JP2008299027A - Microscope apparatus, its control program and control method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microscope apparatus designed so as to dispense with a bothersome operation of moving a specimen movable stage when changing an observation magnification and so as to speedily and appropriately select an observation magnification. <P>SOLUTION: The microscope device stores: a first magnification error and a first coordinate error, obtained in advance under conditions using a low magnification objective lens and a predetermined zoom magnification; and a second magnification error and a second coordinate error, obtained based on a combination of each of high magnification objective lenses and each of zoom magnifications. In such a microscope apparatus, if a user specifies a specific area of a low magnification observation image, the apparatus captures the center coordinate of the specified area and corrects the first coordinate error. In addition, the apparatus calculates a magnification required for magnifying and corrects the magnification error by using the first magnification error. Subsequently, among actual magnifications obtained by combinations of the objective lens magnifications and the zoom magnifications, the apparatus specifies a combination that yields an actual magnification closest to the corrected magnification. Then, by using the second coordinate error, the apparatus further corrects the corrected center coordinate. Based on the contents of the correction, the apparatus adjusts the objective lens, zoom magnification, and the stage position. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a map function on an image captured by a microscope.

  Conventionally, a microscope is capable of capturing and recording a magnified image of a fine sample, and thus has been widely used in the industrial field including research and inspection in the biological field. Such a microscope generally includes a plurality of objective lenses having different magnifications attached to a rotary revolver. Therefore, the microscope changes the observation magnification by rotating the revolver to switch the objective lens and inserting an arbitrary objective lens into the observation optical path. In addition to the plurality of objective lenses, the microscope also has a zoom magnification mechanism, and can continuously change the observation magnification.

  In changing the observation magnification, normally, with the switching of the objective lens from the low magnification to the high magnification, a phenomenon occurs in which the target position of the site to be observed is shifted and deviates from the observation field. For this reason, the target position is moved in advance to the center of the visual field at a high magnification to prevent the target position from deviating from the observation visual field.

  In microscope observation, if such an operation is manually performed every time the magnification is changed, a lot of time is wasted in the microscope operation. Also, manual microscope operation may not be able to handle samples with a fast changing speed. Further, if too much time is taken for the microscope operation, the fluorescent sample may be faded.

  In order to solve such a problem, for example, in Patent Document 1, a monitor unit that displays an observation image of a sample, an instruction unit that specifies a desired region of the observation image displayed on the monitor unit, and this instruction Based on the objective magnification necessary for displaying the designated area of the observation image by the means on almost the entire screen of the monitor means, the computing means for computing the position of the sample moving stage, and the data computed by the computing means A microscope magnifying device including an objective switching mechanism that inserts a lens into an observation optical path and a driving unit that drives a sample moving stage is disclosed. Accordingly, in Patent Document 1, an objective lens corresponding to a magnification corresponding to a desired area specified in an observation image is automatically inserted into the observation optical path, and the specified area is automatically set to substantially the center of the observation field. It is trying to be located in.

Patent documents 2 and 3 are patent documents related to the present application.
JP-A-3-296011 JP-A-11-250238 JP 2002-197451 A

  However, in the conventional microscope as described above, the correction of the deviation distribution of the coordinates in the observation visual field due to the distortion (distortion aberration) of the microscope image or the magnification deviation distribution in the observation visual field due to the distortion is not sufficient. . Further, the zoom magnification that continuously changes the magnification cannot be corrected. In particular, a method for acquiring concentric shift correction data based on component accuracy of an objective lens and a rotating revolver has not been clarified.

  11A and 11B show an example in which an arbitrary area on a conventional navigation map image is enlarged and displayed. An entire image (hereinafter referred to as a macro observation image) 101 of an observation sample photographed with a low-magnification objective lens with a microscope is acquired as a navigation map image. While referring to the monitor of the personal computer, a region to be subjected to detailed observation in the macro observation image 101 is dragged with a mouse, and is enclosed in a rectangle with a ROI (Region Of Interest) 103. Then, the zoom magnification is changed corresponding to the size of the range surrounded by the ROI 103, and the sample moving stage moves to the center position of the ROI 103. Then, the high magnification objective lens is switched to display a detailed image (micro live image 102) of the ROI 103.

  In FIG. 11A, the observation area around the center of the macro observation image 101 is surrounded by the ROI 103, and the micro live image 102 surrounded by the ROI 103 is displayed. In FIG. 11B, the observation area near the upper right of the macro observation image 101 is surrounded by the ROI 103, and the micro live image 102 surrounded by the ROI 103 is displayed.

  At this time, the displacement distribution of the coordinates in the observation field due to the distortion of the low-magnification objective lens and the displacement distribution of the magnification, the slash of the zoom center of the zoom lens (the zoom center shifts every time the zoom is performed), the zoom magnification error, and Based on the deviation of the center coordinate of the high-magnification objective lens and the magnification error, there is an error in the coordinate value and the magnification value between the range of the ROI 103 designated by the macro observation image 101 and the micro live image 102 displayed based on the ROI 103. Occurs. Therefore, the range specified by the ROI 103 cannot be correctly displayed on the micro live image 102. In such a case, it is necessary for the user to finely adjust the position of the sample moving stage and adjust the zoom again while confirming the micro live image 102.

  In view of the above problems, in the present invention, when a predetermined region of the low-magnification observation image displayed on the display device is specified, a troublesome sample at the time of changing the observation magnification in the microscope device that displays the high-magnification observation image of the predetermined region There is provided a microscope apparatus that does not require a moving operation of a moving stage and can select an observation magnification quickly and appropriately.

  A microscope apparatus according to the present invention includes a sample moving stage on which an observation sample is placed and movable in a direction perpendicular to the observation optical axis, and a revolver that switches a plurality of objective lenses inserted in the observation optical axis and having different magnifications. A zoom magnification changing means for changing a zoom magnification, an imaging means for picking up an optical image of the observation sample via the objective lens, and a first of the objective lenses based on an instruction operation from a user. Based on the first objective lens and the first zoom magnification, an area designating means capable of designating a predetermined area of the first observation image captured by the objective lens at the first zoom magnification as the designated area. The first error information storage means storing the first coordinate error and the first magnification error of the observation image to be obtained, the second objective lens among the objective lenses, and zooming within a range where zooming is possible Before letting The second coordinate information of the second observation image obtained based on the combination with the zoom magnification, the second error information storage means in which the second magnification error is made, and the designated center coordinate which is the center coordinate of the designated area Is corrected with the first coordinate error to be a first corrected center coordinate, and a magnification for enlarging the designated area is calculated based on the size of the designated area with respect to the first observation image, and the magnification Is corrected with the first magnification error to obtain a first correction magnification, and the first correction among the magnification based on the combination of the magnification of the second objective lens and the zoom magnification. A second error correction unit that identifies a combination that has a magnification closest to the magnification, corrects the first correction center coordinate with the second coordinate error, and sets the second correction center coordinate; and the specified objective Lens magnification and zoom magnification Based on the combination, the revolver is controlled to switch the objective lens, the zoom magnification unit is controlled to vary the zoom magnification, and based on the second correction center coordinate, And a control means for controlling the movement of the sample moving stage.

