Microscope system

Abstract

A microscope system 1 is provided with a microscope 100 and a control device 200 that controls the microscope 100. The microscope 100 comprises: a gel immersion objective lens 130 with a gel attached to the distal end; and a stage 110 that moves a sample in a direction orthogonal to the optical axis. The control device 200 restricts the movement of the stage 110 within a predetermined range, in a contact state in which the gel is in contact with the sample or a sample holder that accommodates the sample.

Description

Microscope system

[0001] The disclosure of this specification relates to a microscope system.

[0002] An objective lens for a microscope (hereinafter referred to as a gel immersion objective lens) having a gel attached to the tip of the lens as a substitute for an immersion liquid such as water or oil is superior in maintainability and workability compared to an immersion objective lens using an immersion liquid, and is expected to improve the efficiency of microscope observation. Such a gel immersion objective lens is described in, for example, Patent Document 1.

[0003] Japanese Patent Application Laid-Open No. 2022-071821

[0004] Observation using a gel immersion objective lens has the above-mentioned advantages compared to observation using an immersion objective lens, but has problems specific to gel immersion objective lenses that do not occur in immersion objective lenses. For example, when the observation position is greatly changed while the gel remains in contact with the sample or the sample holder, the field of view is missing, which is an example of such a problem.

[0005] Based on the above circumstances, an object according to one aspect of the present invention is to provide a technique for dealing with missing fields of view in observation using a gel immersion objective lens.

[0006] A microscope system according to one aspect of the present invention includes a microscope including a gel immersion objective lens having a gel attached to the tip, and a stage that moves a sample in a direction orthogonal to the optical axis, and a control device that controls the microscope. The control device restricts the movement of the stage within a predetermined range in a contact state where the gel is in contact with the sample or a sample holder that houses the sample.

[0007] According to the above aspect, it is possible to deal with missing fields of view in observation using a gel immersion objective lens.

[0008] This is a diagram illustrating the configuration of a microscope system according to one embodiment. This is a diagram illustrating the configuration of a gel immersion objective lens. This is a diagram illustrating a map image. This is an example of a table showing the specifications of the objective lens. This is an example of a table showing the settings for each mounting hole of the revolving nosepiece. This is a diagram explaining the focusing operation. This is a diagram explaining the bubble removal operation. This is a diagram explaining the state of the gel when no field of view defects occur. This is a diagram explaining the state of the gel when field of view defects occur. This is a diagram showing the relationship between the field of view and the XY movable range. This is an example of displaying a map image showing the XY movable range. This is a flowchart of the processing according to the first embodiment. This is a flowchart of an example of stage movement processing. This is a flowchart of an example of objective lens return processing. This is a flowchart of an example of bubble removal processing. This is a diagram illustrating the history of bubble detection processing results. This is a flowchart of an example of notification judgment processing. This is a diagram explaining the retraction and return operation of the objective lens when field of view movement is instructed on the application window. This is an example of displaying the map image before field of view movement when field of view movement is instructed on the application window. This is an example of displaying the map image after field of view movement when field of view movement is instructed on the application window. This is another example of how the map image will be displayed after the field of view has been moved, when the field of view movement is instructed on the application window. This is a diagram illustrating the retraction and return operation of the objective lens when the field of view movement is instructed on the JOG controller. This is an example of how the map image will be displayed just before the objective lens retracts when the field of view movement is instructed on the JOG controller. This is an example of how the map image will be displayed immediately after the objective lens returns to its original position when the field of view movement is instructed on the JOG controller. This is a flowchart of the process according to the second embodiment. This is a flowchart of the process according to the third embodiment. This is an example of how the map image will be displayed when generating a stitched image. This is an example of how the map image will be displayed when generating a stitched image. This is an example of how the map image will be displayed when generating a stitched image. This is a flowchart of the process according to the fourth embodiment. This is a flowchart of the process for determining the shooting order. This is a diagram illustrating the size of the continuous shooting range. This is a diagram illustrating the minimum number of retractions. This is a diagram illustrating the ground contact point and the shooting point.This diagram illustrates the order of movement of the grounding point. This diagram illustrates the order of movement of the shooting point for each continuous shooting range. This diagram illustrates the order of movement of the shooting point within the target range. This is a flowchart of the process according to the fifth embodiment. This is a flowchart of the process according to the sixth embodiment. This diagram illustrates the hardware configuration of a computer for realizing the control device.

[0009] Figure 1 is a diagram illustrating the configuration of a microscope system according to one embodiment. Figure 2 is a diagram illustrating the configuration of a gel immersion objective lens. Figure 3 is a diagram illustrating a map image. The microscope system 1 will be described below with reference to Figures 1 to 3.

[0010] The microscope system 1 shown in Figure 1 comprises a microscope 100 having a gel immersion objective lens 130 and a control device 200 for controlling the microscope 100. As shown in Figure 1, the microscope system 1 may also include a plurality of drive controllers (drive controller 301, drive controller 302) that control each motorized part of the microscope 100 according to instructions from the control device 200, and may further include a display device 401, an input device 402, and a jog controller 403 connected to the control device 200.

[0011] The microscope 100 is a magnifying observation device that allows observation of an object 10 at a magnified level, and includes a stage 110 on which the object 10 is placed, and a revolving nosepiece 120 to which multiple objective lenses, including a gel immersion objective lens 130, can be attached. The microscope 100 may further include, as shown in Figure 1, a light source unit 140 for transmitted illumination, a light source unit 150 for reflected illumination, an eyepiece 160 for visual observation, and an imaging device 170 for imaging. The imaging device 170 is, for example, a digital camera equipped with a CMOS image sensor, a CCD image sensor, etc., and acquires an image of the sample 11 based on light from the sample 11 (see Figure 2) that constitutes the object 10, which is incident on the sample 11 through the gel immersion objective lens 130.

[0012] The stage 110 and the revolving nosepiece 120 are both motorized units whose operation is controlled by the control device 200. The stage 110 is controlled by the control device 200 via the drive controller 301. The revolving nosepiece 120 is controlled by the control device 200 via the drive controller 302. The light source unit 140, the light source unit 150, and the imaging device 170 may also be controlled by the control device 200, and the control device 200 may perform light emission control and imaging control by controlling these units.

[0013] The microscope system 1 shown in Figure 1 supports both visual observation, where the user looks through the eyepiece 160 to observe the object 10, and image acquisition, where the imaging device 170 captures images of the object 10. The image of the object 10 acquired by the imaging device 170 is output to the control device 200. The control device 200 displays the image of the object 10 acquired by the imaging device 170 on the display device 401, allowing the user to observe the object 10 on the display device 401. Alternatively, the user may directly observe the object 10 by looking through the eyepiece 160.

[0014] The object to be observed 10, placed on the stage 110, includes a sample 11 and a sample holder 12 for containing the sample 11, as shown in Figure 2. The sample 11 is not particularly limited, but for example, it could be cultured cells. The sample holder 12 is not particularly limited, but for example, it could be a glass-bottom dish. The stage 110 is an electrically powered stage that moves in the XY direction perpendicular to the optical axis of the objective lens used for observation. The control device 200 controls the rotation of the motor 181 of the stage 110 via the drive controller 301 to move the stage 110 in the XY direction, thereby changing the position of the sample 11 relative to the objective lens in the XY direction and thus moving the field of view of the microscope 100.

[0015] The revolving nosepiece 120 is an electric revolving nosepiece with multiple mounting holes for attaching multiple objective lenses. The revolving nosepiece 120 is a switching unit that switches the objective lens used for observation, and by rotating around its axis, it switches the objective lens positioned on the observation optical path from among the multiple objective lenses attached to the multiple mounting holes. The control device 200 controls the rotation of the motor 182 of the revolving nosepiece 120 via the drive controller 302 to rotate the revolving nosepiece 120 and insert the objective lens to be used for observation into the optical path, thereby changing the observation magnification of the microscope 100.

[0016] Furthermore, the revolving nosepiece 120 also functions as a focusing device, changing the distance in the direction of the optical axis between the position of the objective lens used for observation and the stage 110. The control device 200 controls the rotation of the motor 183 of the revolving nosepiece 120 via the drive controller 302, moving the revolving nosepiece 120 in the Z direction, thereby changing the position of the sample 11 relative to the position of the objective lens in the Z direction, and thus moving the focus position of the microscope 100. Alternatively, the stage 110 may function as a focusing device instead of the revolving nosepiece 120. In this case, the distance in the direction of the optical axis between the position of the objective lens used for observation and the stage 110 may be changed by moving the stage 110 in the Z direction instead of the revolving nosepiece 120.

