position of the colony, the bacterial solution produced thereby is not adjusted to a predetermined turbidity. . Therefore, if the suspension is less than the predetermined turbidity, colonies must be added. Also, if the suspension is thicker than the predetermined turbidity, the suspension must be diluted with saline until the predetermined turbidity is reached. In the pretreatment process for bacterial inspection, the turbidity adjustment of the bacterial solution must be repeated many times, which takes time.

  It is an object of the present invention to provide a fishing fungus device that fishes a necessary amount of colonies in a short time and produces a suspension with a predetermined turbidity.

  The fishing fungus device of the present invention can obtain in advance the area of a colony necessary for creating a suspension with a predetermined turbidity by obtaining in advance information on the group to which the colony belongs by user input. Furthermore, it is possible to switch the fishing operation of the fishing tool according to the area obtained in advance.

  The present invention switches the fishing fungus operation according to the information of the group to which the colony belongs. Thereby, a repetitive operation for adjusting the turbidity can be omitted, and a necessary amount of colonies can be caught in a short time, and a suspension having a predetermined turbidity can be produced.

Schematic of a bacterial colony fishing device. Schematic of the fishing fungus operation | movement part of a bacteria colony fishing fungi apparatus. The graph which shows the relationship between the area of a colony of a cocci and a gonococcus, and turbidity. The flowchart which shows the step until producing a fungal liquid. Schematic of a fishing fungus tool. An example of the display screen of a display part. The figure which shows the dimension measurement of one colony by image processing. Schematic diagram of fishing fungus action by stamp method. The schematic diagram of the fishing fungus operation by a horizontal slide system. The flowchart which shows the step until Example 2 produces bacterial liquid. Experimental results of the number of fishing bacteria by the stamp method required for preparation of a bacterial solution with a predetermined turbidity. The schematic diagram which shows arrangement | positioning of the colony on a culture medium. The flowchart which shows the steps until Example 4 produces bacterial liquid.

  A first embodiment of the present invention will be described.

[Explanation of bacterial colony fishing device]
FIG. 1 is a block diagram of a bacterial colony fishing device. The bacterial colony fishing device includes an input unit 101, a display unit 102, an imaging unit 103, a fishing fungus operation unit 104, and a fungus solution adjustment unit 105. The input unit 101 and the display unit 102 that are interfaces with the user may be integrally formed. The imaging unit 103 includes a camera 106. The fishing fungus operation unit 104 includes a fishing fungus tool 107. Further, a petri dish 109 is provided between the imaging unit 103 and the fishing fungus action unit 104 to support the petri dish 108 and to transport the petri dish 108 as indicated by an arrow (the movement of the petri dish) in the figure. The bacterial solution adjusting unit 105 includes a turbidity adjusting mechanism 111 including a turbidimeter 110 and a saline dispensing mechanism.

  The general flow of fishing fungi operation will be described. A colony on the petri dish 108 is photographed with the camera 106, the petri dish 108 is moved to the fishing fungus operation unit 104, and the colony is fished using the fishing fungus tool 107. Thereafter, the colony that has been fished is put into the container 112 and mixed with a suspension preparation solvent such as saline to prepare a suspension. Further, the container 112 is moved to the bacterial liquid adjustment unit 105 as indicated by an arrow (movement of the container) in the figure, and the turbidity of the suspension is measured by the turbidimeter 110. If the turbidity is high, the turbidity is turbid. When dilution is performed using the degree adjusting mechanism 111 and the turbidity is low, a colony is added again by the fishing fungus operating unit 104 to adjust the turbidity of the suspension to a predetermined value.

