JP6253509B2 - Image display method, control device, microscope system - Google Patents

Description

  The present invention relates to an image display method, a control device, and a microscope system, and more particularly, to an image display method when aligning a biological sample, a control device that executes the method, and a microscope system including the control device.

  When observing a specimen with a microscope system, a preparatory work (hereinafter referred to as alignment) is performed in which the stage is moved to position the region of interest in the specimen within the field of view of the microscope system. This alignment is usually performed while displaying a live image of the sample on a monitor or the like.

  However, when the specimen is a fluorescent specimen, the fluorescent specimen is faded by excitation light irradiated for visual or live display during alignment, and the fluorescence intensity generated from the fluorescent specimen is reduced. For this reason, in fluorescence observation performed after alignment, fluorescence image data with a deteriorated S / N ratio is generated.

  Various techniques for suppressing the fading of the fluorescent specimen generated in the preparation work have been proposed. For example, Patent Document 1 describes a technique for reducing the intensity of excitation light in the preview mode than in the image recording mode. Patent Document 2 describes a technique for acquiring a macro phase difference image and a macro fluorescence image at the preparation stage and displaying them in a superimposed manner. Patent Document 3 describes a technique for blocking or dimming illumination light when the relative position between the sample and the objective lens is not changed. Patent Document 4 describes a technology that measures the irradiation time of excitation light and warns when the irradiation time exceeds a predetermined reference value.

JP 2008-304613 A JP 2007-328134 A JP 2009-15301 A JP 2005-331889 A

Various techniques as described above have been proposed, but there is a need for a technique that further suppresses damage to a biological specimen during observation preparation work.
In light of the above circumstances, an object of the present invention is to provide a technique for suppressing damage to a biological specimen during observation preparation work.

  A first aspect of the present invention is a method for displaying an image of a biological specimen, and an imaged area that has already been imaged in a display target area selected from the area of the biological specimen as an area to be displayed is still imaged. When an unimaged area is included, the imaging target area including the unimaged area and not including the imaged area is irradiated with illumination light to capture an image of the imaged area, and the imaged area An image display method for displaying an image of the display target area including the imaged area and the imaged imaging target area based on the image data of the imaging target area and the image data of the imaging target area is provided.

  According to a second aspect of the present invention, in the image display method according to the first aspect, a second unimaged area that has not yet been imaged is included in a spare area of a predetermined size adjacent to the display target area. Sometimes, an illumination light is irradiated to a second imaging target area that includes the second non-imaging area and does not include the captured area and the imaging target area, and captures an image of the second imaging target area. An image display method is provided.

  According to a third aspect of the present invention, in the image display method according to the second aspect, the predetermined size is from a display target area selected before the current display target area to the current display target area. An image display method that depends on the moving speed of the display target area is provided.

  According to a fourth aspect of the present invention, in the image display method according to the first aspect, the display target area from the display target area selected before the current display target area to the current display target area is displayed. The movement provides an image display method that is performed in steps smaller than the size of the display target area.

  According to a fifth aspect of the present invention, there is provided a control device for a microscope system including an imaging device and a display device, and the imaging device has already imaged a display target region selected from a region of a biological specimen as a region to be displayed. When an imaged region that has been captured and an unimaged region that has not yet been imaged by the imaging device are included, illumination light is irradiated to an imaging target region that includes the unimaged region and does not include the imaged region, Based on an imaging control unit that causes the imaging device to capture an image of the imaging target region, image data of the captured region that is data of an image captured by the imaging device, and image data of the imaging target region, There is provided a control device comprising: a display control unit that causes the display device to display an image of the display target region including the imaged region and the imaged target region.

  According to a sixth aspect of the present invention, in the control device according to the fifth aspect, the imaging control unit is a second non-imaging area that has not yet been imaged in a spare area of a predetermined size adjacent to the display target area. Is included, the second imaging target area including the second non-imaging area and not including the imaging target area and the imaging target area is irradiated with illumination light, and the second imaging target area is irradiated to the imaging apparatus. A control device that captures an image of an imaging target region is provided.

  According to a seventh aspect of the present invention, in the control device according to the sixth aspect, the predetermined size is from the display target area selected before the current display target area to the current display target area. Provided is a control device that depends on a moving speed of a display target area.

  According to an eighth aspect of the present invention, in the control device according to the fifth aspect, the display target area is moved from the display target area selected before the current display target area to the current display target area. Provides a control device that is performed in steps smaller than the size of the display target area.

