JP2016009163A - Microscope system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a microscope system that allows a simple configuration to accurately correct a deviation of a specified position of a sample due to a thermal drift regardless of a kind of the sample.SOLUTION: A microscope system 1 comprises: a deviation-amount calculation unit 625 that calculates an amount of deviation of an index indicative of a motion of a stage 211 arising from at least a thermal drift on the basis of a reference image corresponding to image data generated by having the index photographed at a prescribed position on the stage 211 by an imaging device 3 before the stage 211 moves to a plurality of specified positions in a sample SP, and a current image corresponding to image data generated by having the index photographed at the prescribed position by the imaging device 3 after one cycle of the number of times when defining photography at the plurality of specified positions by the imaging device 3 in the sample SP as one cycle is over; and a correction unit 626 that corrects the plurality of specified positions in the sample SP to be recorded by a photography information recording unit 611 on the basis of the amount of deviation calculated by the deviation-amount calculation unit 625.

Description

  The present invention relates to a microscope system that displays an image corresponding to image data generated by imaging a specimen placed on a stage.

  Conventionally, as an observation method for observing a specimen such as a living cell with a microscope, a specified position of the specimen is photographed at predetermined time intervals to generate specimen image data, and a series of image data is time-sequentially after photographing is completed. There is a method of observing a change in the shape of a temporal specimen as a moving image (hereinafter referred to as “time-lapse observation”) by connecting and reproducing the images along the display.

  When such lime lapse observation is performed for a long time, the relative positional relationship between the objective lens and the designated position of the sample is shifted due to the occurrence of thermal drift (expansion) due to the heat of the motorized part of the microscope and the light source. There is a problem that a moving image is reproduced and displayed as if the sample has moved.

  For this reason, the microscope is provided with a two-dimensional photoelectric converter for detecting specular reflection and scattered light from the cover glass of the sample, and the detection result detected by the two-dimensional photoelectric converter is used to shift the designated position of the sample. Is known, and based on the detection result, a technique for correcting the deviation of the designated position of the sample is known (see Patent Document 1).

  In addition, a temperature sensor is provided in the microscope, and based on the temperature detected by this temperature sensor, the amount of deviation is estimated from related data that correlates the amount of temperature change and the amount of deviation (thermal drift amount) of the microscope. Based on the above, a technique for correcting the deviation of the designated position of the sample is known (see Patent Document 2).

International Publication No. 2011/007768 JP2013-8030A

  However, since the technique of Patent Document 1 described above requires a two-dimensional photoelectric converter, the configuration of the apparatus is complicated, and the reflected light and scattered light from the cover glass of the specimen are used, so that the specimen that can be observed can be observed. There was a problem that the types were limited.

  In addition, since the technique of Patent Document 2 described above requires a temperature sensor, the configuration of the apparatus is complicated, and the amount of thermal drift is estimated based on the temperature detected by the temperature sensor. There was a problem of lacking.

  The present invention has been made in view of the above, and provides a microscope system capable of accurately correcting a deviation of a designated position of a specimen due to thermal drift with a simple configuration regardless of the kind of specimen. With the goal.

  In order to solve the above-described problems and achieve the object, a microscope system according to the present invention is a stage on which a specimen is placed and which can be moved in a horizontal direction, and is arranged to face the stage, and observe the specimen. An objective lens, a stage drive unit that moves the stage relative to the objective lens, an imaging device that images the specimen through the objective lens and generates image data of the specimen, A plurality of designated positions for designating observation locations on the specimen, a photographing order of each of the plurality of designated positions by the imaging device, a photographing time of each of the plurality of designated positions by the imaging device, and the imaging device by the imaging device A shooting information setting unit for setting shooting information in association with the number of cycles indicating the number of times of repeating shooting at each of a plurality of designated positions; A stage control unit that drives the stage driving unit to sequentially move the stage to the plurality of designated positions based on the imaging information set by the unit; and the imaging at the imaging time set by the imaging information setting unit An imaging control unit for imaging the sample by the apparatus, and the stage of the stage at least due to thermal drift at the predetermined position on the stage before the stage moves to the plurality of designated positions. A reference image corresponding to the image data generated by shooting the index shown, and a current image corresponding to the image data generated by shooting the index at the predetermined position after the first cycle is completed Based on the deviation amount calculation unit for calculating the deviation amount of the index, and based on the deviation amount calculated by the deviation amount calculation unit, the plurality of specified positions Characterized in that the provided correction unit for correcting each of the.

  In the microscope system according to the present invention as set forth in the invention described above, the stage control unit drives the stage driving unit to position the index on the optical axis of the objective lens every time one cycle is completed. The stage is moved as described above.

  The microscope system according to the present invention is the microscope system according to the above aspect, wherein the stage control unit is configured so that the stage is turned on every time when the predetermined number of cycles ends or every time a predetermined time elapses after the imaging device images the sample. The drive unit is driven to move the stage to the predetermined position so that the index is positioned on the optical axis of the objective lens.

  The microscope system according to the present invention further includes a recording unit that records the image data sequentially generated by the imaging device and the shift amount in association with each other in the above-described invention, and the correction unit has the number of cycles After completion, correction is performed so that the center in the image corresponding to the image data becomes the specified position based on the shift amount associated with the image data.

  In the microscope system according to the present invention as set forth in the invention described above, the index is a fluorescent substance that emits fluorescence with respect to light having a predetermined wavelength band.

  Moreover, the microscope system according to the present invention is characterized in that, in the above invention, the index is provided in a container for storing the specimen.

  In the microscope system according to the present invention as set forth in the invention described above, the index is provided on a placement surface on which the specimen is placed on the stage.

  The microscope system according to the present invention is characterized in that, in the above invention, a plurality of the indicators are provided in the same plane.

  According to the microscope system of the present invention, before the shift amount calculation unit moves to a plurality of designated positions of the specimen by the stage, the imaging apparatus performs at least a stage movement caused by thermal drift at a predetermined position on the stage. Based on the reference image corresponding to the image data generated by shooting the indicated index and the current image corresponding to the image data generated by shooting the index at a predetermined position after the first cycle is completed. Then, the deviation amount of the index is calculated, and the correction unit corrects each of the plurality of designated positions indicating the observation location on the sample set by the imaging information setting unit based on the deviation amount calculated by the deviation amount calculation unit. . As a result, regardless of the type of the sample, the displacement of the designated position of the sample due to the thermal drift can be accurately corrected with a simple configuration.

FIG. 1 is a schematic configuration diagram showing an example of a configuration of a microscope system according to Embodiment 1 of the present invention. FIG. 2 is a schematic diagram schematically showing the configuration of the microscope system according to the first embodiment of the present invention. FIG. 3 is a plan view of a container containing a plurality of specimens. FIG. 4 is a flowchart showing an outline of processing executed by the microscope system according to Embodiment 1 of the present invention. FIG. 5 is a flowchart showing an outline of the setting process of FIG. FIG. 6A is a flowchart showing an overview of the time lapse process of FIG. FIG. 6B is a flowchart showing an overview of the time lapse process of FIG. FIG. 6C is a diagram illustrating a positional deviation of the designated position coordinates on the sample surface. FIG. 6D is a time chart showing an overview of the time lapse process of FIG. FIG. 7 is a flowchart showing an outline of setting processing executed by the microscope system according to Embodiment 2 of the present invention. FIG. 8A is a flowchart showing an overview of time-lapse processing executed by the microscope system according to Embodiment 2 of the present invention. FIG. 8B is a flowchart showing an overview of time-lapse processing executed by the microscope system according to Embodiment 2 of the present invention. FIG. 8C is a time chart illustrating an outline of a time lapse process executed by the microscope system 1 according to the second embodiment. FIG. 9 is a schematic diagram schematically showing the configuration of the microscope system according to the third embodiment of the present invention. FIG. 10A is a flowchart showing an overview of time-lapse processing executed by the microscope system according to Embodiment 3 of the present invention. FIG. 10B is a flowchart showing an overview of time-lapse processing executed by the microscope system according to Embodiment 3 of the present invention. FIG. 10C is a diagram showing an outline of time-lapse moving image file creation in the time-lapse process executed by the microscope system according to Embodiment 3 of the present invention. FIG. 11A is a flowchart showing an overview of time-lapse processing executed by the microscope system according to Embodiment 4 of the present invention. FIG. 11B is a flowchart showing an overview of time-lapse processing executed by the microscope system according to Embodiment 4 of the present invention. FIG. 12 is a top view showing a configuration of a stage according to Modification 1 of Embodiments 1 to 4 of the present invention. FIG. 13 is a top view of a container for storing a specimen according to Modification 2 of Embodiments 1 to 4 of the present invention. FIG. 14 is a diagram schematically showing a correction method for correcting the shift amount of the designated position of the sample according to the second modification of the first to fourth embodiments of the present invention. FIG. 15A is a diagram showing concentric markers fixed in a container for accommodating a specimen according to Modification 3 of Embodiments 1 to 4 of the present invention. FIG. 15B is a diagram showing circular markers with different diameters that are fixed in a container that accommodates a specimen according to Modification 3 of Embodiments 1 to 4 of the present invention.

  DESCRIPTION OF EMBODIMENTS Hereinafter, embodiments for carrying out the present invention (hereinafter referred to as “embodiments”) will be described with reference to the drawings. The present invention is not limited to the embodiments described below. The drawings referred to in the following description only schematically show the shape, size, and positional relationship so that the contents of the present invention can be understood. That is, the present invention is not limited only to the shape, size, and positional relationship illustrated in each drawing.

(Embodiment 1)
[Configuration of microscope system]
FIG. 1 is a schematic configuration diagram showing an example of a configuration of a microscope system according to Embodiment 1 of the present invention. FIG. 2 is a schematic diagram schematically showing the configuration of the microscope system according to the first embodiment of the present invention. 1 and 2, the plane on which the microscope system 1 is placed will be described as an XY plane, and a direction perpendicular to the XY plane will be described as a Z direction.

  A microscope system 1 shown in FIGS. 1 and 2 includes a microscope apparatus 2 for observing a specimen SP, an imaging apparatus 3 for imaging the specimen SP through the microscope apparatus 2, and generating image data of the specimen SP, and the microscope system 1. An input unit 4 that receives input of various operations of the above, a display unit 5 that displays an image corresponding to image data generated by the imaging device 3, a microscope control device 6 that comprehensively controls each unit of the microscope system 1, and Is provided.

[Configuration of microscope device]
First, the configuration of the microscope apparatus 2 will be described. The microscope apparatus 2 includes a main body unit 20 that forms a base, a stage mechanism 21 that is mounted on the upper surface of the main body unit 20, and a specimen SP that is placed on the stage 211. Part 22, objective lens 23 for taking in observation light from specimen SP on stage mechanism 21, revolver 24 holding a plurality of objective lenses 23 having different magnifications interchangeably, holding revolver 24, and optical path L1 The focusing unit 25 that moves the objective lens 23 arranged above in the vertical direction along the optical path L1 to focus the objective lens 23 on the specimen SP, and the epi-illumination light source unit 26 that emits epi-illumination light, have mutually different characteristics. Mirror turret 27 holding a plurality of different optical systems, and observation light of specimen SP transmitted through mirror turret 27 is imaged and emitted to lens barrel 29 It includes a system 28, among the side surfaces of the main body portion 20, and an eyepiece is provided a lens barrel portion 29 provided on the front side (see FIG. 1) is a side facing the user microscope apparatus 2, a.

  The stage mechanism 21 includes a stage 211 configured to be movable in the horizontal direction within the XY plane, a stage driving unit 212 that moves the stage 211 within the XY plane, a position detection unit 213 that detects the position of the stage 211, Have

  The stage 211 is configured by sequentially laminating a plate-shaped first member, second member, and third member. The stage 211 uses the first member as a reference (fixed), and the stage driving unit 212 moves the second member and the third member on a plane whose plane is the plate surface of the first member. In this case, the specimen SP is placed on the third member via the container D1. At this time, the second member and the third member move in directions orthogonal to each other on a plane parallel to the main surface. The stage 211 has an opening R1. The opening R1 is formed with a size including the optical path L1 regardless of the movement of the second member and the third member.