[Description of the fishing fungus operation unit 104]
Next, the fishing fungus operation unit 104 will be described. FIG. 2 is a diagram illustrating the fishing fungus operation unit 104. The fishing fungus action unit 104 grips the fishing fungus tool 107 at the tip of the fishing fungus tool rack 201, the petri dish 109 that supports the petri dish 108, the horizontal movement shaft 202, the X-axis body 203, the Y-axis body 204, the Z-axis body 205. A fishing fungus tool gripping part 206, a container 112 for containing bacteria, a mounting part 207 on which the container 112 is mounted, a syringe 208, a flow path 209, a tank 210 for storing saline, a three-way valve 211, and a saline solution for dividing the saline into the container 112 A dispensing pipettor 212 for pouring is provided. A syringe 208, a flow path 209, a tank 210 for storing saline, a three-way valve 211, and a pipetting pipetter 212 for dispensing saline into a container have the same configuration as the turbidity adjusting mechanism 111 shown in FIG. A thing may be shared by the fishing fungus operation unit 104 and the fungus solution adjustment unit 105.

  A plurality of fishing fungus tools 107 are accommodated in the fishing fungus tool rack 201. The horizontal movement shaft 202 is connected to one end of a rod-shaped Z-axis body 205, and moves the Z-axis body 205 in the X direction in the figure together with the X-axis body 203, the Y-axis body 204, and the fishing fungus tool gripping portion 206. Let The horizontal movement shaft 202 allows the fishing fungus tool gripping portion 206 to attach the fishing fungus tool 107 with the rack 201 and move between the petri dish 108 at the fishing fungus position and the container 112 at the suspension preparation position. It becomes. Further, the X-axis body 203, the Y-axis body 204, and the Z-axis body 205 move the fishing fungus tool gripping portion 206 in the X direction, the Y direction, and the Z direction in the drawing, respectively.

  The three-way valve 211 switches (i) a flow path 209 connecting the syringe 208 and the tank 210, and (ii) a flow path 209 connecting the syringe 208 and the container 112. By switching these flow paths 209 and operating the syringe 208 as shown by the arrows in the figure, the saline in the tank 210 can be fed into the container 112 via the dispensing pipettor 212.

[Control method for fishing fungi]
The present invention is particularly characterized by a method for controlling the operation of the fishing fungus operation unit 104.

The bacterial colony fishing device has a control unit (not shown) and a database (not shown) for controlling the fishing fungus operation.
First, some functions of the control unit are shown.
(1) Information is mutually communicated with the input unit 101, the display unit 102, the imaging unit 103, the fishing fungus operation unit 104, the fungus solution adjustment unit 105, and the database.
(2) A colony amount (colony area, volume, number, etc.) required to achieve a predetermined turbidity is determined based on the database.
(3) The area of each colony is obtained by image processing of an image photographed by the camera 106. (The method will be described later.)
(4) Find which colony or which colony range should be collected to achieve the number and amount of colonies found in (2) above. When there are a plurality of colonies to be fished and there are a plurality of combinations (patterns), they are also obtained.
(5) When one or more colonies or a range (area) of colonies are given, the number of times of fishing and the time taken for fishing are obtained.
(6) When one or more colonies or a range (area) of colonies are given, turbidity is obtained when the fungi are picked based on the database. Here, the colony range (area) refers to a region on the surface of the culture medium, in which at least one colony exists in the region.

  Next, the contents held in the database will be described. The bacterial colony fishing device has, for example, a database relating to the relationship between the colony area and turbidity. Information stored in the database is generally known as information used to calculate the quantitative conditions of colonies to be fished when given a predetermined turbidity value of the suspension. Any known fact or information that can be obtained in advance by experiments may be used. In addition to the relationship between the area and turbidity shown as examples, the relationship between volume and turbidity, or the relationship between the number of fishing bacteria and turbidity may be used. Further, instead of turbidity, absorbance (which can be converted to turbidity) may be used. As an example, the relationship between the area of staphylococci and Neisseria gonorrhoeae colonies and turbidity is shown in FIG. Details of FIG. 3 will be described later.