[0017] The revolving nosepiece 120 is equipped with multiple objective lenses, including a gel immersion objective lens 130. As shown in Figure 2, the gel immersion objective lens 130 is an objective lens in which a gel 134 is attached to the tip 131a of the objective lens body 131, and is used for observation with the gel 134 in contact with the object to be observed 10. More specifically, the gel immersion objective lens 130 includes an objective lens body 131 and an attachment 132 attached to the tip 131a of the objective lens body 131. The attachment 132 further includes a frame 133 fixed to the tip 131a and a gel 134 held by the frame 133. The gel immersion objective lens 130 is configured such that the attachment 132 is attached to the tip 131a, so that the gel 134 is in gapless contact with the tip lens 131b exposed from the tip 131a. In this example, the microscope 100 is an inverted microscope, and observation is performed with the gel 134 of the gel immersion objective lens 130 in contact with the bottom surface of the sample holder 12. The attachment 132 of the gel immersion objective lens 130 is detachable, and if the gel 134 deteriorates, the gel 134 can be replaced by removing and replacing the attachment 132.

[0018] The control device 200 is, for example, a computer including a processor and memory. The control device 200 controls the operation of the microscope 100 by having the processor execute a program of control software 210 stored in memory. Users of the microscope system 1 may input commands to the control device 200 by operating the input device 402 or Jog controller 403 while viewing the application screen displayed on the display device 401, and the control device 200 may control the microscope 100 according to the commands input by the user.

[0019] The microscope system 1 also has a function to display a map image 501. The map image 501 is displayed, for example, in a window 500 of an application screen displayed on the display device 401, as shown in Figure 3. A bounding box C indicating the current field of view may be displayed on the map image 501. The map image is an image of the object being observed 10, which is an image that captures a wider area than the field of view of the microscope 100. For example, the map image is an image that captures a wide area of ​​the object being observed 10 using an objective lens with a lower magnification than the objective lens used for observation. The map image may also be created by stitching together multiple images.

[0020] In the microscope system 1 configured as described above, when observation is performed using a gel immersion objective lens, a control specific to the gel immersion objective lens is performed, which differs from the control performed when observation is performed using a dry objective lens or a liquid immersion objective lens.

[0021] Figure 4 is an example of a table showing the specifications of the objective lens. Figure 5 is an example of a table showing the settings for each mounting hole of the revolving nosepiece. Figure 6 is a diagram illustrating the focusing operation. Figure 7 is a diagram illustrating the bubble elimination operation. Figure 8 is a diagram illustrating the state of the gel when no field of view defects occur. Figure 9 is a diagram illustrating the state of the gel when field of view defects occur. Figure 10 is a diagram showing the relationship between the field of view range and the XY movable range. Figure 11 is an example of a map image displaying the XY movable range. Hereinafter, the specific controls performed during observation using gel immersion objective lenses will be explained with reference to Figures 4 to 11.

[0022] To perform control specific to gel immersion objective lenses, the control device 200 has the necessary parameter values ​​stored in advance. For example, as shown in Figure 4, the necessary parameter values ​​may be stored in table T1 for each objective lens, in which case it is desirable that the necessary parameter values ​​be stored for more objective lenses, including at least multiple objective lenses attached to the revolving nosepiece 120.

[0023] Figure 4 shows information corresponding to the objective lens with identification ID 001 in Table T1, which stores the values ​​of parameters necessary for control specific to gel immersion objective lenses as the specifications of the objective lens. The following are specific examples of parameters necessary for control specific to gel immersion objective lenses in Table T1 shown in Figure 4: first return speed Vr1, second return speed Vr2, return speed change distance Dr, bubble elimination distance Db, allowable movement distance Da, retraction speed Ve, retraction distance De, first movement speed Vm1, second movement speed Vm2, movement speed change distance Dm, contact count threshold (specified value) THc, bubble detection rate threshold (specified value) THr, bubble detection period (specified value) THt, contact count threshold (last replacement) THcp, bubble detection rate threshold (last replacement) THrp, and bubble detection period (last replacement) THtp.

[0024] The type in table T1 shown in Figure 4 is a parameter that classifies objective lenses into dry objective lenses, liquid immersion objective lenses, and gel immersion objective lenses. The parameter values ​​necessary for control specific to gel immersion objective lenses, as described above, may be stored only when the type value is gel. Furthermore, the parameter values ​​necessary for control specific to gel immersion objective lenses, as described above, may be common to any objective lens whose type is gel (i.e., any gel immersion objective lens), or they may differ for each gel immersion objective lens.

[0025] Furthermore, the control device 200 stores settings for each mounting hole of the revolving nosepiece 120 in order to achieve appropriate control according to the objective lens used for observation. For example, as shown in Figure 5, the ID, start date and time, number of groundings, grounding threshold, bubble detection rate threshold, bubble detection period, grounding conditions, and bubble detection conditions may be set in table T2 for each mounting hole (revolving nosepiece number) of the revolving nosepiece. Of these, the number of groundings, grounding threshold, bubble detection rate threshold, bubble detection period, grounding conditions, and bubble detection conditions are settings specific to gel immersion objective lenses. These may be set only when a gel immersion objective lens is mounted in that mounting hole (i.e., when the ID of that revolving nosepiece number indicates a gel immersion objective lens).

[0026] The number of times table T2 has been grounded is incremented and updated each time the gel of the gel immersion objective lens corresponding to its mounting hole comes into contact with the object being observed 10 (i.e., grounds). The grounding count threshold for table T2 is set to either the grounding count threshold (default value) or the grounding count threshold (value at the time of the last replacement) of table T1 corresponding to the gel immersion objective lens identified by ID. The bubble detection rate threshold and bubble detection period are set similarly to the grounding count threshold. The grounding count condition for table T2 is set to enabled or disabled. When set to enabled, the control device 200 performs notification processing to recommend the user to replace the gel when, for example, the number of groundings reaches the grounding count threshold. The bubble detection rate condition for table T2 is set to enabled or disabled. When set to enabled, the control device 200 performs notification processing to recommend the user to replace the gel when, for example, the bubble detection rate within the bubble detection period reaches the bubble detection rate threshold. The notification processing will be described in detail later.

[0027] In the microscope system 1, the control device 200 uses the information stored in the tables shown in Figures 4 and 5 to perform specific controls for observation using a gel immersion objective lens. Below, as specific examples of the specific controls performed for observation using a gel immersion objective lens, the case of using a gel immersion objective lens 130 will be explained, including (1) controls to suppress the inclusion of air bubbles in the field of view, (2) controls to eliminate air bubbles that have entered the field of view, and (3) controls to address field defects.

[0028] Referring to Figure 6, (1) control to suppress the inclusion of air bubbles in the field of view will be explained. The gel used in the gel immersion objective lens 130 has a higher viscosity than immersion liquids such as water or oil, so air bubbles are likely to form between the gel and the sample holder 12 when they come into contact. Typically, air bubbles are likely to be introduced into the field of view during the focusing operation in which the gel immersion objective lens 130 is moved to the observation position by the revolving nosepiece 120, which is the focusing device. Therefore, in the microscope system 1, when the gel immersion objective lens 130 is used, the control device 200 performs control specific to the gel immersion objective lens 130 during the focusing operation which involves contact with the gel to suppress the inclusion of air bubbles.

[0029] Specifically, when moving the gel immersion objective lens 130 from a retracted position Z1 sufficiently far from the sample holder 12 towards the stage 110, the control device 200 first causes the revolving nosepiece 120 to move the gel immersion objective lens 130 towards the stage 110 at a first return speed Vr1 (first state in Figure 6). Here, the return speed refers to the speed at which the focusing device (revolving nosepiece 120) moves the gel immersion objective lens 130 toward the observation position in the Z direction, that is, the speed at which the gel immersion objective lens 130 moves toward the stage 110. Note that the first return speed Vr1 is relatively fast and is faster than the second return speed Vr2, which will be described later. Subsequently, when the gel immersion objective lens 130 reaches position Z2, where the distance in the optical axis direction between the position of the gel immersion objective lens 130 and the stage 110 becomes the return speed change distance Dr, the control device 200 reduces the return speed and causes the revolving nosepiece 120 to move the gel immersion objective lens 130 closer to the stage 110 at a second return speed Vr2, which is slower than the first return speed Vr1 (second state in Figure 6). Then, when the gel 134 comes into contact with the sample holder 12 (object to be observed 10) and the gel immersion objective lens 130 reaches the observation position Z3, the control device 200 causes the revolving nosepiece 120 to stop the gel immersion objective lens 130 (third state in Figure 6).

