JPH03219898A - Method for automatically transplanting microorganism or the like - Google Patents

Abstract

PURPOSE:To improve the automatic control of colonies by disposing one or more AGP for index outside the rows of agar pieces(AGP) which are agar cylindrical media arranged in the lower dish of a Schale. CONSTITUTION:5X5 Pieces of AGP 5 for transplantation are arranged on the inside bottom surface of the lower dish 1 of a Schale. AGP 7a and 7b for indexing are disposed at the upper and lower positions (positions outside the rotation symmetrical positions of the pattern image of the AGP group), respectively, which are outside the rows of the AGP 5 and are eccentric from the center of the whole AGP 5. A camera is disposed above the Schale 1, reads the index pieces 7a, 7b and treats the read images to recognize the index pieces 7a, 7b. An order for transplanting the AGP 5 is automatically determined from a relation between the positions of the recognized index pieces 7a, 7b and the AGP 5, and microorganisms, etc., are automatically transplanted.

Description

DETAILED DESCRIPTION OF THE INVENTION [Technical Field of the Invention] The present invention relates to a method for automatically transplanting microorganisms, etc. More specifically, the present invention provides a method for specifying the order of transplantation when a single piece (agar cylinder medium) is used as a medium for the transfer of microorganisms or microbial cultures (e.g., colonies). Regarding the automatic porting method that can be used.

[Prior Art] Conventionally, new bacterial species are generally treated and cultured through the following nine steps.

(+) Collection of soil, (2) Dilution with water.

(3) Planting on agar medium in culture dish, (4)
Cultivate for a certain period of time in a warmer, (5) Visually judge the colonies of various bacteria that have grown on the agar medium, and (6) collect only colonies with the desired color using a platinum loop.

(7) Transferring a test dish or test tube to an agar medium;
(8) If the bacterium is cultivated and shows promise, it is examined through repeated tests, and (9) it is applied to medicine, brewing, and other biotechnology fields.

In the second step, the visual identification, collection, and transplantation of colonies of specific bacteria that appear promising are performed manually, which is tedious and inefficient work. Therefore, in order to eliminate these drawbacks, we have developed an operating device that can sequentially observe the culture medium in a petri dish using an X-Y stage, or an alternative to observing the culture medium in a culture dish directly with the naked eye or through a microscope. Therefore, automatic devices have been proposed and developed that process images using a color TV monitor and visually observe the color TV, thereby increasing the efficiency of the work and reducing fatigue.

The automatic colony transplantation device developed as an automatic device of this type selects colonies of a predetermined color from the culture medium of a source petri dish in which various colonies are propagated in a mixed state, and automatically selects colonies according to the selection. In addition, the cells are collected one after another and transplanted into the medium of the object (culture) Nyare L one after another.

[Problems to be Solved by the Invention] A plate-shaped agar medium formed by pouring agar into the inside of a Nyare lower plate is widely used for culturing microorganisms and the like. However, when a plurality of types of microorganisms are transferred to this culture medium, since there are no boundaries on the surface of the culture medium, the grown colonies come into contact with each other, which may lead to incorrect test results.

For this reason, recently, a plurality of cylindrical agar plates (agar pieces) of a predetermined height are arranged at predetermined intervals on the inner bottom surface of a petri dish lower plate. In this form, the media are independent and separated from each other, so there is extremely little risk of colonies coming into contact with each other even when cultured.

However, since the petri dish is almost perfectly round, the agar pieces are
When Xn pieces are arranged, if the petri dish rotates when being transported to the automatic transfer device, it becomes extremely difficult to specify the transfer order.

Therefore, an object of the present invention is to provide an automatic transplantation method that can specify the transplantation order even when using agar pieces.

[Means for Solving the Problems] In order to achieve the above object, in the present invention, a plurality of cylindrical agar pieces of a predetermined height are arranged at a predetermined interval on the inner bottom surface of a lower plate of a petri dish, and each of the agar pieces is arranged at a predetermined interval. In the method of transferring microorganisms or microbial cultures to the upper surface, at least one indexing agar piece is arranged outside the row of agar pieces at a position that is not rotationally symmetrical to a pattern image formed by a plurality of arranged agar pieces. Provided is a method for transplanting microorganisms, etc. characterized by the following.

