JPH0549467A - Apparatus for observing cultured cell - Google Patents

Abstract

PURPOSE:To constantly observe a chosen cell at the center of a frame by controlling the observation image of a culture cell vessel having displayed coordinate axes by stage-control and performing the coordinates transformation according to the change of the position of the culture cell vessel. CONSTITUTION:Stage is shifted in such a manner as to keep a chosen cell at the center O1 of a monitor frame and the coordinates P1 (X, Y) of the chosen cell P1 on the coordinate axes 14, 15 of the vessel is stored in a memory. When the vessel is mounted on a microscope, the coordinate axes of the vessel are detected, the coordinates P2 (X', Y') of the chosen cell on the present detection coordinates are calculated by coordinates transformation based on the shifts X0, Y0 and rotation angle Q of the present detected coordinates relative to the former detected coordinates and the image of the chosen cell is moved to the center based on the transformation result.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus for observing cultured cells in which an image observed by a microscope is captured by a CCD camera or the like and is observed or recorded on a monitor.

[0002]

2. Description of the Related Art Conventionally, for example, a device for observing cultured cells of this type is shown in FIG. In FIG. 6, reference numeral 1 denotes a microscope, on which a cell sample of a plant or an animal placed in a container 3 such as a petri dish or a slide glass is placed on a stage 2, captured by an objective lens 4, and observed by an eyepiece lens 5. ..

A CCD is provided on the eyepiece 5 side of the microscope 1.
The camera 6 is mounted so that the image captured by the CCD camera 6 can be displayed on the monitor television 7 or can be recorded and recorded by the video recorder 8. A controller 9 is provided for the video recorder 8 to set a recording period or time and control the time to enable long-time recording of an observation image.

[0004]

However, in such a conventional cultured cell observation apparatus, when the container 3 such as a petri dish or a slide glass containing a sample is moved,
The position of the sample being observed cannot be known, and it becomes difficult to know which cell was observed. In particular, cells such as plant and animal cells that slowly grow and divide need to be cultured in an incubator or the like, and therefore, the operation of observing with a microscope and then returning to the incubator is repeated. For this reason, it is difficult to know which cells have been observed until now, and it has been difficult to observe specific cells or record for a long time.

The present invention has been made in view of the above-mentioned conventional problems, so that even if the culture cell container is replaced with a microscope, the first designated cell can always be caught and observed at a predetermined screen position. An object of the present invention is to provide a cultured cell observation device.

[0006]

To achieve this object, the present invention is constructed as follows. The reference numerals in the drawings of the embodiments are also shown. That is, in the cultured cell observation device of the present invention, the cultured cell container 3 such as a petri dish displaying coordinate axes is used.
And a microscope 1 for observing the cultured cells in the cultured cell container 3.
And an imaging device 6 that captures an observation image including the coordinate axes of the microscope 1, a frame memory 16 that stores the image of the imaging device 6, and an observation cell designation unit that designates cells to be observed in the observation image of the monitor 7. 17 and observation cell designation unit 17
A stage control unit 18 for moving the stage 2 of the microscope 1 so that the designated cells of No. 1 are at a predetermined observation position on the screen,
A coordinate detection unit 19 that detects the coordinates and the coordinate position of the designated cell from the coordinate axes of the cell observation container 3 in the state where the designated cell is positioned at the observation position, and the designation at the previously detected coordinate when the cultured cell container 3 is replaced. A coordinate conversion unit 20 for converting the coordinate position of the cell to the coordinate position of the detected coordinate this time and moving the designated cell to the predetermined observation position by the stage control unit 18 is provided.

Here, the stage controller 18 centers the stage of the microscope 1 so that the designated cell is located at the center of the screen. Further, the coordinate conversion unit 20 detects the movement amounts Xo, Yo and the rotation angle θ of the current detection coordinates with respect to the previous detection coordinates, and the stage control unit 18 detects the movement amounts Xo, Y.
It is also possible to move the stage 2 so as to return it to the original position by o and the rotation angle θ and correct it to the previously detected coordinates.