  According to a ninth aspect of the present invention, there is provided a microscope system including the control device according to any one of the fifth to eighth aspects.

  ADVANTAGE OF THE INVENTION According to this invention, the technique which suppresses the damage which a biological sample receives during the preparation work of observation can be provided.

1 is a diagram illustrating a configuration of a microscope system according to Embodiment 1. FIG. 6 is a flowchart illustrating processing performed during preliminary observation in the microscope system according to the first embodiment. It is the figure which showed the example judged that imaging is unnecessary. It is the figure which showed the example judged to be imaging. It is the figure which showed another example judged that imaging was required. FIG. 2 is a diagram illustrating a hardware configuration of a control device according to the first embodiment. FIG. 6 is a diagram illustrating a configuration of a microscope system according to a second embodiment. It is the figure which showed the GUI screen displayed on a monitor during preparatory observation. It is the figure which showed the table regarding the setting at the time of the preparatory observation displayed on a condition setting screen. It is the figure which showed the table regarding the setting at the time of this observation displayed on a condition setting screen. It is the figure which showed the table regarding the other setting displayed on a condition setting screen. FIG. 10 is a diagram illustrating a configuration of a microscope system according to a third embodiment. It is the figure which showed the relationship between a frame area | region and a reserve area | region. It is the figure which showed the relationship between a frame area | region, the imaged area | region, and the 2nd imaging object area | region. FIG. 10 is a diagram illustrating a configuration of a microscope system according to a fourth embodiment. It is the figure which showed the relationship between a frame area | region and an image pick-up element area | region. FIG. 10 is a diagram for explaining a modification of the imaging device of the microscope system according to the fourth embodiment. It is a modification of the flowchart which shows the process performed during preparatory observation.

  Examples of the present invention will be described below. In each example, a case where fluorescence observation is mainly performed will be described as an example. However, the present invention is not limited to the application to the fluorescence observation method, but can be applied to any microscopic method for observing a biological specimen. Can do. Further, hereinafter, observations performed during alignment are referred to as preparatory observations, and observations performed after alignment are referred to as main observations as necessary, and are distinguished from each other.

  FIG. 1 is a diagram illustrating a configuration of a microscope system 100 according to the present embodiment. The microscope system 100 includes a microscope apparatus main body 110, a control apparatus 120, a monitor 130, and an input apparatus 140.

  The microscope apparatus main body 110 is a fluorescent microscope, and includes a light source 111, an optical fiber 112, a shutter 113, a fluorescent cube 114, an objective lens 115, an imaging lens 116, a CCD camera 117, and a specimen S that is a biological specimen. Is provided, and a driving device 119 for driving the stage 118 is provided.

  The light source 111 is, for example, a mercury lamp. The optical fiber 112 is, for example, a liquid fiber having a liquid core. The shutter 113 is a light shielding means, and is controlled to be open during the exposure period of the CCD camera 117 and closed during the non-exposure period of the CCD camera 117. The fluorescent cube 114 is an optical path branching unit that splits the illumination optical path and the detection optical path, and stores therein a fluorescent filter set including a dichroic mirror, an excitation filter, and a barrier filter. The CCD camera 117 is an imaging device that has a two-dimensional image sensor and captures an image of the specimen S. The drive device 119 is, for example, a stepping motor.

  The control device 120 is a device that controls the microscope system 100, and includes an imaging control unit 120a and a display control unit 120b. The imaging control unit 120a mainly controls the operations of the shutter 113, the CCD camera 117, and the driving device 119 to cause the CCD camera 117 to capture an image of the sample S. The display control unit 120 b causes the monitor 130 to display an image of the display target area based on image data (image data) captured by the CCD camera 117. Note that the display target area is an area selected from the area of the sample S as an area to be displayed.

  The monitor 130 is a display device that displays a fluorescent image of the specimen S, and is, for example, a liquid crystal display device, an organic EL display device, a CRT display device, or the like. The input device 140 is a device for selecting a display target area. The input device 140 is provided with a handle for moving the display target region in the X direction and the Y direction, such as a stage handle provided in a conventional microscope system.

  FIG. 2 is a flowchart showing processing performed during preliminary observation in the microscope system 100. When the processing shown in FIG. 2 is started, the control device 120 performs initial setting (step S1). Here, for example, various parameters such as the magnification of the objective lens to be used, the excitation wavelength, and the fluorescence wavelength are set.