  Here, the container D1 that houses the specimen SP will be described in detail. FIG. 3 is a plan view of the container D1 containing a plurality of specimens SP.

  As shown in FIG. 3, the container D1 used for time-lapse observation is provided with a position information acquisition marker M1 (hereinafter simply referred to as “marker M1”) that is an index indicating at least the movement of the stage 211 due to thermal drift. It has been. Here, the index is for detecting a shift in the relative positional relationship between the specimen SP and the objective lens 23 due to the thermal drift of the microscope apparatus 2, and the index of the stage 211 at a predetermined position in advance. This is for detecting the position. The marker M1 is a fluorescent material that emits light with respect to light having a predetermined wavelength band. The marker M1 is smaller than the field of view of the objective lens 23. The container D1 is configured using a transparent member, and for example, a glass bottom dish, a well plate, a slide glass, a beaker, or the like is used. Furthermore, a plurality of specimens SP are accommodated in the container D1. The number of specimens SP can be changed as appropriate according to the observation.

  The stage driving unit 212 is configured using a stepping motor, an ultrasonic motor, or the like. The stage drive unit 212 moves the observation region of the specimen SP by moving the stage 211 in the XY plane under the control of the microscope control device 6.

  The position detection unit 213 is configured using a linear scale, a glass scale, a photo interrupter, or the like. The position detection unit 213 detects a predetermined origin position in the XY plane by an XY position origin sensor (not shown), and uses the origin position as a reference to determine the position (XY coordinate) regarding the X position and the Y position of the stage 211 as a microscope control device. 6 is output. Note that the position detection unit 213 may detect the position of the stage 211 based on the drive amount of the stage drive unit 212.

  The transmitted illumination unit 22 includes a transmitted illumination column 221 that is attached to the main body unit 20 and extends upward, an arm unit 222 that extends from the upper end of the transmitted illumination column 221 in a direction orthogonal to the direction in which the transmitted illumination column 221 extends, and the transmitted illumination column. A first lamp house portion 223 having a transmissive light source 223a that emits transmissive illumination light, and a transmissive illumination light emitted from the first lamp house portion 223 near the upper end of 221 on the side opposite to the side where the arm portion 222 extends. The condenser lens 224 that focuses and forms an image on the specimen SP, the condenser holder 225 that holds the condenser lens 224, and the condenser holder 225 provided on the side surface of the transmission illumination column 221 are moved up and down to focus the condenser lens 224. And a condenser focusing operation unit 226 that performs a semi-operation.

  Inside the arm portion 222, at least the condensing lens 222 a that condenses the transmitted illumination light emitted from the transmitted light source 223 a, and the transmitted illumination light emitted from the condensing lens 222 a is reflected to the direction of the optical axis of the condenser lens 224. And a mirror 222b that reflects in the direction of the optical path L1.

  The transmissive light source 223a is configured by a halogen lamp, a xenon lamp, an LED (Light Emitting Diode), or the like. The transmitted light source 223a emits transmitted illumination under the control of the microscope control device 6.

  As the objective lens 23, a plurality of objective lenses having different magnifications, for example, objective lenses having relatively high magnifications of 10 times, 20 times, and 50 times, objective lenses having low magnifications of 2 times and 5 times, and the like can be freely attached to and detached from the revolver 24. Attached to.

  The revolver 24 is rotatably provided, and the objective lens 23 is disposed below the sample SP. The revolver 24 changes the magnification of the image in the field of view by selectively switching the objective lens 23 used for observation of the specimen SP arranged on the optical path L1 by rotating.

  The focusing unit 25 transmits the observation image of the sample SP incident through the objective lens 23 to the mirror turret 27, while reflecting the transmissive mirror 251 that reflects to the AF unit 252 described later, and the sample reflected by the transmissive mirror 251. An AF unit 252 that generates an AF signal for adjusting the focus of the objective lens 23 by performing phase difference AF processing or contrast AF processing using the SP observation image, and an AF signal input from the AF unit 252 The focusing unit 253 adjusts the focal point of the objective lens 23 by moving the revolver 24 up and down.

  The epi-illumination light source unit 26 includes a second lamp house unit 261 having an epi-illumination light source 261 a that emits epi-illumination light, a floodlight tube 262 that collects the epi-illumination light emitted by the epi-illumination light source 261 a and emits it to the mirror turret 27, Have The light projection tube 262 is provided with at least a condenser lens 262a for condensing the epi-illumination light emitted from the epi-illumination light source 261a, a diaphragm, a filter, and the like.

  The mirror turret 27 includes a reflected light optical system 271 for fluorescence that switches the optical path of the reflected light or transmitted light of the specimen SP incident through the objective lens 23 or the reflected illumination light emitted from the second lamp house unit 261, and the reflected light optics. A mirror unit 272 that holds the system 271, a mirror cassette 273 that can accommodate a plurality of mirror units 272 that respectively hold the incident optical systems 271 having different characteristics, and a mirror drive unit 274 that rotates the mirror cassette 273.

  The epi-illumination optical system 271 reflects an excitation filter 271a that transmits only light having a predetermined wavelength as epi-illumination light (excitation light), and irradiates the sample SP with light having a wavelength corresponding to the excitation light. A dichroic mirror 271b that transmits light having a wavelength corresponding to the observation light, and an absorption filter 271c that transmits only a predetermined fluorescent component of the observation light of the specimen SP that has passed through the dichroic mirror 271b.

  The mirror cassette 273 is rotatably arranged, and is rotated by the mirror driving unit 274, thereby arranging a desired mirror unit 272 on the optical path L1.

  The observation optical system 28 transmits part of the observation light (fluorescence) from the specimen SP transmitted through the mirror turret 27 and branches the remaining light to the imaging device 3 to the imaging device 3. The first mirror 282 that reflects the observation light of the specimen SP transmitted through the half mirror 281, the second mirror 283 that reflects the observation light of the specimen SP reflected by the first mirror 282, and the specimen SP that is reflected by the second mirror 283. And a third mirror 284 that reflects the observation light to the lens barrel portion 29.

  The lens barrel 29 includes an imaging lens 291 that forms an image of the observation light of the sample SP reflected by the third mirror 285, an eyepiece lens 292 that collects the observation light of the sample SP formed by the imaging lens 291; Have

[Configuration of imaging device]
The image pickup apparatus 3 receives the observation light of the specimen SP incident from the half mirror 281 and performs photoelectric conversion, whereby a CCD in which a plurality of pixels that convert light into an electric signal (analog signal) is two-dimensionally arranged. (Charge Coupled Device) or CMOS (Complementary Metal Oxide Semiconductor) or the like and a digital sample SP by performing signal processing such as amplification on the electrical signal output from the image sensor and performing A / D conversion And a signal processing unit that converts the image data and outputs the image data to the microscope control device 6. The imaging device 3 images the specimen SP under the control of the microscope control device 6.

[Configuration of input section]
The input unit 4 is configured using input devices such as a keyboard, a mouse, a joystick, and various switches. The input unit 4 outputs an instruction signal corresponding to the operation received by the various input devices to the microscope control device 6.

[Configuration of display section]
The display unit 5 displays an image corresponding to the image data generated by the imaging device 3 via the microscope control device 6 and various operation information of the microscope system 1. The display unit 5 is configured using a display panel made of liquid crystal or organic EL (Electro Luminescence). A touch panel that detects a contact position of an object from the outside and outputs a position signal corresponding to the detected position to the microscope control device 6 may be provided on the display screen of the display unit 5 in an overlapping manner.

[Configuration of Microscope Control Device]
The microscope control device 6 includes a recording unit 61 that records various information related to the microscope system 1 and a control unit 62 that comprehensively controls the operation of each unit constituting the microscope system 1.

  The recording unit 61 is configured using a semiconductor memory such as a flash memory and a RAM (Random Access Memory). The recording unit 61 records various programs to be executed by the microscope system 1, various data used during the execution of the programs, and image data generated by the imaging device 3. The recording unit 61 temporarily records information being processed by the control unit 62. Furthermore, the recording unit 61 includes an imaging information recording unit 611 regarding imaging information when performing time-lapse observation of the specimen SP in the microscope system 1.

  The control unit 62 is configured using a CPU or the like, and controls the operation of the microscope system 1 by transferring data, instructing, and the like corresponding to each unit constituting the microscope system 1 in accordance with an operation signal received by the input unit 4. Control.

  Here, a detailed configuration of the control unit 62 will be described. The control unit 62 includes a stage control unit 621, a focusing control unit 622, an image processing unit 623, a shooting information setting unit 624, a deviation amount calculation unit 625, a correction unit 626, a shooting control unit 627, and a display. And a control unit 628.

  The stage control unit 621 drives the stage driving unit 212 and moves the stage 211 in accordance with the instruction signal input from the input unit 4. In addition, when the microscope system 1 performs time-lapse observation of the specimen SP, the stage control unit 621 drives the stage 211 based on the photographing information recorded by the photographing information recording unit 611 and causes the stage 211 to be observed on the specimen SP. Are sequentially moved to a plurality of designated positions. In addition, the stage control unit 621 moves the stage 211 to place the marker M1 on the optical axis of the objective lens 23 before starting the time-lapse observation of the specimen SP.

  The focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the revolver 24 in the vertical direction (Z direction), thereby the objective lens 23 for the specimen SP. Adjust the focus.

  The image processing unit 623 performs predetermined image processing on the image data generated by the imaging device 3. Specifically, the image processing unit 623 performs white balance adjustment processing and development processing on the image data generated by the imaging device 3.

  The imaging information setting unit 624 responds to an instruction signal from the input unit 4 to specify a plurality of specified positions for specifying observation positions on the specimen SP, the imaging order of each of the plurality of specified positions by the imaging device 3, and the imaging device 3. Shooting information in which the shooting time (intermittent time) of each of the plurality of designated positions by the image capturing apparatus 3 is associated with the number of cycles indicating the number of times the imaging apparatus 3 repeats shooting at the plurality of designated positions is set, and the set shooting Information is recorded in the photographing information recording unit 611.

  The shift amount calculation unit 625 includes a reference image corresponding to the image data of the marker M1 generated by imaging using the imaging device 3 and the first time-lapse observation before the stage 211 moves to a plurality of designated positions of the specimen SP. After one cycle is completed, pattern matching is performed with the current image corresponding to the image data generated by photographing the marker M1 with the imaging device 3 at a predetermined position, and the deviation amount of the marker M1 is calculated.

  The correction unit 626 sets each of a plurality of designated positions (coordinate information on the stage 211) for designating observation positions on the specimen SP recorded by the imaging information recording unit 611 based on the deviation amount calculated by the deviation amount calculation unit 625. to correct.

  The imaging control unit 627 causes the imaging device 3 to image the sample SP at the imaging time recorded by the imaging information recording unit 611. In addition, the imaging control unit 627 controls imaging of the imaging device 3.

  The display control unit 628 causes the display unit 5 to display an image corresponding to the image data on which the image processing unit 623 has performed image processing. Further, the display control unit 628 causes the display unit 5 to display an operation screen for instructing various operations of the microscope system 1.

  In the microscope system 1 having the above configuration, the user sets the observation method of the microscope apparatus 2 as the transmission observation method or the epi-illumination observation method via the input unit 4 while viewing the operation screen displayed on the display unit 5. Thereafter, when the user performs time-lapse observation via the input unit 4, the user moves to a predetermined registration point of the marker M1 and the specimen SP, and designates each of a plurality of designated positions that designate the observation location on the marker M1 and the specimen SP. The position, shooting order, shooting time (intermittent time), and number of cycles are registered (recorded) in the shooting information recording unit 611. In this case, when the observation method by the microscope apparatus 2 is the epi-illumination observation method and the excitation wavelength of the sample SP and the marker M1 is not the same, the microscope control apparatus 6 rotates the mirror turret 27 and the mirror unit 272 suitable for each. And the designated position of the specimen SP and the position of the marker M1 are recorded in the photographing information recording unit 611. Subsequently, in accordance with an instruction signal from the input unit 4, the imaging information setting unit 624 captures imaging information as imaging information in which each designated position of the specimen SP in time-lapse observation, the imaging time of the designated position, the imaging order, and the number of cycles are set. Records in the recording unit 611.