[0008]

According to the cultured cell observation apparatus of the present invention having such a configuration, when a cell to be observed is designated on the monitor screen, the stage is moved, that is, centering, so that the designated cell comes to the center of the screen, for example. Is done. After this centering, coordinates are detected from the coordinate axes provided on the container, and the coordinate position of the designated cell at the centering position in the detected coordinates is detected and stored.

After observing with a microscope and returning the container to an incubator or the like and culturing until the next observation time, when the container is set again in the microscope, the coordinate axes displayed on the container are detected, and the coordinates detected at the previous time are detected. The coordinate position (X ′, Y ′) of the designated cell at the current detected coordinate is obtained by coordinate conversion based on the movement amounts Xo, Yo and the rotation angle θ of the designated coordinate of the designated cell (X ′, Y
′) Is stage-controlled and centered so that it becomes the center of the screen.

Therefore, even if the position or orientation of the container changes, the first designated cell can always be captured in the center of the screen, and the designated cell will not be lost.

[0011]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 is a block diagram of an embodiment showing an embodiment of the apparatus of the present invention. In FIG. 1, 1 is a microscope, and a container 3 for storing cultured cells such as a petri dish and a slide glass is placed on a stage 2. A stage drive unit 11 is provided on the stage 2 so that the container 3 placed on the stage 2 can be moved in the biaxial directions of the X axis and the Y axis. Further, the stage driving unit 11 can be provided with a function of driving the container 3 to rotate.

An objective lens 4 is provided below the stage 2, and an image of the cultured cells in the container 3 captured by the objective lens 4 is enlarged by an internal optical system to be imaged by an eyepiece lens 5. Further, the microscope 1 is provided with a focus drive unit 12 that also has a function as a focus adjustment knob, and the focus position by the focus drive unit 12 can be detected by the rotary encoder 13.

A CCD camera 6 is mounted as an image pickup device on the eyepiece 5 side of the microscope 1 so that the same image as the observation image by the eyepiece 5 can be picked up. 10
Is a computer to which an observation image taken by the CCD camera 6 is input, and a stage control signal based on the observation image is supplied to the stage drive unit 11 to position and move the container 3 on the stage 2. A monitor television 7 is connected to the computer 10 so that an observation image captured by the CCD camera 6 can be displayed.

FIG. 2 is an explanatory view of the container 3 used in the present invention. In FIG. 2, a petri dish is taken as an example of the container 3. In this container 3, for example, coordinate axis displays 14 and 15 are drawn orthogonally outside the glass surface of the bottom of the container. Further, a marker 21 that determines the orientation when the container 3 is placed on the stage 2 of the microscope 1 is also drawn. In the cultured cell observation apparatus of the present invention, the coordinate axis displays 14 and 15 drawn on the container 3 shown in FIG. 2 are detected, and even if the container 3 is replaced with the microscope 1, the observation is always performed at the center of the observation screen. Stage control is performed so that the target cells come.

FIG. 3 is a block diagram of an embodiment showing the control configuration of the cultured cell observation apparatus of the present invention realized by the computer 10 of FIG. In FIG. 3, the observation image captured by the CCD camera 6 is converted into binary data and is converted into the frame memory 16
Memorized in. The image capturing period from the CCD camera 6 to the frame memory 16 can be appropriately determined as needed. That is, the frame memory 1 in real time
6, or may be loaded into the frame memory 16 at preset time intervals.

The observation image stored in the frame memory 16 can be read by the display control unit 21 and displayed on the monitor television 7. Of course, in addition to the monitor TV 7, a video recorder for long-time recording may be connected. In the computer 10, an observation cell designation unit 17, a stage control unit 18, a coordinate detection unit 19, and a coordinate conversion unit 20.
Is provided.

