WO2019012573A1 - Cell image acquisition device and cultured cell management method - Google Patents

Abstract

The purpose of the present invention is to identify cells in a culture vessel without confusion and without the need to deploy a special device for identifying culture vessels such as a barcode reader, while also dispensing with the chore of assigning identification information for identifying cells. The cell image acquisition device (1) is provided with: a condensing lens (7) which is disposed below the bottom surface (2a) of a culture vessel (2) for culturing cells (X) and collects light that passes through the bottom surface; an image acquisition unit for detecting light collected by the condensing lens and acquiring an image; a focus adjustment mechanism (9) for vertically moving the focal position of the condensing lens; a vessel identification unit for extracting identification information based on features of the culture vessel itself from an image acquired by the image acquisition unit with the condensing lens focused on the bottom surface by the focus adjustment mechanism; and a storage unit for storing, in association with the identification information extracted by the vessel identification unit, the cell image acquired by the image acquisition unit with the condensing lens focused on the bottom surface by the focus adjustment mechanism.

Description

Cell image acquisition apparatus and cultured cell management method

The present invention relates to a cell image acquisition apparatus and a cultured cell management method.

A method of attaching identification information such as a bar code to a culture container or attaching identification information to a place where the culture container is disposed in order to observe cells cultured in a plurality of culture containers without mistaking them. Are known (see, for example, Patent Document 1 and Patent Document 2).

JP 2017-23055 A JP 2007-330144 A

However, according to the method of Patent Document 1, it is necessary to separately prepare a barcode reader for reading identification information or to attach barcodes to all the culture vessels, and every time when observing cells in the culture vessel. There is a disadvantage that the reading operation by the bar code reader has to be performed in order to associate the observation result with the culture vessel.
In addition, the method of Patent Document 2 has a disadvantage that if the place where the culture vessel is placed is wrong, the cells in the culture vessel may be mixed up.

The present invention has been made in view of the above-described circumstances, and can save time and effort for attaching identification information for identifying cells, and a special device such as a barcode reader for identifying a culture container. It is an object of the present invention to provide a cell image acquisition apparatus and a cultured cell management method which do not need to be prepared and can identify cells in a culture container without mistake.

In one aspect of the present invention, a condensing lens is disposed below the bottom surface of a culture vessel for culturing cells, and the condensing lens that condenses the light transmitted through the bottom surface, and the light collected by the condensing lens is detected An image acquisition unit for acquiring an image, a focusing mechanism for moving a focal position of the focusing lens in the vertical direction, and a focusing mechanism for focusing the focusing lens on the bottom surface A container identification unit that extracts identification information based on the characteristics of the culture container itself from the captured image, and the focus adjustment mechanism focuses the cell on the cell by the focus adjustment mechanism to obtain the cell It is a cell image acquisition device provided with a storage part which matches and memorizes an image and the above-mentioned discernment information extracted by the above-mentioned container discernment part.

According to this aspect, by adjusting the focus position of the condensing lens in the vertical direction by the focusing mechanism to align with the cells in the culture vessel, an image of the cell is acquired by the image acquisition unit, and the focusing mechanism is condensed. By aligning the focal position of the lens to the bottom surface, an image of the bottom surface is acquired.

Then, based on the acquired bottom surface image, the container identification unit extracts identification information based on the characteristics of the culture container itself, and the extracted identification information is associated with the cell image and stored in the storage unit.
That is, since the identification information based on the characteristics of the culture container itself is extracted from the image of the bottom surface of the culture container, it is possible to save the trouble of attaching identification information such as a barcode to the culture container and special The identification information can be acquired and the cells can be managed by using an image acquisition unit that acquires an image of the cells without preparing a suitable reader.

In the above aspect, the identification information may be a flaw on the inner surface, the outer surface or the inner surface of the bottom surface of the culture vessel.
By doing this, identification information based on the characteristics of the culture container itself can be extracted from the bottom surface flaw that can be detected from below the culture container, and cells cultured in the culture container can be managed.