[Microscope system processing]
Next, the process performed by the time lapse observation which the microscope system 1 performs is demonstrated. FIG. 4 is a flowchart showing an outline of processing executed by the microscope system 1.

  As shown in FIG. 4, first, the microscope system 1 moves to a predetermined registration point of the marker M1 and the specimen SP, and designates a designated position for each of a plurality of designated positions that designate observation points on the marker M1 and the specimen SP. A setting process for registering the shooting order, the shooting time (intermittent time) and the number of cycles in the shooting information recording unit 611 is executed (step S101). Details of the setting process will be described later.

  Subsequently, when an instruction signal for starting time-lapse observation is input from the input unit 4 (step S102: Yes), the microscope system 1 performs time-lapse processing for observing the marker M1 and the specimen SP according to the contents set in the setting processing. Execute (Step S103). Details of the time lapse process will be described later. After step S103, the microscope system 1 ends this process.

  In step S102, when the instruction signal for starting the time-lapse observation is not input from the input unit 4 (step S102: No), the microscope system 1 ends this process.

[Setting processing]
Next, details of the setting process described in step S101 in FIG. 4 will be described. FIG. 5 is a flowchart showing an outline of the setting process.

  As shown in FIG. 5, the imaging information setting unit 624 is a place where the user operates the focusing unit 25 and the stage 211 via the input unit 4 and moves the specimen SP to a predetermined registration point to perform time-lapse observation. The registration point (designated position) is recorded (registered) in the photographing information recording unit 611 (step S201). Specifically, the shooting information setting unit 624 records X, Y, and Z coordinate information of the registration point in the shooting information recording unit 611. More specifically, the imaging information setting unit 624 detects the X and Y coordinates detected by the position detection unit 213 and the height of the stage 211 in the vertical direction with respect to a predetermined registration point of the specimen SP. The Z coordinate detected by (not shown) is recorded in the photographing information recording unit 611. For example, the imaging information setting unit 624 registers the registration point A (Xa0, Ya0, Za0), the registration point B (Xb0, Yb0, Zb0), and the registration point for each of the predetermined registration points A, B, and C of the specimen SP. C (Xc0, Yc0, Zc0) is recorded in the photographing information recording unit 611.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 and switches the marker M1 to a marker photographing wavelength (step S202). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the wavelength for photographing the marker M1.

  Thereafter, the imaging control unit 627 operates the focusing unit 25 and the stage 211 via the input unit 4 by the user to move the marker M1 that has been moved within the field of view of the imaging device 3 (for example, the center of the field of view of the imaging device 3). The image pickup apparatus 3 is caused to take a picture (step S203). In this case, the shooting information setting unit 624 records the X and Y coordinates detected by the position detection unit 213 and the Z coordinate detected by the height detection unit (not shown) in the shooting information recording unit 611 with respect to the marker M1. Let For example, the shooting information setting unit 624 causes the shooting information recording unit 611 to record the registration point (Xm0, Ym0, Zm0) of the marker M1. Furthermore, since the shooting information setting unit 624 records the position of the marker M1 in the image corresponding to the image data shot by the imaging device 3, the image coordinates (Px0) of the marker M1 in the image detected by the image processing unit 623 are recorded. , Py0) is recorded in the photographing information recording unit 611.

  Subsequently, the shooting information setting unit 624 records the time lapse time interval, the shooting time, and the number of shootings input via the input unit 4 in the shooting information recording unit 611 (step S204). For example, the shooting information setting unit 624 sets the shooting interval t for the first to second shots of the point A input via the input unit 4 and the shooting information recording unit 611 for the shooting interval t for T hours or the number of repetitions n. To record. After step S204, the microscope system 1 returns to the main routine of FIG.

[Time-lapse processing]
Next, details of the time lapse process described in step S103 of FIG. 4 will be described. FIG. 6A is a flowchart showing an overview of the time lapse process in step S103 of FIG. FIG. 6B is a flowchart showing an overview of the time lapse process in step S103 of FIG.

  As shown in FIGS. 6A and 6B, first, the stage control unit 621 moves the stage 211 to the marker position by driving the stage driving unit 212 based on the position information recorded by the imaging information recording unit 611. (Step S301). Specifically, the stage control unit 621 moves the stage 211 to the registration point (Xm0, Ym0, Zm0) of the marker M1 recorded by the imaging information recording unit 611.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) based on the AF signal input from the AF unit 252 (step S302). Specifically, the focusing control unit 622 detects the position ΔZm1 in the vertical direction of the focusing unit 25 based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focus position in order to execute the AF process for focusing the focus of 23 on the marker M1 (step S303). Specifically, the focusing control unit 622 moves the focusing unit 25 to the focal position Zm1 (Zm1 = Zm0−ΔZm1) based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 and switches the marker M1 to a marker photographing wavelength (step S304). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the wavelength for photographing the marker M1.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the marker M1 in the visual field region of the imaging device 3 (step S305). In this case, in the imaging control unit 627, the position detection unit 213 detects the marking image (reference image) corresponding to the marking image data generated by the imaging device 3 imaging the marker M1 accommodated in the container D1. The position information (Xm0, Ym0) of the stage 211 and the position information (Zm1) of the focus position of the focusing unit 25 are associated with each other and recorded in the photographing information recording unit 611.

  Subsequently, the deviation amount calculation unit 625 calculates the in-image coordinates of the marker M1 in the marking image generated by the imaging device 3, and records it in the recording unit 61 (step S306). Specifically, the deviation amount calculation unit 625 calculates the in-image coordinates (Px1, Py1) of the marker M1 from the marking image generated by the imaging device 3 and records it in the recording unit 61.

Thereafter, the deviation amount calculation unit 625 calculates the deviation amount of the marker M1 (step S307). Specifically, the deviation amount calculation unit 625 calculates a movement amount (drift amount) by which the marker M1 has moved from the marking image generated by the imaging device 3. More specifically, the shift amount calculation unit 625 calculates the shift amount (ΔPx1, ΔPy1) of the marker M1 on the marking image in pixel conversion by the following equation.
ΔPx1 = Px1−Px0 (1)
ΔPy1 = Py1−Py0 (2)

Subsequently, the deviation amount calculation unit 625 calculates the deviation amount on the sample SP surface (step S308). Specifically, the shift amount calculation unit 625 calculates the shift amount on the specimen SP surface based on the shift amount on the marking image (shift amount on the pixel) calculated in step S307 described above. More specifically, the deviation amount calculation unit 625 calculates the deviation amount (ΔX1, ΔY1) on the specimen SP surface by the following equation.
ΔX1 = ΔPx1 × size of one pixel of the image sensor in the imaging device 3 / objective magnification
... (3)
ΔY1 = ΔPy1 × size of one pixel of the image pickup device in the image pickup apparatus 3 / objective magnification
... (4)
Specifically, the shift amount (ΔX1, ΔY1) of the designated position on the specimen SP surface is illustrated as shown in FIG. 6C (FIG. 6 (a) → FIG. 6 (b)). As described above, the deviation amount calculation unit 625 calculates the deviation amount of the designated position on the specimen SP surface after the stage 211 is moved.

Thereafter, the correction unit 626 corrects the coordinate information of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 (step S309). Specifically, the correction unit 626 corrects the XY coordinates (Xm0, Ym0) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625. More specifically, the correction unit 626 calculates the XY coordinates (Xm0, Ym0) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 according to the following expression. to correct.
Xm1 = Xm0−ΔX1 (5)
Ym1 = Ym0−ΔY1 (6)
Here, Xm1 indicates the X coordinate of the marker M1 after correction, and Ym1 indicates the Y coordinate of the marker M1 after correction. In this case, the correction unit 626 captures the XY coordinates (Xm1, Ym1) of the marker M1 corrected by the equations (5) and (6) and the Z coordinate (Zm1) of the marker M1 detected by the focusing control unit 622, respectively. It is recorded in the recording unit 611 and the coordinate information of the marker M1 is updated ((Xm0, Ym0, Zm0) → (Xm1, Ym1, Zm1)).

Subsequently, the correcting unit 626 corrects the coordinate information of the registration point (designated position) based on the coordinate information of the marker M1 recorded by the imaging information recording unit 611 (step S310). Specifically, the correction unit 626 corrects the coordinate information of the registered points (for example, registered points A, B, and C) based on the coordinate information of the marker M1 corrected in step S309 described above. More specifically, the correction unit 626 corrects the coordinate information of the registration points A to C by the following formula.
Xa1 = Xa0−ΔX1 (7)
Ya1 = Ya0−ΔY1 (8)
Xb1 = Xb0−ΔX1 (9)
Yb1 = Yb0−ΔY1 (10)
Xc1 = Xc0−ΔX1 (11)
Yc1 = Yc0−ΔY1 (12)
Here, Xa1 indicates the X coordinate of the registration point A after correction, Ya1 indicates the X coordinate of the registration point A after correction, Xb1 indicates the X coordinate of the registration point B after correction, and Yb1 indicates The X coordinate of the registration point B after correction is indicated, Xc1 is the X coordinate of the registration point C after correction, and Yc1 is the X coordinate of the registration point C after correction.

  Thereafter, the stage control unit 621 moves the stage 211 to the registration point recorded by the imaging information recording unit 611 (step S311). For example, the stage control unit 621 moves the stage 211 to the registration point A (Xa1, Ya1, Za0) corrected by the correction unit 626.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) with respect to the registration point based on the AF signal input from the AF unit 252 (step S312). For example, the focusing control unit 622 detects the position ΔZa1 in the vertical direction of the focusing unit 25 with respect to the registration point A based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF process for focusing the focal point 23 on the marker M1 (step S313). For example, the focusing control unit 622 moves the focusing unit 25 to the focal position Za1 (Za1 = Za0−ΔZa1) with respect to the registration point A based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 so as to switch to the wavelength at which the specimen SP can be photographed (step S314). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the observation wavelength.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the sample SP in the visual field region of the imaging device 3 (step S315). For example, the imaging control unit 627 captures the registration point A of the sample SP in the visual field region of the imaging device 3 and generates image data of the registration point A.

  Subsequently, the microscope control device 6 determines whether or not the photographing of all the registration points for the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611 (step S316). When the microscope control device 6 determines that the imaging of all the registration points for the specimen SP has been completed (step S316: Yes), the microscope system 1 proceeds to step S317 described later. On the other hand, when the microscope control device 6 determines that the imaging of all the registration points for the specimen SP has not been completed (step S316: No), the microscope system 1 returns to the above-described step S311.

  In step S317, the microscope control device 6 determines whether or not the registration time has elapsed since all of the registration points for the sample SP were captured based on the imaging information recorded by the imaging information recording unit 611. When it is determined that the registration time has elapsed since the microscope control device 6 has photographed all the registration points for the specimen SP (step S317: Yes), the microscope system 1 proceeds to step S318 described later. On the other hand, when the microscope control device 6 determines that the registration time has not elapsed since the imaging of all the registration points for the specimen SP (step S317: No), the microscope system 1 passes the registration time. Continue this judgment until.