  In the microscope apparatus, the first error information storage means stores, as the first coordinate error, a distance moved by a predetermined position on the sample moving stage through the first objective lens, and the sample A ratio between a distance between two predetermined points on the moving stage and a distance between the two points expanded and contracted through the first objective lens is stored as the first magnification error. .

  In the microscope apparatus, the first error information storage unit divides the first observation image into a plurality of regions concentrically from the center of the first observation image, and the first coordinate error and the first error for each of the divided regions. The magnification error is calculated and stored.

  In the microscope apparatus, the second error information storage unit includes a combination of the second objective lens and each zoom magnification with respect to the first observation image captured at the first objective lens and the first zoom magnification. Is stored as the second coordinate error, and the ratio of the actual magnification to the display magnification is obtained for each magnification obtained by the combination of the second objective lens and each zoom magnification. It is stored as the second magnification error.

  In the microscope apparatus, the second error information storage means includes coordinates in the second observation image corresponding to the first correction center coordinates in the first observation image, and the second observation image. The difference from the center coordinate of the observation field is stored as the second coordinate error, and the ratio between the total display magnification and the total real magnification for each zoom magnification in the second objective lens is used as the second magnification error. It is characterized by storing.

The microscope apparatus further includes a reference chart that is detachable with respect to the observation optical axis and has a grid pattern on the sample moving stage.
A sample moving stage on which an observation sample is placed and movable in a direction perpendicular to the observation optical axis, a revolver for switching a plurality of objective lenses having different magnifications inserted into the observation optical axis, and a zoom according to the present invention A zoom magnification changing mechanism for changing magnification, an imaging device for taking an optical image of the observation sample via the objective lens, and a first objective lens among the objective lenses based on an instruction operation from a user Is obtained on the basis of the area designating function capable of designating the predetermined area of the first observation image captured at the first zoom magnification as the designated area, the first objective lens, and the first zoom magnification. The first error information storage device storing the first coordinate error and the first magnification error of the observation image, and the second objective lens among the objective lenses, are scaled within a range where scaling is possible. Zoom A computer for processing to control the operation of the microscope apparatus comprising: a second coordinate error of the second observation image obtained based on the combination with the rate; and a second error information storage device in which the second magnification error has occurred. The microscope control program to be executed causes the designated center coordinate, which is the center coordinate of the designated area, to be corrected with the first coordinate error to be the first corrected center coordinate, and the designated area for the first observation image. A first error correction process for calculating a magnification for enlarging the designated area based on the size of the first area and correcting the magnification with the first magnification error to obtain a first correction magnification; and the second objective. Among the magnifications based on the combination of the lens magnification and the zoom magnification, a combination that is closest to the first correction magnification is specified, and the first correction center coordinate is corrected by the second coordinate error, and the first coordinate is corrected. Correction of 2 Based on the combination of the second error correction processing to be the center coordinates and the magnification of the specified objective lens and the zoom magnification, the revolver is controlled to switch the objective lens, and the zoom magnification mechanism is controlled. And a drive control process for controlling the movement of the sample moving stage based on the second correction center coordinates and changing the zoom magnification.

  A sample moving stage on which an observation sample is placed and movable in a direction perpendicular to the observation optical axis, a revolver for switching a plurality of objective lenses having different magnifications inserted into the observation optical axis, and a zoom according to the present invention A zoom magnification changing mechanism for changing magnification, an imaging device for taking an optical image of the observation sample via the objective lens, and a first objective lens among the objective lenses based on an instruction operation from a user Is obtained on the basis of the area designating function capable of designating the predetermined area of the first observation image captured at the first zoom magnification as the designated area, the first objective lens, and the first zoom magnification. The first error information storage device storing the first coordinate error and the first magnification error of the observation image, and the second objective lens among the objective lenses, are scaled within a range where scaling is possible. Zoom And a second error information storage device in which a second coordinate error of the second observation image and a second magnification error obtained based on the combination with the ratio are controlled. Is based on the size of the designated area with respect to the first observation image, while correcting the designated center coordinates, which are the center coordinates of the designated area, with the first coordinate error to obtain first corrected center coordinates. A magnification for enlarging the designated area is calculated, the magnification is corrected by the first magnification error to be a first correction magnification, and a magnification based on a combination of the magnification of the second objective lens and the zoom magnification A combination having a magnification closest to the first correction magnification is specified, the first correction center coordinate is corrected by the second coordinate error to be a second correction center coordinate, and the magnification of the specified objective lens And the zoom magnification Based on the combination, the revolver is controlled to switch the objective lens, the zoom magnification mechanism is controlled to change the zoom magnification, and based on the second correction center coordinate, Control is performed to move the sample moving stage.

  By using the microscope apparatus according to the present invention, it is not necessary to perform a troublesome moving operation of the sample moving stage when changing the observation magnification, and the observation magnification can be selected promptly and appropriately.

  The microscope apparatus according to the present invention can display a high-magnification observation image of the predetermined area on the display device when a predetermined area of the low-magnification observation image displayed on the display device is designated. A microscope apparatus according to the present invention includes a sample moving stage, a revolver, a zooming / magnifying unit, an imaging unit, a region specifying unit, a first error information storage unit, a second error information storage unit, a first error correction unit, a first error correction unit, 2 error correction means and control means.

  The sample moving stage is a stage on which an observation sample is placed and is movable in a direction perpendicular to the observation optical axis. In this embodiment, the sample moving stage is, for example, the sample moving stage 3.

The revolver switches a plurality of objective lenses having different magnifications inserted into the observation optical axis. The revolver is, for example, the revolver 6 in this embodiment.
The zoom scaling unit is a lens driving mechanism that continuously varies the magnification. In the present embodiment, the zoom scaling unit is, for example, the zoom scaling mechanism 9.

The imaging means captures an optical image of the observation sample through the objective lens. The imaging means is, for example, the TV camera 10 in this embodiment.
The area designating means is a first observation image (macro observation image) imaged at a first zoom magnification by a first objective lens (low magnification objective lens) of the objective lenses based on an instruction operation from a user. The predetermined area can be designated as the designated area. In this embodiment, the area designating unit is, for example, the ROI 103 used when designating a predetermined area of the low-magnification observation image displayed on the TV monitor 20 by the drag operation of the PC mouse 21.

  The first error information storage means stores a first coordinate error and a first magnification error of the observation image obtained based on the first objective lens and the first zoom magnification. In the present embodiment, the first error information storage means is, for example, a macro observation image correction table or a complementary expression corresponding to this table.