First, the observation cell designating unit 17 first sets the CCD.
Coordinate axis display 14, 15 drawn on the container 3 captured by the camera 6
In a state in which the observation image including is displayed on the monitor television 7, a marker formed of a rectangular frame is displayed on the monitor television 7 via the display control unit 21, and the marker is displayed as an observation target displayed on the monitor screen. When the designated operation is performed on the cells to be designated, the stage control unit 18 causes the stage drive unit 11 to position the designated cells at the center of the monitor screen.
The so-called centering display of a so-called designated cell for driving is performed.

Of course, the designated cell may not be centered in the center of the screen, but an appropriate position on the screen may be designated and the stage control may be performed so that the designated cell comes to that position. When the observed cells are designated by the observed cell designating unit 17 and the stage is centered in accordance with the designation of the cells, from the image stored in the frame memory 16 in this centering state, the coordinate detecting unit 19 causes the container 3 shown in FIG. The coordinate axis displays 14 and 15 are detected. In addition, the coordinate detection unit 1
In 9, the coordinate position (X, Y) of the designated cell at the current detection coordinate, that is, the center of the observed image is detected.

The coordinate conversion unit 20 performs coordinate conversion based on the previously detected coordinates and the currently detected coordinates in the coordinate detection unit 19, and determines the coordinate position (X, Y) of the designated cell in the previously detected coordinates as the currently detected coordinates. At the coordinate position (X ',
Y '). That is, if the movement amount of the current detected coordinate with respect to the previous detected coordinate is Xo, Yo and the rotation angle of the coordinate axis is θ, the coordinate position (X ′, Y ′) of the current designated cell in the last detected coordinate. ) Can be calculated by the following equation.

[0020]

[Equation 1]

Here, as shown in FIG. 1, when the container 3 is not moved, the movement amounts Xo and Yo of the present detected coordinates with respect to the previously detected coordinates in the above equation (1) are both 0, and the containers are rotated. Since the angle θ is also 0, the coordinate conversion unit 20 outputs to the stage control unit 18 the coordinate position of the designated cell that has the same detected coordinate and the same coordinate position as the previous time.

Therefore, once the cells to be observed are designated and centered by the observation cell designation unit 17, the stage control unit 18 does not drive the stage unless the container 3 is moved. On the other hand, when the once-observed container 3 is returned to the incubator or the like for further culturing, and is placed on the stage 2 of the microscope 1 again after the lapse of a predetermined culturing time, the coordinates detected by the coordinate detecting unit 19 are the same as those previously detected. There is a deviation between the detected coordinates of this time, and the coordinate position of the designated cell at the detected coordinates of this time is calculated from the equation (1) based on the movement amount (Xo, Yo) and the rotation angle θ indicating the coordinate deviation at this time. Desired.

Since the coordinate position of the designated cell at this time is displaced from the center of the screen due to the displacement of the coordinate system, the stage control unit 18 determines the coordinate position (X ', Y) of the designated cell obtained by the coordinate conversion for the detected coordinates at this time. The stage drive unit 11 is driven by the stage control unit 18 so that ′) comes to the center of the screen. FIG. 4 is an explanatory diagram showing a processing operation of coordinate conversion by the computer 10 of FIG.

First, when the marker 3 is moved on the monitor screen by the observation cell designating section 17 while the container 3 is set in the microscope 1 and displayed on the monitor TV 7, the cells to be observed are captured, as shown in FIG. As shown in, the stage control is performed so that the designated cell captured by the marker comes to P ₁ at the center of the screen. In FIG. 4 (a), the solid-line coordinate axes are coordinate axis displays 14, 15 drawn on the container 3 shown in FIG.
And the coordinates of the CCD camera 16 are Xi, Y
i, and the designated cell P ₁ is centered so as to come to the origin Oi of the coordinates Xi, Yi of the CCD camera 6. The coordinate position of the designated cell P1 in the coordinate system of the coordinate axis displays 14 and 15 at this time is detected as (X, Y).