  In step S318, the stage control unit 621 moves the stage 211 to the marker position by driving the stage driving unit 212 based on the position information of the marker M1 recorded by the imaging information recording unit 611. Specifically, the stage control unit 621 moves the stage 211 to the registration point (Xm1, Ym1, Zm1) of the marker M1 recorded by the imaging information recording unit 611.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction based on the AF signal input from the AF unit 252 (step S319). Specifically, the focusing control unit 622 detects the position ΔZm2 of the focusing unit 25 in the vertical direction based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF processing for focusing the focal point 23 on the marker M1 (step S320). Specifically, the focusing control unit 622 moves the focusing unit 25 to the focal position Zm2 (Zm2 = Zm1−ΔZm2) based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 and switches the marker M1 to a marker photographing wavelength (step S321). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the wavelength for photographing the marker M1.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the marker M1 in the visual field region of the imaging device 3 (step S322). In this case, in the imaging control unit 627, the position detection unit 213 detects the marking image (reference image) corresponding to the marking image data generated by the imaging device 3 imaging the marker M1 accommodated in the container D1. The position information (Xm1, Ym1) of the stage 211 and the position information (Zm2) of the focus position of the focusing unit 25 are recorded in the photographing information recording unit 611 in association with each other.

  Subsequently, the deviation amount calculation unit 625 calculates the in-image coordinates of the marker M1 in the marking image generated by the imaging device 3 and records it in the imaging information recording unit 611 (step S323). Specifically, the deviation amount calculation unit 625 calculates the in-image coordinates (Px2, Py2) of the marker M1 from the marking image generated by the imaging device 3, and records it in the recording unit 61.

Thereafter, the deviation amount calculation unit 625 calculates the deviation amount of the marker M1 (step S324). Specifically, the deviation amount calculation unit 625 calculates a movement amount (drift amount) by which the marker M1 has moved from the marking image generated by the imaging device 3. More specifically, the deviation amount calculation unit 625 calculates the deviation amount (ΔPx2, ΔPy2) of the marker M1 on the marking image in pixel conversion by the following equation.
ΔPx2 = Px2−Px1 (13)
ΔPy2 = Py2−Py1 (14)

Subsequently, the deviation amount calculation unit 625 calculates the deviation amount on the specimen SP surface (step S325). Specifically, the shift amount calculation unit 625 calculates the shift amount on the specimen SP surface based on the shift amount on the marking image (shift amount on the pixel) calculated in step S324 described above. More specifically, the deviation amount calculation unit 625 calculates the deviation amount (ΔX2, ΔY2) on the sample SP surface by the following equation.
ΔX2 = ΔPx2 × size of one pixel of the image sensor in the imaging device 3 / objective magnification
... (15)
ΔY2 = ΔPy2 × size of one pixel of the image sensor in the image pickup apparatus 3 / objective magnification
... (16)

Thereafter, the correction unit 626 corrects the coordinate information of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 (step S326). Specifically, the correction unit 626 corrects the XY coordinates (Xm2, Ym2) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625. More specifically, the correction unit 626 calculates the XY coordinates (Xm2, Ym2) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 according to the following expression. to correct.
Xm2 = Xm1−ΔX2 (17)
Ym2 = Ym1−ΔY2 (18)
Here, Xm2 indicates the X coordinate of the corrected marker M1, and Ym2 indicates the Y coordinate of the corrected marker M1. In this case, the correction unit 626 captures the XY coordinates (Xm2, Ym2) of the marker M1 corrected by the equations (17) and (18) and the Z coordinate (Zm2) of the marker M1 detected by the focusing control unit 622, respectively. It is recorded in the recording unit 611 and the coordinate information of the marker M1 is updated ((Xm1, Ym1, Zm1) → (Xm2, Ym2, Zm2)).

Subsequently, the correction unit 626 corrects the coordinate information of the registration point based on the coordinate information of the marker M1 recorded by the imaging information recording unit 611 (step S327). Specifically, the correction unit 626 corrects the coordinate information of the registered points (for example, registered points A, B, and C) based on the coordinate information of the marker M1 corrected in step S326 described above. More specifically, the correction unit 626 corrects the coordinate information of the registration points A to C by the following formula.
Xa2 = Xa1-ΔX2 (19)
Ya2 = Ya1-ΔY2 (20)
Xb2 = Xb1-ΔX2 (21)
Yb2 = Yb1−ΔY2 (22)
Xc2 = Xc1-ΔX2 (23)
Yc2 = Yc1−ΔY2 (24)
Here, Xa2 indicates the X coordinate of the registration point A after correction, Ya2 indicates the X coordinate of the registration point A after correction, Xb2 indicates the X coordinate of the registration point B after correction, and Yb2 Xc of the registration point B after correction, Xc2 indicates the X coordinate of the registration point C after correction, and Yc2 indicates the X coordinate of the registration point C after correction.

  Thereafter, the stage control unit 621 moves the stage 211 to a registration point recorded by the imaging information recording unit 611 (step S328). For example, the stage control unit 621 moves the stage 211 to the registration point A (Xa2, Ya2, Za1) corrected by the correction unit 626.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) with respect to the registration point based on the AF signal input from the AF unit 252 (step S329). For example, the focusing control unit 622 detects the position ΔZa2 in the vertical direction of the focusing unit 25 with respect to the registration point A based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focus position in order to execute the AF process for focusing the focus of the marker 23 on the marker M1 (step S330). For example, the focusing control unit 622 moves the focusing unit 25 to the focal position Za2 (Za2 = Za1-ΔZa2) with respect to the registration point A based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 to switch the sample SP to a wavelength at which the specimen SP can be photographed (step S331). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the observation wavelength.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the specimen SP in the visual field region of the imaging device 3 (step S332). For example, the imaging control unit 627 captures the registration point A of the sample SP in the visual field region of the imaging device 3 and generates image data of the registration point A.

  As described above, the microscope system 1 repeats steps S328 to S332 in the same manner for the registration points B and C as shown in FIG. 6D.

  Subsequently, the microscope control device 6 determines whether or not the photographing of all the registration points for the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611 (step S333). When the microscope control device 6 determines that the imaging of all the registration points for the sample SP has been completed (step S333: Yes), the microscope system 1 proceeds to step S334 described later. On the other hand, when the microscope control device 6 determines that the imaging of all the registration points for the specimen SP has not been completed (step S333: No), the microscope system 1 returns to the above-described step S328.

  In step S334, the microscope control device 6 determines whether or not the registration time has elapsed since the imaging of all the registration points for the specimen SP based on the imaging information recorded by the imaging information recording unit 611. When it is determined that the registration time has elapsed since the microscope control device 6 photographed all the registration points for the specimen SP (step S334: Yes), the microscope system 1 proceeds to step S335 described later. On the other hand, when the microscope control apparatus 6 determines that the registration time has not elapsed since the imaging of all the registration points for the specimen SP (step S334: No), the microscope system 1 passes the registration time. Continue this judgment until.

  In step S335, the microscope control device 6 determines whether or not the designated cycle for photographing all the registration points for the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611. When the microscope control device 6 determines that the designated cycle for imaging all the registration points for the specimen SP has ended (step S335: Yes), that is, when it is determined that the total imaging time has ended, the microscope system 1 Then, the process proceeds to step S336 described later. On the other hand, when the microscope control device 6 determines that the designated cycle for photographing all the registration points for the specimen SP has not been completed (step S335: No), the microscope system 1 returns to step S318 described above.

  In step S336, the image processing unit 623 generates a time-lapse moving image in which a plurality of pieces of image data recorded by the recording unit 61 are sequentially connected for each designated position of the sample SP and records the generated time-lapse moving image in the recording unit 61. After step S336, the microscope system 1 returns to the main routine of FIG.

  According to the first embodiment of the present invention described above, it is possible to accurately correct the deviation of the designated position of the specimen due to the thermal drift with a simple configuration regardless of the kind of specimen.

  Furthermore, according to the first embodiment of the present invention, since a plurality of designated positions that designate observation locations on the specimen SP are corrected for each cycle, more accurate time-lapse observation can be performed.

(Embodiment 2)
Next, a second embodiment of the present invention will be described. The microscope system according to the second embodiment has the same configuration as that of the first embodiment described above, and the processing to be executed is different. Specifically, the setting process and the time lapse process are different. For this reason, below, the process which the microscope system which concerns on this Embodiment 2 performs is demonstrated. In addition, the same code | symbol is attached | subjected to the structure same as the microscope system 1 which concerns on Embodiment 1 mentioned above, and description is abbreviate | omitted.

[Setting processing]
First, details of the setting process executed by the microscope system 1 according to the second embodiment will be described. FIG. 7 is a flowchart showing an outline of setting processing executed by the microscope system 1 according to the second embodiment.

  As shown in FIG. 7, the imaging information setting unit 624 is a place where the user operates the focusing unit 25 and the stage 211 via the input unit 4 to move to a predetermined registration point of the sample SP and perform time-lapse observation. The registration point is recorded (registered) in the photographing information recording unit 611 (step S401). Specifically, the shooting information setting unit 624 records X, Y, and Z coordinate information of the registration point in the shooting information recording unit 611. More specifically, the imaging information setting unit 624 detects the X and Y coordinates detected by the position detection unit 213 and the height of the stage 211 in the vertical direction with respect to a predetermined registration point of the specimen SP. The Z coordinate detected by (not shown) is recorded in the photographing information recording unit 611. For example, the imaging information setting unit 624 registers the registration point A (Xa0, Ya0, Za0), the registration point B (Xb0, Yb0, Zb0), for each of the predetermined registration points A, B, C, and D of the specimen SP. The registration point C (Xc0, Yc0, Zc0) and the registration point D (Xd0, Yd0, Zd0) are recorded in the imaging information recording unit 611.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 to switch the marker M1 to a marker photographing wavelength (step S402). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the wavelength for photographing the marker M1.

  Thereafter, the imaging control unit 627 operates the focusing unit 25 and the stage 211 via the input unit 4 by the user to move the marker M1 that has been moved within the field of view of the imaging device 3 (for example, the center of the field of view of the imaging device 3). The imaging device 3 is caused to take an image (step S403). In this case, the shooting information setting unit 624 records the X and Y coordinates detected by the position detection unit 213 and the Z coordinate detected by the height detection unit (not shown) in the shooting information recording unit 611 with respect to the marker M1. Let For example, the shooting information setting unit 624 causes the shooting information recording unit 611 to record the registration point (Xm0, Ym0, Zm0) of the marker M1. Furthermore, since the shooting information setting unit 624 records the position of the marker M1 in the image corresponding to the image data shot by the imaging device 3, the image coordinates (Px0) of the marker M1 in the image detected by the image processing unit 623 are recorded. , Py0) is recorded in the photographing information recording unit 611.

  Subsequently, the shooting information setting unit 624 records the time lapse time interval, shooting time, shooting count, and correction count input via the input unit 4 in the shooting information recording unit 611 (step S404). For example, the shooting information setting unit 624 inputs the first to second shooting intervals t at the point A input via the input unit 4, the number n of repetitions of the shooting interval t for T time, and the time in one cycle. The number of corrections θ for correcting the shift amount due to thermal drift to be corrected in the tuple observation is recorded in the imaging information recording unit 611. After step S404, the microscope system 1 returns to the main routine of FIG.

[Time-lapse processing]
Next, details of the time lapse process executed by the microscope system 1 according to the second embodiment will be described. FIG. 8A is a flowchart showing an outline of a time lapse process executed by the microscope system 1 according to the second embodiment. FIG. 8B is a flowchart illustrating an overview of a time lapse process executed by the microscope system 1 according to the second embodiment. FIG. 8C is a time chart illustrating an outline of a time lapse process executed by the microscope system 1 according to the second embodiment.

  As shown in FIGS. 8A, 8B, and 8C, first, the stage control unit 621 drives the stage driving unit 212 based on the position information recorded by the imaging information recording unit 611, thereby setting the stage at the marker position. 211 is moved (step S501). Specifically, the stage control unit 621 moves the stage 211 to the registration point (Xm0, Ym0, Zm0) of the marker M1 recorded by the imaging information recording unit 611.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) based on the AF signal input from the AF unit 252 (step S502). Specifically, the focusing control unit 622 detects the position ΔZm1 in the vertical direction of the focusing unit 25 based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF process for focusing the focal point 23 on the marker M1 (step S503). Specifically, the focusing control unit 622 moves the focusing unit 25 to the focal position Zm1 (Zm1 = Zm0−ΔZm1) based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 and switches the marker M1 to a marker photographing wavelength (step S504). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the wavelength for photographing the marker M1.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the marker M1 within the visual field region of the imaging device 3 (step S505). In this case, in the imaging control unit 627, the position detection unit 213 detects the marking image (reference image) corresponding to the marking image data generated by the imaging device 3 imaging the marker M1 accommodated in the container D1. The position information (Xm0, Ym0) of the stage 211 and the position information (Zm1) of the focus position of the focusing unit 25 are associated with each other and recorded in the photographing information recording unit 611.