  The second error information storage means includes a second objective lens that is obtained based on a combination of the second objective lens (high-magnification objective lens) of the objective lenses and the zoom magnification that has been changed within a variable range. A second coordinate error and a second magnification error of the observation image (micro live image) are set. In the present embodiment, the second error information storage means is, for example, a micro live image correction table or a complementary expression corresponding to this table.

  The first error correction means corrects the specified center coordinate, which is the center coordinate of the specified area, with the first coordinate error to be the first corrected center coordinate, and also specifies the specified area for the first observation image. A magnification for enlarging the designated area is calculated based on the size of the first magnification, and the magnification is corrected with the first magnification error to be a first correction magnification. In the present embodiment, the first error correction unit corresponds to, for example, the processing of S40 to S42 in FIGS. 10A and 10B executed by the control device 25.

  The second error correction means specifies a combination that is the closest to the first correction magnification among the magnifications based on the combination of the magnification of the second objective lens and the zoom magnification, and the first correction center. The coordinates are corrected with the second coordinate error to obtain second corrected center coordinates. In the present embodiment, the second error correction unit corresponds to, for example, the processing of S44 to S45 of FIGS. 10A and 10B executed by the control device 25.

  The control means controls the revolver to switch the objective lens based on the combination of the specified objective lens magnification and the zoom magnification, and controls the zoom scaling means to change the zoom magnification. In addition, control for moving the sample moving stage is performed based on the second correction center coordinates. In this embodiment, the control means corresponds to, for example, the processing of S43 and S45 in FIGS. 10A and 10B executed by the control device 25 and the drive circuit 28.

With this configuration, it is not necessary to perform a troublesome operation of moving the sample moving stage when changing the observation magnification, and the observation magnification can be selected promptly and appropriately.
The first error information storage means stores, as the first coordinate error, a distance traveled by a predetermined position on the sample moving stage through the first objective lens, and stores the predetermined error on the sample moving stage. The ratio of the distance between the two points and the distance between the two points expanded and contracted through the first objective lens is stored as the first magnification error.

With this configuration, it is possible to save the coordinate error and magnification error caused by the low-magnification objective lens acquired at the time of calibration.
The first error information storage means divides a plurality of regions concentrically from the center of the first observation image, and the first coordinate error and the first magnification are divided into the divided regions. The error is calculated and stored.

With this configuration, the first coordinate error and the first magnification error can be set as a representative value of the coordinate error and a representative value of the magnification error for the region, respectively.
The second error information storage means captures the first objective lens and the first objective image captured at the first zoom magnification with respect to the first observation image and the second objective lens and the respective zoom magnifications. 2 is stored as the second coordinate error, and the ratio of the actual magnification to the display magnification for each magnification obtained by the combination of the second objective lens and each zoom magnification is the second magnification. Stored as error.

  Further, the second error information storage means includes coordinates in the second observation image corresponding to the first correction center coordinates in the first observation image and an observation field of view of the second observation image. The difference from the center coordinate of the second objective lens is stored as the second coordinate error, and the ratio between the total display magnification and the total actual magnification for each zoom magnification of the second objective lens is stored as the second magnification error. Yes.

The microscope apparatus may further include a reference chart that is detachable with respect to the observation optical axis and has a grid pattern on the sample moving stage.
With this configuration, calibration can be easily performed every time the power of the microscope zooming device is turned on.

Hereinafter, embodiments of the present invention will be described in detail.
FIG. 1 shows an overall configuration of a microscope system according to the present embodiment. The microscope system mainly includes a microscope main body 1 and a personal computer (hereinafter referred to as a PC) 19.

  The microscope main body 1 includes an illumination device 2, a sample moving stage 3, a focusing device 4, an objective lens 5 (low magnification objective lens 5a, high magnification objective lens 5b), a revolver 6, a zoom magnification mechanism 9, a TV camera 10, and a high luminance. A mercury lamp 11, a fluorescence observation device 12, a lens barrel 18, a control device 25, and a TV camera control device 26 are provided.

  The illumination device 2 is a device that irradiates the observation sample 7 placed on the sample moving stage 3 with illumination light. The illumination device 2 includes a condenser lens and an FS (Field Stop) mechanism (not shown). The condenser lens can be selected for critical illumination or telecentric illumination. The FS mechanism blocks illumination light outside the range imaged by a CCD (Charge Coupled Device) in the TV camera 10 so that useless light is not applied to the observation sample 7.

  The sample moving stage 3 can move in the XY directions orthogonal to each other in a plane orthogonal to the illumination light (observation optical axis 8) from the illumination device 2. The focusing device 4 performs focus adjustment by moving the sample moving stage 3 in the direction of the observation optical axis 8. The revolver 6 can replace the objective lens 5 and inserts an objective lens used for observation into the observation optical axis 8.

  The zoom magnification mechanism 9 is a mechanism for continuously enlarging or reducing an enlarged image captured using the objective lens 5 inserted in the observation optical axis 8. The zoom magnifying mechanism 9 includes an AS (Aperture Stop) mechanism (not shown) that limits the numerical aperture (NA) of the observation image and increases the depth of focus.

  In the lens barrel 18, the observation light is branched to the eyepiece lens side and the TV camera 10 side. The eyepiece 18a is for making an observation image with the naked eye. The TV camera 10 converts an observation image from the observation sample 7 into an electric signal. The TV camera control device 26 is a device that controls the operation of the TV camera 10.

  The high-intensity mercury lamp 11 is an illumination light source used during fluorescence observation. The fluorescence observation apparatus 12 holds a turret mechanism 17 that enables switching between the fluorescent cubes 16 and 16a. The fluorescent cube 16 holds the excitation filter 13, the absorption filter 14, and the dichroic mirror 15 together.

  The excitation filter 13 transmits only excitation light having a specific wavelength among irradiation light emitted from the high-intensity mercury lamp 11. The absorption filter 14 transmits the wavelength of fluorescence emitted when the observation sample 7 is irradiated with excitation light. The dichroic mirror 15 aligns excitation light and fluorescence with the observation optical axis 8. The fluorescence observation apparatus 12 further incorporates an epi-illumination FS mechanism (not shown) having the same function as the FS mechanism.

Each of the various mechanical units and driving units constituting the microscope main body 1 has an electric driving device and is connected to the control device 25. Further, the control device 25 is connected to the PC 19.
The PC 19 controls a GUI (graphical user interface) on which a user actually performs a microscope operation. The PC 19 includes a TV monitor 20 and a PC mouse 21. The TV monitor 20 displays the GUI image and an image captured by the TV camera 10. The PC mouse 21 can perform a GUI operation and a click operation or a drag operation for designating an observation range on the image.

  FIG. 2 shows an example of the configuration of the control device 25 of FIG. The TV camera control device 26 controls input of an image signal from the TV camera, input of control data from the TV camera 10, and output of a drive signal for driving the TV camera 10.