In the above aspect, the identification information may be a strain on the bottom surface of the culture vessel.
By doing so, identification information based on the characteristics of the culture container itself can be extracted from the strain on the bottom surface that can be detected from below the culture container, and the cells cultured in the culture container can be managed.

In another aspect of the present invention, there is provided a cell image acquisition step of focusing on the cells and acquiring an image of the cells below the bottom surface of the culture container for culturing the cells, and below the bottom surface of the culture container. Identifying the information based on the characteristics of the culture vessel from the bottom image acquisition step of focusing on the bottom surface to acquire an image of the bottom surface and the image of the bottom surface acquired in the bottom image acquisition step A cultured cell management method comprising an identification information extraction step, and a storage step for storing the identification information extracted in the identification information extraction step and the image of the cell acquired in the cell image acquisition step in association with each other. is there.

According to the present invention, it is possible to save time and effort for attaching identification information for identifying cells, and it is not necessary to prepare a special device such as a barcode reader for identifying a culture container, and it is possible to It has the effect that cells can be identified without confusion.

BRIEF DESCRIPTION OF THE DRAWINGS It is a whole block diagram which shows the cell image acquisition apparatus which concerns on one Embodiment of this invention. It is a block diagram explaining the control part of the cell image acquisition apparatus of FIG. It is a flowchart which shows the cultured cell management method using the cell image acquisition apparatus of FIG.

A cell image acquisition apparatus 1 and a cultured cell management method according to an embodiment of the present invention will be described below with reference to the drawings.
As shown in FIG. 1, the cell image acquisition apparatus 1 according to the present embodiment includes a stage 3 for placing the culture vessel 2 having a transparent bottom surface 2 a for accommodating and culturing the cell X together with the medium Y; And a control unit 5 that controls the imaging unit 4.

The stage 3 has a window 6 for transmitting light in at least a part of the portion on which the culture vessel 2 is placed.
The imaging unit 4 includes a condenser lens 7 for condensing light transmitted downward through the window 6 of the stage 3 from the side of the culture vessel 2 placed on the stage 3 and the condenser lens 7. Imaging device (image acquisition unit) 8 that detects an incident light and acquires an image, a focusing mechanism 9 that vertically adjusts the focal position of the condensing lens 7, the condensing lens 7, the imaging device 8, and the focal point A slider 10 on which the adjusting mechanism 9 is mounted, and a moving mechanism 11 for moving the slider 10 in the horizontal two-dimensional direction.

As shown in FIG. 2, the control unit 5 includes an image processing unit 12 that processes an image acquired by the imaging device 8, a storage unit 13 that stores an image, an imaging device 8, a focusing mechanism 9, and a moving mechanism. 11 includes an operation control unit (container identification unit) 14 that controls the image processing unit 12 and the storage unit 13.

The operation control unit 14 first operates the moving mechanism 11 to move the imaging range of the imaging device 8 to a predetermined position, and operates the focusing mechanism 9 to focus the focusing lens 7 on the bottom surface 2 a of the culture vessel 2. Match to Then, in this state, the image pickup device 8 is operated to obtain an image of the bottom surface 2 a of the culture container 2.

Thereafter, the operation control unit 14 causes the image processing unit 12 to process the image of the bottom surface 2 a to extract identification information based on the features of the culture container 2 itself included in the image of the bottom surface 2 a.
Here, as identification information based on the characteristics of the culture vessel 2, a scratch on the inner surface side of the bottom surface 2 a of the culture container 2, a scratch on the outer surface side of the bottom surface 2 a, a distortion of the bottom surface 2 a, or the material constituting the bottom surface 2 a It means the pattern of air bubbles present in.