  Thereafter, the control device 120 detects a display target area (step S2). Here, immediately after the initial setting, for example, a preset area such as the vicinity of the reference coordinates of the stage is detected as the display target area. When an operation for designating a region by the user is detected, the display target region is detected based on a signal output from the input device 140 according to the user's operation.

  When the display target area is detected, the control device 120 determines whether or not imaging is necessary (step S3). Here, if the display target area detected in step S2 includes an unimaged area that has not yet been imaged by the CCD camera 117, it is determined that imaging is necessary. That is, the control device 120 needs to image not only when the entire display target area is an unimaged area but also when the display target area includes an already imaged area and an unimaged area. judge. On the other hand, when an unimaged area is not included, it is determined that imaging is unnecessary. This point will be specifically described with reference to FIGS. 3A to 3C.

  As shown in FIG. 3A, the control device 120 displays the imaged area A (the imaged unit area A1, the imaged unit area A2, the imaged unit) of which the display target area (frame area F) detected in step S2 is the sample S. If it is located in the area A3, the imaged unit area A4), it is determined that imaging is unnecessary. Further, as shown in FIGS. 3B and 3C, the control device 120 is located when the display target area (frame area F) detected in step S <b> 2 protrudes from the imaged area A (or is located outside the imaged area A). If it is determined that imaging is necessary.

  When it is determined that imaging is necessary, the control device 120 determines an imaging target area B that is an area to be imaged (step S4). Here, the control device 120 determines, as the imaging target area B, an area that includes the non-imaging area in the display target area and does not include the captured area A. The imaging target area B is preferably determined as an area adjacent to the imaged area A.

  For example, as shown in FIG. 3B, the imaging target area B may be composed of a plurality of areas (imaging target unit area B1 and imaging target unit area B2) each having a size corresponding to the real field of view. As shown in FIG. 3C, it may be composed of a single region having a size corresponding to the real field of view.

  Next, the control device 120 moves the stage 118 to the driving device 119 so that the imaging target region B determined in step S4 is located in the field of view. Thereafter, the control device 120 controls the shutter 113 to the open state, so that the excitation light is irradiated onto the imaging target region B (step S5). At the same time, the control device 120 causes the CCD camera 117 to which the fluorescence generated from the imaging target area B enters to capture an image of the imaging target area B (step S6). After the imaging is completed, the control device 120 controls the shutter 113 to be closed. The image data of the imaging target area B generated by the CCD camera 117 is output to the control device 120 and stored in the memory of the control device 120.

  Steps S5 and S6 are performed under the control of the imaging control unit 120a. As illustrated in FIG. 3B, when the imaging target area B includes a plurality of imaging target unit areas, the imaging control unit 120a repeats the processes of step S5 and step S6 for each imaging target unit area. Control.

  In step S7, the control device 120 causes the monitor 130 to display an image of the display target area detected in step S2. Here, the control device 120 determines the display target area based on the data of the image already captured by the CCD camera 117 (the image data of the captured area A and the image data of the imaging target area B generated in step S6). The image is displayed on the monitor 130. Specifically, the control device 120 extracts a portion corresponding to the display target area from these image data. Then, based on the image data of the part, the image of the display target area is displayed. The process in step S7 is performed by the display control unit 120b. Note that the display target area moves in steps (for example, in units of one pixel) smaller than the size of the display target area. For this reason, it is normal that the display target area includes both the imaged area and the imaging target area except immediately after the start of the processing shown in FIG. Therefore, in step S <b> 7, the control device 120 causes the monitor 130 to display an image of the display target area including the captured area and the captured imaging target area except immediately after the start of processing.

  Finally, the control device 120 determines whether or not there is an end instruction (step S8), and if there is no end instruction, redefines the imaging target area B determined in step S4 as a part of the imaged area A, The microscope system 100 is controlled so that the processing from step S2 to step S7 is repeated.

  In the microscope system 100 operating as described above, it is possible to prevent the excitation light from being irradiated a plurality of times to the same region in the sample S during the preparatory observation for observation. Since the excitation light is irradiated only during the exposure period of the CCD camera 117, the irradiation time can be suppressed. Since irradiation of excitation light to a region unnecessary for image display is limited, the irradiation range can be suppressed. Therefore, according to the microscope system 100, it is possible to greatly suppress damage to the specimen S during preparation for observation. For this reason, it is possible to align the specimen while observing a bright image without excessively suppressing the intensity of the excitation light irradiated during the preparatory observation. However, it is desirable to set the intensity of the excitation light irradiated during the preliminary observation to be lower than the intensity of the excitation light irradiated during the main observation.