Next, it is assumed that the observation screen when the container 3 of the microscope 1 is returned to the incubator or the like and cultured and then set again in the microscope 1 is shown in FIG. 4 (b), for example. In FIG. 4B, 14 'and 15' are shown in FIG.
It is the coordinate axis display of the previously detected coordinates shown in (a), and the solid axis coordinate axis displays 14 and 15 are the current detected coordinates. When the detected coordinates of this time are viewed on the basis of the previously detected coordinates, Xo in the X axis direction. It moves, moves Yo in the Y-axis direction, and rotates clockwise by θ.

In the detection of the movement amounts Xo, Yo and the rotation angle θ of the previously detected coordinates and the currently detected coordinates, the coordinate conversion unit 20 compares the coordinate axes of the previously detected coordinates and the currently detected coordinates. You can ask for it. In this way, if the movement amounts Xo and Yo and the rotation angle θ of the current detected coordinates with reference to the previously detected coordinates are obtained, the expression (1) of the designated cell of the present time at the last detected coordinates is obtained. The coordinate position P ₂ (X ′, Y ′) is obtained.

Since the coordinate position P ₂ (X ', Y') of the designated cell obtained by this coordinate conversion is displaced from the center of the screen due to the displacement of the coordinate system, the coordinate position P ₂ (X ', Y') is displayed on the screen. The stage controller 18 drives the stage so that it comes to the center.
As shown in FIG. 4C, a centering state in which the designated cell comes to P ₁ at the center of the screen is created. In this centering state, although the coordinate directions are different with respect to the coordinate systems of the displaced coordinate axis displays 14 and 15 this time, the coordinate position P ₁ (X, Y) is the same as in the initial state of FIG. 4A. When the coordinate conversion processing for centering the designated cell is completed after the replacement of the container 3 in this way, the detected coordinates obtained in the centering state shown in FIG. 4C are stored and prepared for the next replacement of the container 3.

Referring again to FIG. 3, in this embodiment, the computer 10 is further provided with a focus control section 23 so that the observation screen detected by the rotary encoder 13 is focused in a focused state. The position detection information is stored in the focus control unit 23, and if the detection value from the rotary encoder 13 when the container 3 is replaced is different from the previously stored value, focus drive is performed so as to return to the first stored value. The part 12 is controlled.

In normal observation of the container 3, once the focus is adjusted, it is not necessary to re-execute the focus control even if the container 3 is replaced. However, for example, the same microscope 1 is used and different containers are used. If the focal position changes due to
Rotary encoder 1 with a code indicating the type of container
The focus value from 3 is stored, and when the container 3 is replaced with another type of container, the operator inputs the code number of the container, so that the focus value stored for each container is automatically set. It is possible to control the focus drive unit 12 effectively.

Of course, in the cultured cell observation apparatus of the present invention, focus control is not essential and may be appropriately provided as needed. FIG. 5 is an explanatory diagram showing a coordinate conversion process in the second embodiment of the present invention. In the image conversion processing as the first embodiment shown in FIG. 4, after the container 3 is replaced, the designated cell after the replacement is centered and controlled without correcting the orientation (rotation) of the coordinate system. However, in the second embodiment of FIG. 5, the stage drive unit 11 is provided with the rotary drive mechanism so that even if the container 3 is placed in a direction different from the initial direction, the correction control is always performed at the same coordinate position as the initial position. Is characterized by.

That is, FIG. 5A shows a state in which the designated cells are caught and centered by the marker on the monitor screen in the cell observation designating unit 17, as in the case of FIG. 4A.
Similar to FIG. 4B, FIG. 4B shows a state in which the container 3 is returned to the incubator, cultured, and then returned to the microscope 1, and the coordinate axes are displaced. The movement amount of the current coordinate axis with respect to the previous coordinate axis in FIG. 5B is Xo in the X-axis direction, Yo in the Y-axis direction, and θ is the rotation angle.

