JPS611378A - Automatic colony transplantater - Google Patents

Abstract

PURPOSE:The titled device that selects desired colonies based on the color phase information which is obtained by subjecting the light reflected from the object colony to spectral diffraction with a grating, thus enabling correct transplantation of desired colonies only by indicating the color phase of the colony at the start. CONSTITUTION:The light reflected from a culture dish is divided into 2 beams with a reflection mirror 22 and one beam is observed with naked eyes 15. Simultaneously, the other beam is subjected to spectral diffraction with a grating and guided through a CCD type photoelectric conversion array into an AD converter 6. When a colony of the desired color tone is found, the spectrum is memorized in memory 7 as the standard color signals and the comparer 8 amplifies the difference between the standard signals and the signals from a scanned colony. When the output from the comparer 8 is almost zero or within a prescribed value all over range of wavelength, the driving system 11 is operated to pick up and transplant it.

Description

[Detailed Description of the Invention] [Field of Industrial Application] The present invention is based on a method in which colonies of a desired color are once selected and determined by human visual observation from a petri dish culture medium in which colonies of various bacteria are grown in a mixed state. Then, the present invention relates to an automatic colony transplantation device that can automatically select and collect only colonies of that color and transplant them onto a culture medium in a separately prepared test dish without any manual effort.

[Conventional technology]

Traditionally, new bacterial species were transplanted and cultured using the following steps. That is, (a) collecting soil, (b) diluting it with water,
(C) Planting on an agar medium in a culture dish, (d) Cultivating in a heat insulator for a certain period of time, (e) Visually determining colonies of various bacteria that have grown on the agar medium, (f) Colonies with the desired color. (g) Transfer to an agar medium in a test dish or test tube; (h) Cultivate and repeat various tests if the bacteria is promising. The process consists of seven steps: application to the field. In the above process, the visual identification, collection, and transplantation of colonies of specific bacteria that appear promising are performed manually.
They were forced to do tedious and complicated manual work.

On the other hand, in the past, the X-Y stage on which the culture dish was placed was automatically moved to scan the surface of the culture dish in a sequential manner so that the surface of the culture dish could be observed without omission, or the culture medium in the culture dish was directly scanned with the naked eye or through a microscope. Instead of observing the top of the culture dish, a color television camera is used to sequentially photograph parts of the top of the culture culture medium, and the greatly enlarged image is played back on a monitor color television receiver, reducing the fatigue caused by visual observation. Humans do everything up to the selection of colonies, but at the stage of selection, when a person gives a command to the control system, the collection means automatically collects the selected colonies and transports them to another location. Attempts have been made to develop devices that partially reduce human labor and fatigue, such as automatically transplanting cells into test petri dishes or test tube culture media. However, even the most advanced conventional devices of this kind cannot select colonies of a desired color by simply using a monitor color television receiver screen to display the observed and sorted color and the memory. Colonies of colors recognized as equivalent are automatically collected by comparing them with color samples, and for delicate color selection, color television cameras and monitor color television reception Machines etc. intervene,
Considering the color expression mechanisms of these devices, there was some concern as to whether colonies of the desired color could be selected accurately.

[Problem that the invention seeks to solve]

An object of the present invention is to provide an automatic colony transplanting device that can accurately select colonies of a desired color, including subtle intermediate colors, and automatically perform transplantation work, without the problems of the prior art. shall be.

[Means for solving problems]

In order to solve the above problems of the conventional technology, in the present invention, a sample cultured with a mixture of various bacteria is treated with a white light source that is correctly controlled to give a predetermined brightness and a predetermined spectral distribution. First, the reflected light is visually observed by a human, and at the same time, it is separated by a diffraction grating to detect the spectral distribution, which is photoelectrically converted using a COD with a large number of pixels in the - dimension, and then A-D conversion. However, when a human continues to visually observe a bacterial colony of a desired color, the reflected light from that colony is reflected as described above.
The result of the A-D conversion is stored in a memory and used as a reference hue signal. Once the reference hue signal is stored in the memory, the control system of the device controls the Petri dish in which the sample was cultured.
The stage on which it is mounted is moved by a small distance, and the color measurement and scanning of each position on the culture medium surface in the Petri dish (including, of course, the colonies that have grown there) is started.