  FIG. 4 is a diagram illustrating a hardware configuration of the control device 120 illustrated in FIG. 1. The control device 120 includes a CPU 121, a memory 122, a storage device 123, a reading device 124, a display IF 125, an input IF 126, and a communication IF 127, which are connected to each other via a bus 128.

  The CPU 121 can provide the functions of the imaging control unit 120a and the display control unit 120b by executing a control program that executes the processing illustrated in FIG.

  The memory 122 is a semiconductor memory, for example, and includes a RAM area and a ROM area. The storage device 123 is a hard disk device, for example, and stores a control program. Note that the storage device 123 may be a semiconductor memory such as a flash memory. The storage device 123 may be an external recording device. The image data generated by the CCD camera 117 is stored in the storage device 123, for example.

  The reading device 124 accesses the portable recording medium 129 in accordance with an instruction from the CPU 121. The portable recording medium 129 includes, for example, a semiconductor device (such as a USB memory), a medium (such as a magnetic disk) through which information is input / output by a magnetic action, and a medium (CD-ROM, etc.) through which optical information is input / output For example, a DVD).

  The display IF 125 is an interface device that outputs data to the monitor 130 in accordance with instructions from the CPU 121. The input IF 126 is an interface device that receives data from the input device 140 and is a device that receives an instruction from a user. The communication IF 127 is an interface device that transmits / receives data to / from an external device (for example, the shutter 113, the CCD camera 117, the driving device 119, etc.) in accordance with an instruction from the CPU 121.

  The control program may be installed in advance in the storage device 123, for example, and may be provided to the control device 120 via the portable recording medium 129 or via a network (not shown).

  FIG. 5 is a diagram illustrating a configuration of the microscope system 200 according to the present embodiment. The microscope system 200 includes a microscope apparatus main body 210, a control apparatus 220, and a monitor 230.

  The microscope apparatus main body 210 is different from the microscope apparatus main body 110 according to the first embodiment in that a transmission illumination unit is provided. In addition, the microscope apparatus main body 210 includes a turret 211 that switches a fluorescent cube arranged on the optical path between a plurality of fluorescent cubes (fluorescent cube 114a and fluorescent cube 114b), and a motor 212 that drives the turret 211. This is different from the microscope apparatus main body 110. Other points are the same as those of the microscope apparatus main body 110.

  The transmission illumination means of the microscope apparatus main body 210 includes a light source 213 that is a halogen lamp, a shutter 214, an illumination lens 215, and optical elements arranged on the optical path of a plurality of optical elements (ring slit 217a, DIC prism 217b). A turret 216 that switches between them, a motor 218 that drives the turret 216, and a condenser lens 219 are provided.

  The microscope apparatus main body 210 can perform phase difference observation, differential interference observation, and bright field observation by illuminating the specimen S with the transmission illumination means. Note that switching between phase difference observation, differential interference observation, and bright field observation is performed by rotating the turret 216 and switching the optical elements arranged on the optical path. For example, at the time of phase difference observation, the control device 220 controls the motor 218 to rotate the turret 216 so that the ring slit 217a is positioned on the optical path. At this time, the control device 220 also rotates the turret 211 so that an optical element corresponding to the observation method is also arranged on the detection optical path. For example, during differential interference observation, the control device 220 controls the motor 218 to rotate the turret 216 so that the DIC prism 217b is positioned on the illumination optical path. Further, the control device 220 controls the motor 212 to rotate the turret 211 so that a DIC prism (not shown) is positioned on the detection optical path.

  The control device 220 is a device that controls the microscope system 200, and includes an imaging control unit 220a and a display control unit 220b. The control device 220 has the same hardware configuration as that of the control device 120 according to the first embodiment, and operates in the same manner as the control device 120.

  The monitor 230 is a touch panel display device that functions not only as a display device but also as an input device. This point is different from the monitor 130 according to the first embodiment.

  FIG. 6 is a diagram showing a GUI screen (screen 231) displayed on the monitor during the preparatory observation. The screen 231 is provided with two image display areas. One is a frame window 232 on which an image of a frame area (display target area) is displayed. The other is a viewfinder window 236, which displays a frame area image and an already generated image of the surrounding area in a reduced size. In the finder window 236, a frame frame 237 indicating the position of the frame region is displayed so as to overlap the image.