  Subsequently, the deviation amount calculation unit 625 calculates the in-image coordinates of the marker M1 in the marking image generated by the imaging device 3 and records it in the recording unit 61 (step S506). Specifically, the deviation amount calculation unit 625 calculates the in-image coordinates (Px1, Py1) of the marker M1 from the marking image generated by the imaging device 3 and records it in the recording unit 61.

  Thereafter, the deviation amount calculation unit 625 calculates the deviation amount of the marker M1 (step S507). Specifically, the deviation amount calculation unit 625 calculates a movement amount (drift amount) by which the marker M1 has moved from the marking image generated by the imaging device 3. More specifically, the shift amount calculation unit 625 calculates the shift amount (ΔPx1, ΔPy1) of the marker M1 on the marking image in pixel conversion using the above-described equations (1) and (2).

  Subsequently, the deviation amount calculation unit 625 calculates the deviation amount on the specimen SP surface (step S508). Specifically, the shift amount calculation unit 625 calculates the shift amount on the specimen SP surface based on the shift amount on the marking image (shift amount on the pixel) calculated in step S307 described above. More specifically, the deviation amount calculation unit 625 calculates the deviation amount (ΔX1, ΔY1) on the specimen SP surface by the above-described equations (3) and (4).

  Thereafter, the correction unit 626 corrects the coordinate information of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 (step S509). Specifically, the correction unit 626 corrects the XY coordinates (Xm0, Ym0) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625. More specifically, the correction unit 626 calculates the XY coordinates (Xm0, Ym0) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 ( Correct by 5) and (6). In this case, the correction unit 626 captures the XY coordinates (Xm1, Ym1) of the marker M1 corrected by the equations (5) and (6) and the Z coordinate (Zm1) of the marker M1 detected by the focusing control unit 622, respectively. It is recorded in the recording unit 611 and the coordinate information of the marker M1 is updated ((Xm0, Ym0, Zm0) → (Xm1, Ym1, Zm1)).

  Subsequently, the correction unit 626 corrects the coordinate information (designated position) of the registration point based on the coordinate information of the marker M1 recorded by the imaging information recording unit 611 (step S510). Specifically, the correction unit 626 corrects the coordinate information of the registered points (for example, registered points A and B) based on the coordinate information of the marker M1 corrected in step S309 described above. More specifically, the correction unit 626 corrects the coordinate information of the registration points A and B by the above-described equations (7) to (10).

  Thereafter, the stage control unit 621 moves the stage 211 to a registration point recorded by the imaging information recording unit 611 (step 511). For example, the stage control unit 621 moves the stage 211 to the registration point A (Xa1, Yb1, Za0) corrected by the correction unit 626, as shown in FIG. 8C.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) with respect to the registration point based on the AF signal input from the AF unit 252 (step S512). For example, the focusing control unit 622 detects the position ΔZa1 in the vertical direction of the focusing unit 25 with respect to the registration point A based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF processing for focusing the focal point 23 on the marker M1 (step S513). For example, the focusing control unit 622 moves the focusing unit 25 to the focal position Za1 (Za1 = Za0−ΔZa1) with respect to the registration point A based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 so as to switch to the wavelength at which the specimen SP can be photographed (step S514). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the observation wavelength.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the specimen SP in the visual field region of the imaging device 3 (step S515). For example, the imaging control unit 627 captures the registration point A of the sample SP in the visual field region of the imaging device 3 and generates image data of the registration point A.

  Subsequently, the microscope control device 6 determines whether or not the photographing of the designated registration point with respect to the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611 (step S516). Specifically, the microscope control device 6 determines whether or not the photographing of the registration points A and B with respect to the specimen SP has been completed. If it is determined that the imaging of the designated registration point for the specimen SP has been completed based on the imaging information recorded by the imaging information recording unit 611 (step S516: Yes), the microscope system 1 proceeds to step S517 described later. On the other hand, when it is determined that the imaging of the designated registration point for the specimen SP has not been completed (step S516: No), the microscope system 1 returns to the above-described step S511.

  In step S517, the stage control unit 621 moves the stage 211 to the marker position by driving the stage driving unit 212 based on the position information of the marker M1 recorded by the imaging information recording unit 611. Specifically, the stage control unit 621 moves the stage 211 to the registration point (Xm1, Ym1, Zm1) of the marker M1 recorded by the imaging information recording unit 611.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction based on the AF signal input from the AF unit 252 (step S518). Specifically, the focusing control unit 622 detects the position ΔZm1 ′ of the focusing unit 25 in the vertical direction based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF processing for focusing the focal point 23 on the marker M1 (step S519). Specifically, the focusing control unit 622 moves the focusing unit 25 to the focal position Zm1 ′ (Zm1 ′ = Zm1−ΔZm1 ′) based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 to switch the marker M1 to a marker photographing wavelength (step S520). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the wavelength for photographing the marker M1.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the marker M1 in the visual field region of the imaging device 3 (step S521). In this case, in this case, the imaging control unit 627 displays the position detection unit on the marking image (reference image) corresponding to the marking image data generated by imaging the marker M1 accommodated in the container D1. The position information (Xm1, Ym1) of the stage 211 detected by 213 and the position information (Zm1 ′) of the focal position of the focusing unit 25 are recorded in association with each other in the imaging information recording unit 611.

  Subsequently, the deviation amount calculation unit 625 calculates the in-image coordinates of the marker M1 in the marking image generated by the imaging device 3, and records it in the imaging information recording unit 611 (step S522). Specifically, the deviation amount calculation unit 625 calculates the in-image coordinates (Px1 ′, Py1 ′) of the marker M1 from the marking image generated by the imaging device 3 and records it in the recording unit 61.

Thereafter, the deviation amount calculation unit 625 calculates the deviation amount of the marker M1 (step S523). Specifically, the deviation amount calculation unit 625 calculates a movement amount (drift amount) by which the marker M1 has moved from the marking image generated by the imaging device 3. More specifically, the shift amount calculation unit 625 calculates the shift amount (ΔPx1 ′, ΔPy1 ′) of the marker M1 on the marking image in pixel conversion by the following equation.
ΔPx1 ′ = Px1′−Px1 (25)
ΔPy1 ′ = Py1′−Py1 (26)

Subsequently, the deviation amount calculation unit 625 calculates the deviation amount on the specimen SP surface (step S524). Specifically, the shift amount calculation unit 625 calculates the shift amount on the specimen SP surface based on the shift amount on the marking image (shift amount on the pixel) calculated in step S523 described above. More specifically, the shift amount calculation unit 625 calculates the shift amount (ΔX1 ′, ΔY1 ′) on the sample SP surface by the following formula.
ΔX1 ′ = ΔPx1 ′ × size of one pixel of the image pickup device in the image pickup apparatus 3 / objective magnification
... (27)
ΔY1 ′ = ΔPy1 ′ × size of one pixel of the image pickup device in the image pickup apparatus 3 / objective magnification
... (28)

Thereafter, the correction unit 626 corrects the coordinate information of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 (step S525). Specifically, the correction unit 626 corrects the XY coordinates (Xm1 ′, Ym1 ′) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625. . More specifically, the correction unit 626 calculates the XY coordinates (Xm1 ′, Ym1 ′) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 as follows. Correct by the formula.
Xm1 ′ = Xm1−ΔX1 ′ (29)
Ym1 ′ = Ym1−ΔY1 ′ (30)
Here, Xm1 ′ represents the X coordinate of the corrected marker M1, and Ym1 ′ represents the Y coordinate of the corrected marker M1. In this case, the correction unit 626 uses the XY coordinates (Xm1 ′, Ym1 ′) of the marker M1 corrected by the equations (29) and (30) and the Z coordinate (Zm1 ′) of the marker M1 detected by the focusing control unit 622, respectively. Is recorded in the imaging information recording unit 611, and the coordinate information of the marker M1 is updated ((Xm1, Ym1, Zm1) → (Xm1 ′, Ym1 ′, Zm1 ′)).

Subsequently, the correction unit 626 corrects the coordinate information of the registration point based on the coordinate information of the marker M1 recorded by the imaging information recording unit 611 (step S526). Specifically, the correction unit 626 corrects the coordinate information of the registered points (for example, registered points C and D) based on the coordinate information of the marker M1 corrected in step S525 described above. More specifically, the correction unit 626 corrects the coordinate information of the registration points C and D by the following formula.
Xc1 = Xc0−ΔX1 ′ (31)
Yc1 = Yc0−ΔY1 ′ (32)
Xd1 = Xd0−ΔX1 ′ (33)
Yd1 = Yd0−ΔY1 ′ (34)
Here, Xc1 indicates the X coordinate of the corrected registration point C, Yc1 indicates the X coordinate of the corrected registration point C, Xd1 indicates the X coordinate of the corrected registration point D, and Yd1 indicates The X coordinate of the registration point D after correction is shown.

  Thereafter, the stage control unit 621 moves the stage 211 to the registration point recorded by the imaging information recording unit 611 as shown in FIG. 8C (step S527). For example, the stage control unit 621 moves the stage 211 to the registration point C (Xc1, Yc1, Za0) corrected by the correction unit 626.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) with respect to the registration point based on the AF signal input from the AF unit 252 (step S528). For example, the focusing control unit 622 detects the position ΔZc1 in the vertical direction of the focusing unit 25 with respect to the registration point C based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF process for focusing the focal point 23 on the marker M1 (step S529). For example, the focusing control unit 622 moves the focusing unit 25 to the focal position Zc1 (Za1 = Za0−ΔZa1) with respect to the registration point C based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 so as to switch to the wavelength at which the specimen SP can be photographed (step S530). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the observation wavelength.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the specimen SP in the visual field region of the imaging device 3 (step S531). For example, the imaging control unit 627 captures the registration point C of the specimen SP in the visual field region of the imaging device 3 and generates image data of the registration point C.

  Subsequently, the microscope control device 6 determines whether or not the photographing of the designated registration point with respect to the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611 (Step S532). Specifically, the microscope control device 6 determines whether or not the photographing of the registration points C and D with respect to the specimen SP has been completed. When it is determined that the imaging of the designated registration point with respect to the sample SP has been completed based on the imaging information recorded by the imaging information recording unit 611 (step S532: Yes), the microscope system 1 proceeds to step S533 described later. On the other hand, when it is determined that the imaging of the designated registration point for the specimen SP has not been completed (step S532: No), the microscope system 1 returns to the above-described step S527.

  In step S533, the microscope control device 6 determines whether or not the registration time has elapsed since the imaging of all the registration points for the specimen SP based on the imaging information recorded by the imaging information recording unit 611. When it is determined that the registration time has elapsed since the microscope control device 6 photographed all the registration points for the specimen SP (step S533: Yes), the microscope system 1 proceeds to step S534 described later. On the other hand, when the microscope control device 6 determines that the registration time has not elapsed since the imaging of all the registration points for the specimen SP (step S533: No), the microscope system 1 passes the registration time. Continue this judgment until.

  In step S534, the microscope control device 6 determines whether or not the designated cycle for photographing all the registration points for the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611. When the microscope control device 6 determines that the designated cycle for photographing all the registration points for the specimen SP has been completed (step S534: Yes), the microscope system 1 proceeds to step S535 described later. On the other hand, when the microscope control device 6 determines that the designated cycle for photographing all the registration points for the specimen SP has not ended (step S534: No), the microscope system 1 returns to step S511 described above.

