JPS63290946A - Cell identification device - Google Patents

Abstract

PURPOSE:To enable easy identification of cells by using ample identification parameters by picking up the microscopic images of the cells via plural freely selectable interface filters, subjecting the respective spectral images to image processing and outputting the information relating to the respective wavelengths, densities and picture elements. CONSTITUTION:A cell group is put into the visual field of a microscope (a) and is displayed on the image plane of a display means (d). A marker is put to the desired cell by a marker applying means (e). An image processing means (f) executes the image processing with the cell image attached with the marker from the position data of this marker. The interference filter b1 are successively changed at this time ans the processing with the spectral images obtd. via the respective filters is executed. The obtd. in association to the respective wavelengths based on the respective interference filters bi; namely, the information, the information relating to the densities picture elements-wavelengths is eventually obtd. The ample identification parameters are thus obtd. and the identification is facilitated.

Description

[Detailed Description of the Invention] <Industrial Application Field> The present invention can be used, for example, to distinguish fused cells from other cells in cell fusion, to distinguish cells after gene introduction from other cells,
Alternatively, the present invention relates to a cell identification device that can be used to identify live cells and dead cells.

<Prior Art> Conventionally, a device called a cell sorter is known as a device for selecting a specific cell from a large number of cells or classifying cells. A cell sorter is a device that stains specific cells in advance with a fluorescent dye and identifies the specific cells based on the amount of fluorescence when irradiated with laser light.

<Problems to be Solved by the Invention> By the way, in the above-mentioned cell sorter, cells are put into a droplet one by one for identification operation, but in order to form this droplet,
It is necessary to inject the cells along with the medium from the nozzle. Upon ejection from this nozzle, the cells are subjected to a pressure shock. Therefore, there is no problem with relatively robust animal cells, but there is a problem with fragile cells such as protoplast-formed plant cells, which are broken by this pressure impact.

Furthermore, it has not been easy to apply a cell sorter to plant cells because plant cells are difficult to stain with fluorescent light.

Furthermore, conventional cell sorters require expensive laser light sources and complicated mechanisms, and are extremely expensive.

Although it is conceivable to use an image analysis method for cell identification as described above, cells have an extremely wide range of variation in their morphology, so it is difficult to identify them from mere morphological information.

The present invention has been made in view of the above, and aims to provide a relatively inexpensive cell identification device that is capable of identifying cells without applying pressure shock or the like to cells and without applying staining or the like.

<Means for Solving the Problems> The configuration for achieving the above object will be explained with reference to the basic conceptual diagram shown in FIG. 1. The present invention comprises a plurality of selectable interference filters b, b2..., an image capturing means C that captures the image of the microscope a through the selected interference filter b, a display means d that displays the image taken by the image capturing means C, and a screen of the display means d. The image processing means f is equipped with a marker applying means e that generates a marker at an arbitrary position, and an image processing means f that inputs image data from the imaging means C and position data of the applied marker in the screen, and this image processing means f:
By configuring the display device d to output information regarding the density of each pixel in each selected interference filter b and spectral image of the cell to which a marker has been added on the screen of the display means d. , characterized.

<Operation> A group of cells is placed within the field of view of the microscope a and displayed on the screen of the display means d, and a marker is attached to the desired cells by the marker attaching means e. From this marker position data, the image processing means f performs image processing on the cell image with the marker attached. At this time, the interference filter b is sequentially changed and the spectral image obtained through each filter is processed. By doing this, information regarding one density pixel is obtained in relation to each wavelength based on each interference filter b, that is, density -
Information related to pixels and wavelengths can be obtained, and identification parameters are abundant and identification becomes easy.

<Example> Embodiments of the present invention will be described below based on the drawings.

FIG. 2 is a block diagram showing the configuration of an embodiment of the present invention.

The optical image of the microscope 1 is obtained by using an interference filter 2a, 2b or 2c.
It becomes a spectral image through the microchannel plate 3.
After being intensified by an image intensifier such as, the image is input to the TV camera 4.

The interference filters 2a, 2b, and 2c are each mounted on a turret stand that is rotationally driven by a turret drive device 5, and the one selected by the filter selection signal from the arithmetic control unit 6 is used for the microscope 1 and the microchannel plate 3. It is configured to be interposed between.

The video signal from the TV camera 4 is sent to an image synthesis circuit 7,
It is distributed to a gate circuit 8 and a switch circuit 9.

The image synthesis circuit 7 combines the video signal from the TV camera 4,
A drawing signal from the light pen 10, which will be described later, and identification data and a character signal, which will be described later, from the arithmetic control section 6 are combined, and the output thereof is supplied to the CRT 124.

The gate circuit 8 receives the above-mentioned video signal and drawing signal, passes only the video signal corresponding to the position corresponding to the drawing position by the light ben 10, and passes the video signal to the switch circuit 9.
supply to.

The switch circuit 9 is a circuit for selecting which of the video signal of the entire image from the TV camera 4 and the above-mentioned partial video signal passed through the gate circuit 8 is to be input to the A-D converter 13. The selection status (A or B) of this switch circuit 9 corresponds to two types of modes of the arithmetic control unit 6, which will be described later.

The A-D converter 13 can digitize the input video signal based on a conversion command from the arithmetic control section 6 and supply it to the arithmetic control section 6 as image data.

Drawing on the CRT12 screen using Lightben 10 is as follows:
It is written into the 1-bit frame memory 11 and is also incorporated into the arithmetic control section 6 as well as the image synthesis circuit 7 and gate circuit 8 described above as a drawing signal representing the drawing position.