  The display mode selection button 240 is a button for selecting an observation method. By tapping the display mode selection button 240, switching to the observation method selected from fluorescence observation (red, blue, green), phase difference observation, differential interference observation, and bright field observation is electrically performed. In the microscope system 200, a plurality of observation methods can be selected. In the screen 231 shown in FIG. 6, phase difference observation and fluorescence observation for blue and green fluorescence are selected. When a plurality of observation methods are selected as shown in FIG. 6, switching to each of the plurality of observation methods is performed in order, and image data is generated by each observation method. Images obtained by a plurality of observation methods are displayed in a different color on the frame window 232 and the finder window 236.

  A movement button 233 is provided around the frame window 232. When the movement button 233 is tapped, the frame area (display target area) moves in a direction corresponding to the movement button 233. Specifically, it moves continuously in steps smaller than the size of the frame area (for example, in units of one pixel). The movement of the frame area may be performed by a flick operation performed on the frame window 232.

  By dragging the zoom slider 239 provided below the finder window 236, the reduction display magnification of the image displayed in the finder window 236 is changed. The reduction display magnification may be changed by a pinch-in / pinch-out operation performed on the finder window 236.

  When the condition setting button 234 is tapped, a condition setting screen including the tables shown in FIGS. 7A to 7C is popped up on the screen 231. FIG. 7A shows a table T1 related to settings during preparatory observation such as alignment, FIG. 7B shows a table T2 related to settings during main observation, and FIG. 7C shows a table T3 related to other settings. . Note that the black circle S1 shown in FIGS. 7A and 7B indicates the observation method being selected, and the white circle S2 indicates the non-selection observation method.

  In the condition setting screen, as shown in FIGS. 7A and 7B, for each observation method, the intensity of illumination light, the exposure time of the image sensor, the gain, the binning, and the like are set for each of the preparatory observation and the main observation. . By setting the gain and binning, a bright image can be obtained with a small amount of excitation light. For this reason, these settings in the preparatory observation are particularly useful.

  In the condition setting screen, as shown in FIG. 7C, the time until the image of the imaged area A is read again, the necessity of the smoothing process when generating the combined image displayed in the finder window 236, and the upper limit of the moving speed of the stage 118 Etc. are also set. The setting of the time until recovery is particularly useful when observing the specimen S with a large temporal change or movement. Moreover, the acceleration applied to the sample S can be limited by setting the upper limit of the moving speed.

  When the clear button 238 is tapped for a long time, the image data of the imaged area A is deleted, and the display of the frame window 232 and the viewfinder window 236 is updated. Thereafter, the image of the frame area is picked up again and displayed on the frame window 232 and the viewfinder window 236.

  The imaging button 235 is a button that is tapped when starting the main observation. When the image capture button 235 is tapped, the preparatory observation is completed, and an image of the frame area is captured regardless of whether or not an unimaged area is included in the frame area. The generated image data is stored in the storage device 123 of the control device 220 in a nonvolatile manner.

  Also with the microscope system 200 configured as described above, similarly to the microscope system 100 according to the first embodiment, it is possible to significantly suppress damage to the specimen S during the preparatory observation for observation. For this reason, it is possible to align the specimen while observing a bright image without excessively suppressing the intensity of the excitation light irradiated during the preparatory observation. In particular, according to the microscope system 200, the same effect as that of the microscope system 100 according to the first embodiment can be obtained by an observation method other than the fluorescence observation.

  Although not shown in FIG. 5, the microscope system 200 may include an autofocus device. Thereby, the temporal change of the focal position with respect to the sample S in the Z direction can be prevented. The microscope system 200 may further include an input device such as a mouse, and various operations may be performed on the GUI screen illustrated in FIG. 6 using the input device.

  FIG. 8 is a diagram illustrating a configuration of the microscope system 300 according to the present embodiment. The microscope system 300 includes a microscope apparatus main body 400, a control device 320, a monitor 130, and an input device 340. In FIG. 8, the electrical connection relationship between the control device 320 and the components in the microscope apparatus main body 400 is not shown.

  Similarly to the microscope apparatus main body 210 according to the second embodiment, the microscope apparatus main body 400 executes means for executing the fluorescence observation method and other observation methods (phase difference observation, differential interference observation, bright field observation). Means. However, the microscope apparatus body 400 is different in that the means for executing the fluorescence observation method is configured as a confocal laser scanning microscope.