If a hue signal (reflected light is separated by a diffraction grating and subjected to photoelectric conversion and A-D conversion) is obtained at a certain position, when compared with the reference hue signal in memory, the difference is within a predetermined range. , it is assumed that a colony of the desired color exists on the surface of the Petri dish medium at that position, and the control system interrupts the scanning and activates the collection means to collect the colony at that position and move it to another position in advance. Transplant the cells onto the culture medium of the test petri dish prepared in advance. During scanning, if the difference between the hue signal obtained at a certain position and the reference hue signal is outside a predetermined range, the control system assumes that a colony of the desired color does not exist at that position and continues scanning. do.

[Effect]

As mentioned above, considering the purpose of color television and the process of improvement, doubts remain regarding the reproduction of faithful primary colors, especially subtle intermediate colors. On the other hand, if the light reflected from the target colony is separated into spectra using a diffraction grating as in the apparatus of the present invention, the most accurate and well-resolved spectral distribution can be obtained. CCD
Currently, the number of pixels in the -type photoelectric conversion array is 20.
00 is commercially available, and is sufficient for accurately and faithfully photoelectrically converting the above spectral distribution. This C again
The processing speed of the OD array is also sufficiently high.

The reason for performing A-D conversion is that subsequent processing, especially storage, becomes easier. It is well known that large-capacity, high-speed memories using semiconductors are now readily available.

Note that if accurate and well-resolved hue information is to be obtained using the diffraction grating, sufficient attention must be paid to the light source that illuminates the sample. But 1. In this field, white lamp light sources have been perfected for a long time, providing light with constant brightness and a constant spectral distribution as long as they are properly controlled. Furthermore, there is no problem because a sufficiently high-performance microcomputer can currently be used for the control system of the entire device.

Note that, in reality, when selecting desired colonies, not only the hue but also the shape is a condition, but the shape can be sufficiently handled with current pattern recognition technology and is not a target of the present invention.

〔Example〕

FIG. 1 shows a schematic basic configuration of an embodiment of the present invention. In the figure,
1 is a sample culture dish, 2 is an X equipped with a culture dish
-Y stage, 3 is a white light source lamp, 4 is a diffraction grating, 5
is a CCD type photoelectric conversion array, 6 is an A-D converter, 7 is a memory, 8 is a comparator, 9 is a control system, 10 is a test dish,
11 is a drive system of the collection means, 12 is a test petri dish conveyor,
13 is a pickup, 14 is an objective lens, 15 is a visual observer's eye, 16 is a translucent mirror, 17 is an eyepiece lens, 18 is a variable aperture, 19 is a condenser lens, 20 is an aperture, 21
2 is a semi-cylindrical lens, and 22 is a prism type semi-transparent reflecting mirror.

Various types of bacteria are cultured on the agar medium in the culture dish 1, and they multiply on the medium to form colonies.

This culture dish 1 is placed on an X-Y stage 2, and can be moved back and forth perpendicularly to the plane of this figure, and left and right parallel to the plane of the figure while accurately positioning it by minute distances. The lamp 3 is controlled by a control means (not shown) so that it emits light of a constant brightness and a constant spectral distribution. The white light from this lamp 3 is reflected by a semi-transparent mirror 16, passes through an objective lens 14, and illuminates a circular area of, for example, 1 to 3 squares in diameter on the culture medium surface of the culture dish 1.