[0030] As described above, in the microscope system 1, the control device 200 slows down the return speed at which the revolving nosepiece 120 brings the gel immersion objective lens 130 closer to the stage 110 during the close-up period, when the distance in the optical axis direction between the position of the gel immersion objective lens 130 and the stage 110 is less than or equal to a predetermined value (return speed change distance Dr). As a result, during the focusing operation, the gel 134 contacts the sample holder 12 at a slow speed (second return speed Vr2), making it difficult for air bubbles to enter between the gel 134 and the sample holder 12, and consequently suppressing the inclusion of air bubbles in the field of view. Furthermore, during periods when there is no risk of the gel 134 contacting the sample holder 12 (outside the close-up period), the gel immersion objective lens 130 is moved at a higher speed to shorten the time required for the focusing operation. Therefore, with the microscope system 1, even when using the gel immersion objective lens 130, it is possible to perform the focusing operation in a short time while suppressing the inclusion of air bubbles. Furthermore, by bringing the gel immersion objective lens 130 into contact with the object to be observed 10 at a slow speed, the deterioration of the gel can also be suppressed.

[0031] The first return speed Vr1, the second return speed Vr2, and the return speed change distance Dr may differ for each gel immersion objective lens used, and the control device 200 may read the values ​​corresponding to the gel immersion objective lens being used from table T1 and use them. That is, the control device 200 may determine the return speed change distance Dr according to the gel immersion objective lens, and the first return speed Vr1 and the second return speed Vr2 may also be determined according to the gel immersion objective lens. More specifically, for example, when the control device 200 determines that the gel immersion objective lens has been placed in the optical path by the revolving nosepiece 120, it reads the parameter values ​​stored in table T1 in association with that gel immersion objective lens, and determines the return speed change distance Dr, etc., based on the parameter values. As a result, the return speed change distance Dr may be set to a longer value the longer the working distance (WD), for example, and the second return speed Vr2 may be set to a speed corresponding to the shape and characteristics of the gel of the gel immersion objective lens being used.

[0032] Compared to the recovery speed of a dry objective lens, the recovery speed of a gel immersion objective lens may have the following characteristics. As described above, in the microscope system 1, when using a gel immersion objective lens, control is performed to reduce the recovery speed during the close-up period when the object to be observed 10 is close. The second recovery speed Vr2 of the gel immersion objective lens, which is the speed after the speed has been reduced, may be slower than the recovery speed of a dry objective lens, and the first recovery speed Vr1 of the gel immersion objective lens, which is the speed before the speed has been reduced, may be faster than the recovery speed of a dry objective lens. This makes it possible to complete the focusing operation in the same or shorter time as when using a dry objective lens, while suppressing the inclusion of air bubbles that tend to occur when using a gel immersion objective lens.

[0033] In contrast to the focusing operation, during the retraction operation, which moves the gel immersion objective lens 130 away from the stage 110 to the retraction position Z1, the control device 200 does not need to limit the movement speed of the gel immersion objective lens 130, and may have the revolving nosepiece 120 retract the gel immersion objective lens 130 from the object of observation 10 at high speed. That is, the control device 200 may set the retraction speed Ve, which moves the gel immersion objective lens 130 away from the stage 110 by the revolving nosepiece 120 (which is the focusing device), to a high speed, and may be at least faster than the second return speed Vr2, which is the focusing speed during the close-up period.

[0034] The retraction speed Ve and the retraction distance De, which is the distance from the stage 110 to the retraction position Z1, may also differ for each gel immersion objective lens, similar to the first return speed Vr1, the second return speed Vr2, and the return speed change distance Dr. The control device 200 can read and use values ​​for the retraction speed Ve and retraction distance De from table T1 that correspond to the gel immersion objective lens being used.

[0035] Next, with reference to Figure 7, (2) Control for eliminating air bubbles that have entered the field of view will be explained. As mentioned above, when using the gel immersion objective lens 130, air bubbles tend to enter the field of view. Therefore, in the microscope system 1, when the gel immersion objective lens 130 is used, the control device 200 performs control specific to the gel immersion objective lens 130 when the gel is in contact with the object to be observed 10 (sample 11 or sample holder 12) to eliminate air bubbles that have entered the field of view.

[0036] Specifically, when the gel 134 is in contact with the object to be observed 10, such as when the gel immersion objective lens 130 is at observation position Z3 (first state in Figure 7), the control device 200 first causes the microscope 100 to perform a compression operation to compress the gel 134. The compression operation is the operation in which the focusing device (revolving nosepiece 120) brings the gel immersion objective lens 130 closer to the stage 110, compressing the gel 134. When the gel immersion objective lens 130 moves from position Z3 to position Z4 by a bubble elimination distance Db and the gel 134 is compressed (second state in Figure 7), the control device 200 then causes the microscope 100 to perform an operation in which the focusing device (revolving nosepiece 120) moves the gel immersion objective lens 130 away from the stage 110, thereby releasing the compression of the gel 134. The action to release the compression is, for example, to move the gel immersion objective lens 130 away from the stage 110 by the same bubble elimination distance Db as the compression action, but in the opposite direction to the compression action. As a result, the gel immersion objective lens 130 is positioned at the same observation position Z3 as before the compression action, as shown in the third state of Figure 7.

[0037] As described above, in the microscope system 1, the control device 200 causes the microscope 100 to perform a bubble-removal operation that includes the operation of the focusing device bringing the gel immersion objective lens 130 closer to the stage 110 to compress the gel 134 while the gel 134 is in contact with the object to be observed 10, and the operation of the focusing device moving the gel immersion objective lens 130 away from the stage 110. As a result, even if, for example, air bubbles get trapped between the gel 134 and the object to be observed 10 during the focusing operation, the compression operation performed from the contact state applies pressure to the air bubbles between the gel 134 and the object to be observed 10, pushing them out of the gel 134. Furthermore, after the compression operation, an operation is performed to release the compression of the gel 134, and the position of the gel immersion objective lens 130 is returned to the same observation position Z3 as immediately after the focusing operation, so that the object to be observed 10 can be observed clearly. Therefore, with the microscope system 1, even if air bubbles get mixed into the field of view during observation using the gel immersion objective lens 130, it is possible to eliminate the air bubbles mixed into the field of view, and the object to be observed 10 can be observed clearly without air bubbles in the field of view.

[0038] Furthermore, the control device 200 may determine whether or not air bubbles have formed between the gel 134 and the object 10 based on the image of the object 10 acquired by the imaging device 170, and if it determines that air bubbles have formed, it may cause the microscope 100 to perform the air bubble removal operation described above. This avoids the air bubble removal operation being performed when no air bubbles have formed, thus eliminating unnecessary operations and allowing the observation preparation to be completed earlier. The presence or absence of air bubbles can be detected by any known image processing method. In addition, when a user observing the object 10 through the eyepiece 160 discovers air bubbles, the control device 200 may cause the microscope 100 to perform the air bubble removal operation described above in response to a predetermined operation (for example, pressing an application button).

[0039] The bubble elimination distance Db may differ for each gel immersion objective lens, and the control device 200 can read the value corresponding to the gel immersion objective lens being used from table T1 and use it. In other words, the control device 200 may determine the bubble elimination distance Db according to the gel immersion objective lens. This allows, for example, the bubble elimination distance Db to be set to a value corresponding to the working distance (WD) of the gel immersion objective lens and the characteristics of the gel.

[0040] Finally, with reference to Figures 8 to 11, (3) Control to address field of view defects will be explained. When using the gel immersion objective lens 130, if the observation position is changed by moving the stage 110 in the XY direction while the gel 134 is in contact with the object to be observed 10, the field of view may be lost. Therefore, in the microscope system 1, when the gel immersion objective lens 130 is used, the control device 200 addresses field of view defects by limiting the movement of the stage 110 within a predetermined range while the gel 134 is in contact with the object to be observed 10 (sample 11 or sample holder 12).

[0041] The gel 134 of the gel immersion objective lens 130 is pre-designed to contact the object to be observed 10 over a wider area than the field of view of the microscope 100 using the gel immersion objective lens 130. Therefore, immediately after the gel 134 contacts the sample holder 12 due to the focusing operation, as shown in Figure 8, the optical path of the on-axis ray L1 from the center of the field of view to the tip lens 131b and the optical path of the off-axis ray L2 from the edge of the field of view to the tip lens 131b are both completely filled with the gel 134, so no field of view defects occur. The process of moving the gel immersion objective lens 130 in the Z direction so that the gel 134 contacts the object to be observed 10 is called grounding.

