JPS621714B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention enables colonies (bacterial species) to be transferred from cultured dishes to test dishes or test tubes without any manual intervention, except for the visual selection of colonies to be transplanted. The present invention relates to a colony transfer device.

In order to develop new species such as antibiotics, methods are used to find effective bacteria from soil etc.
For this purpose, colonies are collected and cultivated from a wide variety of heterogeneous soils, all of which involve colony transfer operations between test dishes or between test dishes and test tubes. . In addition, research in genetic engineering, which has been rapidly progressing in recent years, involves selecting and culturing specific varieties of bacteria and recombining their genes in hopes of creating new organisms. is carried out frequently.

Conventionally, colony transplantation was performed manually using a specially shaped "platinum loop" while observing the colony group with the naked eye or with an optical projection device. There are various methods of transplantation, but one example is to select a colony that has grown to a diameter of 1 to 2 mm from among the colonies growing in a culture medium made of agar of cultured Sheare, and to contact this with a platinum loop. They are collected, arranged and transplanted in designated positions in a separate test shear medium, and this process requires considerable skill and effort.

Next, to describe the different aspects of the transplant destination container, the test tube is suitable for transplanting many types of bacterial colonies, and the test tube is used to culture a large amount of a single type of bacteria. A method of widely distributed transplantation is adopted.

During transplantation work, it is important to prevent contamination by other germs, so the work must be done in a safety cabinet, and all instruments used must be disinfected and sterilized (hereinafter simply referred to as sterilization) in a timely manner.

To further summarize the points mentioned above, in the case of research and development, initially, from among a variety of miscellaneous bacterial species being cultured, promising and useful new bacterial strains are cultivated as pure as possible into a single variety. The process of extracting and culturing pure bacteria is repeated, and in cases where it is already at the stage of industrial mass production, it is necessary to remove problematic bacteria and contaminant bacteria that should not be present among the cultured bacteria. It is a process to eliminate such things as
In reality, it is often necessary to handle thousands or more specimens every day, and the seemingly simple colony transplantation process described above involves identifying the target colony within the field of view of a microscope over a long period of time, and only then can it be done accurately. Repeated harvesting and transplanting work is extremely tiring for workers, and requires a large number of workers with some level of skill.

The purpose of the present invention is to automatically and mechanically perform processes other than the specific colony selection work, which currently has to rely on humans. An object of the present invention is to provide a colony transfer device that does not cause physical fatigue.

In recent years, with the remarkable progress of so-called mechatronics, although it is still difficult to produce a device that can continuously repeat positioning with an accuracy of 1 to 2 μm, it is relatively easy to obtain a device that can position with much higher accuracy than humans, for example, about 10 μm, and with microscopes. Instead of looking directly into the microscope, a television camera is connected to the microscope, and an enlarged image is played back on a large receiver.When selecting the position on this screen, an XY stage ( Instead of reading the scale by slightly moving the stage), for example, if you specify an object on the enlarged playback image with a cursor, the position information of the object (the position information itself can be in the μm order) is immediately stored in the storage device. I was able to remember things. Furthermore, a series of long and complex sequential controls, which were impossible in the past when cams were used, can now be performed extremely easily and accurately using computers, and it is also easy to change control procedures. It is well known that prices of computer-based control equipment are decreasing rapidly. The present invention optimally combines the above-mentioned high-precision mechatronics technology and computer-based control technology in a colony transfer device, so that only a relatively small number of workers can perform visual selection of target colonies and perform other tasks. The aim is to make the entire colony transplanting process as automatic as possible and to make it as efficient as possible. Currently, there are many technologies and parts that can be used for automatic wire bonders, etc., which are widely used in semiconductor manufacturing, and because they have already been used in many cases, they are known to be highly reliable and inexpensive.

The present invention will be explained in more detail below using the drawings. In addition, the side from which specific colonies on the culture medium are selected and collected is called the culture shear, and the side to which the colonies are transplanted is called the test shear.