[Operation] As described above, according to the method of the present invention, the index agar piece is placed at a rotationally asymmetric position of the pattern image formed by the transplant agar piece, so if this index agar piece is used as a landmark, , no matter how the Petri dish is rotated, the transplant starting point can always be easily identified.

[Example] Hereinafter, the method of the present invention will be explained in detail with reference to the drawings.

Generally, the Nyare consists of a lower plate with a predetermined depth and an upper plate that is placed over the opening of the F plate. A plurality of cylindrical agar pieces are arranged at predetermined intervals on the F side of the petri dish as a culture medium. Is there a relationship between the plate culture medium and the agar piece culture culture? It is widely used in large-scale culture tests of microorganisms because it does not cause staining. However, when agar pieces are arranged in a nearly perfectly circular Petri dish, it is extremely difficult to identify the reference points such as the transplant start point and transplant end point, and it is extremely difficult to identify the points that should be used as reference points, such as the transplant start point and transplant end point. If this occurs, the reference point for transplantation will be completely unknown. In cases where the culture medium is poured over the entire surface of the petri dish, it is possible to clearly mark the reference point for transplantation by marking the surface of the culture medium with a mark such as a scratch. It is not possible to perform marking treatment on the medium itself.

Therefore, when 5 x 5 agar pieces 5 for transplantation are arranged on the inner bottom surface 3 of the lower dish l as shown in FIG. Place one index agar piece 7 on the top and bottom at a position eccentric from the overall center of the index agar piece 7.
Place a + 7 b.

A camera is placed above the petri dish to read the agar pieces and index pieces in the petri dish, process the images, and recognize the index pieces. The order of transplanting the agar pieces is automatically determined from the recognized position of the index piece and the positional relationship of the agar pieces.

In the index piece recognition process, as shown in Figure 2, the enlargement process continues until all the pieces become one image.
Thereafter, as shown in FIG. 3, the reduction process is continued until the image is divided into three pieces, and the index pieces are recognized.

When the index piece is recognized, it is determined which side the index piece is on with respect to the center of the agar piece group, and the order of transplanting the agar pieces is automatically determined as shown in FIG.

The number of agar pieces provided is not limited to n×n as illustrated. For example, an arrangement of NnX(n-1) is naturally possible.

Furthermore, the number of index pieces provided is not limited to the two illustrated examples. As long as the order of transplantation can be determined, the number of index pieces may be one. What is important is that the index piece is located at a rotationally asymmetric position of the pattern image of the agar piece group. Even if two index pieces are used, if they exist at rotationally symmetrical positions in the pattern image of one piece group, the implantation order cannot be determined. For example, if the index pieces are placed in the positions shown in FIGS. 5, 6, and 7, the implantation order cannot be determined because the positions are rotationally symmetrical to the pattern image of the agar piece group.

[Effects of the Invention] As explained above, according to the method of the present invention, it is possible to specify the order in which agar pieces are transplanted, so it is possible to determine the position on which agar pieces were transplanted from another culture dish. Automatic colony management and automatic transplantation become possible.

[Brief explanation of drawings]

FIG. 1 is a plan view showing an example of the arrangement of an agar piece and an index piece for carrying out the method of the present invention, FIG. 2 is an enlarged image thereof, and FIG. 3 is a reduced image thereof. Fig. 4 is a schematic plan view showing the transplantation order specified by the method of the present invention, and Figs. FIG. 3 is a plan view showing an arrangement example. 1... Nyare. 3... Inner bottom of petri dish. 5... Agar peas. 7a, 7b... Index piece Figure 1

Claims (2)

[Claims] (1) A method in which a plurality of cylindrical agar pieces of a predetermined height are arranged at a predetermined interval on the inner bottom surface of a lower dish of a petri dish, and microorganisms or microbial cultures are transferred to the top surface of each of the agar pieces. 1. A method for automatically transplanting microorganisms, etc., characterized in that at least one index agar piece is placed at a position that is not rotationally symmetrical to a pattern image formed by a plurality of arranged agar pieces. (2) The method for automatically transplanting microorganisms, etc. according to claim 1, characterized in that two index pieces are arranged.