JP2006038977A - Optical microscopic system and sample dynamic image forming method using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an optical microscopic system capable of positioning and photographing many samples existing in a plurality of different observation fields by one-step operation. <P>SOLUTION: The optical microscopic system 100 is equipped with a sample chamber 10, a two-dimensional moving stage 12 and a microscopic device 30. The sample chamber 10 is placed on the two-dimensional moving stage 12 while holding a well plate 1 having a plurality of wells 1a in which the samples S are housed, and is moved in an X-Y direction by the two-dimensional moving stage 12. A stage control part 51 controls the two-dimensional moving stage 12, so as to move the sample S to a predetermined photographing position along a moving path extended to a plurality of observation ranges. The X-Y position data of the two-dimensional moving stage 12 is used for the movement of the sample on the moving path. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an optical microscope system and a sample moving image generation method using the same.

  When a biological sample such as a living cell is observed with a microscope, it is an effective method to observe the reaction or change of the cell with respect to the reagent over time. For this purpose, a plurality of microscopic images of specific cells are photographed at regular time intervals, and after the photographing is completed, these microscopic images are arranged in time series and observed as a moving image. By developing this method, after selecting a plurality of microscope field of view, selecting a plurality of shooting ranges while observing the microscope within the microscope field of view, positioning a plurality of shooting ranges, and sequentially shooting repeatedly, 2. Description of the Related Art A microscopic photographing apparatus that obtains the same number of moving images as the number of photographing ranges is known (see, for example, Patent Document 1).

JP 2002-277754 A (2nd page, FIGS. 3 and 4)

  In the technique of the above-mentioned patent document 1, since a plurality of imaging ranges are selected and positioned within the same microscope field range after selecting the microscope field range, there is a problem that the operation until imaging is two-stage and lack of startability. is there.

  (1) In order to solve the above problem, the optical microscope system according to the first aspect of the present invention forms a microscope image of the observation sample and a holding means for holding a container in which the observation sample is accommodated in each of a plurality of chambers. An observation means, a relative movement means for relatively moving the holding means and the observation means so as to form respective microscope images of the observation sample by the observation means, an imaging means for imaging a microscope image by the observation means, and an imaging means. Storage means for storing image data of the captured microscopic image, and drive control of the relative movement means so that each of the observation samples is imaged by the imaging means at imaging positions in a predetermined order based on the position information of the chamber. A moving image that generates each moving image of the observation sample by arranging the image data of the observation sample stored in the control means and the storage means in time series for each observation sample Characterized in that it comprises a generation unit.

(2) The optical microscope system according to the second aspect of the present invention is the optical microscope system according to the first aspect, wherein the environment around the container in which the observation sample is accommodated in each chamber is maintained at a predetermined temperature, humidity and carbon dioxide concentration. It is preferable to further comprise an apparatus.
The optical microscope system according to a third aspect of the present invention is preferably the optical microscope system according to the first or second aspect, further comprising an environment changing means for adding, removing, and replacing a reagent in each small chamber.
(3) The optical microscope system of the invention according to claim 4 is the optical microscope system according to any one of claims 1 to 3, wherein the control means is present on a mark formed on the bottom surface of each small chamber or on the bottom surface of each small chamber. It is preferable that a morphological feature is input in advance as position information, and a predetermined photographing position is corrected using the mark or morphological feature as a position reference.
An optical microscope system according to a fifth aspect of the present invention may further include an autofocus mechanism in the optical microscope system according to any one of the first to fourth aspects.

(4) The sample moving image generating method of the invention according to claim 6 is obtained by imaging microscopic images of biological samples at imaging positions in a predetermined order based on positional information of a plurality of wells. The image data of the microscopic image is stored together with the position information of the plurality of wells, and after a predetermined time has passed, the imaging process and the storage process are performed, and each stored image data is stored for each biological sample based on the position information of the plurality of wells. The moving image data of each biological sample is generated by arranging them in time series.
The sample moving image generating method of the invention according to claim 7 is the sample moving image generating method of claim 6,
After the imaging step and the storage step, an environment changing step by adding or removing a reagent is further performed on one or more wells of the plurality of wells.

  According to the present invention, many operations in a plurality of different observation fields can be performed by one-step operation by the control means without performing a complicated operation of selecting a plurality of imaging ranges while performing microscopic observation in each observation field. The subject can be photographed by positioning the subject.