  The control device 25 controls the TV camera control device 26 and various circuits. The TV camera control device 26 controls the shutter speed, ISO sensitivity, white balance, exposure, pixel shift, image processing, and the like of the TV camera 10. The image signal output from the TV camera 10 can be obtained as a captured image or a live image.

  The input circuit 27 receives illumination light ON / OFF data, critical illumination / telecent illumination selection data, FS opening / closing amount data, stage XY position data, focusing position data, revolver position data, and observation optical axis from the microscope body 1. Objective type data, zoom magnification data, AS opening / closing amount data, mercury lamp ON / OFF data, epi-illumination FS opening / closing amount data, fluorescent cube type data on the observation optical axis, and the like are input. In addition, PC pointer coordinate data, user operation data on the PC 19, and the like are input to the input circuit 27 from the PC 19. The input circuit 27 passes the input data in response to a request from the control device 25.

  The drive circuit 28 turns the illumination device 2 and the high-intensity mercury lamp 11 on / off, switches the condenser lens, the FS mechanism, the sample moving stage 3, the focusing device 4, the revolver 6, the zoom magnification mechanism 9, and the AS. A control signal (for example, the number of rotations of the motor, an ON / OFF signal) for driving the mechanism, the epi-illumination FS mechanism, and the mechanism such as the fluorescent cube turret is output according to a request from the control device.

  The storage circuit 29 stores a coordinate correction value and a magnification correction value for each region of the visual field in a specific low-magnification observation image obtained by calibration described in detail later in a data table or a complementary expression. Further, the storage circuit 29 stores the difference between the display magnification and the actual magnification for each objective lens 5 and for each of a plurality of magnification ranges of the zoom magnifying mechanism 9 in a table or a complementary manner.

  Furthermore, the storage circuit 29 stores correction data relating to the absolute position of the stage for correcting an error in the amount of movement that occurs due to a rotation angle error of the stage drive motor of the sample moving stage 3 in a table or a complementary expression. . The storage circuit 29 outputs these table data in response to a request from the control device 25.

  The arithmetic circuit 30 executes predetermined arithmetic processing in response to a request from the control device 25. For example, when the correction data is stored in the storage circuit 29 as a complementary expression, the control device 25 passes the complementary expression to the arithmetic circuit 30, and the arithmetic circuit 30 generates data from the complementary expression, and the data Is output to the control device 25.

  The image output circuit 31 receives an image signal of a captured image and a live image captured by the TV camera 10 and subjected to image processing by the TV camera control device 26 in response to a request from the control device 25. The image output circuit 31 outputs the image signal to the TV monitor 20.

  FIG. 3 shows an overall flow of the present embodiment. First, the microscope body 1 is calibrated (step 1. Hereinafter, the step is referred to as “S”). Then, the control device 25 detects the deviation of the coordinates in the observation field and the deviation of the magnification in the observation field due to the distortion of the low-magnification objective lens, the blur and zoom magnification error of the zoom center of the zoom lens, and the deviation of the center coordinate of the high-magnification objective lens A correction table related to the error between the coordinate value and the magnification value based on the magnification error is created.

  That is, the control device 25 acquires an error of “coordinate values of the entire field of view” and an error of “magnification value of each part of the field of view” of the macro observation image captured using the low-magnification objective lens 5a, and stores the memory circuit. 29 in the macro observation image correction table. Further, the control device 25 acquires an error of “magnification value near the center” and an error of “field center coordinate value” of each zoom magnification in a state where the high-magnification objective lens 5b is inserted in the observation optical axis 8, and a storage circuit 29 is stored in the micro live image correction table 29.

  Next, the user performs microscopic observation with the macro observation image and the micro live image (S2). That is, when the user surrounds an arbitrary area in a rectangle with the ROI 103 on the macro observation image 101 displayed on the TV monitor 20 by a drag operation of the PC mouse 21, the low magnification objective lens 5a is switched to the high magnification objective lens 5b. The zoom magnification corresponding to the range surrounded by the ROI 103 is changed, the sample moving stage 3 moves to the center position of the ROI 103 range, and the micro live image 102 of the ROI 103 range is displayed.

  This will be described in more detail. When the ROI 103 is set by the user, the center coordinates of the range surrounded by the rectangle by the ROI 103 are recognized, and the true center coordinates (designated center coordinates) are calculated based on a macro observation image correction table described later.

  Further, the magnification value is recognized from the range surrounded by the rectangle in the ROI 103, and the true magnification value (specified magnification value) corresponding to the coordinates of the ROI range is calculated based on the macro observation image correction table described later.

  Then, when the objective lens is switched to the high-magnification objective lens 5b, the zoom magnification is determined based on the micro live image correction table so that the magnification closest to the designated magnification value is obtained. Therefore, the control device 25 controls the sample moving stage 3 so that the designated center coordinates are located at the center position of the observation visual field, and changes the magnification to the determined zoom magnification.

  In this way, the ROI range designated by the user with the macro observation image can be displayed with the micro live image. This eliminates the troublesome work for the user when changing the observation magnification, and allows the observation magnification to be selected quickly and appropriately. Hereinafter, S1 and S2 will be described in detail.

<S1: Creation of correction table by calibration>
Calibration is performed using a reference chart. When the reference chart 40 having a grid pattern is placed on the sample moving stage 3 instead of the observation sample 7, and the objective lens 5 (low magnification objective lens 5a, high magnification objective lens 5b) is switched, the TV monitor 20 is switched to FIG. 4A. And the image of the reference chart shown in FIG. 4B is taken.

  FIG. 4A shows a display example of a reference chart imaged under the conditions of the low magnification objective lens 5a and the minimum magnification zoom magnification. A reference chart 40 is displayed in the display area 41 of the TV monitor 20. The reference chart 40 has a grid pattern (squares) indicated by reference numeral 42. The two intersecting lines indicated by reference numeral 43 are diagonal lines of the display area 41 displayed by predetermined jig software. Reference numeral 44 indicates the center of the basic chart 40. In this embodiment, it is assumed that the macro observation image 101 is captured under the conditions of the low magnification objective lens 5a and the minimum magnification zoom magnification.

  FIG. 4B shows a display example of a reference chart imaged under the condition of the high magnification objective lens 5b and the minimum magnification zoom magnification. The micro live image 102 is captured at each zoom magnification for each high-magnification objective lens. In this embodiment, for convenience of explanation, the high magnification objective lens to be used is fixed to one and the zoom magnification is changed.

  As shown in FIGS. 4A and 4B, the grid of the reference chart 40 displayed on the TV monitor 20 spreads in a pincushion shape due to the influence of distortion or the like. In addition, the spreading method may indicate a shrinking method called a barrel shape depending on whether the distortion is positive or negative.