Next, the operation | movement using this fishing fungus operation part 104 is demonstrated using the flowchart shown in FIG.
(S1) From the input unit 101, a group to which bacteria such as cocci or bacilli belong and a predetermined turbidity (target turbidity) are input.
(S2) Petri dishes and colony images captured by the camera 106 are displayed on the display unit 102. Colonies may be automatically identified from image information such as color and shape by image processing and displayed to the user in an easy-to-understand manner with illustrations and characters.
(S3) When the user selects one colony to be picked from the input unit 101, the control unit obtains the area of the colony.
(S4) From the information input in (S1) above and the information in the database, the total area of colonies necessary for producing a suspension with a predetermined turbidity is obtained.
(S5) The control unit can calculate the number of colonies to be the total area by dividing the total area of colonies obtained in (S4) above by the area of one colony obtained in (S3). In general, since one colony is fished by one fishing fungus operation, if the number of colonies is known, the number of times of fishing to activate the fishing fungus tool can also be known. These are displayed on the display unit 102 (the display method will be described later). If there are multiple colony patterns (how to combine) to be fished, they are also displayed. Here, the user may select a colony to be fished, an area (range) of the colony, or the like from the input unit 101, and in that case, the number of fungi and the number of colonies are recalculated and redisplayed.
(S6) The colony selected by the user is fished a predetermined number of times.
(S7) The container 112 is filled with saline using the syringe 208, the tank 210, and the dispensing pipetter 212.
(S8) The fishing fungus tool is immersed in the saline solution in the container 112 to prepare a suspension.

  This is the operation of the fishing fungus operation unit 104. After that, the container 112 is moved to the bacterial solution adjusting unit 105, and the turbidity of the suspension is measured using the turbidimeter 110, and adjusted to a predetermined turbidity. For example, when the turbidity is high, it is diluted with a saline solution, and when it is thin, colonies are added.

  Here, an example of information held in the database will be described with reference to FIG. In the experiment, the present inventors investigated the relationship between the area of a colony in which six types of cocci and gonococci were collected and the turbidity of a suspension prepared by fishing the colonies. As a result, it was found that the turbidity of the bacteria is proportional to the area, and that the slope varies depending on the bacterial species, and that there is a three-fold difference in turbidity between the cocci and bacilli even in the same area. FIG. 3 is a graph showing the relationship between the area and turbidity of the 6 bacterial species tested, which are the least prone to obtain turbidity in each of the cocci and bacilli, that is, those having the lowest fishing fungus efficiency. As can be seen from this graph, the total area of colonies to be fished is different between cocci and bacilli even though the turbidity is the same. The fact that the total colony areas are different may mean that the number of colonies to be fished and the number of fish are different. Therefore, in order to produce a suspension having a certain turbidity, it is necessary to change the number of fishing bacteria depending on whether the bacterium is cocci or bacilli. By taking this point into consideration before fishing, it is possible to fish efficiently. That is, it is possible to prevent a large amount of fish from being wasted and a small amount of fish from being caught.

  Furthermore, if the user selects neither cocci or gonococcus, or other, the database of gonococcus can be selected, so that the bacterial solution can be either cocci or gonococci After moving to the adjustment unit 105, it is possible to prevent a situation where the amount of fishing bacteria is insufficient and fished again, and it is possible to cope with preparation of a bacterial solution having a predetermined turbidity only by dilution.

  As described above, by using the fact that the area varies depending on whether it is a cocci or a gonococcus, it is possible to catch a lot of fungi in vain by knowing in advance which colony should be fished and how many times. It is possible to prevent the amount from being small, and it is possible to omit unnecessary adjustment of the bacterial solution.