[0042] When the stage 110 moves in the XY direction while the gel 134 remains in contact with the object 10, the portion of the gel 134 in contact with the object 10 moves with the stage 110 in the XY direction, but the portion of the gel 134 in contact with the tip lens 131b remains in its original position, causing shear deformation in the gel 134 as shown in Figure 9. The shear-deformed gel 134 becomes asymmetrical with respect to the optical axis of the gel immersion objective lens 130. Therefore, if the amount of movement of the stage 110 is too large, even if the gel 134 was designed in advance to be in contact with the object 10 over a wider area than the field of view, a portion of the field of view will extend beyond the contact area between the gel 134 and the object 10. As a result, as shown in Figure 9, although the optical path of the on-axis ray L1 from the center of the field of view to the tip lens 131b is completely filled with gel 134, a portion of the optical path of the off-axis ray L2 from the edge of the field of view to the tip lens 131b is not filled with gel 134. In such cases, the off-axis light ray L2 is not properly imaged by the imaging device 170, resulting in a field of view defect.

[0043] The distance at which the stage 110 must be moved in the XY direction after grounding will cause a field of view defect is predetermined by the gel immersion objective lens 130. This is because the field of view defect occurs when light rays pass through areas where the gel 134 and the object to be observed 10 are not in contact, and the maximum distance Da that light rays from within the field of view can travel within the contact area between the gel 134 and the object to be observed 10 (hereinafter referred to as the allowable travel distance) is determined by the size of the contact area between the gel 134 and the object to be observed 10 and the size of the field of view. That is, as shown on the map image 501 in Figure 10, the predetermined XY movable range R1, which is the range in which no field of view defect occurs, can be calculated from the field of view (bounding box C) at grounding and the allowable travel distance Da. The XY movable range R1 roughly corresponds to the contact area between the gel 134 and the object to be observed 10.

[0044] More specifically, the center position of the XY movable range R1 can be determined according to the position of the stage 110 when it is grounded, and the size of the XY movable range R1 can be determined according to the allowable movement distance Da, that is, according to the immersion gel objective lens 130. Since the size of the XY movable range R1 does not change unless the objective lens inserted on the optical path is changed, when the control device 200 determines that the immersion gel objective lens 130 is arranged on the optical path by the revolver 120, based on the value of the allowable movement distance Da which is the value of the parameter stored in association with the immersion gel objective lens 130, the size of the XY movable range R1 may be determined. After that, each time the stage is grounded, the control device 200 may determine the XY movable range R1 based on the position of the stage 110 when newly grounded and the size of the XY movable range R1 determined in advance.

[0045] Regarding the allowable movement distance Da, for example, the values measured by experiments (experiments for checking the range without missing visual fields) performed in advance using the immersion gel objective lens are stored in the table T1 for each immersion gel objective lens. Since the size of the contact area varies depending on the compression state of the gel 134, it may also change depending on the observation depth. Therefore, it is desirable that the allowable movement distance Da for each immersion gel objective lens stored in the table T1 is the value measured under the condition where the visual field is most likely to be missing and the observation depth is the shallowest (that is, the condition where the size of the contact area is the smallest). If the size of the contact area between the gel 134 and the observation object 10 is known, the values obtained by calculation or simulation may be stored in the table T1.

[0046] Based on the above, in the microscope system 1, the control device 200 reads the permissible movement distance Da of the gel immersion objective lens 130 which is stored in advance in the table T1, and restricts the movement of the stage 110 in the contact state to within the XY movable range R1 calculated based on the read permissible movement distance Da. More specifically, when the control device 200 detects a movement instruction to move within the XY movable range R1 calculated based on the permissible movement distance Da, it moves the stage 110 to the indicated position in the XY direction while the gel 134 and the object to be observed 10 are in contact. On the other hand, when the control device 200 detects a movement instruction to move outside the XY movable range R1, it uses the revolving nosepiece 120, which is a focusing device, to separate the gel 134 and the object to be observed 10 before moving the stage 110 outside the XY movable range R1. Then, after the control device 200 moves the stage 110 outside the XY movable range R1, it brings the gel 134 and the object to be observed 10 into contact with the revolving nosepiece 120, which is the focusing device. Therefore, with the microscope system 1, it is possible to quickly change the observation range by moving the stage 110 to a range where no field of view is lost, without the need for retraction and return operations, while preventing field of view loss due to movement of the stage 110 beyond the allowable movement distance Da while maintaining contact.

[0047] As described above, the XY movable range R1 is determined based on the position of the stage 110 when it makes contact with the ground. Therefore, it is desirable for the control device 200 to calculate and determine the XY movable range R1 each time it makes contact with the ground, based on the position of the stage 110 when it makes contact with the ground. By updating the XY movable range R1 as needed, it becomes possible to correctly recognize movements that cause field of view obstruction and movements that do not, thereby avoiding the execution of unnecessary retraction and return operations.

[0048] In order to enable the user to recognize the XY movable range R1 calculated by the control device 200, the control device 200 may display the XY movable range R1 on the map image 502 displayed within the window 500 of the display device 401 so as to be distinguishable, as shown in FIG. 11. In FIG. 11, in the map image 502 showing the current visual field range (bounding box C), the XY movable range R1 is displayed as distinguishable from the range R2 by displaying the range R2 outside the XY movable range R1 in a shaded manner. However, it is only necessary for the user to be able to recognize the XY movable range R1, and the control device 200 may cause the display device 401 to display the XY movable range R1 on the map image 502 in a distinguishable manner by other methods without relying on the shaded display.

[0049] In the following embodiments, specific examples of various processes performed by the microscope system 1 in response to various commands from the user will be described.

[0050] (First Embodiment) FIG. 12 is a flowchart of the process according to this embodiment. FIG. 13 is a flowchart showing an example of the stage movement process. FIG. 14 is a flowchart showing an example of the objective lens return process. FIG. 15 is a flowchart showing an example of the bubble elimination process. FIG. 16 is a diagram illustrating the history of the bubble detection process results. FIG. 17 is a flowchart showing an example of the notification determination process. Hereinafter, referring to FIGS. 12 to 17, an example of the movement process performed by the microscope system 1 when a movement command for the stage 110 is input from the user during the observation of the observation object 10 will be described.

[0051] When a movement command is input from the user during the observation of the observation object 10, the control device 200 executes a program, and the microscope system 1 performs the process shown in FIG. 12. First, the control device 200 determines whether the objective lens inserted on the optical path is a gel immersion objective lens (step S1). The control device 200 can detect, for example, the hole number of the revolver 120 arranged on the optical path by a sensor attached to the revolver 120, and based on the ID of the objective lens stored in the table T2 corresponding to the detected hole number, determine whether the objective lens inserted on the optical path is a gel immersion objective lens.

[0052] If the control device 200 determines that the objective lens inserted in the optical path is not a gel immersion objective lens (step S1NO), it performs the stage movement process shown in Figure 13 (step S11). In this case, during the stage movement process shown in Figure 13, the control device 200 determines that the objective lens inserted in the optical path is not a gel immersion objective lens (step S21NO), and moves the stage 110 to the destination at a predetermined movement speed (step S23). The predetermined movement speed is, for example, the maximum speed of the stage 110.

[0053] When the stage movement process is completed, the control device 200 updates the map image (step S12) and terminates the movement process. In step S12, the control device 200 updates, for example, the position of the bounding box C on the map image that indicates the current field of view.

[0054] As described above, when the objective lens is not a gel immersion objective lens, the control device 200 can complete the movement process in a short time by moving the stage 110 at high speed during the movement process.

[0055] If the control device 200 determines that the objective lens inserted in the optical path is a gel immersion objective lens (step S1YES), it further determines whether the destination specified by the movement command is outside the XY movable range R1 (step S2). If the control device 200 determines that the destination is within the XY movable range R1 (step S2NO), it performs the stage movement process shown in Figure 13 (step S11). In this case, during the stage movement process shown in Figure 13, the control device 200 determines that the objective lens inserted in the optical path is a gel immersion objective lens (step S21YES), and further determines whether the position of the objective lens (Z position) is less than or equal to the movement speed change distance Dm from the stage 110 (step S22).

[0056] The movement speed change distance Dm is a parameter value pre-stored in table T1 that indicates the maximum distance at which the gel may come into contact with the object to be observed 10, and is determined for each gel immersion objective lens. In step S22, the control device 200 reads the movement speed change distance Dm corresponding to the gel immersion objective lens inserted in the optical path from table T1 and performs a determination process. During observation when a movement command is input, the gel is in contact with the object to be observed 10, so the Z position is less than or equal to the movement speed change distance Dm from the stage 110. Therefore, the control device 200 determines that the Z position is less than or equal to the movement speed change distance Dm from the stage 110 (step S22 YES), and moves the stage 110 to the destination at the second movement speed Vm2 (step S24).