  FIG. 5 shows the shape of the distortion that changes depending on the combination of the objective lens and the zoom magnification of the zoom magnifying mechanism. The vertical axis shows the distortion amount, and the horizontal axis shows the image height. FIG. 5A shows a distortion shape under a setting condition of a low magnification objective lens and a zoom magnification of 0.5 ×. FIG. 5B shows a distortion shape under a setting condition of a high magnification objective lens and a zoom magnification of 0.5 ×. FIG. 5C shows a distortion shape under a setting condition of a high magnification objective lens and a zoom magnification of 2 ×. FIG. 5D shows a distortion shape under a setting condition of a high magnification objective lens and a zoom magnification of 6 ×.

  Thus, it can be seen that the distortion changes for each objective lens and for each zoom magnification. Suppose that the designated central coordinates of the ROI 103 designated on the macro observation image 101 are shifted from a five times macro observation image to a 100 times micro live image including an error of 1% due to distortion. . Then, unless the error is corrected, the center coordinates specified by the ROI 103 are shifted by 20% or more from the total length of the field of view of the micro live image. Furthermore, the specified magnification value enclosed by the ROI also needs error correction.

  In order to correct such an error between the designated center coordinates and the designated magnification, it is necessary to calibrate the microscope apparatus 1 according to the flow shown in FIGS. 6A and 6B and create a correction table.

  6A and 6B show a flow of calibration in the present embodiment. A basic chart 40 is placed in advance on the sample moving stage 3. Further, the coordinates of the cells of the reference chart 40 are stored in advance in the storage circuit 29 as XY coordinate values of absolute values on the sample moving stage 3.

  First, when the user performs a predetermined operation, the control device 25 controls the revolver 6 and the zoom magnification changing mechanism 9 of the microscope main body 1 so as to find the origin of the revolver 6 and the zoom magnification changing mechanism 9 (S11, S12). .

  Next, the revolver 6 is rotated under the control of the control device 25, and the high-magnification objective lens 5b is inserted into the observation optical axis 8 (S13). Then, under the control of the control device 25, the zoom magnifying mechanism 9 moves the zoom lens so that the maximum zoom is achieved (S14). The control device 25 drives the TV camera 10 to capture an image of the basic chart 40 under the high magnification objective lens 5b and the maximum magnification zoom condition.

  Next, the user moves the sample moving stage 3 while referring to the TV monitor 20 so that the center position 44 of the reference chart 40 coincides with the intersection of the diagonal lines 43 displayed by the jig software. At the same time, the user rotates the reference chart 40 on the sample moving stage 3 so that the intersections of the meshes of the reference chart 40 are aligned in the diagonal direction of the visual field to align the intersections of the meshes (S15). The reference chart 40 is fixed on the sample moving stage 3 with this position and orientation.

  Next, the revolver 6 is rotated under the control of the control device 25, and the low-magnification objective lens 5a is inserted into the observation optical axis 8 (S16). Then, under the control of the control device 25, the zoom magnifying mechanism 9 moves the zoom lens so as to achieve the minimum zoom (S17). The control device 25 drives the TV camera 10 to capture an image of the basic chart 40 under the low magnification objective lens 5a and the minimum magnification zoom condition. Then, the screen shown in FIG. 4A is displayed on the TV monitor 20.

  The user refers to the TV monitor 20 and confirms whether the intersection of the diagonal lines 43 and the center position 44 of the reference chart 40 match (S18). If the intersection of the diagonal lines 43 and the center position 44 of the reference chart 40 do not match (go to “No” in S18), the process is repeated again from S13.

  If the intersection of the diagonal lines 43 and the center position 44 of the reference chart 40 coincide with each other, it is confirmed whether the intersections of the squares of the reference chart 40 are substantially aligned in the diagonal direction of the visual field (S19). If the intersections of the squares of the reference chart are not aligned in the diagonal direction of the field of view (go to “No” in S19), the process is repeated again from S13.

  When the intersections of the grids of the reference chart are aligned in the diagonal direction of the field of view, the PC mouse 21 is used to refer to the diagonal direction of the reference chart 40 while referring to the TV monitor 20, and the intersections of the grids in a certain direction from the center (for example, In the upper right direction), a plurality of points (for example, about 7 points) are pointed (S20). For example, in the case of FIG. 4A, the points P1, P2, and P3 are pointed from the center side.

  The intersection points P1, P2, and P3 are points actually clicked on the reference chart 40 displayed on the TV monitor, and these are affected by the distortion, so that they deviate from the true intersection positions P1a, P2a, and P3a. Yes. The intersection position of the true square is the absolute coordinates of the reference chart 40 stored in the storage circuit 29 in advance.

  The difference between the absolute coordinates of the reference chart 40 and the apparent coordinates pointed on the TV monitor 20 (difference between P1 and P1a, difference between P2 and P2a, difference between P3 and P3a), that is, the deviation amount is a coordinate error. As a correction amount of coordinates. In addition, since the length of the diagonal of the true square can be calculated based on the absolute coordinates of the reference chart 40, the distance between points (between P1-P2 and between P2-P3) with respect to the length of the diagonal of the true square. The interval is a magnification correction amount as a magnification error.

  In general, the distortion has a uniform distribution from the center of the visual field to the image height direction. Therefore, the n points pointed above are set to n strip-shaped correction regions concentrically with respect to the center of the visual field. An example of this will be described with reference to FIG.

  FIG. 7 shows an example of dividing the correction area when three points are pointed under the low magnification objective lens and the minimum magnification zoom condition in the present embodiment. In the example shown in the figure, the three points pointed at P1, P2, and P3 are divided into three strip-shaped correction regions A, B, and C concentrically with respect to the center of the visual field.

The control device 25 stores the coordinate error and the magnification error with respect to the coordinates of the region in the macro observation image correction table in the storage circuit 29 (S21).
FIG. 8 shows an example of a macro observation image correction table in the present embodiment. Here, description will be made using a macro observation image correction table corresponding to FIG. The macro observation image correction table 50 includes data items of “region range” 51, “coordinate error (X coordinate, Y coordinate)” 52, and “magnification error” 53. The “area range” 51 stores coordinates for specifying each area in FIG. The “coordinate error (X coordinate, Y coordinate)” 52 and “magnification error” 53 store the coordinate error and magnification error at each point pointed above.

  In this embodiment, a plurality of points are sampled to obtain a coordinate error and a magnification error, and these are used as representative values for each coordinate region. The macro observation image correction table may be created by acquiring the coordinate error and the magnification error for the point.

The error information about the macro observation image has been obtained through the steps so far. Next, error information related to the micro live image is acquired.
First, the revolver 6 is rotated by the control of the control device 25 to switch the objective lens, and the high-magnification objective lens 5b is inserted into the observation optical axis 8 (S22). Then, under the control of the control device 25, the zoom magnifying mechanism 9 moves the zoom lens so that the minimum zoom is achieved. The control device 25 drives the TV camera 10 to capture an image of the basic chart 40 under the conditions set to the high magnification objective lens 5b and the minimum magnification zoom. Then, the screen shown in FIG. 4B is displayed on the TV monitor 20.