Conventionally, when manually harvesting and transplanting colonies, a device called a "platinum loop" with a tip shape suitable for collection and transplantation is used, which is washed and sterilized, and the tip is brought into contact with the colony on the culture medium of the culture shear. As mentioned above, after attachment, the cells were brought into contact with the culture medium in a test shear or test tube again and transplanted.
In this case, washing and sterilization are omitted during continuous transplantation of the same colony, and are performed when restarting work after an interruption or when transplanting a different colony, but it is necessary to perform the same in the case of automation. Although the platinum loop can be held by a suitable drive mechanism and used in conjunction with cleaning and sterilization means, it is also possible to create a pick-up section with a harvesting and transplanting end that is more suitable for automation. The main structure of such a pickup section is shown in FIG. As shown in Figure a, a thin wire 2 made of gold, aluminum, glass fiber, etc. is wound around a wire reel 1, fed out by a feeder 3, fixed by a clamper 4, and the tip is cut off by a cutter 5. Next, as shown in Figure b, the cut end of the thin wire 2 is heated and melted using a torch flame 6 to form a ball 7. Using this ball 7, colonies are collected and transplanted as shown in Figure c. If cleaning or sterilization is required, return to Figure a and cut off the end including the ball 7 that was previously used, and heat and melt the cut end with a torch to make a new ball 7. In this way, the molten ball is sterilized at the same time as it is made, and washing is not necessary. The process of creating the molten ball 7 is carried out under computer control while moving the XY stage. As the torch flame 6, a discharge torch or an oxyhydrogen flame (direction controlled by leaving it on) may be used. Instead of using a molten ball, the tip is cut off using a cutting tool 8 as shown in FIG. 2a, and at the same time the end is bent and formed as shown in FIG. Heat sterilization is performed using a high frequency heating type heater 9. In this case as well, the washing step is not necessary because the previously used portion is discarded.

Next, regarding the automation of transplant operations, we will first discuss the case of test shears. FIG. 3a shows the main parts of the pick-up section 10, and shows the culture medium 12 of the culture tray 11.
The position of the visually selected colony 13-1 among the colonies of various bacterial species distributed in the area is stored in the storage device at the same time as the selection, as described above.
Move the XY stage based on this memory information,
After the colony 13-1 is positioned directly below the ball 7 (see FIGS. 1a and b), the ball 7 is lowered to allow the colony 13-1 to adhere to the ball and to be collected. At this time, the height of the ball 7 is controlled by means 14 for measuring the distance (height) to the surface of the culture medium 12 based on the principle of triangulation, which is equipped with an LED, an optical fiber, a photodiode, etc., and the ball 7 is smoothly connected to the colony. Next, the ball 7 in the pick-up section is pulled up, and the ball 7 in the pick-up section is brought to a designated position on the culture medium 12 of the test tray 15-1 as shown in FIG. 3b by the transfer mechanism described later. The ball 7 is lowered and brought into contact with the culture medium to transplant the colony 13-1. To adjust the relative position of the ball 7 and the culture shear dish and test shear dish, move the XY stage side on which the shear dish is mounted. All of the above series of porting operations can be easily performed under computer control. In the case shown in FIG. 3b, colonies 13-1 of one variety are transplanted in a plurality of locations on the test shear 15-1, but as shown in FIG.
-2, colonies 13 of multiple different bacterial species
-2,13-3 may be transplanted. ball 7
The transplantation procedure is the same, except that a new one is made each time in response to changes in the bacterial species.

FIG. 4 shows an example of transplanting a colony of one bacterial species into a test tube 16. In this case, medium 1 in the tube
It is necessary that the surface 12' of the tube 2 be sloped with respect to the tube axis, and colonies 13-1 of one species of bacteria should be dispersed and transplanted onto this surface 12'. The reason why the surface is sloped is that it is desirable for culture to widen the effective area of the medium surface. In this case, the test tube 16 is rotated by the turntable 17, the height of the surface 12' is detected by the distance measuring means 14, and the test tube 16 is stopped at, for example, the lowest position to fix the surface 12' in a fixed direction. Thereafter, the ball 7 is brought into contact with the sloped surface 12' while using the distance measuring means 14 to spread the bacteria.

FIG. 5 is a block diagram showing the entire configuration of an embodiment of the present invention, in which a culture shear 11 and a test shear 15 are shown.
Alternatively, the test tubes 16 are mounted on XY stages 19-1 and 19-2 installed near both ends of the common stage 18, and the pick-up section 10 is moved between the culture shear 11 and the test shear by the transfer mechanism 20. 15 or between the test tubes 16, the memory 24,
Microcomputer 2 with operation panel 25
Move under control of 3. As described above, the height to the culture medium surface is detected by the distance measuring means 14 attached to the pick-up section 10, and the height of the end of the pick-up section for collection and transplantation is controlled.
The XY stages 19-1 and 19-2 can be slightly moved in two orthogonal directions, X and Y, by driving a motor such as a pulse motor. A pick-up processing section 21 is provided between the XY stages 19-1 and 19-2 for reprocessing and sterilizing the end of the pick-up section for collection and transplantation, such as manufacturing the molten ball 7 described with reference to FIG. This process is necessary when the transplant target colony selected via the monitor unit 22 (described later) is different from the one that has been handled up to that point, or when starting up after work has been interrupted, and the process is necessary. is controlled by the microcomputer 23 in conjunction with the movement of the XY stage. The collection and transplantation of colonies by the pick-up section are as already described in connection with FIGS. 1 to 4. The monitor section 22 consists of a television camera and a monitor television combined with a microscope if necessary, and the television camera is mounted on a common stand 18.
It is attached so that the surface of the culture medium of the culture shear plate 11 can be clearly photographed. FIG. 6a is a plan view of the main part of the embodiment of the present invention, centering on the transfer mechanism 20 described with reference to FIG.
As a result of this rotation, the pickup section 10 makes a circumferential movement as indicated by arrow D and stops at a desired position. FIG. 6b is a front (partially sectional) view of the above embodiment, in which a television camera 22-1 is attached above the culture dish 11, and a monitor television 22-2 is placed at an appropriate location where it can be easily seen by the operator. ing. Monitor TV 2 by operating the operation panel 25
2-2 You can select the colony to be transplanted by moving the cursor on the screen. By eliminating the need for the worker to look directly into the microscope, the physical fatigue of the worker is greatly reduced. In addition, 26 in the figure is a cover for antibacterial and light shielding.

