JP2006017489A - Cell identification device, cell identification program, cell analysis device, and cell analysis program - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To identify each cell from a picked up image of a cell. <P>SOLUTION: Edge extraction processing is applied to a region including any one cell existing in an image specified by a user in a phase difference image of the cell (step S20), and a profile of the cell is obtained (step S30). Then, the profile of the extracted cell is identified as one cell (step S40). <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention captures a cell identification device, a cell identification program, and a cell identification program for identifying a cell by extracting a cell outline from the captured cell image, and processing and analyzing the captured image The present invention relates to a cell analysis device and a cell analysis program.

  Patent Document 1 discloses a cell image processing method that extracts an outline of a cell by binarizing an image obtained by imaging a cell.

JP-A-5-274422

  However, the cell image processing method described in Patent Document 1 has a problem that two or more cells connected to each other on the image may be erroneously extracted as one cell.

According to the first aspect of the present invention, in a cell identification device for identifying individual cells existing in an image obtained by imaging an observation target, display means for displaying an image of the captured cell, and display on the display means The image processing apparatus includes: designation means for designating an area in the cell image, and identification means for identifying a cell by assuming that one cell exists in the area designated by the designation means.
According to a second aspect of the present invention, in the cell identification device according to the first aspect, the identification unit extracts an outline of one cell by performing edge extraction on the image in the region designated by the designation unit. The extracted cell outline is distinguished from one cell.
According to a third aspect of the present invention, in the cell identification device according to the first or second aspect, an image obtained by imaging an observation target is a phase difference image or differential interference image of a cell.
The cell analysis device according to claim 4, the cell identification device according to any one of claims 1 to 3, and a plurality of images of cells at predetermined time intervals, and the plurality of images captured And analyzing means for executing time-series image processing and analyzing time-series changes of individual cells.
The invention according to claim 5 is a cell identification program for executing, by a computer, a cell identification process for identifying individual cells existing in an image obtained by imaging an observation target. Display processing for displaying the image, drawing processing for drawing a region in the displayed cell image, and identification processing for identifying a cell by assuming that one cell exists in the region drawn by the drawing processing It is characterized by having.
According to a sixth aspect of the present invention, in the cell identification program according to the fifth aspect, the identification processing is performed by extracting an edge of an image in the drawn region to extract an outline of one cell. The outline of the formed cell is distinguished from one cell.
The invention according to claim 7 is the cell identification program according to claim 5 or 6, wherein the image obtained by imaging the observation object is a phase difference image or differential interference image of the cell. .
The cell analysis program according to claim 8 is a plurality of images obtained by capturing a plurality of images of the cell identification program according to any one of claims 5 to 7 and a cell image at a predetermined time interval. And an analysis process for performing an image process on a single image and analyzing a time-series change of individual cells.

  According to the present invention, one cell can be accurately extracted, and a plurality of cells can be prevented from being mistakenly extracted as one cell.

  FIG. 1 is a block diagram showing a configuration of an embodiment of a cell identification device according to the present invention. The cell identification device 100 includes an input device 110, a control device 120, a monitor 130, and an image memory 140. The cell identification device 100 is connected to a microscope 200 with a camera that captures an enlarged image of a cell. An image of an arbitrary cell observed by the user and captured by the microscope 200 is transferred to the control device 120. Are stored in the image memory 140 and displayed on the monitor 130. The input device 110 is configured so that the user can input a region surrounded by a closed curve in the cell image displayed on the monitor 130. For example, the user can drag the region surrounded by the closed curve. It can be input.

  The microscope 200 includes a CCD 210 as an imaging device, and includes a phase difference observation optical system using transmitted light and a fluorescence observation optical system using epifluorescence. In the present embodiment, the cells are identified based on the phase difference image captured by the phase difference observation optical system, and the identified cells are observed by the fluorescence observation optical system.

  Cell identification device 100 in the present embodiment identifies individual cells included in the cell image captured by microscope 200. When imaging the cells with the microscope 200, the cells are stained with a fluorescent dye, for example, DiBAC, in order to fluorescently stain the cells. Then, an enlarged image of the stained cells is taken with the microscope 200. At this time, by changing the wavelength of the excitation light irradiated to the illumination optical system or the cell, either the phase difference image or the fluorescence image obtained by fluorescently staining the cell with the fluorescent dye can be captured.