Then, the operation control unit 14 determines whether or not the extracted identification information matches any of the identification information stored in the storage unit 13, and in the case of agreement, the image of the cell X to be photographed from now is It is determined to be the cell X in the culture container 2 stored. On the other hand, when the extracted identification information is not stored in the storage unit 13, the operation control unit 14 determines that the image of the cell X to be captured from now is the cell X in the new culture container 2. It is designed to judge.

Next, the operation control unit 14 operates the moving mechanism 11 to move the imaging element 8 to a position where the cell X is included in the imaging range of the imaging element 8 and operates the focusing mechanism 9 to focus the focusing lens 7. Match cell X. Then, in this state, the image pickup device 8 is operated to obtain an image of the cell X in the culture container 2.

The operation control unit 14 adds the image of the cell X acquired by the imaging device 8 to the image of the cell X stored in the storage unit 13 in association with the same identification information, and associates the image with the identification information. Or stored in the storage unit 13 in association with new identification information.

A cultured cell management method using the cell image acquisition device 1 according to the present embodiment configured as described above will be described below.
The cultured cell management method according to the present embodiment is a cell image in which the operation control unit 14 operates the focus adjustment mechanism 9 to focus the cell X from below the bottom surface 2a of the culture container 2 and acquire an image of the cell X Based on the acquisition steps S8, S9, S10, the bottom image acquisition step S3 of focusing on the bottom surface 2a of the culture vessel 2 to acquire an image of the bottom surface 2a, and the acquired image of the bottom surface 2a Storage step for storing identification information extraction step S4 for extracting identification information based on the extracted identification information and the image of cell X acquired at cell image acquisition steps S8, S9 and S10 in the storage unit 13 And S11.

Specifically, as shown in FIG. 3, first, the operation control unit 14 operates the moving mechanism 11 to move the slider 10 to a predetermined first position (step S1), and operates the focusing mechanism 9 The focus position of the condenser lens 7 is made to coincide with the bottom surface 2a of the culture vessel 2 (step S2), and the image pickup device 8 is operated to acquire an image of the bottom surface 2a (bottom surface image acquisition step S3). Then, the image processing unit 12 processes the acquired image to extract identification information based on the characteristics of the culture container 2 itself (identification information extraction step S4).

The operation control unit 14 determines whether or not the extracted identification information is stored in the storage unit 13 by searching the inside of the storage unit 13 (step S5). Is identified as the same existing cell X as the acquired and stored cell X (step S6). If not stored, it is determined whether or not the slider 10 is to be moved (step S13). If the slider 10 is not moved, it is determined that the cell X is a new cell X for which an image is newly obtained. A storage area is generated in the new identification information (step S7).

In addition, when moving the slider 10, the process from step S1 is repeated. By moving the slider 10 to change the imaging position of the bottom surface of the culture vessel 2 and repeating the process a plurality of times, a new scratch, dirt, or the like adheres to the scratch already stored and is already stored. In the case where it is difficult to discriminate a scratch or the like at the first imaging position, the discrimination can be made based on the scratch or the like at the next imaging position.

Then, the operation control unit 14 operates the focusing mechanism 9 to make the focal position of the condenser lens 7 coincide with the cell X in the culture vessel 2 (step S8), and operates the moving mechanism 11 to change the slider 10 While sequentially moving to the imaging position (step S9), the image pickup device 8 is operated to acquire an image of the cell X (step S10). When the identification information is already stored in the storage unit 13, the acquired image is added to the storage area of the same identification information in the storage unit 13 and stored, and the identification not existing in the storage unit 13 is performed. If it is the information, the acquired image is stored in the storage area of the newly generated identification information (storage step S11). It is determined whether all the images of the cell X have been acquired (step S12), and if it has not ended, the process from step S9 is repeated.