  Specifically, the microscope apparatus main body 400 includes a laser combiner 401, a single-mode optical fiber 402, a collimator 403, and a plurality of dichroic mirrors arranged on the optical path as means for executing the fluorescence observation method. A turret 404 that switches between the dichroic mirrors, a motor 405 that drives the turret 404, a galvano mirror 406 that scans the specimen S, a drive device 407 that drives the galvano mirror 406, a pupil relay lens 408, and an optical path. A removable mirror 409, an objective lens 410, a stage 411 on which the specimen S is arranged, a turret 412 for switching a barrier filter arranged on the optical path among a plurality of barrier filters, and a motor 413 for rotating the turret 412 And a lens 414, Focal aperture is provided with a photomultiplier tube (PMT) 415 which is disposed immediately in front of the light receiving surface.

  In the microscope apparatus main body 400, image data of a fluorescence image (confocal image) is generated by the control device 320 based on a signal from the PMT 415 and information on the scanning position of the galvano mirror 406. That is, in the microscope system 300, the photodetector (PMT 415), the scanner (galvano mirror 406, driving device 407), and the control device 320 function as an imaging device.

  Further, the microscope apparatus main body 400 includes, as means for executing another observation method, a light source 416 that is a halogen lamp, a shutter 417, a lens 418, and an optical element disposed on the optical path. A turret 419 for switching between a ring slit 419a and a DIC prism 419b, a motor 420 for rotating the turret 419, a condenser lens 421, an imaging lens 422, and a CCD camera 423 having a two-dimensional image sensor. ing.

  The control device 320 is a device that controls the microscope system 300, and includes an imaging control unit 320a and a display control unit 320b. The control device 320 has the same hardware configuration as that of the control device 120 according to the first embodiment.

  Also with the microscope system 300, the same effects as those of the microscope system 100 according to the first embodiment and the microscope system 200 according to the second embodiment can be obtained. In the microscope system 300, the imaging control unit 320a can arbitrarily set a scanning range (that is, an imaging target region) by controlling the driving device 407 when imaging a fluorescent image. For this reason, it is possible to determine only the non-imaging area included in the frame area (display target area) designated by the input device 340 (operation stick) as the imaging target area. Thereby, the irradiation range of excitation light can be suppressed as compared with the microscope system 100 and the microscope system 200. Therefore, according to the microscope system 300, it is possible to further suppress damage to the specimen S during preparation observation for observation.

  In the microscope system 300, the imaging control unit 320a may set a spare area C having a predetermined size adjacent to the frame area F outside the frame area F, as shown in FIG. In this case, the imaging control unit 320a determines that imaging is necessary even in the case where the preliminary area C includes an area that has not been captured in step S3 illustrated in FIG. Hereinafter, an area that has not yet been imaged included in the spare area C is referred to as a second unimaged area.

  Furthermore, when the second unimaged area is included in the spare area C, the imaging control unit 320a includes the second unimaged area and does not include the imaged area A and the imaging target area B in step S4. The region is determined as the second imaging target region D. FIG. 10 shows an example in which the second unimaged area itself is determined as the second imaging target area D.

  In step S5 and step S6, the imaging control unit 320a irradiates the second imaging target region D with the excitation light in addition to the imaging target region B, and in addition to the imaging target region B, the second imaging target region. An image of D is taken. In step S5 and step S6, the imaging target region B and the second imaging target region D may be separately irradiated with excitation light to capture each image separately. Alternatively, these regions may be irradiated with excitation light at the same time, and respective images may be captured simultaneously (that is, through a series of scans). The process in step S7 is the same as in the first and second embodiments.

  The predetermined size related to the spare area C may be a predetermined size such as a size corresponding to five pixels of the CCD camera 423, for example. The predetermined size may be set dynamically. For example, the higher the movement speed of the stage 411, that is, the higher the movement of the frame area from the frame area selected before the current frame area (display target area) to the current frame area, the larger the setting. It may be made to depend on the moving speed.

  As described above, by providing the spare area C, the microscope system 300 can capture an image of an area that is likely to be set as a frame area in the future. For this reason, the delay of the image display of a frame area | region can be suppressed.

  FIG. 11 is a diagram illustrating a configuration of the microscope system 500 according to the present embodiment. The microscope system 500 includes a microscope apparatus main body 510, a control apparatus 520, and a monitor 230. In FIG. 11, illustration of the electrical connection relationship between the control device 520 and the components in the microscope apparatus main body 510 is omitted.