Hereinafter, an optical microscope system according to an embodiment of the present invention will be described with reference to FIGS.
FIG. 1 is an overall configuration diagram schematically showing the configuration of an optical microscope system according to an embodiment of the present invention. The optical microscope system 100 according to the present embodiment includes a sample chamber 10, a two-dimensional moving stage 12, a sealed container 20, and a microscope apparatus 30. The sealed container 20 is a container that houses a part of the sample chamber 10, the two-dimensional moving stage 12, and the microscope apparatus 30.

  The sample chamber 10 is opened such that the ceiling portion is indicated by the open surface 10 </ b> A, and the opening 11 </ b> A is formed in the bottom plate 11. The culture vessel 1 is placed on the bottom plate 11 so as to close the opening 11A. The culture vessel 1 is, for example, a well plate as shown in the figure, and has a plurality of small chambers (wells) 1a, and each of the small chambers contains a biological sample and a reagent (an aqueous solution such as a culture solution and a staining solution). The The sample chamber 10 is placed on the two-dimensional movement stage 12 and moved by the two-dimensional movement stage 12 in the X direction and the Y direction that are perpendicular to the optical axis AX. That is, the culture vessel 1 can move in the X direction and the Y direction along the horizontal plane. An opening 21A is formed in the upper plate 21 of the sealed container 20, and a shutter mechanism 22 for opening and closing the opening 21A is provided.

  The microscope device 30 includes an objective lens 31, a focus adjustment mechanism 32, an excitation light illumination device 33, a fluorescent filter device 34, a second objective lens 35, a half prism 36, an imaging device 37, and a transmission illumination device 40. The objective lens 31 is held by a focus adjustment mechanism (vertical movement stage) 32 and is moved in the direction of the optical axis AX, that is, the Z direction by the focus adjustment mechanism 32. The transmitted illumination device 40 is disposed above the sealed container 20 and includes a light source 41 and a collimator lens 42.

  The stage control unit 51 is electrically connected to the two-dimensional movement stage 12 via wiring and is connected to a personal computer (PC) 53. Further, the stage control unit 51 includes a position correction unit 51a. The autofocus unit 52 is electrically connected to the focus adjustment mechanism 32 via wiring and is also connected to the PC 53. The PC 53 sends data relating to the movement path, movement distance, movement direction, and the like of the two-dimensional movement stage 12 to the stage control unit 51, and inputs XY position data of the two-dimensional movement stage 12 from the stage control unit 51. Remember. The moving range of the two-dimensional moving stage 12 is not limited to the observation field of view of the microscope apparatus 30, but covers a wide range including a plurality of observation fields. Further, the PC 53 receives the focal position data of the objective lens 31 from the autofocus unit 52 and stores it.

  Further, the PC 53 includes an image data storage unit 54 and an image data editing unit 55. The image data storage unit 54 stores an image signal of a microscope image from the image sensor 37a of the image pickup device 37 as image data. The image data editing unit 55 inputs and edits image data from the image data storage unit 54 to generate a continuous image (moving image). The display unit 56 displays the moving image generated by the image data editing unit 55. In the PC 53, the XY position data, the focus position data, and the image data can be associated.

Hereinafter, microscope observation, photographing, and moving image editing using the optical microscope system 100 of the present embodiment will be described.
FIG. 2 is a plan view schematically showing the structure of the culture vessel (well plate) 1. The well plate 1 is a transparent plate in which a total of 96 wells 1a in 8 rows and 12 rows are formed. Each well 1a contains an observation sample (hereinafter referred to as sample S) in which at least one of a biological sample and a reagent is different, and microscopic images are taken at predetermined time intervals for all the wells 1a. The positional information such as the movement path, movement distance, and movement direction of the two-dimensional movement stage 12 described above is data based on the position of each well 1a. The imaging order of the sample S in each well 1a will be described later in connection with the moving image editing process shown in FIG. In each well 1a, a mark for fine adjustment of the photographing position may be formed in advance.

  First, microscopic observation and photographing of the sample S will be described with reference to FIG. A well placed on the two-dimensional moving stage 12 by a control signal from the stage control unit 51 so that the sample S accommodated in the first well 1a enters a desired observation field, that is, at a predetermined imaging position. The plate 1 is moved on the XY plane by a predetermined moving distance and moving direction. This XY position data can be expressed by, for example, two-dimensional coordinates with a specific well 1a position as the origin. In the observation visual field, that is, in the position in the XY direction, the focus adjustment mechanism 32 is driven by the autofocus unit 52 to move the objective lens 31 in the Z direction, and the focal position of the microscope image of the sample S is determined. This focal position data can be expressed by, for example, one-dimensional coordinates with the position of the sample S in a specific well 1a as the origin.