  FIG. 4B shows a state in which the diagonal line 43 is displayed by the jig software in a state where the zoom is the minimum magnification with the high-magnification objective lens 5b. In this state, the center point P0 of the reference chart 40 is pointed (S23). The difference between the center P0 of the reference chart 40 and the intersection of the diagonal line 43 is stored in the micro live image correction table 60 in the recording circuit 29 as a coordinate error of the high magnification objective lens 5b and the high magnification center coordinate with respect to the lowest zoom.

  Next, the intersection point P11 of the mesh close to the center P0 of the reference chart 40 is pointed (S24). The control device 25 calculates the magnification value M1 from the distance between the point P0 and the point P11 and the distance of the intersection of the grids of the reference chart 40, and calculates the magnification value M1 with respect to the magnification value M2 possessed by the zoom magnification changing mechanism 9. Is stored in a micro live image correction table to be described later.

  This operation is performed, for example, about 10 points up to the maximum zoom, and the correction amount of the coordinate of the center P0 and the correction amount of the zoom magnification are stored in the micro live image correction table for each zoom magnification (S25, S26). The calibration is completed when the correction amount of the coordinate of the center P0 and the correction amount of the zoom magnification are stored in the micro live image correction table for each zoom magnification of the maximum magnification.

  FIG. 9 shows an example of the micro live image correction table in the present embodiment. The micro live image correction table 60 includes data items of “zoom magnification” 61, “coordinate error of high magnification center coordinates (X coordinate, Y coordinate)” 62, and “magnification error of high magnification magnification” 63. The “zoom magnification” 61 stores a zoom magnification that can be set by the zoom magnification mechanism 9.

  In the “coordinate error (X coordinate, Y coordinate) of high magnification center coordinate” 62, a coordinate error corresponding to the “zoom magnification” 61 acquired in S23 of FIG. 6B is stored. In the “magnification error of high magnification” 63, a magnification error corresponding to the “zoom magnification” 61 acquired in S24 of FIG. 6B is stored.

  In this embodiment, only one type of high-magnification objective lens is used, so there is only one micro live image correction table 60. However, when a plurality of types of high-magnification objective lenses are used, the micro-magnification corresponding to each high-magnification objective lens is used. There is a live image correction table.

  In the present embodiment, the correction data is stored in the storage circuit 29 as a data table as described above, but the storage format of the correction data is not limited to the data table. For example, as shown in FIG. 5, the distortion can be expressed by an n-order function according to each objective lens and each zoom magnification. Therefore, instead of the macro observation image correction table and the micro live image correction table, the objective can be expressed. An n-order function indicating a distortion curve for each lens and each zoom magnification may be stored in the storage circuit 29 as a complementary expression. Note that calibration is performed at the time of shipment of the product, and calibration can be performed again when the objective lens is replaced or broken.

<Microscopic observation by macro observation image and micro live image (S2)>
In S2, first, when the user surrounds an arbitrary place with a ROI in the navigation map image, the center coordinates of the ROI range are recognized and corrected, and stored as designated center coordinates. Further, the magnification value is recognized from the ROI range, the magnification is corrected in accordance with the coordinates of the ROI range, and stored as the designated magnification value. Next, the zoom lens is switched to a high magnification objective lens, and the magnification is changed to a correction zoom magnification corresponding to the designated magnification value. The corrected zoom magnification is a magnification corrected for each zoom by calibration. The correction center coordinate value is also a center coordinate corrected for each zoom by calibration.

  In this way, the coordinate error between the macro observation image and the micro live image and the magnification error are corrected based on the macro observation image correction table and the micro live image correction table generated in the calibration of S1. Can do.

  FIG. 10A and FIG. 10B show an operation flow of the microscope system that corrects the coordinate error and the magnification error when performing the microscope observation using the macro observation image and the micro live image in the present embodiment. First, the power source of the microscope body 1 is turned on (S31). Then, the control apparatus 25 controls each electric drive part of the microscope main body 1, and performs origin search of each drive part (S32). Then, each drive unit stops at the origin position.

  Next, the macro observation image 101 used when the user designates a range in which detailed observation is desired is acquired (S33). Here, the revolver 6 is driven by the control of the control device 25, and the low-magnification objective lens 5 a of the two objective lenses 5 is inserted on the observation optical axis 8. Then, the zoom magnifying mechanism 9 moves the zoom lens so that the minimum zoom is achieved. Then, an imaging magnification for capturing the macro observation image 101 can be obtained.

  The TV camera control device 26 controls the TV camera 10 to optimally set the exposure amount, white balance, and the like, and acquires the macro observation image 101. The macro observation image 101 is displayed on the TV monitor 20 via the image output circuit 31 (S34).

  Next, as described with reference to FIGS. 11A and 11B, the user performs a drag operation with the PC mouse 21 to enclose a region to be observed in detail in the macro observation image 101 on the TV monitor 20 with a ROI 103 (S35). ).

  Here, since the ROI 103 enclosed by the user is usually different from the aspect ratio of the CCD, the portion that is not displayed in the micro live image when the screen is shifted to the micro live image 102 corresponding to the image of the area specified by the ROI 103 as it is. Occurs. Therefore, in the present embodiment, when the user releases the button of the PC mouse 21 after enclosing the rectangle with the ROI 103 by a drag operation, the lower right position of the ROI 103 is adjusted by the control device 25, and the selected ROI 103 has the CCD aspect ratio. The ratio is changed to the same ratio (S36).

  When the size of the ROI 103 is large enough to exceed the magnification range obtained by the combination of the magnification range of the high magnification objective lens 5b and the zoom magnification mechanism 9, the control device 25 obtains the magnification obtained by the combination of the size of the ROI 103. The range is converted into the size of the ROI 103 having the lowest magnification. Conversely, if the size of the ROI 103 is small enough to exceed the magnification range obtained by the combination of the magnification range of the high-magnification objective lens 5b and the zoom magnification mechanism 9, the control device 25 can obtain the size of the ROI 103 by that combination. It converts to the size of ROI of the highest magnification in the magnification range. (S37).

  If the position and size of the ROI 103 displayed on the macro observation image 101 are acceptable, the user confirms the range of the ROI 103 by performing a predetermined operation (proceed to "Yes" in S38). If the position or size of the ROI 103 is changed, the ROI 103 is enclosed again (proceed to “No” in S38).

  After determining the range of the ROI 103, the control device 25 calculates the diagonal center coordinate of the ROI 103 on the macro observation image 101, and sets the calculated diagonal center coordinate as the designated center coordinate C1 (S39).