[About the fishing fungus tool 107]
FIG. 5 is a diagram showing an outline of the fishing fungus tool 107. The fishing fungus tool 107 includes a fishing fungus tool tip 501 and a fishing fungus tool support 502. As the fishing fungus tool tip 501, urethane foam may be used. Further, the fishing fungus tool 107 can be a disposable one or can be used while carrying out a sterilization process in order to produce a bacterial solution without contamination. The shape of the cross-sectional area of the fishing fungus tool tip 501 may be a triangle, a rectangle, a star, an ellipse, or a circle. The size of the cross-sectional area of the fishing fungus tool tip 501 varies, and what the user wants can be selected.

[Display on display unit 102]
Information displayed on the display unit 102 in the above (S5) will be described with reference to FIG. FIG. 6 is a diagram showing the display unit 102. The display unit 102 displays a colony image 601 on the petri dish, and displays the colonies to be collected necessary for the suspension with a predetermined turbidity in the figure and the characters “A, B, and C for the required amount of bacteria. Please select a colony ". In addition, the user can arbitrarily select the colonies that he / she wants to fish, and can display the predicted number of fish and the number of colonies when assuming that these are fished, as well as the predicted turbidity of the suspension. Yes (character information is omitted in the figure). In addition, when the required amount of bacteria does not exist on the petri dish, it can be displayed that the amount of bacteria does not exist.

[About image processing]
Explain how to determine the area of the colony. FIG. 7 shows a colony outline 701 taken by the camera 101. By the image processing, the colony size 704 obtained by connecting the center of gravity 702 of the colony photographed by the camera 101 and the end 703 having the shortest distance from the center of gravity is measured. You may assume the area of the circle (broken line) which makes this dimension a radius as an area of a colony. The area of the colony may be obtained by counting the total number of pixels.

[About input information to the input unit 101]
In the above, the information to be first input to the input unit 101 is turbidity, whether it is a cocci or a bacilli, but is not limited to this, information on the name of the bacterium, what type of fishing bacteria tool is used, what type of medium May be input. The medium herein includes a selective medium that is a medium for growing only the bacterial species to be examined.

  In the above, the database of turbidity and area was used for each of cocci and bacilli, but it is not limited to the classification of cocci and bacilli, and if there is information about known bacteria in advance, classification such as the group is used. May be.

[About fishing fungus action]
The fishing fungus operation of the present invention includes not only the number of fungi and the number of colonies to be fished as shown in Example 1, but also the direction of operation of the fishing fungus tool. The present invention is a fishing device capable of switching between a “stamp method” in which a fishing fungus tool is moved up and down in a direction perpendicular to the medium surface and a “lateral slide method” in which the fishing fungus tool is operated in a horizontal direction with respect to the medium surface. .

  FIG. 8 is a schematic diagram of the action of the fishing fungus by the stamp method. It is the figure which looked at the culture medium 802 which the fishing fungus tool which has the fishing fungus tool front-end | tip part 501 and the fishing fungus tool support body 502, and the colony 801 grew from the horizontal direction. In the stamp method, the colony 801 is aimed from above, the fishing fungus tool is lowered (FIG. 8 (a)), the fishing fungus tool tip 501 is brought into contact with the colony (FIG. 8 (b)), and fishing is performed again. The fungus tool is moved upward (FIG. 8C), and the colony 801 is collected. Basically, one colony is collected by this series of operations. When there are different types of colonies on the medium, the stamp method, which can be picked up by targeting only one colony, is excellent in that it is less likely to be contaminated by one fishing fungus.

  FIG. 9 is a schematic diagram of the action of the fishing fungus by the lateral slide method. In the horizontal slide method, the fishing fungus tool is lowered and brought close to or in contact with the medium surface (FIG. 9 (a)), and from that state, it is moved horizontally with respect to the medium surface (FIG. 9 (b)). Is contacted and attached to the tip portion 501 of the fishing fungus tool (FIGS. 9C and 9D), the fishing fungus tool is moved upward (FIG. 9E), and colonies are collected. That is, when there are a plurality of colonies 801, the colony 801 is continuously moved from the horizontal direction in a series of movements in the horizontal direction without moving the fishing fungus tool upward for each colony. Can be scraped off. Furthermore, even when colonies are adjacent to each other as shown in FIG. 9 (f), these continuous colonies can be collected using the horizontal slide method. Therefore, when you want to collect many colonies in a short time, the horizontal slide method is appropriate. In addition, the direction of sliding may be a direction of drawing a straight line or a curve on the medium as long as it is horizontal to the medium surface. The sliding distance and area are also variable depending on the colony and area shape of the fungus.