  In the case of transmission image observation, the illumination light from the transmission illumination device 40 irradiates the sample S in the well 1a through the opening 21A of the upper plate 21 of the sealed container 20. The light transmitted through the sample S enters the objective lens 31 through the bottom plate of the well plate 1. The light incident on the objective lens 31 passes through the fluorescent filter device 34 and the second objective lens 35 and is branched by the half prism 36. One light passes through the half prism 36 and enters the imaging device 37, and forms an image on the imaging surface of the imaging element 37a. The other light is reflected by the half prism 36 and guided to an eyepiece (not shown). Microscopic image data of the sample S is sent from the image sensor 37 a to the PC 53 and stored in the image data storage unit 54.

  During transmission observation, the mark formed on the bottom surface of the well 1a can be recognized in the observation field of the well 1a, and the photographing position can be finely adjusted based on the position of this mark. That is, the mark position data in the observation visual field is input to the PC 53, and this position data is sent to the position correction unit 51a to correct the XY position data. Due to mechanical errors during driving of the two-dimensional moving stage 12, the position where the well 1a is first set, and the position where the well 1a is reset after the driving of the two-dimensional moving stage 12 (second and subsequent setting) Then, a slight deviation may occur. By finely adjusting the photographing position, the well 1a can be accurately moved to a predetermined photographing position. Further, instead of the mark, morphological features such as scratches and dents existing on the bottom surface of each well 1a may be used for fine adjustment of the photographing position.

  Similarly, when observing and photographing a sample S ′ accommodated in another well 1a, the well plate 1 is moved by the two-dimensional moving stage 12 and the focus of the objective lens 31 is adjusted by the focus adjustment mechanism 32, and the sample S ′ is thus obtained. The XY position data and the focal position data are stored by the PC 53. The focus of the sample S ′ is also adjusted because the focus position may be shifted depending on the movement of the well plate 1 or the state of the sample S in each well 1a. This focus adjustment may be performed only once for each well 1a or may be performed each time.

  Microscope image data of the sample S ′ is also sent from the image sensor 37 a to the PC 53 and stored in the image data storage unit 54. In this way, the image data storage unit 54 stores image data for the samples S in all the wells 1 a of the well plate 1. The above photographing and recording are performed for the number of samples S, and this series of photographing and recording is repeatedly performed with a predetermined waiting time. When all photographing and recording are finished, all the microscope image data are edited by the image data editing unit 55, and a moving image is generated. The moving image generation by the image data editing unit 55 will be described in detail later.

  In the case of fluorescence image observation, the illumination light emitted from the excitation light illumination device 33 passes through the light control filter 33 a and the fluorescence filter device 34 and enters from below the objective lens 31. The light incident on the objective lens 31 passes through the bottom plate of the well plate 1 and irradiates the sample S. Fluorescence is emitted from the sample S by this excitation light. The fluorescence passes through the well plate 1, the objective lens 31, the fluorescence filter device 34, and the second objective lens 35, and is branched by the half prism 36. One light passes through the half prism 36 and enters the imaging device 37, and forms an image on the imaging surface of the imaging element 37a. The other light is reflected by the half prism 36 and guided to an eyepiece (not shown). The transmission and storage of the microscope image data of the sample S, the movement of the two-dimensional moving stage 12, the focus adjustment, and the like are the same in both the transmission image observation and the fluorescence image observation.

  FIG. 3 is a flowchart showing a process from photographing of the sample S to editing of the moving image in the optical microscope system according to the present embodiment. Now, assume that the number of wells 1a, that is, the number of samples S, is m, and the number of times that each sample S is repeatedly performed at predetermined time intervals is n. In step S1, the position of the well 1a of the sample S to be photographed first is determined, in step S2, the sample S is photographed, and in step S3, a microscope image of the sample S is stored. In step S4, it is checked whether or not the microscope images have been stored for all the samples S. If “No”, the process proceeds to step S5, and position setting, imaging (step S2), and image storage (step S3) are performed for the next sample S.