  The control device 25 reads the macro observation image correction table 50 from the storage circuit 29, and records the record R1 having the “region range” 51 including the designated center coordinate C1 from the macro observation image correction table 50 using the designated center coordinate C1 as a key. Extract. The control device 25 acquires the “coordinate error” 52 from the extracted record R1. Then, the control device 25 adds the “coordinate error” 52 to the designated center coordinate C1 in the arithmetic circuit 30, and obtains the added value as the center coordinate correction value C2 (S40).

Next, the control device 25 calculates a designated magnification M3 from the ratio of the long side of the ROI 103 to the long side of the macro observation image 101 (S41).
Then, the control device 25 acquires the “magnification error” 53 from the record R1 extracted above. The control device 25 multiplies the designated magnification M3 by the “magnification error” 53 in the arithmetic circuit 30 to obtain a magnification correction value M4 (S42).

Next, the revolver 6 is rotated under the control of the control device 25, and the high-magnification objective lens 5b is inserted into the observation optical axis 8 (S43).
Then, the control device 25 reads the micro live image correction table 60 from the storage circuit 29, and uses the magnification correction value M4 as a key, and among the records included in the micro live image correction table 60, the “magnification error of high magnification” 63 is the magnification. The record R2 having the “high-magnification magnification error” 63 closest to the correction value M4 is extracted. The control device 25 acquires the “zoom magnification” 61 from the extracted record R2, and drives the zoom scaling mechanism 9 to set the zoom magnification (S44). Thereby, in order to realize a high magnification correction value equivalent to the magnification correction value M2, it is possible to set the zoom magnification determined by the combination of the high magnification objective lens 5b and the zoom magnification changing mechanism 9.

  Next, the control device 25 acquires the “coordinate error of the high magnification center coordinates” 62 from the record R2 extracted above. Then, the control device 25 adds the “coordinate error of the high-magnification center coordinate” 62 to the center coordinate correction value C2 in the arithmetic circuit 30 to obtain the center coordinate correction value C3. The control device 25 drives the sample moving stage 3 so that the center coordinate correction value C3 becomes the center of the visual field (S45). The storage circuit 29 stores correction data relating to the absolute position of the stage for correcting a movement amount error caused by a rotation angle error of the stage drive motor of the sample moving stage 3, and therefore the sample moving stage. When driving 3, the movement amount error is also corrected.

  Next, the TV camera control device 26 acquires the micro live image 102 by optimally setting the shooting conditions. As shown in FIGS. 11A and 11B, the image output circuit 31 displays the acquired micro live image 102 on the TV monitor 20 (S46).

  When the image of the micro live image 102 displayed on the TV monitor 20 is stored, the process proceeds to an imaging flow (not shown) (proceed to “Yes” in S47). Note that the imaging flow is not directly related to the present embodiment, and a description thereof will be omitted.

  In addition, when the micro live image 102 is obtained at a site different from the ROI 103 selected above or at a different magnification (proceed to “No” in S48 and proceed to “Yes” in S49), it is desired to observe on the macro observation image 101 in S35. The process is repeated from the process of enclosing the area in a rectangle with the ROI 103 (proceed to “Yes” in S50). Or it moves to the flow which designates other micro live images which are not illustrated (it progresses to "No" at S50).

When all the processes are completed (proceed to "Yes" in S48), the power is turned off (S51), and the observation is terminated.
According to the present embodiment, simply by designating the region to be observed using the ROI using the PC mouse 21, the observation magnification is automatically determined accordingly, and the sample moving stage 3 moves, and the range designated by the ROI. The same magnification and center position can be displayed. As a result, an enlargement magnification selection error does not occur as in the prior art. Further, the troublesome operation of moving the sample moving stage when changing the magnification is not necessary. In addition, since the ROI designation is made in the macro observation image, the observation position of the sample is clear. In addition, tolerances that cannot be removed by lens design or mechanical adjustment can be corrected.

  In the above embodiment, the microscope main body 1 is not provided with a reference chart. However, a reference chart that can be inserted into and removed from the observation optical axis 8 may be held on the sample moving stage 3. Thereby, calibration can be easily performed every time the power of the microscope zooming device is turned on. Note that the number of calibrations can be changed. Therefore, in addition to the above embodiment, effects such as self-correction of correction values and automatic failure determination can be obtained.

  According to the present embodiment, the following can be realized. When a region to be observed on the macro observation image is surrounded by ROI, the center coordinates of the ROI range are recognized and corrected, and stored as designated center coordinates. Further, the magnification value is recognized from the ROI range, the magnification is corrected according to the coordinates of the ROI range, and stored as the designated magnification value. Then, the zoom lens is switched to a high magnification objective lens, and the magnification is changed to a correction zoom magnification corresponding to the designated magnification value. The corrected zoom magnification is a magnification corrected for each zoom by calibration. The corrected center coordinate value is also a center coordinate corrected for each zoom by calibration.

  As described above, the ROI designated by the user in the navigation map image can be displayed as a live image. This eliminates the troublesome work for the user when changing the observation magnification, and allows the observation magnification to be selected quickly and appropriately.

1 shows an overall configuration of a microscope system in the present embodiment. An example of a structure of the control apparatus 25 of FIG. 1 is shown. An overall outline flow of the present embodiment is shown. The example of a display of the reference | standard chart imaged on the conditions of the low magnification objective lens 5a and the zoom magnification of the minimum magnification is shown. The example of a display of the reference | standard chart imaged on the conditions of the high magnification objective lens 5b and the zoom magnification of the minimum magnification is shown. The example of a display of the reference | standard chart imaged on the conditions of the high magnification objective lens 5b and the zoom magnification of the minimum magnification is shown. A distortion shape that changes depending on the combination of the objective lens and the zoom magnification of the zooming mechanism is shown. The flow (the 1) of the calibration in this embodiment is shown. The flow (the 2) of the calibration in this embodiment is shown. An example of dividing a correction area when three points are pointed under the low magnification objective lens and the minimum magnification zoom condition in the present embodiment is shown. An example of the macro observation image correction table in this embodiment is shown. An example of the micro live image correction table in this embodiment is shown. An operation flow (part 1) of the microscope system for correcting the coordinate error and the magnification error when performing the microscope observation using the macro observation image and the micro live image in the present embodiment is shown. An operation flow (part 2) of the microscope system for correcting the coordinate error and the magnification error when performing microscopic observation with the macro observation image and the micro live image in the present embodiment is shown. An example (part 1) in which an arbitrary area on a conventional navigation map image is enlarged and displayed is shown. An example (part 2) in which an arbitrary area on a navigation map image in the related art is displayed in an enlarged manner is shown.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microscope main body 2 Illuminating device 3 Sample moving stage 4 Focusing device 5 Objective lens 5a Low magnification objective lens 5b High magnification objective lens 6 Revolver 9 Zoom magnification mechanism 10 TV camera 11 High brightness mercury lamp 12 Fluorescence observation device 13 Excitation filter 14 Absorption Filter 15 Dichroic mirror 16, 16a Fluorescent cube 17 Turret mechanism 18 Lens barrel 18a Eyepiece 19 PC
20 TV monitor 21 PC mouse 25 Control device 26 TV camera control device 27 Input circuit 28 Drive circuit 29 Memory circuit 30 Arithmetic circuit 31 Image output circuit