  Therefore, it is possible to fish in high accuracy and in a short time by switching between the stamp method and the horizontal slide method by receiving information on the group to which the colony belongs and instructions from the user.

[About input information to the input section]
In addition to those shown in the first embodiment, a threshold value may be input to the input unit 101. The threshold is the number of fishing bacteria necessary to create a suspension with a predetermined turbidity (the number of stamping operations of the fishing fungus tool), the number of colonies to be fished, the time taken for the fungi, It is an index such as total area, total volume, individual colony size (diameter, area, volume, etc.), distance between colonies (density), and the like, and is a value that defines a limit value for separating the stamp method and the horizontal slide method.

  In addition to the user input from the input unit 101, the threshold is set by a clinical test information system (LIS) used for information communication in a clinical laboratory, a host computer, or a method that is input to the apparatus via a network such as the Internet. But you can. Further, it may be information stored in advance in a database or apparatus, and the setting method is not limited.

  In addition, the conditions of fishing fungi can be input from the input unit 101. The conditions of the fishing fungus are the colony of the fungus itself, the area of the colony to catch the fungus, the type of fishing fungi tool if there are multiple types of fungus tools, and the force and fishing force that makes the fungus tool contact the medium or colony. It is the descent distance of the fungus tool.

  Below, the example which used the number of times of fishing bacteria as a threshold is shown. However, the threshold value is not limited to the number of fishing bacteria, the number of colonies to be fished, the time taken for fishing fungi, the total area and total volume of the colonies to be fished, and the size (diameter, area, volume, etc.) of each colony The distance (density) between the colonies is not particularly limited as long as it is an index that can be used to discriminate between the stamping method and the horizontal sliding method.

[Control method for fishing fungi]
The bacterial colony fishing device has a control unit for controlling the fishing fungus operation, and this control unit has the following functions in addition to (1) to (6) shown in the first embodiment. Also good.
(7) Compare the fishing conditions under which a suspension with a predetermined turbidity is produced by the fishing fungus action by the stamp method and the threshold value, and determine the fishing action to be executed.

Next, the flow of producing a suspension by separating the stamp method and the horizontal slide method according to the threshold will be described with reference to the flowchart shown in FIG.
(S11) From the input unit 101, either cocci or gonococci, turbidity, threshold value, and the like are input. Here, as an example, the maximum number of fishing bacteria N, which is the upper limit of the number of fishing bacteria by the stamp method, is input as the threshold.
(S12) A colony image is taken by the camera 106.
(S13) Based on the image and database, the colony area that becomes the total bacterial amount (bacterial amount necessary for producing a suspension with a predetermined turbidity) is obtained in the same manner as in Example 1, and the number of colonies and the number of colonies by the stamp method are obtained. And calculate patterns.
(S14) The calculated information is displayed on the display unit 102. Here, the user may select a colony or an area for executing the fungus from the input unit 101. In this case, the number of times of the fungus, the number of colonies, and the like are recalculated and redisplayed.
(S15) The control unit compares the number of fishing fungus n, which is the total amount of bacteria determined from the image, with the threshold value N. When [the number of fishing fungus n ≦ threshold N], it is determined that the vertical operation (stamp method) of the fishing fungus tool is selected. When [the number of fishing fungus n> threshold N], it is determined that the horizontal direction (lateral slide method) operation of the fishing fungus tool is selected. The determination may be displayed on the display unit 102. In this case, the user may determine and change the operation selected by the control unit.
(S16) The colony is caught by the determined operation of the fishing fungus tool 107.
(S17) The container 112 is filled with saline using the syringe 208, the tank 210, and the dispensing pipetter 212.
(S18) The fishing fungus tool is immersed in a saline solution in the container 112 to prepare a suspension.