  In step S535, the image processing unit 623 generates a time-lapse moving image in which a plurality of pieces of image data recorded by the recording unit 61 are sequentially connected for each designated position of the specimen SP and records the time-lapse moving image in the recording unit 61. After step S535, the microscope system 1 returns to the main routine of FIG.

  According to the second embodiment of the present invention described above, the deviation of the designated position of the sample due to thermal drift can be accurately corrected with a simple configuration regardless of the type of sample.

  Further, according to the second embodiment of the present invention, it is possible to reduce the influence of thermal drift that occurs during one cycle of time-lapse observation.

(Embodiment 3)
Next, a third embodiment of the present invention will be described. The microscope system according to the third embodiment is different in the configuration of the microscope system 1 according to the first embodiment described above and the time lapse process to be executed. For this reason, below, after demonstrating the structure of the microscope system which concerns on this Embodiment 3, the process which the microscope system which concerns on this Embodiment 3 performs is demonstrated. In addition, the same code | symbol is attached | subjected to the structure same as the microscope system 1 which concerns on Embodiment 1 mentioned above, and description is abbreviate | omitted.

[Configuration of microscope system]
FIG. 9 is a schematic diagram schematically showing the configuration of the microscope system according to the third embodiment of the present invention. A microscope system 1a illustrated in FIG. 9 includes a microscope device 2, an imaging device 3, an input unit 4, a display unit 5, and a microscope control device 6a.

  The microscope control device 6a includes a recording unit 61 and a control unit 62a that comprehensively controls each unit of the microscope system 1a.

  The control unit 62a includes a stage control unit 621, a focusing control unit 622, an image processing unit 623, a shooting information setting unit 624, a deviation amount calculation unit 625, a correction unit 626a, a shooting control unit 627, and a display. And a control unit 628.

  After the number of cycles designated by the time lapse observation is completed, the correction unit 626a is based on the shift amount associated with the image corresponding to each image data recorded by the recording unit 61 in the image corresponding to the image data. The designated position (registered position) corresponding to the observation location of the specimen SP is corrected.

[Time-lapse processing]
Next, details of the time lapse process executed by the microscope system 1 according to the third embodiment will be described. FIG. 10A is a flowchart showing an overview of a time lapse process executed by the microscope system 1 according to the third embodiment. FIG. 10B is a flowchart illustrating an overview of a time lapse process executed by the microscope system 1 according to the third embodiment.

  As shown in FIGS. 10A and 10B, first, the stage control unit 621 moves the stage 211 to the marker position by driving the stage driving unit 212 based on the position information recorded by the imaging information recording unit 611. (Step S601). Specifically, the stage control unit 621 moves the stage 211 to the registration point (Xm0, Ym0, Zm0) of the marker M1 recorded by the imaging information recording unit 611.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) based on the AF signal input from the AF unit 252 (step S602). Specifically, the focusing control unit 622 detects the position ΔZm1 in the vertical direction of the focusing unit 25 based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF process for focusing the focal point 23 on the marker M1 (step S603). Specifically, the focusing control unit 622 moves the focusing unit 25 to the focal position Zm1 (Zm1 = Zm0−ΔZm1) based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 to switch the marker M1 to a marker photographing wavelength (step S604). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the wavelength for photographing the marker M1.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the marker M1 in the visual field area of the imaging device 3 (step S605). In this case, in the imaging control unit 627, the position detection unit 213 detects the marking image (reference image) corresponding to the marking image data generated by the imaging device 3 imaging the marker M1 accommodated in the container D1. The position information (Xm0, Ym0) of the stage 211 and the position information (Zm1) of the focus position of the focusing unit 25 are associated with each other and recorded in the photographing information recording unit 611.

  Subsequently, the deviation amount calculation unit 625 calculates the in-image coordinates of the marker M1 in the marking image generated by the imaging device 3 and records it in the recording unit 61 (step S606). Specifically, the deviation amount calculation unit 625 calculates the in-image coordinates (Px1, Py1) of the marker M1 from the marking image generated by the imaging device 3 and records it in the recording unit 61.

  Thereafter, the deviation amount calculation unit 625 calculates the deviation amount of the marker M1 (step S607). Specifically, the deviation amount calculation unit 625 calculates a movement amount (drift amount) by which the marker M1 has moved from the marking image generated by the imaging device 3. More specifically, the shift amount calculation unit 625 calculates the shift amount (ΔPx1, ΔPy1) of the marker M1 on the marking image in pixel conversion using the above-described equations (1) and (2).

  Subsequently, the stage control unit 621 moves the stage 211 to a registration point recorded by the shooting information recording unit 611 (step S608). For example, the stage control unit 621 moves the stage 211 to the registration point A (Xa1, Ya1, Za0) corrected by the correction unit 626.

  Thereafter, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) with respect to the registration point based on the AF signal input from the AF unit 252 (step S609). For example, the focusing control unit 622 detects the position ΔZa1 in the vertical direction of the focusing unit 25 with respect to the registration point A based on the AF signal input from the AF unit 252.

  Subsequently, the focusing control unit 622 drives the focusing driving unit 253 on the basis of the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction). The focusing unit 25 is moved to the focal position in order to execute the AF process for focusing the lens 23 on the marker M1 (step S610). For example, the focusing control unit 622 moves the focusing unit 25 to the focal position Za1 (Za1 = Za0−ΔZa1) with respect to the registration point A based on the AF signal input from the AF unit 252.

  Thereafter, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 to switch to the wavelength at which the specimen SP can be photographed (step S611). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the observation wavelength.

  Subsequently, the imaging control unit 627 causes the imaging device 3 to image the sample SP in the visual field region of the imaging device 3 (step S612). For example, the imaging control unit 627 captures the registration point A of the sample SP in the visual field region of the imaging device 3 and generates image data of the registration point A.

  Thereafter, the imaging control unit 627 assigns the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 to the image corresponding to the image data generated by the imaging device 3, and records it in the recording unit 61 (step S613). For example, the imaging control unit 627 captures the registration point A of the sample SP in the field of view of the imaging device 3 and creates an image corresponding to the image data of the registration point A, and the deviation amount calculation unit 625 calculates the marker calculated in step S607. A shift amount (ΔPx1, ΔPy1) of M1 is assigned (corresponding) and recorded in the recording unit 61.

  Subsequently, the microscope control device 6 determines whether or not the photographing of all the registration points for the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611 (Step S614). When the microscope control device 6 determines that the photographing of all the registration points for the specimen SP has been completed (step S614: Yes), the microscope system 1a proceeds to step S615 described later. On the other hand, when the microscope control device 6 determines that the imaging of all the registration points for the specimen SP has not been completed (step S614: No), the microscope system 1a returns to step S608 described above.

  In step S615, the microscope control device 6 determines whether or not the registration time has elapsed since the imaging of all the registration points for the specimen SP based on the imaging information recorded by the imaging information recording unit 611. When it is determined that the registration time has elapsed since the microscope control device 6 photographed all the registration points for the specimen SP (step S615: Yes), the microscope system 1a proceeds to step S616 described later. On the other hand, when the microscope control device 6 determines that the registration time has not elapsed since the imaging of all the registration points for the specimen SP (step S615: No), the microscope system 1a passes the registration time. Continue this judgment until.

  In step S616, the stage control unit 621 moves the stage 211 to the marker position by driving the stage driving unit 212 based on the position information of the marker M1 recorded by the imaging information recording unit 611. Specifically, the stage control unit 621 moves the stage 211 to the registration point (Xm1, Ym1, Zm1) of the marker M1 recorded by the imaging information recording unit 611.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction based on the AF signal input from the AF unit 252 (step S617). Specifically, the focusing control unit 622 detects the position ΔZm2 of the focusing unit 25 in the vertical direction based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF processing for focusing the focal point 23 on the marker M1 (step S618). Specifically, the focusing control unit 622 moves the focusing unit 25 to the focal position Zm2 (Zm2 = Zm1−ΔZm2) based on the AF signal input from the AF unit 252.

  Subsequently, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 and switches the marker M1 to a marker photographing wavelength (step S619). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the wavelength for photographing the marker M1.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the marker M1 within the visual field region of the imaging device 3 (step S620). In this case, in the imaging control unit 627, the position detection unit 213 detects the marking image (reference image) corresponding to the marking image data generated by the imaging device 3 imaging the marker M1 accommodated in the container D1. The position information (Xm0, Ym0) of the stage 211 and the position information (Zm2) of the focus position of the focusing unit 25 are associated with each other and recorded (updated) in the imaging information recording unit 611.

  Subsequently, the deviation amount calculation unit 625 calculates the in-image coordinates of the marker M1 in the marking image generated by the imaging device 3, and records it in the imaging information recording unit 611 (step S621). Specifically, the deviation amount calculation unit 625 calculates the in-image coordinates (Px2, Py2) of the marker M1 from the marking image generated by the imaging device 3, and records it in the recording unit 61.

  Thereafter, the deviation amount calculation unit 625 calculates the deviation amount of the marker M1 (step S622). Specifically, the deviation amount calculation unit 625 calculates a movement amount (drift amount) by which the marker M1 has moved from the marking image generated by the imaging device 3. More specifically, the shift amount calculation unit 625 calculates the shift amount (ΔPx2, ΔPy2) of the marker M1 on the marking image in pixel conversion using the above-described equations (13) and (14).

  Subsequently, the stage control unit 621 moves the stage 211 to a registration point recorded by the imaging information recording unit 611 (step S623). For example, the stage control unit 621 moves the stage 211 to the registration point A (Xa0, Ya0, Za1) recorded by the imaging information recording unit 611.

  Thereafter, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) with respect to the registration point based on the AF signal input from the AF unit 252 (step S624). For example, the focusing control unit 622 detects the position ΔZa2 in the vertical direction of the focusing unit 25 with respect to the registration point A based on the AF signal input from the AF unit 252.

  Subsequently, the focusing control unit 622 drives the focusing driving unit 253 on the basis of the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction). The focusing unit 25 is moved to the focal position in order to execute the AF process for focusing the lens 23 on the marker M1 (step S625). For example, the focusing control unit 622 moves the focusing unit 25 to the focal position Za2 (Za2 = Za1-ΔZa2) with respect to the registration point A based on the AF signal input from the AF unit 252.

  Thereafter, the microscope control device 6 drives the mirror driving unit 274 to rotate the mirror cassette 273 to switch to the wavelength at which the specimen SP can be photographed (step S626). Specifically, the microscope control device 6 controls the mirror turret 27 to switch the illumination light to the observation wavelength.

  Subsequently, the imaging control unit 627 causes the imaging device 3 to image the specimen SP in the visual field region of the imaging device 3 (step S627). For example, the imaging control unit 627 captures the registration point A of the sample SP in the visual field region of the imaging device 3 and generates image data of the registration point A.

  Thereafter, the imaging control unit 627 assigns the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 to the image corresponding to the image data generated by the imaging device 3, and records it in the recording unit 61 (step S628). For example, the imaging control unit 627 captures the registration point A of the specimen SP in the field of view of the imaging device 3 and creates an image corresponding to the image data of the registration point A, and the deviation amount calculation unit 625 calculates the marker calculated in step S622. A shift amount (ΔPx2, ΔPy2) of M1 is assigned (corresponding) and recorded in the recording unit 61.

  Subsequently, the microscope control device 6 determines whether or not the photographing of all the registration points for the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611 (step S629). When the microscope control device 6 determines that the imaging of all the registration points for the specimen SP has been completed (step S629: Yes), the microscope system 1a proceeds to step S630 described later. On the other hand, when the microscope control device 6 determines that imaging of all the registration points for the specimen SP has not been completed (step S629: No), the microscope system 1a returns to step S616 described above.