  The microscope apparatus body 510 includes an illumination mask 511 and a driving device 512 that drives the illumination mask 511 instead of the shutter 113, a point that includes a light source 111 a instead of the light source 111, and instead of the CCD camera 117. The microscope system 200 is the same as the microscope system 200 according to the second embodiment except that a CCD camera 117a is provided.

  The illumination mask 511 is a mask that changes the irradiation range of the excitation light on the specimen S by changing the shape of the opening through which light passes. The illumination mask 511 is, for example, a spatial light modulator (SLM) such as LCOS (trademark) or DMD (trademark). In addition, a diaphragm (an illumination field diaphragm) that can arbitrarily change the shape of the opening may be used.

  The light source 111a is a laser light source capable of emitting pulses, for example. The light source 111a is controlled so as to emit pulses in accordance with the exposure period of the CCD camera 117a.

  The CCD camera 117a is an imaging device including a two-dimensional image sensor, and this is the same as the CCD camera 117 according to the second embodiment. However, in the microscope system 500, as shown in FIG. 12, the frame region F is positioned inside the region on the specimen surface (image sensor region P) where the image of the image sensor of the CCD camera 117a is formed. Is set. That is, the frame area F is set smaller than the image sensor area P.

  The control device 520 is a device that controls the microscope system 500, and includes an imaging control unit 520a and a display control unit 520b. The control device 520 has the same hardware configuration as that of the control device 120 according to the first embodiment.

  In the microscope system 500, the imaging control unit 520a controls the driving device 512 to change the shape of the opening of the illumination mask 511, whereby the illumination range of the excitation light can be arbitrarily set. This functions in substantially the same manner as the control of the scanning range in the third embodiment. In addition, the image capturing control unit 520a sets an area that protrudes from the frame area F in the image sensor area P illustrated in FIG. 12 as a spare area, so that an image of an area that is likely to be set as the future frame area F. Can be imaged in advance. Therefore, according to the microscope system 500, the same effect as the microscope system 300 according to the third embodiment can be obtained.

  Further, the microscope system 500 may include an imaging device in which a two-dimensional image sensor is surrounded by a licensor instead of the CCD camera 117. In this case, as shown in FIG. 13, a line sensor region (line sensor region L1, line sensor region L2, line sensor region L3, line sensor region L4) is provided around the image sensor region P. For this reason, the imaging control unit 520a can set the frame area F to the same size as the imaging element area P, and can set the line sensor area as a spare area. Thereby, a spare area can be provided without reducing the frame area F. When the movement of the stage 118 is limited to a constant speed, the line sensor is preferably a TDI (Time Delay Integration) sensor.

  The embodiments described above are specific examples for facilitating the understanding of the invention, and the present invention is not limited to these embodiments. The image display method, the control device, and the microscope system described in each embodiment can be variously modified and changed without departing from the spirit of the present invention defined by the claims.

  In the above-described embodiment, the example in which the image of the second imaging target region is captured in step S6 in FIG. 2 performed before the display target region display process is shown. In this case, the image may be captured after the display process of the display target area. Specifically, for example, as shown in FIG. 14, after displaying the display target area in step S <b> 7, the control apparatus may determine whether additional imaging is necessary or not based on the presence / absence of the second non-imaging area ( Step S11). Then, when it is determined that it is necessary, the second imaging target area is determined (step S12), the determined second imaging target area is irradiated with excitation light (step S13), and the second imaging target is determined. An image of the area may be taken.

  Moreover, although the example which has a handle | steering-wheel, a touchscreen, and an operating rod as an input device for selecting a display object area was shown in the Example mentioned above, an input device is a dial useful in the point that a fine adjustment is easy, and quick You may have a joystick etc. which can move. In addition, the relationship between the operation direction and the movement direction of the display target area can be arbitrarily set. For example, the display target area may be set to move in the left direction by operating in the right direction.

  In the above-described embodiment, the example in which the excitation light is irradiated in accordance with the exposure period by the opening / closing control of the shutter or the light emission control of the laser light source has been described, but may be realized by other methods. Further, the light emission of the laser light may be controlled so as to always emit light during the exposure period, or may be controlled so as to repeat the pulse light emission during the exposure period.