  In the present embodiment, an inverted microscope is used as the microscope 200, and light of a predetermined wavelength is irradiated from the upper illumination system to capture a phase difference image of the cell. Then, the user designates a region including any one cell existing in the phase difference image of the cells displayed on the monitor 130 using the input device 110. FIG. 2 is a diagram illustrating a specific example when an area including an arbitrary cell existing in a cell phase difference image is designated by the user. In FIG. 2, the user designates the region 2 a including one cell by a drag operation of the input device 110. The control device 120 sets an area specified by the user as a cell identification area, performs an edge extraction process on the target image in the cell identification area, and linearly connects adjacent extracted edges. Extract cell outlines by tying.

  FIG. 3 shows a specific example of an algorithm for extracting the outline of a cell based on the result of the edge extraction process. Each point in FIG. 3 represents the edge of the cell extracted by the edge extraction process. In FIG. 3A, any two edges A and B are connected by a straight line 4a, and the edge C farthest from the straight line 4a in the vertical direction and the end points of the straight line 4a, that is, the edges A and B, are respectively connected to the straight line 4b and Tie with 4c.

  Next, as shown in FIG. 3B, the edges D and E that are furthest away from each of the straight lines 4b and 4c in the vertical direction and the respective end points of the straight lines 4b and 4c are connected by straight lines. As a result, the edges A, D, C, E, and B are connected by straight lines, and the contours are extracted. Then, in FIG. 3C, the process shown in FIG. 3B described above is performed, and then the process is repeated until all the points are connected by a straight line, whereby the user designates as shown in FIG. It is possible to extract the outline of one cell existing in the defined area. In FIG. 4, the outline 4a of one cell existing in the region 2a shown in FIG. 3 is extracted.

  As described above, by extracting the outline of a cell existing in a region including one arbitrary cell designated by the user, individual cells existing in the captured phase difference image are identified. Each identified cell is given an ID (identification code) for uniquely identifying the individual cell, and the given ID indicates the outline of the identified cell, that is, the shape of the cell in the image. The position, for example, the coordinate value of the center of gravity of the cell when the inside of the image is an XY coordinate system, is stored in a memory (not shown) as identification data of each cell. Then, the observed images of the identified individual cells can be image-processed in units of cells based on the cell identification data.

  In the present embodiment, for example, as shown in FIGS. 5 and 6, image processing is performed on observation images of individual cells using a microscope 200 to obtain the observation results. That is, in the microscope 200, the wavelength of the excitation light applied to the cell is changed to a wavelength that excites a dye (for example, DiBAC) that fluorescently stains the cell, and the fluorescence image of the cell shown in FIG. Take an image. Then, the fluorescence intensity of each cell in the fluorescence image is measured, and a graph showing the time series change is displayed on the monitor 130 as shown in FIG.

  FIG. 6 is a graph showing a time-series change in the average value of the fluorescence intensity in the contours of the two cells 5a and 5b identified in FIG. In FIG. 6, it can be seen that when the reagent is added at the time point t1, the average value of the fluorescence intensity in the contours of the cells 5a and 5b increases. For example, when the dye that fluorescently stains cells is DiBAC, it can be seen that the membrane potential of the cells changes.

  FIG. 7 is a flowchart showing the operation of the cell identification device 100 in the present embodiment. When starting observation of cells using the microscope 200, the user sets a petri dish containing cells that have been fluorescently stained in advance by adding the above-described fluorescent dye to the cells to be observed and sets the observation magnification, etc. Make preparations. The process shown in FIG. 7 is executed by the control device 120 as a program to be started when these preparations are completed and the user instructs the execution of the cell identification process.

  In step S <b> 10, an enlarged image captured by the microscope 200 is stored in the image memory 140 and displayed on the monitor 130. Thereafter, the process proceeds to step S20, and it is determined whether or not an area including any one cell existing in the phase difference image of the cell is designated by the user via the input device 110. If it is determined that the area has been designated by the user, the process proceeds to step S30.