Claims (8)

A sample moving stage on which an observation sample is placed and movable in a direction perpendicular to the observation optical axis;
A revolver for switching a plurality of objective lenses with different magnifications inserted into the observation optical axis;
Zoom scaling means for changing the zoom magnification;
An imaging means for imaging an optical image of the observation sample through the objective lens;
An area designating unit capable of designating, as a designated area, a predetermined area of a first observation image captured at a first zoom magnification by the first objective lens among the objective lenses based on an instruction operation from a user; ,
First error information storage means for storing a first coordinate error and a first magnification error of an observation image obtained based on the first objective lens and the first zoom magnification;
The second coordinate error and the second magnification of the second observation image obtained based on the combination of the second objective lens among the objective lenses and the zoom magnification that has been changed within a variable range. Second error information storage means in which an error has occurred;
The designated center coordinate, which is the center coordinate of the designated area, is corrected with the first coordinate error to be the first corrected center coordinate, and the designation is made based on the size of the designated area with respect to the first observation image. A first error correction unit that calculates a magnification for enlarging the region, and corrects the magnification with the first magnification error to obtain a first correction magnification;
Of the magnifications based on the combination of the magnification of the second objective lens and the zoom magnification, a combination that is closest to the first correction magnification is specified, and the first correction center coordinate is set as the second coordinate error. A second error correction unit that corrects the second correction center coordinate to
Based on the combination of the magnification of the specified objective lens and the zoom magnification, the revolver is controlled to switch the objective lens, the zoom magnification changing unit is controlled to vary the zoom magnification, and Control means for performing control to move the sample moving stage based on second correction center coordinates;
A microscope apparatus comprising: The first error information storage means stores, as the first coordinate error, a distance traveled by a predetermined position on the sample moving stage through the first objective lens, and stores the predetermined error on the sample moving stage. The ratio between the distance between the two points and the distance between the two points expanded and contracted through the first objective lens is stored as the first magnification error. Microscope equipment. The first error information storage means includes
A plurality of regions are concentrically divided from the center of the first observation image, and the first coordinate error and the first magnification error are calculated and stored for each of the divided regions. The microscope apparatus according to claim 1. The second error information storage means captures the first objective lens and the first objective image captured at the first zoom magnification with respect to the first observation image and the second objective lens and the respective zoom magnifications. 2 is stored as the second coordinate error, and the ratio of the actual magnification to the display magnification for each magnification obtained by the combination of the second objective lens and each zoom magnification is the second magnification. The microscope apparatus according to claim 1, wherein the microscope apparatus stores the error. The second error information storage means includes coordinates in the second observation image corresponding to the first correction center coordinates in the first observation image and the center of the observation field of the second observation image. The difference from the coordinates is stored as the second coordinate error, and the ratio between the total display magnification and the total actual magnification for each zoom magnification in the second objective lens is stored as the second magnification error. The microscope apparatus according to claim 4. The microscope apparatus further includes:
The microscope apparatus according to claim 1, further comprising a reference chart that can be inserted into and removed from the observation optical axis and has a grid pattern on the sample moving stage. A sample moving stage on which an observation sample is placed and movable in a direction perpendicular to the observation optical axis;
A revolver for switching a plurality of objective lenses with different magnifications inserted into the observation optical axis;
A zoom magnification mechanism that varies the zoom magnification;
An imaging device that captures an optical image of the observation sample via the objective lens;
An area designation function capable of designating the predetermined area of the first observation image captured at the first zoom magnification with the first objective lens among the objective lenses as the designated area based on an instruction operation from the user. When,
A first error information storage device storing a first coordinate error and a first magnification error of an observation image obtained based on the first objective lens and the first zoom magnification;
The second coordinate error and the second magnification of the second observation image obtained based on the combination of the second objective lens among the objective lenses and the zoom magnification that has been changed within a variable range. A second error information storage device having an error;
A microscope control program for causing a computer to execute a process for controlling the operation of a microscope apparatus comprising:
The designated center coordinate, which is the center coordinate of the designated area, is corrected with the first coordinate error to be the first corrected center coordinate, and the designation is made based on the size of the designated area with respect to the first observation image. A first error correction process for calculating a magnification for enlarging the region and correcting the magnification with the first magnification error to obtain a first correction magnification;
Of the magnifications based on the combination of the magnification of the second objective lens and the zoom magnification, a combination that is closest to the first correction magnification is specified, and the first correction center coordinate is set as the second coordinate error. A second error correction process that is corrected with the second correction center coordinates,
Based on the combination of the magnification of the specified objective lens and the zoom magnification, the revolver is controlled to switch the objective lens, the zoom magnification mechanism is controlled to vary the zoom magnification, and A drive control process for performing control to move the sample moving stage based on the second correction center coordinates;
A microscope control program for causing a computer to execute. A sample moving stage on which an observation sample is placed and movable in a direction perpendicular to the observation optical axis;
A revolver for switching a plurality of objective lenses with different magnifications inserted into the observation optical axis;
A zoom magnification mechanism that varies the zoom magnification;
An imaging device that captures an optical image of the observation sample via the objective lens;
An area designation function capable of designating the predetermined area of the first observation image captured at the first zoom magnification with the first objective lens among the objective lenses as the designated area based on an instruction operation from the user. When,
A first error information storage device storing a first coordinate error and a first magnification error of an observation image obtained based on the first objective lens and the first zoom magnification;
The second coordinate error and the second magnification of the second observation image obtained based on the combination of the second objective lens among the objective lenses and the zoom magnification that has been changed within a variable range. A second error information storage device having an error;
A microscope control method for controlling the operation of a microscope apparatus comprising:
The designated center coordinate, which is the center coordinate of the designated area, is corrected with the first coordinate error to be the first corrected center coordinate, and the designation is made based on the size of the designated area with respect to the first observation image. Calculating a magnification for enlarging the region, and correcting the magnification with the first magnification error to obtain a first correction magnification;
Of the magnifications based on the combination of the magnification of the second objective lens and the zoom magnification, a combination that is closest to the first correction magnification is specified, and the first correction center coordinate is set as the second coordinate error. To correct the second correction center coordinates,
Based on the combination of the magnification of the specified objective lens and the zoom magnification, the revolver is controlled to switch the objective lens, the zoom magnification mechanism is controlled to vary the zoom magnification, and Control to move the sample moving stage based on the second correction center coordinates,
The microscope control method characterized by the above-mentioned.

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