  When imaging of all m samples S is completed in step S4, the process proceeds to step S6, and a predetermined waiting time T is set. When the number of times of photographing n = 2, the XY position data of each sample S when the number of times of photographing n = 1 is used to drive the two-dimensional moving stage 12 to set the photographing position. The same is true when the number of times n is increased. Accordingly, each sample S is always photographed at the same photographing position. In addition, when the focus adjustment is performed every time at the time of photographing, the focus position data obtained when the number of times of photographing n = 1 for each of the m samples S is used to perform the focus adjustment after the second number of times of photographing. Can do.

  In step S7, it is checked whether or not a predetermined number of photographing times has been reached for each sample S. If “No”, the process proceeds to step S8, the number of times of photographing n is increased, and step S1 to step S6 are repeated. In step S7, when the predetermined number of photographing times n is reached, the process proceeds to step S9, and the moving image editing is performed. When the edited moving image data is stored in step S10, the series of operations is terminated.

  FIG. 4 is a diagram showing a moving image editing process in the optical microscope system according to the present embodiment. FIG. 4A is a sequence of captured microscopic images arranged in the order of imaging, and FIG. 4B is a moving image in which only the images of the sample S at the position (a, 3) are arranged in time series. is there. From t11 to tnm represents the time when each microscopic image was taken.

  Here, with reference to FIG. 2, the imaging sequence of the sample S in each well 1a will be described from the third to seventh wells 1a of the well plate 1. FIG. Starting from the well 1a at the first position (a, 3), in the figure, proceed downward according to the arrow to the well 1a at the position (h, 3) and proceed upward from the well 1a at the position (h, 4). The position is sequentially advanced to the position (a, 4), and when reaching the well 1a at the final position (h, 7), the first imaging is finished. This series of photographing corresponds to the number of photographing times n = 1 in FIG. 4 and photographs m microscopic images.

  As shown in FIG. 4A, after the standby time T has elapsed, the process of shooting number n = 2 is entered. In FIG. 2, the well plate 1 is moved from the final position (h, 7) to the first position (a, 3). Even when the number of times of photographing n = 2, the same position setting and photographing as in n = 1 are performed, and m microscopic images are photographed. This series of operations is repeated n times. That is, m × n microscope images are taken from time t11 to tnm.

  As shown in FIG. 4B, a moving image is obtained when n images of the sample S at the position (a, 3) are arranged in time series. The time interval of each image is a time required to move m well images to wait time T and to move the well plate 1 from the final position (h, 7) to the first position (a, 3). It is the time which added stage movement time. In this way, a moving image composed of n images with respect to m positions, that is, m samples S is obtained.

  In the optical microscope system 100 of the present embodiment, the stage control unit 51 can drive the two-dimensional moving stage 12 not only in the observation field of the microscope apparatus 30 but also in a wide range including a plurality of observation fields. There is no need to determine the range, and it is possible to position and photograph many samples S in a plurality of different observation fields by one-step operation. Since this positioning is accurately performed by the stage control unit 51, a moving image with smooth motion with little blur is obtained. Further, by providing the position correction unit 51a in the stage control unit 51, a moving image with smooth motion with less blur can be obtained. It should be noted that the position correction unit 51a is not provided, and the image is shifted so that the mark in the image data, the scratch of the well 1a, etc. are at the same position on each screen at the moving image editing stage after all the imaging is completed. You may edit it.

  In the present embodiment, the two-dimensional moving stage 12 is driven to move each sample S to its imaging position. However, the stage on which the well plate 1 is placed is fixed, and the optical microscope 30 is moved in the XY directions. The two-dimensional moving stage 12 and the optical microscope 30 may be moved in the XY direction. Further, the observed image of the sample S may be a dark field image or a phase difference image.

Hereinafter, the environmental adjustment method in the sample chamber 10 will be described with reference to FIG.
FIG. 5 is a block diagram schematically showing the overall configuration when the environment control device 60 is added to the optical microscope system 100 shown in FIG. 1, and the same components as those in FIG. Is omitted. Further, the stage control unit 51, the autofocus unit 52, the PC 53, the image data storage unit 54, the image data editing unit 55, the display unit 56, and the transmitted illumination device 40 are not shown.

  The environment control device 60 is a device that generates a gas having a desired temperature, humidity, and composition and circulates the gas into the sample chamber 10. The environment control device 60 includes a humidifier 61, a heater 62, a blower 63, a temperature / humidity sensor 64, a CO2 gas sensor 65, and a reserve tank 66. The blower 63 is connected to the upper plate 21 of the sealed container 20 by a gas delivery tube 67. The humidifier 61 is connected to the upper plate 21 of the sealed container 20 by a gas suction tube 68. A pipe connection location of the upper plate 21 communicates with the sample chamber 10. The humidifier 61 is connected to the reserve tank 66 by piping. A solenoid valve 69 is provided in a path that branches from between the heater 52 and the blower 53 so that gas can be introduced from a CO2 gas cylinder.