The reflected light from the irradiated position passes through the semi-transparent mirror 16 and reaches the prism-type semi-transparent reflecting mirror 22, where it is divided into two. Using one of the lights, a human observes with the eye 15 through the eyepiece 17 and selects bacteria with a desired hue and shape.
Adjust the variable apertures 18. The other half of the light is
The light is converted into parallel light by a condenser lens 19 and reaches the diffraction grating 4. The light that has been separated and spread out is formed into a linear image by a semi-cylindrical lens 21 and enters a CCD type photoelectric conversion array 5, where it is converted into a voltage proportional to the spectral intensity. This CCD type photoelectric conversion array 5 corresponds to approximately 1400 pixels and operates at high speed. The memory 7 uses a diffraction grating 4 to separate the reflected light from a colony of a promising color discovered through visual observation by the operator, converts it into a voltage output using a photoelectric conversion array 5, and further converts it into a voltage output using an A-D converter 6. The digital value of the output voltage obtained is stored as a reference hue signal. Once the reference hue signal is stored in the memory 7, the operator thereafter causes the control system 9 to automatically start measuring and scanning the color of the medium surface of the culture dish 1. A comparator 8 uses the reference hue signal stored in the memory 7 (this signal value remains constant until the transplantation of colonies of the selected hue is completed) and the signal from the position during scanning of the culture medium in the culture dish. The difference with the hue signal (which typically changes each time the stage moves and scanning continues) is amplified. here,
The reference hue signal input from the memory 7 to the comparator 8 and the hue signal from a position where no colonies of the desired color are present are usually
It is as shown in Figure 2. Therefore, for reflected light from a colony of a desired color, the output of the comparator 8 is almost non-existent or small, but the hue signal from a position where a colony of a desired color does not exist has a wavelength that happens to be There may be cases where the difference is small, but when viewed over the entire wavelength range, the difference in signals is large and falls outside the predetermined range. The control system 9 of this entire device determines that if the output of the comparator 8 is almost non-existent over the entire wavelength range or has a small value within a predetermined range, a colony of the desired color exists at a position that provides such an output. The scanning is temporarily interrupted, the drive system 11 of the collecting means is activated, the pink up 13 (equipped with a platinum loop, etc.) is lowered to collect colonies from the medium surface of the culture dish 1, and the next step is to Then, the pickup is raised, rotated, and lowered to the center of the medium in the test Petri dish 10, where this colony is transplanted. After that, the control system 9
restarts scanning and moves the XY stage 2 slightly to the next position.

Further, if the output of the comparator 8 is large in the entire wavelength range and is outside the predetermined range, the control system 9 assumes that a colony of the desired color does not exist at that position and continues scanning.
Command the X-Y stage 2 to move slightly to the next position. The test dish 10 to which colonies of a desired color have been transplanted by the Binoqua knob 13 is carried away by the carrier 12, and another test dish is prepared at the same position. In addition, the part of the pickup 13 where the colony is directly collected or transplanted (for example, the end of the platinum loop) is sterilized and disinfected by means not shown after the transplant operation is finished and before it returns to the collection position again. It is intended to be manipulated. The above operations are repeated until the entire surface of the culture dish is scanned.

〔Effect of the invention〕

As explained above, according to the present invention, a person can initially select and designate a specific desired color from a culture dish in which various bacteria have proliferated to form colonies, and the device can then automatically select the desired color. Colonies of that color are selected and transplanted with great precision.

[Brief explanation of the drawing]

FIG. 1 is a schematic basic configuration diagram of an embodiment of the present invention, and FIG. 2 is a diagram showing the spectral distribution (hue signal converted into voltage) of reflected light from various colonies. - Culture dish, 2-.-XY stage, 3- White light source lamp, 4- Diffraction grating, 5-- CCD type photoelectric conversion array, 6- A-D converter, 7-, Memory, 8- Comparison 9-control system, 10...test petri dish, 11-drive system, 13-pickup, 15-operator's eyes, 16-
- Semi-transparent mirror, 21- Semi-cylindrical lens.

Claims (1)

[Claims] The petri dish in which various samples were grown on the medium was
, placed on a stage that can be moved in the Y direction and precisely positioned.First, while visually observing the culture medium surface, move the petri dish along with the stage to select colonies of the desired color, and then collect the reflected light of the colonies using a diffraction grating. After spectroscopy, the photoelectric conversion and A-D conversion are performed and stored in the memory as reference hue signal values.Then, the control system of this device automatically moves the stage by small distances in order to The color of the surface of each part of the culture medium is measured and scanned, and the color is measured at each position.
Compare the measured hue signal value and the reference hue signal value above,
If the difference is within a predetermined range, it is assumed that a colony of the desired color exists at that position, the scanning is interrupted, the collection means is activated, the colony is collected from that position, and the colony is placed in a test dish prepared at another position. A colony is automatically transplanted onto a culture medium, and if the above-mentioned difference is outside a predetermined range, it is assumed that no colony of the desired color exists at that position, and scanning continues as it is. Automatic transplant device.