  This is the operation of the fishing fungus operation unit 104. After that, the container 112 is moved to the bacterial solution adjusting unit 105, and the turbidity of the suspension is measured using the turbidimeter 110, and adjusted to a predetermined turbidity.

  Here, the above-mentioned number of fishing bacteria will be described. The inventors of the present invention investigated the number of fishing fungi operations by the stamp method required for producing a suspension with a predetermined turbidity through experiments. The result is shown in FIG. Here, it is assumed that one colony is collected by a single fungus action by the stamp method. As the turbidity of the suspension required for the inspection, McF2.0 was used for B. Pumilus and McF0.5 was used for other bacterial species. Assuming that the number of times of fishing by the stamp method is 10 times, in order to achieve a predetermined turbidity in B. Pumilus, S. dysgalactiae, S. Salivarius species, more than the number of times of fishing is required. . This is because there are differences in the required turbidity, colony size, and colony area for achieving the predetermined turbidity as described in Example 1, depending on the bacterial species.

  Therefore, in the case of a bacterial species that requires a large number of fishing times using the stamp method, such as when a highly turbid bacterial solution is required or when the colony is small, by collecting multiple colonies using the horizontal slide method, the number of operations can be reduced. Can fish.

  As described above, efficient colonization can be performed by grasping in advance which colony should be fished how many times and comparing the colony with a threshold value.

  The colonies grown on the medium are densely arranged or separated from each other, and in various cases. Here, an example of dividing the method of fishing fungus operation suitable for the arrangement of colonies on the medium is shown. In Example 2, a method of selecting either the stamp method or the horizontal slide method and fishing the colonies on one medium was shown, but here, both the stamp method and the horizontal slide method are used. An example of collecting colonies from one medium is also shown.

  The example of proper use of each system by arrangement | positioning of the colony on a culture medium is demonstrated using the schematic diagram of the colony of Fig.12 (a)-(d).

  12A to 12D respectively represent large and small colonies. The large colonies are colonies collected by the stamp method, and the small colonies are colonies collected by the horizontal slide method. The user can determine whether the colony is large or small from the image displayed on the display unit. However, as described in the second embodiment, the colony size can be set as a threshold value and automatically determined. is there. For example, the size of the colony to be fished and calculated by performing image processing on the image acquired by the imaging unit 103 is compared with a threshold value, and the large and small segmentation of each colony is displayed on the display unit 102. FIGS. 12A to 12D are schematic diagrams of a medium 802 and a colony 801 grown on the medium as viewed from the upper surface direction of the medium. Symbols A to D are colonies and colony areas (represented by circles) required for a suspension having a predetermined turbidity. In addition, it is assumed that A to D all indicate colonies of the same bacterial species, and there is no contamination by fishing these.

  In the case of FIG. 12A, a suspension having a predetermined turbidity can be prepared by collecting large colonies A to D by a stamp method.

  In the case of FIG. 12B, a suspension with a predetermined turbidity can be prepared by sliding the fishing fungus tool 107 in the area A and collecting small colonies.

  In the case of FIG. 12C, since there are two areas of colonies to be collected by the horizontal slide method, A and B, the slide is performed in two places. The determination can be made by setting the distance (density) between colonies, the time required for the fishing fungus operation, and the like as thresholds. In this manner, the number of times of the horizontal slide method is divided according to the arrangement of the area of the colony to be collected.

  Further, in the case of FIG. 12 (d), the stamp method and the horizontal slide method are combined in one medium. Colonies A and B are collected by a stamp method, and colonies in area C are further collected by a horizontal slide method. The information can be divided by inputting in advance information on the fishing time required for each of the stamp method and the horizontal slide method, and using this information as a threshold value.