On the basis of the detection information indicating the deviation of the coordinate system, in the second embodiment, the coordinate axis displays 14 and 15 of the present detected coordinates are detected movement amounts Xo and Yo.
Then, Xo and Yo are moved in the opposite direction to the stage moving state shown in FIG. Next, the coordinate axis display 1 shown by the solid line
By driving the stages 4 and 15 so as to rotate counterclockwise to the detected rotation angle θ, that is, counterclockwise by θ, the coordinate axis display 14
Return to the state of ', 15'. This means returning to the coordinate position in the centering state of the first designated cell in FIG. 5 (a).

The advantage of the embodiment shown in FIG. 5 is that the direction of the designated cell can be returned to the same state as at the beginning, so that it is advantageous, for example, when directionality is a problem in cell observation. As another embodiment of the present invention, the stage 2 shown in FIG. 4 or FIG. 5 is constructed by actually converting the image obtained from the CCD camera 6 on the frame memory 16 without performing the driving of the stage 2 and writing the image. It is possible to obtain exactly the same function as in the case of coordinate conversion by movement.

[0034]

As described above, according to the present invention, even if the position of a sample observed under a microscope is returned to an incubator or the like and cultured and then returned to the microscope for observation, the first designated cell is changed. Is always displayed, for example, at the center of the screen, so that even if the position of the sample changes, it is possible to easily carry out long-term observation while always catching the object.

[Brief description of drawings]

FIG. 1 is a configuration diagram of an embodiment showing a device configuration of the present invention.

FIG. 2 is an explanatory view of a culture cell container used in the present invention.

FIG. 3 is a block diagram of an embodiment showing a control processing field configuration of the present invention.

FIG. 4 is an explanatory diagram showing coordinate conversion according to the first embodiment of the present invention.

FIG. 5 is an explanatory diagram showing coordinate conversion according to a second embodiment of the present invention.

FIG. 6 is an explanatory view of a conventional device.

[Explanation of symbols]

 1: Microscope 2: Stage 3: Container (culture cell container) 4: Objective lens 5: Eyepiece 6: CCD camera (imaging device) 7: Monitor TV 10: Computer 11: Stage drive unit 12: Focus drive unit 13: Rotary Encoders 14 and 15: Coordinate axis display 16: Frame memory 17: Observation cell designation unit 18: Stage control unit 19: Coordinate detection unit 20: Coordinate conversion unit 21: Display control unit 22: Marker 23: Focus control unit

Claims (3)

[Claims] 1. A culture cell container such as a petri dish displaying coordinate axes, a microscope for observing cultured cells in the culture cell container, an imaging device for capturing an observation image including the coordinate axes by the microscope, and the imaging device. A frame memory for storing the image of the observation image, an observation cell designation section for designating cells to be observed in the observation image on the monitor, and the designated cell of the observation cell designation section so as to be at a predetermined observation position on the screen. A stage control unit that moves a stage of a microscope, a coordinate detection unit that detects coordinates and coordinate positions of designated cells from coordinate axes of the cell observation container in a state where the designated cells are positioned at the observation position, and the cultured cell container. When replaced, the coordinate position of the designated cell in the previously detected coordinate is converted into the coordinate position of the detected coordinate this time, and the designated cell is observed by the stage control unit in a predetermined manner. An apparatus for observing cultured cells, comprising: a coordinate conversion unit that moves the cell to a position. 2. The apparatus for observing cultured cells according to claim 1, wherein the stage controller centers the stage of the microscope so that the designated cell is located at the center of the screen. 3. The cultured cell observation apparatus according to claim 1, wherein the coordinate conversion unit detects the movement amounts Xo, Yo and the rotation angle θ of the present detected coordinate with respect to the previously detected coordinate, and controls the stage. An apparatus for observing cultured cells, characterized in that the stage moves the stage so as to reverse the detected movement amounts Xo and Yo and the rotation angle θ to correct the display state of the previously detected coordinates.