[0057] The second movement speed Vm2 is a parameter value pre-stored in table T1 that indicates the movement speed at which the stage 110 moves the object 10 in the XY direction when in contact, and is determined for each gel immersion objective lens. The second movement speed Vm2 is slower than the first movement speed Vm1, which is a parameter value that indicates the movement speed at which the stage 110 moves the object 10 in the XY direction when in a non-contact state, as described later.

[0058] When the stage movement process is completed, the control device 200 updates the map image (step S12) and terminates the movement process. In step S12, the control device 200 updates, for example, the position of the bounding box C on the map image that indicates the current field of view.

[0059] As described above, when the objective lens is a gel immersion objective lens and the destination is within the XY movable range R1, the control device 200 slowly moves the stage 110 in the XY direction without retracting the gel immersion objective lens. This allows the movement process to be completed in a relatively short time without spending time retracting and returning the gel immersion objective lens, while also suppressing the deterioration of the gel caused by rapid shear deformation.

[0060] If the control device 200 determines in step S2 that the destination is outside the XY movable range R1 (step S2YES), it first controls the revolving nosepiece 120 to retract the gel immersion objective lens, and then performs the stage movement process shown in Figure 13 (step S4). That is, the stage movement process is performed in a non-contact state. In this case, during the stage movement process shown in Figure 13, the control device 200 determines that the objective lens inserted in the optical path is a gel immersion objective lens (step S21YES). The control device 200 further determines that the Z position exceeds the movement speed change distance Dm from the stage 110 (step S22NO), and moves the stage 110 to the destination at a first movement speed Vm1 (step S25). The first movement speed is faster than the second movement speed, and may be, for example, the maximum speed of the stage 110.

[0061] Once the stage movement process is complete, the control device 200 performs the objective lens return process shown in Figure 14 (step S5). In the objective lens return process shown in Figure 14, the control device 200 first determines whether the position of the objective lens (Z position) is less than or equal to the return speed change distance Dr from the stage 110 (step S31). If it is not less than or equal to the return speed change distance Dr (step S31 NO), that is, if it is outside the proximity period, the control device 200 determines that there is no risk of the gel coming into contact with the object to be observed 10, and moves the gel immersion objective lens in the Z direction at a first return speed Vr1 until it is less than or equal to the return speed change distance Dr (step S32). In other words, it moves the gel immersion objective lens closer to the stage 110 at high speed. Then, when the Z position is less than or equal to the return speed change distance Dr from the stage 110 (step S31 YES), the control device 200 moves the gel immersion objective lens in the Z direction at a relatively low second return speed Vr2 (step S33). In other words, the gel immersion objective lens is moved slowly towards stage 110.

[0062] Subsequently, the control device 200 repeatedly determines whether the Z position has reached the observation position Z3 (step S34). When the control device 200 determines that the gel has made contact with the object to be observed 10 and the Z position has reached the observation position Z3 (step S34 YES), it stops the stage 110, increments the number of contacts stored in the table T2 (step S35), and ends the objective lens return process.

[0063] As described above, when the objective lens is a gel immersion objective lens and the destination is outside the XY movable range R1, the control device 200 temporarily retracts the gel immersion objective lens, then rapidly moves the stage 110 in the XY direction to the destination, and then returns the gel immersion objective lens. This allows observation by freely moving the field of view of the microscope 100 while avoiding the occurrence of field of view defects. In addition, the control device 200 adjusts the return speed of the objective lens in two stages to slowly bring the gel into contact with the object to be observed 10. This makes it difficult for air bubbles to enter between the gel and the object to be observed 10, and suppresses the inclusion of air bubbles in the field of view.

[0064] Once the objective lens return process is complete, the control device 200 determines whether or not bubbles have been detected (step S6). Here, the control device 200 determines whether or not bubbles have formed between the gel and the object to be observed 10 based on the image acquired by the imaging device 170. If it is determined that bubbles have formed (step S6 YES), the control device 200 performs the bubble elimination process shown in Figure 15 (step S7). That is, the control device 200 moves the gel immersion objective lens closer to the stage 110 by the bubble elimination distance Db to compress the gel (step S41), and moves the gel immersion objective lens further away from the stage 110 by the bubble elimination distance Db to release the compression of the gel (step S42).

[0065] As described above, if air bubbles enter the field of view, the control device 200 operates to push the air bubbles out of the gel. This eliminates the air bubbles from the field of view of the microscope 100.

[0066] If no bubbles are detected in step S6, or if the bubble removal process in step S7 is completed, the control device 200 records the bubble detection result as history (step S8). In step S8, the control device 200 only needs to record at least the grounding time and whether or not bubbles were detected at grounding as history H, as shown in Figure 16, for example. History H is recorded for each hole number of the revolving nosepiece.

[0067] Subsequently, the control device 200 performs the notification determination process shown in Figure 17 (step S9). In the notification determination process shown in Figure 17, the control device 200 determines whether the bubble detection rate condition is valid or not (step S51). If the bubble detection rate condition corresponding to the hole number of the revolver located on the optical path set in table T2 is valid, the control device 200 calculates the bubble detection rate (step S52). In step S52, the control device 200 first extracts records from the history H that were recorded after the start date and time set in table T2 and during the bubble detection period set in table T2 (for example, the last two weeks). Subsequently, the control device 200 calculates the bubble detection rate as the proportion of records corresponding to bubble detection among the extracted records.

[0068] Once the bubble detection rate is calculated, the control device 200 further determines whether the grounding count condition is valid (step S53). If the grounding count condition corresponding to the hole number of the revolving nose located on the optical path set in table T2 is invalid (step S53NO), the control device 200 determines whether the bubble detection rate is above a threshold (step S55). Here, the control device 200 determines whether the bubble detection rate calculated in step S52 is above a bubble detection rate threshold corresponding to the hole number of the revolving nose located on the optical path set in table T2. If the bubble detection rate is below the threshold (step S55NO), the control device 200 terminates the notification determination process. On the other hand, if the bubble detection rate is above the threshold (step S55YES), the control device 200 notifies to recommend gel replacement (step S58).

[0069] If the bubble detection rate condition is valid and the grounding count condition is also valid (step S53 YES), the control device 200 determines whether both the grounding count and the bubble detection rate are above a threshold (step S54). Here, the control device 200 determines whether the bubble detection rate calculated in step S52 is above a bubble detection rate threshold corresponding to the hole number of the revolving nose located on the optical path set in table T2, and whether the number of groundings corresponding to the hole number of the revolving nose located on the optical path set in table T2 is above a grounding count threshold corresponding to the same hole number of the revolving nose. If either the grounding count or the bubble detection rate is below the threshold (step S54 NO), the control device 200 terminates the notification determination process. On the other hand, if both the grounding count and the bubble detection rate are above the threshold (step S54 YES), the control device 200 notifies the user to recommend replacing the gel (step S58).

[0070] If the bubble detection rate condition is invalid, the control device 200 further determines whether the grounding count condition is valid (step S56). If the grounding count condition corresponding to the hole number of the revolving nose located on the optical path set in table T2 is invalid (step S56 NO), the control device 200 terminates the notification determination process. On the other hand, if the grounding count condition is valid (step S56 YES), the control device 200 determines whether the number of groundings is equal to or greater than a threshold (step S57). Here, the control device 200 determines whether the number of groundings corresponding to the hole number of the revolving nose located on the optical path set in table T2 is equal to or greater than the grounding count threshold corresponding to the same hole number of the revolving nose. If the number of groundings is less than the threshold (step S57 NO), the control device 200 terminates the notification determination process. On the other hand, if the number of groundings is equal to or greater than the threshold (step S57 YES), the control device 200 notifies to recommend gel replacement (step S58).

[0071] As described above, the control device 200 recommends that the user replace the gel based on the number of times the gel, which has a strong correlation with the degree of gel degradation, has come into contact with the object being observed 10. The control device 200 also recommends that the user replace the gel based on the percentage of air bubbles that enter the field of view of the object being observed 10 when the gel, which has a strong correlation with the degree of gel degradation, comes into contact with the object being observed 10 (air bubble detection rate). Furthermore, the control device 200 recommends that the user replace the gel based on the number of times the gel has come into contact with the object being observed 10 and the percentage of air bubbles that enter the field of view of the object being observed 10 when the gel comes into contact with the object being observed 10 (air bubble detection rate). In this way, the degree of gel degradation can be appropriately communicated to the user, prompting them to replace the gel.

[0072] Once the notification determination process is complete, the control device 200 recalculates the XY movable range R1 based on the position of the stage 110 at ground contact, which was restored in step S5 (step S10), and updates the map image based on the recalculated XY movable range R1 (step S12). Here, the control device 200 updates, for example, the position of the bounding box C, which indicates the current field of view, the XY movable range R1, and the range R2, which is the area outside the XY movable range R1, on the map image.