  In this way, the threshold is not limited to the number of fishing fungi. The number of colonies, the area of the colonies, the volume, etc., including the size of each colony, the time taken for the fungus action, the distance (density) between colonies, etc. Can be set. As a result, not only the stamp method or the horizontal slide method, but also the combination of both methods or the horizontal slide operation can be divided into several times according to the colony layout and area. Colonies can be collected.

  It is said that there are tens of thousands to millions of bacteria on the earth, including those that have not been discovered. When an unexpected colony such as an unknown bacterium or a bacterium with no information in the database is fished, the turbidity of the prepared suspension may not achieve a predetermined turbidity. When the prepared suspension does not reach the predetermined turbidity, it is necessary to further colonize the colony and add it to the suspension. A fishing fungus device that can efficiently add colonies even if it is an unexpected group of fungi is shown as Example 4 below.

[Control method for fishing fungi]
The bacterial colony fishing device has a control unit that controls the fishing fungus operation and a turbidimeter 110, and this control unit may further have the following functions.
(8) After fishing, the prepared suspension is measured with the turbidimeter 110, and the turbidity is received by the control unit.
(9) The turbidity received by the control unit is compared with the predetermined turbidity, and when the predetermined turbidity is not achieved, the deficiency turbidity is calculated.
(10) From the turbidity received by the control unit, the relationship between the area of the previously fished bacteria and the measured turbidity is calculated.
(11) Using the relationship of (10) above, obtain the area of additional colonies that make up for the turbidity deficiency calculated in (9) above, and the number of fishing bacteria.

Next, as an example, if the turbidity of the suspension produced by fishing with the stamp method is thinner than the predetermined turbidity, the flow of additionally colonizing the suspension and preparing the suspension with the predetermined turbidity This will be described with reference to the flowchart shown in FIG.
(S21) From the input unit 1, either cocci or bacilli, turbidity, threshold values, and the like are input.
(S22) The fungus is fished by a stamp method to prepare a suspension. It is assumed that the number of fishing bacteria by the stamp method executed here is m times. This is the same step as S5 to S8 in the first embodiment.
(S23) The turbidity c of the prepared suspension is measured.
(S24) The turbidity measured in S23 is compared with a predetermined turbidity.
(S25) If the predetermined turbidity is not achieved, the deficiency turbidity d is calculated.
(S26) The relationship between the turbidity c measured in the above S23 and the number m of fish caught by the stamp method is obtained. For example, the fungus solution production efficiency e is calculated by dividing the measured turbidity by the number of fishing bacteria (c / m). In other words, the fungus production efficiency is the turbidity of the suspension obtained by the fishing fungus operation by one stamp method.
(S27) The number of fishing fungus n that compensates for the deficiency turbidity d calculated in S25 is determined based on the relationship determined in S26. This is obtained by dividing the deficiency of turbidity by the efficiency of producing the bacterial solution (n = d / e).
(S28) The operation is determined from the threshold value in the same manner as in S14 to S16 of Example 2, and additional colonies are additionally fished.
(S29) A colony is added by immersing a fishing fungus tool in the suspension of S22 to prepare a suspension.

  In S26 to S27, the fungus liquid production efficiency was determined by the number of times of fishing using the stamp method, but information such as the area and volume of the colony may be used. For example, when the suspension prepared by the horizontal slide method is thinner than the predetermined turbidity, the relationship between the area of colonies already caught and the obtained turbidity, that is, the turbidity obtained per fishing colony area (fishing efficiency) ) Is calculated, the area of the colony that compensates for the turbidity of the deficiency is obtained therefrom, and the colony is efficiently added.

  As described above, even if the group information is not available in advance or the turbidity is less than the predetermined turbidity after the initial suspension preparation, the deficiency is calculated without repeating the measurement of fishing bacteria and turbidity many times. Then, a suspension with a predetermined turbidity can be prepared by adding a colony only once more.