  Moreover, in the Example mentioned above, the example which does not irradiate excitation light to the same XY area | region in principle is shown. However, when the focal plane moves in the Z direction, the control device may perform control so as to capture an image of the frame region even when the frame region is included in the captured region. Thereby, for example, a plurality of z stack images having different Z positions can be captured, and three-dimensional image data can be generated. Even when the observation conditions (observation magnification, excitation wavelength, fluorescence wavelength, observation method) are different, the control device captures an image of the frame region even when the frame region is included in the captured region. You may control to do. Further, even under the same viewing condition, for the purpose of improving the S / N of the image of the imaged area, the image of the frame area may be imaged and the image data may be added to the imaged image data.

100, 200, 300, 500 Microscope system 110, 210, 400, 510 Microscope apparatus main body 111, 111a, 213, 416 Light source 112, 402 Optical fiber 113, 214, 417 Shutter 114, 114a, 114b Fluorescent cube 115, 410 Objective lens 116 422 Imaging lens 117, 117a, 423 CCD camera 118, 411 Stage 119, 407, 512 Driving device 120, 220, 320, 520 Control device 120a, 220a, 320a, 520a Imaging control unit 120b, 220b, 320b, 520b Display Control unit 121 CPU
122 Memory 123 Storage device 124 Reading device 125 Display IF
126 Input IF
127 Communication IF
128 Bus 129 Portable recording medium 130, 230 Monitor 140, 340 Input device 211, 216, 404, 412, 419 Turret 212, 218, 405, 413, 420 Motor 215 Illumination lens 217a, 419a Ring slit 217b, 419b DIC prism 219 , 421 Condenser lens 231 Screen 232 Frame window 233 Move button 234 Condition setting button 235 Imaging button 236 Viewfinder window 237 Frame frame 238 Clear button 239 Zoom slider 240 Display mode selection button 401 Laser combiner 403 Collimator 406 Galvano mirror 408 Pupil relay lens 409 Mirror 414, 418 Lens 415 PMT
511 Illumination mask A Imaged area A1, A2, A3, A4 Imaged unit area B Imaging object area B1, B2 Imaging object unit area F Frame area C Preliminary area D Second imaging object area P Imaging element area L1, L2, L3, L4 Line sensor area T1, T2, T3 Table S Sample

Claims (9)

A method of displaying an image of a biological specimen,
When the display target region selected from the region of the biological specimen as the region to be displayed includes the already captured region and the uncaptured region that has not yet been captured, the uncaptured region is included, and Illuminating an imaging target area that does not include an imaged area, and capturing an image of the imaging target area,
An image in which the image of the display target area including the imaged area and the imaged imaging target area is displayed based on the image data of the imaged area and the image data of the imaging target area. Display method. The image display method according to claim 1, further comprising:
When a second unimaged area that has not yet been imaged is included in a spare area of a predetermined size that is adjacent to the display object area, the imaged area and the imaged object area include the second unimaged area, An image display method, comprising: illuminating illumination light onto a second imaging target region that is not included, and capturing an image of the second imaging target region. The image display method according to claim 2,
The image display method according to claim 1, wherein the predetermined size depends on a moving speed of the display target area from a display target area selected before the current display target area to the current display target area. The image display method according to claim 1,
The movement of the display target area from the display target area selected before the current display target area to the current display target area is performed in steps smaller than the size of the display target area. Image display method. A control device for a microscope system including an imaging device and a display device,
When the display target region selected from the region of the biological specimen as the region to be displayed includes the imaged region that has already been imaged by the imaging device and the unimaged region that has not yet been imaged by the imaging device, An imaging control unit that illuminates an imaging target region that includes an unimaged region and does not include the captured region, and causes the imaging device to capture an image of the imaging target region;
Based on the image data of the imaged area and the image data of the imaging target area, which are data of the image captured by the imaging apparatus, the captured area and the imaging target area imaged on the display device And a display control unit that displays an image of the display target area including the control device. The control device according to claim 5,
The imaging control unit includes the second unimaged area and the imaged area when a second unimaged area that has not yet been imaged is included in a spare area of a predetermined size adjacent to the display target area. And a second imaging target area that does not include the imaging target area, and causes the imaging apparatus to capture an image of the second imaging target area. The control device according to claim 6,
The control apparatus according to claim 1, wherein the predetermined size depends on a moving speed of the display target area from a display target area selected before the current display target area to the current display target area. The image display method according to claim 5,
The movement of the display target area from the display target area selected before the current display target area to the current display target area is performed in steps smaller than the size of the display target area. Control device. A microscope system comprising the control device according to any one of claims 5 to 8.