  In step S630, the microscope control device 6 determines whether or not the registration time has elapsed since the imaging of all the registration points for the specimen SP based on the imaging information recorded by the imaging information recording unit 611. When it is determined that the registration time has elapsed since the microscope control device 6 photographed all the registration points for the specimen SP (step S630: Yes), the microscope system 1a proceeds to step S631 described later. On the other hand, when the microscope control apparatus 6 determines that the registration time has not elapsed since the imaging of all the registration points for the specimen SP (step S630: No), the microscope system 1a passes the registration time. Continue this judgment until.

  In step S <b> 631, the microscope control device 6 determines whether or not the designated cycle for photographing all the registration points for the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611. When the microscope control device 6 determines that the designated cycle for photographing all the registration points for the specimen SP has ended (step S631: Yes), the microscope system 1a proceeds to step S632 described later. On the other hand, when the microscope control device 6 determines that the designated cycle for photographing all the registration points for the specimen SP has not been completed (step S631: No), the microscope system 1a returns to step S616 described above.

  In step S632, the image processing unit 623 corrects the position of each image based on the amount of deviation given to each image recorded by the imaging information recording unit 611, and a plurality of pieces of image data at each registration point (observation point). Generate a time-lapse movie that connects videos in sequence.

  Subsequently, as illustrated in FIG. 10C, the image processing unit 632 includes all the images (in which the imaging region of the imaging device 3 overlaps with the time-lapse movie at each registration point (the dotted frame region W10 in FIG. 10C). ) To create a time-lapse moving image file of each registered point (step S633) For example, the image processing unit 632 overlaps the imaging area of the imaging device 3 with the image A1 and the image A2. Trimming processing is performed to leave the area W10, and after step S633, the microscope system 1a returns to the main routine of FIG.

  According to the third embodiment of the present invention described above, it is possible to correct the deviation of the designated position of the specimen due to the thermal drift regardless of the kind of specimen.

(Embodiment 4)
Next, a fourth embodiment of the present invention will be described. The microscope system according to the fourth embodiment has a configuration similar to that of the above-described first embodiment, and a substance having an opaque marker M1 as an index is used. That is, it is a microscope system for transmission observation. For this reason, the microscope system according to the fourth embodiment is different from the time lapse process executed by the microscope system 1 according to the first embodiment described above. Therefore, in the following, a time lapse process executed by the microscope system according to the fourth embodiment will be described. In addition, the same code | symbol is attached | subjected to the structure same as the microscope system 1 which concerns on Embodiment 4 mentioned above, and description is abbreviate | omitted.

[Time-lapse processing]
FIG. 11A is a flowchart illustrating an overview of a time lapse process performed by the microscope system 1 according to the fourth embodiment. FIG. 11B is a flowchart illustrating an outline of a time lapse process executed by the microscope system 1 according to the fourth embodiment. In the following, it is assumed that a marker M1 (opaque substance) as an index is given to the specimen SP.

  As shown in FIGS. 11A and 11B, the stage control unit 621 first moves the stage 211 to the marker position by driving the stage driving unit 212 based on the position information recorded by the imaging information recording unit 611. (Step S701). Specifically, the stage control unit 621 moves the stage 211 to the registration point (Xm0, Ym0, Zm0) of the marker M1 recorded by the imaging information recording unit 611.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) based on the AF signal input from the AF unit 252 (step S702). Specifically, the focusing control unit 622 detects the position ΔZm1 in the vertical direction of the focusing unit 25 based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF processing for focusing the focal point 23 on the marker M1 (step S703). Specifically, the focusing control unit 622 moves the focusing unit 25 to the focal position Zm1 (Zm1 = Zm0−ΔZm1) based on the AF signal input from the AF unit 252.

  Subsequently, the imaging control unit 627 causes the imaging device 3 to image the marker M1 in the visual field region of the imaging device 3 (step S704). In this case, in the imaging control unit 627, the position detection unit 213 detects the marking image (reference image) corresponding to the marking image data generated by the imaging device 3 imaging the marker M1 accommodated in the container D1. The position information (Xm0, Ym0) of the stage 211 and the position information (Zm1) of the focus position of the focusing unit 25 are associated with each other and recorded in the photographing information recording unit 611.

  Thereafter, the deviation amount calculation unit 625 calculates the in-image coordinates of the marker M1 in the marking image generated by the imaging device 3, and records the calculated coordinates in the recording unit 61 (step S705). Specifically, the deviation amount calculation unit 625 calculates the in-image coordinates (Px1, Py1) of the marker M1 from the marking image generated by the imaging device 3 and records it in the recording unit 61.

  Subsequently, the deviation amount calculation unit 625 calculates the deviation amount of the marker M1 (step S706). Specifically, the deviation amount calculation unit 625 calculates a movement amount (drift amount) by which the marker M1 has moved from the marking image generated by the imaging device 3. More specifically, the shift amount calculation unit 625 calculates the shift amount (ΔPx1, ΔPy1) of the marker M1 on the marking image in pixel conversion using the above-described equations (1) and (2).

  Thereafter, the deviation amount calculation unit 625 calculates the deviation amount on the specimen SP surface (step S707). Specifically, the shift amount calculation unit 625 calculates the shift amount on the specimen SP surface based on the shift amount on the marking image (shift amount on the pixel) calculated in step S706 described above. More specifically, the deviation amount calculation unit 625 calculates the deviation amount (ΔX1, ΔY1) on the specimen SP surface by the above-described equations (3) and (4).

  Subsequently, the correction unit 626 corrects the coordinate information of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 (step S708). Specifically, the correction unit 626 corrects the XY coordinates (Xm0, Ym0) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625. More specifically, the correction unit 626 calculates the XY coordinates (Xm0, Ym0) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 ( Correct by 5) and (6). In this case, the correction unit 626 captures the XY coordinates (Xm1, Ym1) of the marker M1 corrected by the equations (5) and (6) and the Z coordinate (Zm1) of the marker M1 detected by the focusing control unit 622, respectively. It is recorded in the recording unit 611 and the coordinate information of the marker M1 is updated ((Xm0, Ym0, Zm0) → (Xm1, Ym1, Zm1)).

  Thereafter, the correction unit 626 corrects the coordinate information of the registration point based on the coordinate information of the marker M1 recorded by the imaging information recording unit 611 (step S709). Specifically, the correction unit 626 corrects the coordinate information of the registered points (for example, registered points A, B, and C) based on the coordinate information of the marker M1 corrected in step S708 described above. More specifically, the correction unit 626 corrects the coordinate information of the registration points A to C by the above-described equations (7) to (12).

  Thereafter, the stage control unit 621 moves the stage 211 to a registration point recorded by the imaging information recording unit 611 (step S710). For example, the stage control unit 621 moves the stage 211 to the registration point A (Xa1, Ya1, Za0) corrected by the correction unit 626.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) with respect to the registration point based on the AF signal input from the AF unit 252 (step S711). For example, the focusing control unit 622 detects the position ΔZa1 in the vertical direction of the focusing unit 25 with respect to the registration point A based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focus position in order to execute the AF process for focusing the focus of the marker 23 on the marker M1 (step S712). For example, the focusing control unit 622 moves the focusing unit 25 to the focal position Za1 (Za1 = Za0−ΔZa1) with respect to the registration point A based on the AF signal input from the AF unit 252.

  Subsequently, the imaging control unit 627 causes the imaging device 3 to image the sample SP in the visual field region of the imaging device 3 (step S713). For example, the imaging control unit 627 captures the registration point A of the sample SP in the visual field region of the imaging device 3 and generates image data of the registration point A.

  Thereafter, the microscope control device 6 determines whether or not the photographing of all the registration points for the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611 (step S714). When the microscope control device 6 determines that the imaging of all the registration points for the specimen SP has been completed (step S714: Yes), the microscope system 1 proceeds to step S715 described later. On the other hand, when the microscope control device 6 determines that the imaging of all the registration points for the specimen SP has not been completed (step S714: No), the microscope system 1 returns to the above-described step S710.

  In step S715, the microscope control device 6 determines whether or not the registration time has elapsed since the imaging of all the registration points for the specimen SP based on the imaging information recorded by the imaging information recording unit 611. When it is determined that the registration time has elapsed since the microscope control device 6 photographed all the registration points for the specimen SP (step S715: Yes), the microscope system 1 proceeds to step S716 described later. On the other hand, when the microscope control apparatus 6 determines that the registration time has not elapsed since the imaging of all the registration points for the specimen SP (step S715: No), the microscope system 1 passes the registration time. Continue this judgment until.

  In step S716, the stage control unit 621 moves the stage 211 to the marker position by driving the stage driving unit 212 based on the position information of the marker M1 recorded by the imaging information recording unit 611. Specifically, the stage control unit 621 moves the stage 211 to the registration point (Xm1, Ym1, Zm1) of the marker M1 recorded by the imaging information recording unit 611.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction based on the AF signal input from the AF unit 252 (step S717). Specifically, the focusing control unit 622 detects the position ΔZm2 of the focusing unit 25 in the vertical direction based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF process for focusing the focal point 23 on the marker M1 (step S718). Specifically, the focusing control unit 622 moves the focusing unit 25 to the focal position Zm2 (Zm2 = Zm1−ΔZm2) based on the AF signal input from the AF unit 252.

  Subsequently, the photographing control unit 627 causes the imaging device 3 to photograph the marker M1 in the visual field region of the imaging device 3 (step S719). In this case, in the imaging control unit 627, the position detection unit 213 detects the marking image (reference image) corresponding to the marking image data generated by the imaging device 3 imaging the marker M1 accommodated in the container D1. The position information (Xm1, Ym1) of the stage 211 and the position information (Zm2) of the focus position of the focusing unit 25 are recorded in the photographing information recording unit 611 in association with each other.

  After that, the deviation amount calculation unit 625 calculates the in-image coordinates of the marker M1 in the marking image generated by the imaging device 3 and records it in the imaging information recording unit 611 (step S720). Specifically, the deviation amount calculation unit 625 calculates the in-image coordinates (Px2, Py2) of the marker M1 from the marking image generated by the imaging device 3, and records it in the recording unit 61.

  Subsequently, the deviation amount calculation unit 625 calculates the deviation amount of the marker M1 (step S721). Specifically, the deviation amount calculation unit 625 calculates a movement amount (drift amount) by which the marker M1 has moved from the marking image generated by the imaging device 3. More specifically, the shift amount calculation unit 625 calculates the shift amount (ΔPx2, ΔPy2) of the marker M1 on the marking image in pixel conversion using the above-described equations (13) and (14).

  Thereafter, the deviation amount calculation unit 625 calculates the deviation amount on the specimen SP surface (step S724). Specifically, the shift amount calculation unit 625 calculates the shift amount on the specimen SP surface based on the shift amount on the marking image (shift amount on the pixel) calculated in step S721 described above. More specifically, the deviation amount calculation unit 625 calculates the deviation amount (ΔX2, ΔY2) on the specimen SP surface by the above-described equations (15) and (16).

  Thereafter, the correction unit 626 corrects the coordinate information of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 (step S724). Specifically, the correction unit 626 corrects the XY coordinates (Xm2, Ym2) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625. More specifically, the correction unit 626 calculates the XY coordinates (Xm2, Ym2) of the marker M1 recorded by the imaging information recording unit 611 based on the deviation amount of the marker M1 calculated by the deviation amount calculation unit 625 ( It is corrected by 17) and (18). In this case, the correction unit 626 captures the XY coordinates (Xm2, Ym2) of the marker M1 corrected by the equations (17) and (18) and the Z coordinate (Zm2) of the marker M1 detected by the focusing control unit 622, respectively. It is recorded in the recording unit 611 and the coordinate information of the marker M1 is updated ((Xm1, Ym1, Zm1) → (Xm2, Ym2, Zm2)).

  Subsequently, the correction unit 626 corrects the coordinate information of the registration point based on the coordinate information of the marker M1 recorded by the imaging information recording unit 611 (step S725). Specifically, the correction unit 626 corrects the coordinate information of the registered points (for example, registered points A, B, and C) based on the coordinate information of the marker M1 corrected in step S724 described above. More specifically, the correction unit 626 corrects the coordinate information of the registration points A to C by the above-described equations (19) to (24).