[0073] The movement process shown in Figure 12 above is applicable whether the view movement is instructed on the application window or on the JOG controller. The differences in operation when the view movement is instructed on the application window and when the view movement is instructed on the JOG controller will be explained below.

[0074] Figure 18 illustrates the retraction and return operation of the objective lens when a field of view movement is instructed on the application window. Figures 19 to 21 are examples of map image displays when a field of view movement is instructed on the application window, with Figure 19 showing an example of the display before field of view movement, and Figures 20 and 21 showing examples of the display after field of view movement. First, the operation when a field of view movement is instructed on the application window will be explained with reference to Figures 18 to 21.

[0075] For example, if a location within range R2 outside the XY movable range R1 is specified as the destination on the map image 502 before field of view movement shown in Figure 19, the control device 200 first moves the gel immersion objective lens 130 from the observation position Z3 (first state in Figure 18) to the retracted position Z1 (second state in Figure 18) at a retraction speed Ve. Then, the control device 200 moves the stage 110 to the specified destination at a first movement speed Vm1, and after the stage 110 has moved to the destination, it returns the gel immersion objective lens 130 from the retracted position Z1 (third state in Figure 18) to the observation position Z3 (fourth state in Figure 18). At this time, by adjusting the return speed in two stages, the gel immersion objective lens 130 can be moved to the observation position in a short time while suppressing the generation of air bubbles. When the gel immersion objective lens 130 returns to its original position, the map image 502 is updated as shown in Figure 20, and the XY movable range R1 and range R2 are displayed on the map image 502 in an identifiable manner, centered on the bounding box C which indicates the field of view after movement. At this time, if the number of contacts exceeds a threshold, a message M prompting gel replacement may be displayed on the window 500 where the map image 502 is displayed, as shown in Figure 21.

[0076] Thus, when a field of view movement instruction is given on the application window, the desired destination (referred to as the final destination) is directly input by the user, allowing the microscope system 1 to move to the final destination with a single retraction and return of the gel immersion objective lens 130.

[0077] Figure 22 illustrates the retraction and return operation of the objective lens when the Jog controller 403 instructs a field of view movement. Figures 23 and 24 are examples of map image displays when the Jog controller 403 instructs a field of view movement, respectively. Figure 23 shows an example of the map image display just before the objective lens retracts, and Figure 24 shows an example of the map image display immediately after the objective lens returns to its original position. The operation when the Jog controller 403 instructs a field of view movement will be explained with reference to Figures 22 to 24.

[0078] For example, when observing the object 10 (first state in Figure 22), if the Jog controller 403 instructs a field of view movement, the control device 200 moves the stage 110 at a second movement speed Vm2 in accordance with the movement instruction from the Jog controller 403, without retracting the gel immersion objective lens 130, as long as the field of view range indicated by the bounding box C on the map image 502 is within the XY movable range R1, thereby moving the field of view (second state in Figure 22). Subsequently, as shown in Figure 23, when the bounding box C reaches the end of the XY movable range R1, the control device 200 stops the movement of the stage 110 and retracts the gel immersion objective lens 130 at a retraction speed Ve (third state in Figure 22). Once the retraction of the gel immersion objective lens 130 is complete, the control device 200 moves the stage 110 a predetermined distance in the direction instructed for field movement at a first movement speed Vm1 which is faster than the second movement speed Vm2 (fourth state in Figure 22), and then returns the gel immersion objective lens 130 at a first return speed Vr1 outside the close-proximity period, and at a second return speed Vr2 within the close-proximity period (fifth state in Figure 22). After the return, as shown in Figure 24, the control device 200 updates the map image 502 by displaying the XY movable range R1 and range R2 on the map image 502, centered on the position of the bounding box C which indicates the current field of view.

[0079] Thus, when the Jog controller 403 issues an instruction to move the field of view, the control device 200 cannot determine the final destination. Therefore, it moves the stage 110 without retracting the gel immersion objective lens 130 until the current field of view (bounding box C) reaches the end of the XY movable range R1. Once it reaches the end of the XY movable range R1, it performs a series of operations: retracting the gel immersion objective lens 130, moving the stage 110, and returning the gel immersion objective lens 130. This process is repeated until the field of view is moved to the final destination. As a result, even with manual field of view movement where the control device 200 cannot determine the final destination, the field of view can be moved efficiently while preventing field of view defects and avoiding the retraction and return of the objective lens as much as possible.

[0080] (Second Embodiment) Figure 25 is a flowchart of the process according to this embodiment. Hereinafter, with reference to Figure 25, an example of the objective lens switching process performed by the microscope system 1 when a command to switch the objective lens is input to the microscope system 1 from the user during observation of the object to be observed 10 will be described.

[0081] When the user inputs a command to switch the objective lens while observing the object 10, the control device 200 executes a program, causing the microscope system 1 to perform the processing shown in Figure 25. First, the control device 200 retracts the objective lens (step S61). Here, the control device 200 first identifies the objective lens mounted on the revolving nosepiece 120 from the ID set in table T2, and then identifies the maximum retraction distance among those objective lenses from table T1. Then, the control device 200 controls the revolving nosepiece 120, which is the focusing device, to retract the objective lens by the maximum retraction distance identified.

[0082] When the objective lens is retracted, the control device 200 rotates the revolving nosepiece 120 to insert the objective lens specified by the switching command into the optical path (step S62). Subsequently, the control device 200 determines whether the objective lens inserted into the optical path is a gel immersion objective lens or not (step S63). This process is the same as the process in step S1 of the movement process shown in Figure 12.

[0083] If the control device 200 determines that the objective lens inserted in the optical path is not a gel immersion objective lens (step S63NO), it instructs the revolving nosepiece 120 to return the objective lens inserted in the optical path to the observation position Z3 at a predetermined return speed (step S70). After that, the control device 200 updates the map image (step S71) and terminates the objective lens switching process. In step S71, the control device 200 updates, for example, the size of the bounding box C on the map image that indicates the current field of view.

[0084] If the control device 200 determines that the objective lens inserted in the optical path is a gel immersion objective lens (step S63 YES), it performs the processing from steps S64 to S69. Note that the processing from steps S65 to S69 is the same as the processing from steps S5 to S10 of the movement processing shown in Figure 12. After that, the control device 200 updates the map image based on the XY movable range R1 recalculated in step S69 (step S71). Here, the control device 200 updates, for example, the size of the bounding box C that indicates the current field of view on the map image, the XY movable range R1, and the range R2 which is the area outside the XY movable range R1.

[0085] By performing the objective lens switching process shown in Figure 25 above, the microscope system 1 can suppress the inclusion of air bubbles into the field of view when switching objective lenses, even when using a gel immersion objective lens, similar to when moving the field of view.

[0086] (Third Embodiment) Figure 26 is a flowchart of the process according to this embodiment. Figures 27 to 29 are examples of map image display during the generation of stitched images. Hereinafter, with reference to Figures 26 to 29, an example of stitched image processing performed by the microscope system 1 when a stitched image command is input to the microscope system 1 from the user during observation of the object to be observed 10 will be described.

[0087] When a user inputs a command to stitch images together while observing the object 10, the control device 200 executes a program, causing the microscope system 1 to perform the processing shown in Figure 26. First, the control device 200 determines the target range for stitching the images together (step S81). The target range for stitching the images together may be, for example, a range specified by the user on a map image, which is a wide-field image acquired using a low-magnification objective lens. In step S81, the control device 200 may determine the range specified on the map image as the target range.

[0088] Once the target area is determined, the control device 200 then determines the shooting order (step S82). Here, the control device 200 determines multiple shooting points based on the target area determined in step S81, and then determines the order in which the shooting will be performed at the multiple shooting points. For example, if it is determined that six shots in a 2x3 grid are necessary from the target area, the control device 200 may determine the shooting order so that each line is photographed using a raster scan method, as shown in Figure 27.

[0089] Once the shooting order is determined, the control device 200 determines whether the objective lens inserted in the optical path is a gel immersion objective lens or not (step S83). This process is the same as the process in step S1 of the movement process shown in Figure 12.

[0090] If the control device 200 determines that the objective lens inserted in the optical path is not a gel immersion objective lens (step S83NO), it repeats the movement of the stage 110 and the imaging device 170 taking images according to the imaging order determined in step S82 until imaging is completed at all imaging points (steps S95 to S97). Finally, the control device 200 combines the multiple images taken to generate a stitched image (step S98) and ends the stitched imaging process.