As described above, according to the cell image acquisition device 1 and the cultured cell management method according to the present embodiment, a pattern such as a scratch, a strain or an air bubble provided on the bottom surface 2a of the culture container 2 is recognized as identification information. There is no need to attach a code or other identification information to each culture vessel 2. Also, in the case of a barcode, it is attached to a place different from the bottom 2a so as not to disturb the observation, so a special device such as a barcode reader different from the imaging unit 4 for observing the cell X from the bottom 2a side Although it is necessary to attach, according to the cell image acquisition device 1 according to the present embodiment, there is an advantage that the imaging unit 4 for acquiring the image of the cell X can be used for acquisition of identification information. . As a result, no special device such as a bar code reader is required.

In the present embodiment, as the identification information based on the characteristics of the culture container 2 itself, the scratch on the inner surface side of the bottom surface 2a of the culture container 2, the scratch on the outer surface side, the distortion of the bottom surface 2a or the air bubble inside the bottom surface are illustrated. All of these patterns may be used as identification information, or one or more may be used in combination as identification information.

In addition, it is judged whether the existing cell X is or not by using a scratch on the inner surface side of the bottom surface 2a, a scratch on the outer surface side, a distortion of the bottom surface 2a or a bubble pattern inside the bottom surface 2a. If it is determined that is, it may be determined again using any other pattern.

In addition, although an illumination method at the time of image acquisition has not been particularly described, any illumination method such as transmissive illumination, epi-illumination, oblique illumination, dark-field illumination can be adopted. In addition, the oblique illumination or phase contrast observation method suitable for observation of transparent cells X is preferable because it can detect flaws, distortions or bubbles of the bottom surface 2a clearly.

In addition, in the storage unit 13, identification information including a flaw on the inner surface side of the bottom surface 2a, a flaw on the outer surface side, a distortion of the bottom surface 2a or a bubble pattern inside the bottom surface 2a is stored in correspondence with the image of the cell X However, in addition to this, easily identifiable identification information such as a user name and a serial number may be stored in association with each other. Then, when identification information having the same pattern is found by searching the inside of the storage unit 13, identification information that is easy to identify may be displayed on a monitor or the like.

Further, in the present embodiment, the image pickup device 8 is horizontally moved by the operation of the moving mechanism 11 to acquire a plurality of images of the partial visual field range of the bottom surface 2a. The moving mechanism 11 may not be necessary when acquiring.

1 Cell Image Acquisition Device 2 Culture Container 2a Bottom 7 Focusing Lens 8 Imager (Image Acquisition Unit)
9 focus adjustment mechanism 13 storage unit 14 operation control unit (container identification unit)
X cell S8, S9, S10 Cell image acquisition step S3 Bottom image acquisition step S4 Identification information extraction step S11 Memory step

Claims (4)

A condenser lens disposed below the bottom surface of a culture vessel for culturing cells, and collecting the light transmitted through the bottom surface;
An image acquisition unit that detects light condensed by the condenser lens and acquires an image;
A focusing mechanism for moving the focus position of the focusing lens in the vertical direction;
A container identification unit that brings the focus of the condensing lens to the bottom surface by the focusing mechanism and extracts identification information based on the characteristics of the culture container itself from the image acquired by the image acquisition unit;
A memory that matches the image of the cell acquired by the image acquisition unit with the focus of the condenser lens on the cell by the focusing mechanism and stores the identification information extracted by the container identification unit in association with each other A cell image acquisition apparatus comprising: The cell image acquisition apparatus according to claim 1, wherein the identification information is a flaw on an inner surface, an outer surface or an inner surface of the bottom surface of the culture vessel. The cell image acquisition apparatus according to claim 1, wherein the identification information is a strain on the bottom surface of the culture vessel. A cell image acquisition step of focusing on the cells and acquiring an image of the cells below a bottom surface of a culture container for culturing the cells;
A bottom image acquisition step of focusing on the bottom surface and acquiring an image of the bottom surface below the bottom surface of the culture vessel;
An identification information extraction step of extracting identification information based on the characteristics of the culture vessel itself from the image of the bottom surface acquired in the bottom surface image acquisition step;
A culture cell management method comprising: storing the identification information extracted in the identification information extraction step in association with the image of the cell acquired in the cell image acquisition step.

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