DESCRIPTION OF SYMBOLS 101 Input part 102 Display part 103 Image pick-up part 104 Fishing fungus action | operation part 105 Bacteria liquid adjustment part 106 Camera 107 Fishing fungus tool 108 Petri dish 109 Petri dish 110 Turbidimeter 111 Turbidity adjustment mechanism 112 Container 201 Fishing fungus tool rack 202 Horizontal movement Shaft 203 X-axis body 204 Y-axis body 205 Z-axis body 206 Fishing fungus tool gripping part 207 Container mounting part 208 Syringe 209 Flow path 210 Tank 211 Three-way valve 212 Dispensing pipettor 501 Fishing fungus tool tip 502 Fishing fungus tool support 601 Colony image 701 Colony outline 702 Colony center of gravity 703 The shortest end 704 from the center of gravity The colony size of one colony obtained by connecting the center of gravity and the shortest end from the center of gravity 801 Colony 802 Medium

Claims (8)

There in picked apparatus having a picked tool for collecting colonies, a driving unit for driving the該釣bacteria tool to vertical and horizontal directions, and the base for mounting a Petri dish, a camera for photographing the colonies, the And
Classify colonies as cocci or bacilli, gram positive or negative, or genus
A database that shows the relationship between the turbidity of the suspension and the total area of the colony, for each categorization of the colony as cocci or bacilli, gram positive or negative, or which genus
It has a control unit that performs image processing to determine the area of the colony based on the colony image taken with the camera,
The movement of the fungus tool by the drive unit is either a vertical direction or a horizontal direction or a combination thereof,
Based on the images taken with the camera and the database, find the colony area that will be the amount of bacteria necessary to make a suspension with the desired turbidity,
Further, the number of times of fishing for fishing the colony area by the vertical movement of the fishing fungus tool is obtained, and when the number of fishing bacteria is less than or equal to a preset threshold value, On the other hand, when the number of times of fishing is larger than a preset threshold, the fishing device is characterized in that the fish is picked up by the operation of the horizontal fishing tool. In claim 1,
A fishing fungus device characterized by displaying the number of colonies to be fished. In claim 1,
A fishing fungus device, wherein an image of a colony on a petri dish is displayed and a colony to be fished is marked. In claim 1,
Furthermore, a fishing fungus device comprising a turbidimeter for determining the turbidity of the suspension. In claim 1,
Furthermore, the fishing-bacteria device provided with the input part which selects the colony and colony area which a user fishes. In claim 1,
A fishing device, characterized in that when a fishing tool is operated in a horizontal direction, at least one of an operating distance, an operating range, or an operating direction is determined, and selected colonies and colonies in an area are fished. Use a picked tool for collecting colonies, a driving unit for driving the該釣bacteria tool to vertical and horizontal directions, and the base for mounting a Petri dish, a camera for photographing the colonies, the picked device provided with A fishing fungus method,
The movement of the fungus tool by the drive unit is either a vertical direction or a horizontal direction or a combination thereof,
Classify colonies as cocci or bacilli, gram positive or negative, or genus
Based on the images taken with the camera and the database showing the relationship between the turbidity of the suspension and the total area of the colony for each of the above classifications, the bacteria required for the preparation of the suspension with the desired turbidity Find the amount of colony area
Further, the number of times of fishing for fishing the colony area by the vertical movement of the fishing fungus tool is obtained, and when the number of fishing bacteria is less than or equal to a preset threshold value, On the other hand, when the number of fish is greater than a preset threshold, the fish is caught by the operation of a horizontal fishing tool. In claim 7,
A fishing method, characterized in that when a fishing tool is operated in a horizontal direction, at least one of an operating distance, an operating range, or an operating direction is determined, and selected colonies and colonies in an area are fished.

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