As explained above, according to the present invention, it is possible to collect accurately selected colonies with much higher precision than when manually culturing a single bacterial species, and the work time is significantly reduced. As a result, microbial research dealing with biological phenomena can be conducted accurately and efficiently, and the physical fatigue associated with conventional manual labor is greatly reduced.

[Brief explanation of the drawing]

Figures 1 a, b, and c are explanatory diagrams of balls formed by heating and melting the cut ends of thin wires used for colony collection and transplantation in an embodiment of the present invention, and Figures 2 a, b, and c are illustrations of balls formed by heating and melting the cut ends of fine wires used for colony collection and transplantation in an embodiment of the present invention. An explanatory diagram showing the bent end of the thin wire cut end used for colony collection and transplantation in the example and heat sterilization, FIG. 3a is a diagram showing the culture shear and pick-up part in the example of the present invention, c is a diagram showing a test shear and transplantation method in an embodiment of the present invention, FIG. 4 is an explanatory diagram of a method of transplanting colonies into a test tube in the present invention, FIG. Figure 6 a, b
1A and 1B are a plan view and a front (partially sectional) view of a main part of an embodiment of the present invention, respectively. 2... thin wire, 3... feeder, 4... clamper, 6... torch flame, 7... ball, 8... cutting tool, 9... heater, 10... pick up part, 11... culture shear, 12...Medium, 13
-1,13-2,13-3... Colony, 14...
... Distance measuring means, 15-1, 15-2 ... Test shear, 16 ... Test tube, 18 ... Common stand, 19
-1,19-2...XY stage, 20...transfer mechanism, 21...pickup processing section, 22...monitor section, 23...microcomputer, 24...
...Memory, 25...Operation panel.

Claims (1)

[Claims] 1. On a common stand, (a) a cultured shear of colonies;
(b) a monitor section for visually selecting a specific colony on the surface of the medium in the culture shear; (c) a monitor section for visually selecting a specific colony on the surface of the medium in the culture shear; (d) a pick-up part for picking the colonies and transplanting them onto the surface of the culture medium in a test dish or test tube as specified;
a pick-up processing section that processes the end of the pick-up section for collection and transplantation, and (e) a transfer section that transfers the pick-up section as specified between the culture shear, test shear or test tube, and the pick-up processing section; (f) A colony transfer system characterized in that, in accordance with colony selection via the monitor unit and associated operation instructions, each of the units described above sequentially operates as specified under the control of a computer. equipment. 2. The pick-up part is equipped with a means for feeding, cutting, and heating a thin wire of gold, aluminum, or glass fiber, and the cut end is heated and melted to form a spherical part as the end for collecting and transplanting colonies. The colony transfer device according to item 1. 3. The pick-up section is equipped with a means for feeding, cutting, and forming a fine wire of gold, aluminum, or stainless steel, and the cut end is processed into a predetermined shape to be used as an end for collecting and transplanting colonies.Claim 1 The colony transfer device described in Section 1. 4. The pick-up section is equipped with a distance measuring means to the culture medium surface consisting of an LED, an optical fiber, and a photodiode, thereby controlling the vertical movement of the end section for colony collection and transplantation. equipment. 5 The monitor unit is equipped with a microscope or television camera and its magnified image reproduction device, and furthermore, when selecting a specific colony, when the XY stage is moved or when instructions are given using the cursor or stick, the position information is stored in the storage device. The colony transfer device according to claim 1, wherein the colony transfer device is configured to store data. 6. The transfer section includes an arm with a pick-up section attached to one end and the other end as the center of rotation, and on the rotational circumference of the end of the arm where the pick-up section is attached,
The colony transfer apparatus according to claim 1, wherein said two sets of XY stage sections and pick-up processing sections are arranged.