  The temperature / humidity sensor 64 is attached to the lower surface of the upper plate 21 of the sealed container 20 in order to monitor the temperature and humidity in the sample chamber 10. The CO 2 gas sensor 65 is attached in the middle of the path of the gas suction tube 67 in order to monitor the CO 2 gas concentration in the sample chamber 10. Each tube is heat-insulated.

  The gas composition, humidity, and temperature of the gas supplied to the sample chamber 10 by the gas delivery tube 67 are adjusted as follows. The gas composition is adjusted by feeding back the measured value of the CO 2 gas sensor 65 and increasing or decreasing the opening amount of the electromagnetic valve 69. Humidity is adjusted by feeding back the measured value of the temperature / humidity sensor 64 and increasing or decreasing the drive voltage to the ultrasonic atomizing element of the humidifier 61. The ultrasonic atomizing element generates water vapor from the water in the humidifier 61 by ultrasonic vibration. Note that a certain amount of water in the humidifier 61 is always secured by supply from the reserve tank 66. The temperature adjustment is performed by feeding back the measured value of the temperature / humidity sensor 54 and increasing or decreasing the current flowing through the electric heater of the heater 51.

  The gas thus adjusted is, for example, 37 ° C.-100% RH, 5% CO 2, supplied from the environmental control device 60 to the sample chamber 10 by the gas delivery tube 67, and from the sample chamber 10 to the gas suction tube 68. To return to the environment control device 60. The adjustment gas on the circulation path is sent again into the sample chamber 10 as air having a predetermined temperature, humidity and gas composition by the environment control device 60. Since the sample chamber 10 is housed in the sealed container 20, there is no possibility that the adjustment gas leaks to the outside. By attaching the environment control device 60 to the optical microscope system 100, the inside of the sample chamber 10 is always maintained in a predetermined environment, so that the sample S can also be maintained in a predetermined condition.

Next, a method for changing the sample S in the well 1a of the well plate 1 arranged in the sample chamber 10 will be described with reference to FIG. As described above, the sample S is obtained by adding a reagent such as a culture solution to a biological sample such as a cell, and is an object in which at least one of the biological sample and the reagent is different.
6 is a block diagram schematically showing the overall configuration when an injection / inhalation mechanism 70 is added to the optical microscope system 100 shown in FIG. 1, and the same reference numerals are given to the same components as in FIG. Description is omitted. Further, the stage control unit 51, autofocus unit 52, PC 53, image data storage unit 54, image data editing unit 55, and display unit 56 are not shown.

  The injection / inhalation mechanism 70 is located above the optical microscope system 100, can be moved in the X, Y, and Z directions by a drive mechanism (not shown), and injects the reagent into the well 1a of the well plate 1 through the pipette 71. Or the reagent in the well 1a can be sucked. At the time of injection and inhalation, the transmission illumination device 40 is removed from the optical axis AX as shown in the figure. The opening 21A of the upper plate 21 of the sealed container 20 is opened by the shutter mechanism 22 so that the inside of the sample chamber 10 is opened to the outside. Thereby, the injection and inhalation of the reagent can be performed freely.

  When the injection, inhalation or exchange of the reagent is completed, the pipette 71 of the infusion / inhalation mechanism 70 is retracted upward, the shutter mechanism 22 is operated, and the opening 21A is closed with, for example, a transparent substrate. The illumination device 40 is returned to the optical axis AX. Thereby, the inside of the sample chamber 10 is sealed with respect to the outside world. By adding the injection / inhalation mechanism 70 to the optical microscope system 100, it is possible to freely inject and inhale the reagent. Before the microscopic observation and photographing of the sample S, the environment in the well 1a (the sample S (Conditions) can be changed, and it is possible to quickly respond to diversifying user demands.

  As a specific example, different reagents are injected into each well 1a containing the same sample S, the sample S in each well 1a is imaged after the injection, and a moving image generated for each well 1a is compared to thereby obtain a reagent. The difference in the effect can be investigated. In addition, to each well 1a containing the same sample S, the injection amount of the same reagent is added in stages in order of imaging, and the sample S of each well 1a is imaged, and a moving image generated for each well 1a is displayed. By comparing, it is possible to know the change state of the sample due to the addition amount of the reagent and the time from the addition until the change appears, and it is possible to find the optimum addition amount that produces the effect of the reagent. In this way, changes with time can be observed for each sample under different conditions. Further, if the injection / inhalation mechanism 70 is configured to control the timing at which the reagent is added to each well 1a and the timing at which the imaging device 37 images the sample S, the time of the sample S in each well 1a is timed. The change can be compared and observed more accurately.