The arithmetic control unit 6 is mainly composed of a computer, and uses image data from the A-D converter 13 and a drawing signal from the 1-bit frame memory 11 to adjust the selection status A or B of the switch circuit 9. Performs image processing to be described later in a corresponding mode, displays the results as identification data, etc. via the image synthesis circuit 7, and outputs the aforementioned filter selection signal, conversion command, etc. to control the entire apparatus. Can be done.

Next, the action will be explained.

When selecting a mode in which data collection for automatically identifying cells and automatic identification after an identification algorithm is determined based on the collected data are performed, the switch circuit 9 is switched to the A side. .

When selecting this mode to collect data,
First, place a group of cells under the field of view of microscope 1 and
While selecting a predetermined interference filter 2a, the operator looks at the display screen and attaches a marker to an arbitrary cell image using the light ben 10.

The arithmetic control unit 6 calculates the cell image to which the markers have been assigned as follows based on the image data for the plan surface on the captured CRTf2 and the image creation signal supplied via the 1-bit frame memory 11. Perform processing. That is, a density histogram of pixels constituting the cell image to which markers have been given is created and stored in the computer's memory. Next, a filter selection signal is outputted to drive the talent driving device 5, which sequentially selects the interference filter 2b.

After selecting 2C and creating a similar density histogram,
For example, data as shown in FIG. 3, that is, wavelength λ8. λ
5 and λ. Data constituted by a density histogram for each spectral image, in other words, information on the number of pixels, density, and wavelength may be displayed on the CRT 12, or may be output to a separately connected X-Y plotter or the like. Note that during this process, morphological information such as the nucleus/cytoplasm area ratio may be obtained for each spectral image and stored and displayed at the same time.

After collecting data for multiple cells as described above, the operator displays the data for each cell on the CRTf2 or looks at the data group printed by an is found and input to the arithmetic control section 6. This identification algorithm is stored in memory and automatic identification operations are subsequently performed in the same mode. This operation automatically collects the same data as above for the cell image displayed on the CRTf2, and uses the stored identification algorithm to automatically determine whether or not it is a specific cell. The results are displayed on the CRT 12 or the like.

When selecting the mode to manually identify cells,
The switch circuit 9 is sent to the B side.

This mode is selected when cells are to be identified from the screen of the CRT 12 based on the operator's judgment, but it is difficult to identify cells by visual inspection of the screen.

In this mode, first, a predetermined interference filter 2a is
As shown in FIG. 4, the operator draws a linear marker M that intersects the desired cell image using the light pen 10 on the screen of the CRT 12. As a result, the arithmetic control section 6 receives this marker M via the gate circuit 8.
Image data of only the video signal of the portion corresponding to the position is sent. The arithmetic control unit 6 uses the image data to determine the relationship between the one-dimensional position along the marker M and the pixel density. Next, the interference filters 2b and 2c are used to find a similar relationship, and as shown in FIG. 5, the position-concentration-wavelength information is displayed on the CR 112 or output to an X-Y plotter or the like. The operator can perform cell identification based on this information.

Note that a mouse can be used in addition to a light pen as a means for applying markers, and it goes without saying that the number and type of interference filters can be changed as desired.

In addition to identifying individual cells as described above, the present invention can also be used for cutting! It can also be used for sorting 1KII textures, identifying specific parts within tissues, etc.

<Effects of the Invention> As explained above, according to the present invention, a microscopic image of a cell is captured through a plurality of selectable interference filters, and each spectral image of a cell is image-processed through each interference filter. Because it is configured to output information related to each wavelength, concentration, and pixel, there is no need to apply harmful staining to cells as with conventional cell sorters, and it is possible to use a rich array of data based on multiple spectral image data that cannot be obtained visually. It is now possible to easily identify them using the appropriate identification parameters. Furthermore, since external forces such as pressure shocks are not applied to the cells, they are not destroyed during identification. Furthermore, since no complicated mechanism is required, it is advantageous in terms of cost.

Furthermore, if the system is configured to perform image processing in two modes as in the embodiment shown in FIG. 2, it can be used for both automatic identification and identification based on human judgment such as in experiments.

[Brief explanation of the drawing]

FIG. 1 is a basic conceptual diagram showing the configuration of the present invention, FIG. 2 is a block diagram showing the configuration of an embodiment of the present invention, and FIGS. 3, 4, and 5 are explanatory diagrams of the functions thereof. 1...Microscopes 2a, 2b, 2c...Interference filter 4...TV camera 6...Arithmetic control section 7...Image synthesis circuit 8...Gate circuit 9...Switch circuit 10...・Light pen 11...1 bit frame memory 12...CRT 13...A-D converter Patent applicant: Shigeo Minami, Shimadzu Corporation

Claims (4)

[Claims] (1) A device for identifying cells placed under the field of view of a microscope, comprising a plurality of selectable interference filters, an imaging means for capturing a microscope image through the selected interference filters, and A display means for displaying an image taken by the imaging means, a marker adding means for producing a marker at an arbitrary position within the screen of the display means, and inputting image data from the imaging means and position data of the attached marker on the screen. The image processing means outputs information relating to concentration-pixel in each image with respect to the spectral image of the cell to which the marker is attached on the screen, obtained by each selected interference filter. A cell identification device, characterized in that it is configured to obtain. (2) The cell identification device according to claim 1, wherein the density-pixel information is a density histogram with the number of pixels as a frequency. (3) The cell identification device according to claim 1, wherein the density-pixel information is information on pixel density with respect to a position within a cell image. (4) a storage means for storing an identification algorithm created based on the information related to the density-pixels; The cell identification device according to claim 1 or 2, characterized in that it comprises arithmetic means for automatically identifying cells.