  In step S30, an area designated by the user is set as a cell identification area, and an edge extraction process for cells existing in the cell identification area is performed. Then, it progresses to step S40 and the extracted edge is connected and the outline of a cell is obtained. Thereafter, the process proceeds to step S50. In step S50, the extracted cell outline is recognized as one cell, an ID number is assigned to each recognized cell, and the above-described cell identification data is stored in the memory. Thereafter, the process ends.

  According to the present embodiment described above, the following effects can be obtained. That is, in the phase difference image of the cell imaged by the microscope 200, an edge extraction process is performed on the region including any one cell existing in the image designated by the user to extract the cell outline, and the extraction is performed. The outline of the obtained cell was identified as one cell. Thereby, it is possible to accurately recognize individual cells existing in the captured phase difference image.

  In the above-described embodiment, the edges extracted by the edge extraction process are connected by the algorithm described in FIG. 3 to obtain the outlines of cells and nuclei. The edges may be joined together.

  In the above-described embodiment, the enlarged image captured by the microscope 200 is taken into the cell identification device 100 and image processing is performed to identify individual cells present in the image. May be taken into a general-purpose PC having a cell identification program, and individual cells existing in the image may be identified on the general-purpose PC.

  In the embodiment described above, the outline of the cell is extracted based on the phase difference image of the cell imaged by the microscope 200. However, a differential interference image of a cell may be captured by the microscope 200, and a cell outline may be extracted based on the captured differential interference image of the cell.

  The correspondence between the constituent elements of the claims and the embodiment will be described. The input device 110 corresponds to designation means, the control device 120 corresponds to identification means, and the monitor 130 corresponds to display means. Note that the present invention is not limited to the configurations in the above-described embodiments as long as the characteristic functions of the present invention are not impaired.

It is a block diagram which shows the structure of one Embodiment of the cell identification device by this invention. It is a figure which shows the specific example when the area | region containing the arbitrary one cell which exists in the phase difference image of a cell is set by the user. The example of the algorithm which extracts the outline of a cell based on the result of having performed the edge extraction process is illustrated in figure. It is a figure which shows the specific example of the result of having extracted the outline of one cell which exists in the area | region designated by the user. It is a 1st figure which shows the specific example of the observation method of a cell. It is a 2nd figure which shows the specific example of the observation method of a cell. It is a flowchart figure which shows the operation | movement of the cell identification device 100 in this Embodiment.

Explanation of symbols

100 Cell Identification Device 110 Input Device 120 Control Device 130 Monitor 140 Image Memory 200 Microscope 210 CCD

Claims (8)

In a cell identification device for identifying individual cells present in an image obtained by imaging an observation target,
Display means for displaying an image of the captured cell;
Designation means for designating an area in the cell image displayed on the display means;
A cell identification apparatus comprising: identification means for identifying a cell on the assumption that one cell exists in an area designated by the designation means. The cell identification device according to claim 1,
The identifying means extracts an outline of one cell by performing edge extraction on an image in an area designated by the designation means, and identifies the extracted cell outline as one cell. Cell identification device. The cell identification device according to claim 1 or 2,
An image obtained by imaging the observation object is a cell phase difference image or a differential interference image. The cell identification device according to any one of claims 1 to 3,
A cell having a plurality of images of a cell at a predetermined time interval, and performing analysis on the captured images to analyze a time-series change of each cell. Analysis device. A cell identification program for executing, by a computer, a cell identification process for identifying individual cells present in an image obtained by imaging an observation target,
Display processing for displaying the image of the captured cell;
A drawing process for drawing an area in the displayed cell image;
A cell identification program, comprising: an identification process for identifying a cell on the assumption that one cell is present in the region rendered by the rendering process. In the cell identification program according to claim 5,
The identification process is characterized in that edge extraction is performed on an image in a drawn region to extract the outline of one cell, and the extracted cell outline is distinguished from one cell. program. In the cell identification program according to claim 5 or 6,
An image obtained by imaging the observation target is a cell phase difference image or a differential interference image. Each process of the program for cell identification as described in any one of Claims 5-7,
A computer having a plurality of images of cells at predetermined time intervals, an image processing being performed on the plurality of captured images, and an analysis process for analyzing a time-series change of each cell. Cell analysis program executed in

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