  The present invention provides a control means for relatively controlling movement of the sample holding means and the observation means so that each observation sample is moved to an imaging position in a predetermined order based on the position information of the chamber in the optical microscope system. There is a feature in providing. The present invention is not limited to the embodiments described above as long as the characteristics are not impaired. The image data editing unit 55 corresponds to a moving image generating unit.

1 is an overall configuration diagram schematically showing a configuration of an optical microscope system according to an embodiment of the present invention. It is a top view which shows typically the structure of the culture container (well plate) used with the optical microscope system which concerns on embodiment of this invention. It is a flowchart which shows the process from imaging | photography of the sample S in the optical microscope system which concerns on embodiment of this invention to animation edit. It is a figure which shows the process of the moving image edit in the optical microscope system which concerns on embodiment of this invention. FIG. 4A is a sequence of captured microscopic images arranged in the order of imaging, and FIG. 4B is a moving image in which only the images of the sample S at the position (a, 3) are arranged in time series. is there. It is a block diagram which shows typically the whole structure at the time of adding an environmental control apparatus to the optical microscope system which concerns on embodiment of this invention. It is a block diagram which shows typically the whole structure at the time of adding an injection / inhalation mechanism to the optical microscope system which concerns on embodiment of this invention.

Explanation of symbols

1: Culture container (well plate)
DESCRIPTION OF SYMBOLS 1a: Well 10: Sample chamber 12: Two-dimensional moving stage 20: Airtight container 22: Shutter mechanism 30: Microscope apparatus 31: Objective lens 32: Focus adjustment mechanism 37: Imaging apparatus 40: Transmitting illumination apparatus 51: Stage control part 52: Autofocus part 53: PC
54: Image data storage unit 55: Image data editing unit 56: Display unit 60: Environmental control device 70: Injection / inhalation mechanism 100: Optical microscope system S: Sample

Claims (7)

Holding means for holding containers each holding an observation sample in a plurality of small chambers;
Observation means for forming a microscopic image of the observation sample;
A relative movement means for relatively moving the holding means and the observation means so as to form respective microscope images of the observation sample by the observation means;
Imaging means for imaging the microscope image by the observation means;
Storage means for storing image data of the microscope image captured by the imaging means;
Control means for driving and controlling the relative movement means so that each of the observation samples is imaged by the imaging means at imaging positions in a predetermined order based on the position information of the chambers;
Moving image generating means for generating respective moving images of the observation sample by arranging the image data of the observation sample stored in the storage means in time series for each observation sample, Optical microscope system. The optical microscope system according to claim 1,
An optical microscope system, further comprising an environment control device for maintaining the periphery of the container in which the observation sample is stored in each of the small chambers at a predetermined temperature, humidity, and carbon dioxide concentration. The optical microscope system according to claim 1 or 2,
An optical microscope system further comprising environment changing means for adding, removing, and replacing a reagent in each of the small chambers. In the optical microscope system according to any one of claims 1 to 3,
The control means acquires in advance, as position information, a mark formed on the bottom surface of each chamber or a morphological feature existing on the bottom surface of each chamber, and the predetermined photographing position using the mark or morphological feature as a position reference. An optical microscope system characterized by correcting the above. In the optical microscope system according to any one of claims 1 to 4,
An optical microscope system further comprising an autofocus mechanism. In a sample moving image generating method for sequentially capturing images of biological samples respectively stored in a plurality of wells of a well plate and generating a moving image of each biological sample,
Each of the microscopic images of the biological sample is captured at a predetermined imaging position based on the position information of the plurality of wells,
Storing image data of the microscopic image obtained by the imaging together with positional information of the plurality of wells,
After a predetermined time has elapsed, the imaging step and the storage step are performed,
The moving image data of each of the biological samples is generated by arranging the stored image data in time series for each biological sample based on the position information of the plurality of wells. Generation method. In the sample moving image generating method according to claim 6,
A method for generating a sample moving image, further comprising performing an environment changing step by adding or removing a reagent to one or more wells of the plurality of wells after the imaging step and the storing step.

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