[0091] If the control device 200 determines that the objective lens inserted in the optical path is a gel immersion objective lens (step S83 YES), it determines whether all imaging points have been captured (step S84). If they have not been captured (step S84 NO), it further determines whether the next imaging point, specified by the imaging order determined in step S82, is outside the XY movable range R1 (step S85).

[0092] As shown in Figure 27, if the control device 200 determines that the next shooting point (the shooting point in the upper middle of Figure 27) is within the XY movable range R1 (step S85NO), it moves to the next shooting point using the stage movement process shown in Figure 13 (step S93), and then performs shooting with the imaging device 170 (step S94).

[0093] On the other hand, as shown in Figure 28, if the control device 200 determines that the next shooting point (the shooting point in the upper right of Figure 27) is outside the XY movable range R1 (step S85 YES), it retracts the gel immersion objective lens and then moves to the next shooting point using the stage movement process shown in Figure 13 (steps S86, S87). After that, the control device 200 performs the processes from steps S88 to S92 and then takes an image with the imaging device 170 (step S94). Note that the processes from steps S88 to S92 are the same as the processes in steps S5, S6, S7, S8, and S10 of the movement process shown in Figure 12. In this case, for example, as shown in Figure 29, the XY movable range R1 is updated to a new range centered on the current field of view.

[0094] The control device 200 repeats the above process until it is determined that all shooting points have been captured. Once it is determined that all shooting points have been captured (step S104 YES), it combines the captured images to generate a stitched image (step S98) and terminates the stitched image capture process.

[0095] By performing the bonded imaging process shown in Figure 26 above, the microscope system 1 can suppress the inclusion of air bubbles in the field of view during bonded imaging, even when using a gel immersion objective lens. Furthermore, it is possible to generate bonded images without causing field defects.

[0096] (Fourth Embodiment) Figure 30 is a flowchart of the process according to this embodiment. Figure 31 is a flowchart of the process for determining the shooting order. Figure 32 is a diagram illustrating the size of the continuous shooting range. Figure 33 is a diagram illustrating the minimum number of retractions. Figure 34 is a diagram illustrating the grounding point and the shooting point. Figure 35 is a diagram illustrating the order of movement of the grounding point. Figure 36 is a diagram illustrating the order of movement of the shooting point for each continuous shooting range. Figure 37 is a diagram illustrating the order of movement of the shooting point within the target range. Hereinafter, with reference to Figures 30 to 37, another example of the stitching imaging process performed by the microscope system 1 when a stitching imaging command is input to the microscope system 1 from the user during observation of the object to be observed 10 will be described.

[0097] When a user inputs a command for image stitching during observation of the object to be observed 10, the control device 200 executes a program, causing the microscope system 1 to perform the processing shown in Figure 30. First, the control device 200 determines the target range of the image stitching (step S101), and then determines whether the objective lens inserted in the optical path is a gel immersion objective lens or not (step S102). Note that the processing in steps S101 and S102 is the same as the processing in steps S81 and S83 of the image stitching process shown in Figure 26.

[0098] If the control device 200 determines that the objective lens inserted in the optical path is not a gel immersion objective lens (step S102NO), it first determines the shooting order (step S115), and then repeats the movement of the stage 110 and shooting with the imaging device 170 according to the determined shooting order until shooting is completed at all shooting points (steps S116 to S118). Finally, the control device 200 combines the multiple captured images to generate a stitched image (step S119), and ends the stitched image processing. Note that the processing from steps S115 to S118 is the same as the processing from steps S82 and S95 to S97 of the stitched image processing shown in Figure 26.

[0099] On the other hand, if the control device 200 determines that the objective lens inserted in the optical path is a gel immersion objective lens (step S102 YES), it performs the shooting order determination process shown in Figure 31 (step S103). The shooting order determination process shown in Figure 31 is a process that determines the shooting order so as to minimize the number of times the objective lens is retracted and returned. The shooting order determination process shown in Figure 31 will be described in detail below with reference to Figures 32 to 37.

[0100] In the shooting order determination process shown in Figure 31, the control device 200 first determines the size of the continuous shooting range (step S121). Here, the continuous shooting range refers to the shooting range per grounding when acquiring images that constitute the stitched image. In order to shoot a range exceeding the XY movable range R1, it is necessary to retract and return the objective lens, so as shown in Figure 32, the size of the continuous shooting range Rc is limited to within the size of the XY movable range R1. The grounding point Pc is a grounding point corresponding to the XY movable range R1, and corresponds to the shooting range that can be captured in one image capture at the grounding position. The size of the continuous shooting range Rc is determined by arranging the shooting ranges Rs that can be captured in one image capture at predetermined intervals within the XY movable range R1. This predetermined interval can be any interval where the shooting ranges Rs partially overlap, and may be set appropriately by the user considering the shooting efficiency. That is, in step S121, the control device 200 determines the size of the continuous shooting range Rc based on the size of the XY movable range R1 and the size of the shooting range Rs. Figure 32 shows an example where the size of the 3x3 shooting range Rs is determined to be the same as the continuous shooting range Rc.

[0101] Next, the control device 200 calculates the minimum number of ground contacts required to photograph the target area based on the size of the target area determined in step S101 and the size of the continuous shooting area determined in step S121 (step S122). The minimum number of ground contacts is the same as the number of continuous shooting areas Rc required to cover the target area Rt. In the example shown in Figure 33, the target area Rt is covered by four continuous shooting areas Rc, so the minimum number of ground contacts is four.

[0102] Once the minimum number of groundings is calculated, the control device 200 determines the grounding points and imaging points (step S123). The grounding points Pc and imaging points within the continuous imaging range Rc are determined by the arrangement of the continuous imaging range Rc relative to the target range Rt. For example, as shown in Figure 34, the control device 200 may determine the arrangement of the continuous imaging range Rc such that the same number of continuous imaging ranges Rc as the minimum number of groundings (four in this case) are contained within the target range Rt, and the target range Rt is covered by the continuous imaging range Rc. Such an arrangement is desirable because the grounding points Pc are located close to each other, and the time required to move between grounding points can be reduced.

[0103] Once the grounding points and shooting points are determined, the control device 200 determines the order in which to visit the grounding points (step S124). The control device 200 may determine the order in which to visit the grounding points in a spiral pattern, for example, as shown in Figure 35. Moving in this manner shortens the total distance traveled between multiple grounding points and reduces the time required for travel. Although Figure 35 shows an example of visiting the grounding points clockwise, the direction of visiting the grounding points can be clockwise or counterclockwise. Furthermore, if the grounding points are dispersed at multiple different distances from the center of the target range Rt, the order in which to visit the grounding points may be determined to be a spiral pattern, moving from the outside to the inside or from the inside to the outside of the target range Rt.

[0104] Once the order in which the ground contact points are visited is determined, the control device 200 determines the order in which the imaging points are visited (step S125). First, the control device 200 determines the order in which the imaging points are visited for each ground contact point. The control device 200 may, for example, determine the order in which the imaging points are visited (e.g., from 1-1 to 1-9) so that they are visited in a spiral pattern within the continuous imaging range Rc, starting from the ground contact point, as shown in Figure 36. It is desirable to determine the order so that the last imaging point visited (e.g., 1-9) is close to the next ground contact point (e.g., 2-1). Once the order in which the imaging points are visited for each ground contact point is determined, the control device 200 determines the order in which the imaging points are visited throughout the entire process, as shown in Figure 37.

[0105] The processing after the shooting order determination process in step S103 (steps S104 to S114) is the same as the processing in steps S84 to S94 of the stitched image processing shown in Figure 26. When shooting is completed at all shooting points, the control device 200 combines the multiple captured images to generate a stitched image (step S119) and terminates the stitched image processing.

[0106] By performing the bonded imaging process shown in Figure 30 above, the microscope system 1 can complete bonded imaging in a shorter time than when performing the bonded imaging process shown in Figure 26. Furthermore, even when using a gel immersion objective lens, the ability to suppress the inclusion of air bubbles during bonded imaging and to generate bonded images without field defects is the same as when performing the bonded imaging process shown in Figure 26.

[0107] (Fifth Embodiment) Figure 38 is a flowchart of the process according to this embodiment. Hereinafter, with reference to Figure 38, an example of multi-point imaging processing performed by the microscope system 1 when a multi-point imaging command is input to the microscope system 1 from the user during observation of the object to be observed 10 will be described. Multi-point imaging refers to the continuous imaging of multiple distant points within the object to be observed 10.

[0108] When a multi-point imaging command is input by the user during observation of the object to be observed 10, the control device 200 executes a program, causing the microscope system 1 to perform the processing shown in Figure 38. First, the control device 200 determines the imaging points for multi-point imaging (step S201). The imaging points for multi-point imaging may be specified by the user on a map image, which is a wide-field image acquired using a low-magnification objective lens. In step S201, the control device 200 may determine the points specified on the map image as the imaging points for multi-point imaging. The subsequent processing is the same as the image stitching process shown in Figure 30, except that image stitching is not performed at the end.