  Thereafter, the stage control unit 621 moves the stage 211 to a registration point recorded by the shooting information recording unit 611 (step S725). For example, the stage control unit 621 moves the stage 211 to the registration point A (Xa2, Ya2, Za1) corrected by the correction unit 626.

  Subsequently, the focusing control unit 622 detects the position of the focusing unit 25 in the vertical direction (Z direction) with respect to the registration point based on the AF signal input from the AF unit 252 (step S726). For example, the focusing control unit 622 detects the position ΔZa2 in the vertical direction of the focusing unit 25 with respect to the registration point A based on the AF signal input from the AF unit 252.

  After that, the focusing control unit 622 drives the focusing driving unit 253 based on the AF signal input from the AF unit 252 to move the focusing unit 25 in the vertical direction (Z direction), whereby the objective lens The focusing unit 25 is moved to the focal position in order to execute the AF process for focusing the focal point 23 on the marker M1 (step S727). For example, the focusing control unit 622 moves the focusing unit 25 to the focal position Za2 (Za2 = Za1-ΔZa2) with respect to the registration point A based on the AF signal input from the AF unit 252.

  Thereafter, the imaging control unit 627 causes the imaging device 3 to image the specimen SP in the visual field region of the imaging device 3 (step S728). For example, the imaging control unit 627 captures the registration point A of the sample SP in the visual field region of the imaging device 3 and generates image data of the registration point A.

  Subsequently, the microscope control device 6 determines whether or not the imaging of all the registration points for the sample SP has been completed based on the imaging information recorded by the imaging information recording unit 611 (step S729). When the microscope control device 6 determines that the imaging of all the registration points for the sample SP has been completed (step S729: Yes), the microscope system 1 proceeds to step S730 described later. On the other hand, when the microscope control device 6 determines that imaging of all the registration points for the specimen SP has not been completed (step S729: No), the microscope system 1 returns to step S725 described above.

  In step S729, the microscope control device 6 determines whether or not the registration time has elapsed since the imaging of all the registration points for the sample SP based on the imaging information recorded by the imaging information recording unit 611. When it is determined that the registration time has elapsed since the microscope control device 6 photographed all the registration points for the specimen SP (step S730: Yes), the microscope system 1 proceeds to step S731 described later. On the other hand, when the microscope control device 6 determines that the registration time has not elapsed since the imaging of all the registration points for the specimen SP (step S730: No), the microscope system 1 passes the registration time. Continue this judgment until.

  In step S <b> 731, the microscope control device 6 determines whether or not the designated cycle for photographing all the registration points for the specimen SP has been completed based on the photographing information recorded by the photographing information recording unit 611. When the microscope control device 6 determines that the designated cycle for photographing all the registration points for the specimen SP has been completed (step S731: Yes), the microscope system 1 proceeds to step S7 described later. On the other hand, when the microscope control apparatus 6 determines that the designated cycle for photographing all the registration points for the specimen SP has not been completed (No at Step S731), the microscope system 1 returns to Step S716 described above.

  In step S732, the image processing unit 623 generates a time-lapse moving image in which a plurality of pieces of image data recorded by the recording unit 61 are sequentially connected for each designated position of the sample SP and records the generated time-lapse moving image in the recording unit 61. After step S732, the microscope system 1 returns to the main routine of FIG.

  In the fourth embodiment described above, since the process of switching the mirror turret 27 can be omitted, the processing time of the time lapse process can be shortened.

  Furthermore, in the fourth embodiment, the deviation of the designated position of the specimen due to the thermal drift can be accurately corrected with a simple configuration regardless of the kind of specimen.

(Modification 1)
Next, Modification 1 according to Embodiments 1 to 4 of the present invention will be described. In the first to third embodiments described above, the marker M1 is provided in the container D1, but in the first modification according to the present embodiment, a marking is provided on the placement surface on which the specimen SP is placed.

  FIG. 12 is a top view showing the configuration of the stage according to the first modification of the present embodiment. The stage 211a shown in FIG. 12 has an opening R10 and a transparent sheet M2 within the interference avoidance range of the objective lens 23. A marker M1 is provided on the sheet portion M2.

  According to Modification 1 of the present embodiment described above, the thermal drift of the microscope apparatus 2 can be corrected even when performing time-lapse observation of a large specimen SP that cannot be held in the container D1.

(Modification 2)
Next, Modification 2 according to the embodiment of the present invention will be described. In Embodiments 1 to 3 described above, there is one marker M1, but in Modification 2 according to the present embodiment, a plurality of markers M1 are provided in the same plane.

  FIG. 13 is a top view of a container that accommodates a specimen according to Modification 2 of the present embodiment. In the container D10 shown in FIG. 13, a plurality of markers M1a, markers M1b, and markers M1c are provided on the same plane. In this case, in the process of performing the time lapse observation described above, all the markers M1a to M1c are sequentially photographed in the step of photographing the marker M1 to generate each marking image. The position information detected by the markers M1a to M1c is associated and recorded in the recording unit 61. The deviation amount calculation unit 625 includes M1a to M1c in the reference image corresponding to each marking image recorded by the recording unit 61, and M1a to M1c in the current material image corresponding to each latest marking recorded by the recording unit 61. Based on the above, the drift amount distribution in the container D10 is calculated.

  Specifically, as shown in FIG. 14, the designated positions of the markers M1a, M1b, and M1c set by the imaging information setting unit 624 are (a, b), (a + 1, b), (a, b−). 1) and when the designated position of the sample SP is (a + 1, b−0.5), the drift amounts of the markers M1a and M1b calculated by the deviation amount calculation unit 625 are (−0.1, −0.1). ), When the drift amount of the marker M1c is (−0.1, −0.2), the X direction is uniformly drifted by −0.1 dots in the container D10. Further, in the container D10, the Y direction is the distance from the marker M1b to the marker M1c, and the drift amount is −0.1 to 0.2. As a result, it is found that the drift amount dy = 0.1 × (Y−b) −0.1. As a result, at the designated position (a + 1, b−0.5) of the sample SP, the deviation amount calculation unit 625 calculates the drift amount as (−0.1.0.15).

  According to the modified example 2 according to the embodiment of the present invention described above, the plurality of markers M1a to M1c are provided in the same plane on which the sample SP is placed. Can be corrected more accurately.

(Modification 3)
Next, Modification 3 according to the embodiment of the present invention will be described. FIG. 15A is a diagram showing concentric markers fixed in a container for accommodating a specimen according to Modification 3 according to the present embodiment. FIG. 15B is a diagram showing circular markers with different diameters that are fixed in a container that accommodates a specimen according to Modification 3 of Embodiments 1 to 4 of the present invention.

  As shown in FIG. 15A, the container D20 is provided with a plurality of markers M10 to M13 formed in a concentric shape having different diameters. Thereby, an optimal marker can be applied among the plurality of markers M10 to M13 for each magnification of the objective lens 23.

  Further, as shown in FIG. 15B, the container D20 is provided with a plurality of markers M14 to M18 having different diameters. Thereby, according to the magnification of the objective lens 23, an optimal marker can be applied among the plurality of markers M10 to M18.

  According to Modification 3 according to the embodiment of the present invention described above, the designated position of the specimen SP can be corrected with the markers M10 to M13 suitable for the magnification of the objective lens 23.

(Other embodiments)
In the present invention, a microscope system including a microscope device, an imaging device, a display unit, an input unit, and a microscope control device has been described as an example. For example, an objective lens for enlarging a specimen, and an imaging function for imaging a specimen via the objective lens The present invention can also be applied to an imaging device having a display function for displaying an image, such as a video microscope.

  In the present invention, the fluorescent material is described as an example of the marker. In this case, in the microscope system, the marker may be imaged by transmission observation, and the sample may be imaged by fluorescence observation. At this time, the microscope control device performs illumination switching by rotating the mirror turret before and after photographing with the marker and the specimen.

  In the present invention, an inverted microscope apparatus has been described as an example of the microscope apparatus. However, the present invention can be applied to, for example, an upright microscope apparatus. Furthermore, the present invention can be applied to various systems such as a line apparatus incorporating a microscope apparatus.

  In the present invention, the display unit, the input unit, and the microscope control device are separately configured. However, for example, a portable terminal in which the display unit, the input unit, and the microscope control device are integrally formed may be used.

  In the description of the flowchart in the present specification, the context of the processing between steps is clearly indicated using expressions such as “first”, “after”, “follow”, etc., in order to implement the present invention. The order of processing required is not uniquely determined by their representation. That is, the order of processing in the flowcharts described in this specification can be changed within a consistent range.

  As described above, the present invention can include various embodiments not described herein, and various design changes and the like can be made within the scope of the technical idea specified by the claims. Is possible.

DESCRIPTION OF SYMBOLS 1,1a Microscope system 2 Microscope apparatus 3 Imaging apparatus 4 Input part 5 Display part 6,6a Microscope control apparatus 20 Main body part 21 Stage mechanism 22 Transmitted illumination part 23 Objective lens 24 Revolver 25 Focusing part 26 Incident light source part 27 Mirror turret 28 Observation optical system 29 Lens barrel unit 61 Recording unit 62, 62a Control unit 211, 211a Stage 212 Stage drive unit 213 Position detection unit 611 Imaging information recording unit 621 Stage control unit 622 Focusing control unit 623 Image processing unit 624 Imaging information setting unit 625 Deviation amount calculation unit 626, 626a Correction unit 627 Imaging control unit 628 Display control unit D1, D10, D20 Container M1, M10, M1a, M1b, M1c Marking SP sample

Claims (8)

A stage on which a specimen can be placed and moved horizontally,
An objective lens arranged opposite to the stage for observing the specimen;
A stage drive unit for moving the stage relative to the objective lens;
An imaging device that images the specimen through the objective lens and generates image data of the specimen;
A plurality of designated positions for designating observation locations on the specimen, a photographing order of each of the plurality of designated positions by the imaging device, a photographing time of each of the plurality of designated positions by the imaging device, and the imaging device by the imaging device A shooting information setting unit for setting shooting information in association with the number of cycles indicating the number of times to repeat shooting at each of a plurality of designated positions;
A stage controller that drives the stage driving unit to sequentially move the stage to the plurality of designated positions based on the imaging information set by the imaging information setting unit;
An imaging control unit for imaging the sample by the imaging device at the imaging time set by the imaging information setting unit;
Before the stage moves to the plurality of designated positions, the imaging device corresponds to image data generated by photographing an index indicating the movement of the stage caused by thermal drift at a predetermined position on the stage. The index shift amount is calculated based on the reference image to be processed and the current image corresponding to the image data generated by capturing the index at the predetermined position after the first cycle is completed. A deviation amount calculation unit to perform,
A correction unit that corrects each of the plurality of designated positions based on the shift amount calculated by the shift amount calculation unit;
A microscope system comprising:   The stage control unit drives the stage driving unit to move the stage so that the index is positioned on the optical axis of the objective lens every time one cycle is completed. The microscope system according to 1.   The stage control unit drives the stage driving unit on the optical axis of the objective lens every time when the predetermined number of cycles is completed or every time the imaging apparatus images the sample and a predetermined time elapses. The microscope system according to claim 1, wherein the stage is moved to the predetermined position so that the index is located. A recording unit that records the image data sequentially generated by the imaging device and the shift amount in association with each other;
The correction unit corrects the center in the image corresponding to the image data to be the specified position based on the shift amount associated with the image data after the number of cycles is completed. The microscope system according to claim 1, characterized in that:   The microscope system according to claim 1, wherein the index is a fluorescent material that emits fluorescence with respect to light having a predetermined wavelength band.   The microscope system according to claim 1, wherein the index is provided in a container that accommodates the specimen.   The microscope system according to claim 1, wherein the index is provided on a placement surface on which the specimen is placed on the stage.   The microscope system according to claim 1, wherein a plurality of the indicators are provided in the same plane.

Images