[0109] By performing the multi-point imaging process shown in Figure 38 above, the microscope system 1 can suppress the inclusion of air bubbles in the field of view during multi-point imaging, even when using a gel immersion objective lens. Furthermore, it can generate stitched images without causing field-of-view defects.

[0110] (Sixth Embodiment) Figure 39 is a flowchart of the process according to this embodiment. Hereinafter, with reference to Figure 39, an example of the stitched multi-point imaging process performed by the microscope system 1 when a stitched multi-point imaging command is input to the microscope system 1 by the user during observation of the object to be observed 10 will be described. Note that stitched multi-point imaging refers to performing stitched imaging at multiple separate points within the object to be observed 10.

[0111] When a user inputs a command for multi-point image stitching during observation of the object to be observed 10, the control device 200 executes a program, causing the microscope system 1 to perform the processing shown in Figure 39. First, the control device 200 determines multiple target areas corresponding to multiple stitched images (step S301). The processing in step S301 is the same as the processing in step S81 of the stitched image stitching process shown in Figure 26, except that there are multiple target areas to be determined.

[0112] Next, the control device 200 determines the order in which the multiple target ranges determined in step S301 move (step S302). The order of movement should be set such that the total travel distance is short, for example, and the control device 200 determines the order in which the target ranges are moved so that they move in a spiral pattern from the outside to the inside or from the inside to the outside.

[0113] Subsequently, the control device 200 temporarily retracts the objective lens (step S303), performs the movement process shown in Figure 13, and moves the stage 110 to the target range according to the movement sequence determined in step S302 (step S304).

[0114] When the stage 110 is moved to the target area, the control device 200 determines whether the objective lens inserted in the optical path is a gel immersion objective lens or not (step S305). Note that the process in step S102 is the same as the process in step S83 of the bonded imaging process shown in Figure 26.

[0115] Subsequently, the control device 200 returns the objective lens (steps S306 and S307). If the objective lens inserted in the optical path is not a gel immersion objective lens, the control device 200 returns the objective lens to the observation position Z3 at the normal return speed (step S306). On the other hand, if the objective lens inserted in the optical path is a gel immersion objective lens, the control device 200 returns the objective lens to the observation position Z3 by the objective lens return process shown in Figure 14 (step S307). The processing after the return is the same as the composite imaging process shown in Figure 30, except that the above-described process is repeated until, finally, image synthesis is performed over the entire target range (step S308 YES).

[0116] By performing the multi-point image stacking process shown in Figure 39 above, the microscope system 1 can suppress the inclusion of air bubbles in the field of view during multi-point image stacking, even when using a gel immersion objective lens. Furthermore, it can generate a stacked image without causing field-of-view defects.

[0117] Figure 40 is a diagram illustrating the hardware configuration of a computer 200a for realizing the control device 200 according to the above-described embodiment. The hardware configuration shown in Figure 40 includes, for example, a processor 201, memory 202, storage device 203, reader 204, communication interface 206, and input / output interface 207. The processor 201, memory 202, storage device 203, reader 204, communication interface 206, and input / output interface 207 are connected to each other, for example, via a bus 208.

[0118] The processor 201 executes the control processes illustrated in Figures 12, 25, 26, 30, 38, 39, etc., by reading the program stored in the storage device 203 into the memory 202 and executing it. The memory 202 is, for example, a semiconductor memory. The storage device 203 is, for example, a semiconductor memory such as a hard disk or flash memory, or an external storage device.

[0119] The reader 204 accesses the removable storage medium 205, for example, according to instructions from the processor 201. The removable storage medium 205 is implemented by, for example, a semiconductor device, a medium through which information is input / output by magnetic action, or a medium through which information is input / output by optical action. The communication interface 206 communicates with other devices, for example, according to instructions from the processor 201. The input / output interface 207 is, for example, an interface between the display device 401, the input device 402, the Jog controller 403, the drive controller 301, and the drive controller 302.

[0120] The programs executed by the processor 201 are provided to the computer in the following forms, for example: (1) pre-installed in the storage device 203; (2) provided via a removable storage medium 205; (3) provided from a server such as a program server.

[0121] Note that the computer hardware configuration for realizing the control device described with reference to Figure 40 is illustrative, and the embodiment is not limited thereto. For example, some of the above configurations may be deleted, or new configurations may be added. In another embodiment, for example, some or all of the functions of the above-described control device may be implemented as hardware such as FPGA (Field Programmable Gate Array), SoC (System-on-a-Chip), ASIC (Application Specific Integrated Circuit), and PLD (Programmable Logic Device).

[0122] The embodiments described above are provided as concrete examples to facilitate understanding of the invention, and the present invention is not limited to the embodiments described above, but should be understood as encompassing various modifications and alternative forms of the embodiments described above. For example, it will be understood that the embodiments described above can be materialized by modifying the components without departing from the spirit thereof. It will also be understood that various embodiments can be implemented by appropriately combining the multiple components disclosed in the embodiments described above. Furthermore, it will be understood by those skilled in the art that various embodiments can be implemented by deleting some components from all the components shown in the embodiments, or by adding some components to the components shown in the embodiments.

[0123] In the embodiments described above, an example was shown where the microscope 100 is an inverted microscope, but the microscope 100 may also be an upright microscope. Furthermore, although an example was shown where the microscope 100 has an eyepiece 160 and an imaging device 170, the microscope 100 only needs to support at least one of visual observation and photography.

[0124] In the embodiment described above, when the gel of the gel immersion objective lens is in contact with the object to be observed 10, an example of an operation to restrict the movement of the stage 110 to a predetermined range was shown, in which the objective lens is automatically retracted and returned when it moves beyond the predetermined range. However, the operation to restrict movement to a predetermined range is not limited to this example. For example, when a command to move the stage 110 beyond the predetermined range is input to the control device 200, the stage 110 may be stopped at the end of the predetermined range and the risk of field of view obstruction may be reported. Alternatively, instead of restricting the movement of the stage 110 to a predetermined range, the control device 200 may report the risk of field of view obstruction when the stage 110 moves outside the predetermined range.

[0125] This application is based on Japanese Patent Application No. 2024-188872, filed on October 28, 2024. All of its contents are included herein.

[0126] 1: Microscope system, 10: Object to be observed, 11: Sample, 12: Sample holder, 100: Microscope, 110: Stage, 120: Revolving nosepiece, 130: Gel immersion objective lens, 131: Objective lens body, 131a: Tip, 131b: Tip lens, 132: Attachment, 133: Frame, 134: Gel, 140, 150: Light source unit, 160: Eyepiece, 170: Imaging device, 181, 182, 183: Motor, 200: Control device, 200a: Computer, 201: Processor, 202: Memory, 203: Storage device, 204: Reader, 205: Removable storage medium, 206: Communication interface, 207: Input / output interface, 208: Bus, 210: Control software, 301, 302: Drive controller, 401: Display device, 402: Input device, 403: Jog controller, 500: Window, 501, 502: Map image, C: Bounding box, Da: Allowable travel distance, Db: Bubble elimination distance, De: Evacuation distance, Dm: Travel speed change distance, Dr: Return speed change distance, H: History, L1: On-axis ray, L2: Off-axis ray, M : Message, Pc: Ground contact point, R1: XY movable range, R2: Range, Rc: Continuous shooting range, Rs: Shooting range, Rt: Target range, T1, T2: Table, Ve: Evacuation speed, Vm1: First movement speed, Vm2: Second movement speed, Vr1: First return speed, Vr2: Second return speed, Z1: Evacuation position, Z2, Z4: Position, Z3: Observation position

Claims

1. A microscope comprising: a gel immersion objective lens with a gel attached to its tip; a stage for moving a sample in a direction perpendicular to the optical axis; and a control device for controlling the microscope, wherein the control device restricts the movement of the stage within a predetermined range when the gel is in contact with the sample or a sample holder containing the sample.

2. The microscope system according to claim 1, wherein the microscope further comprises a focusing device for changing the distance in the direction of the optical axis between the position of the gel immersion objective lens and the stage, and the control device, when it detects an instruction to move the stage out of a predetermined range, causes the focusing device to separate the gel from the sample or the sample holder before moving the stage out of the predetermined range, and causes the focusing device to bring the gel from the sample or the sample holder into contact with the stage after moving the stage out of the predetermined range.

3. A microscope system according to claim 1 or claim 2, wherein the control device determines the predetermined range each time the gel and the sample or the sample holder come into contact, based on the position of the stage when the gel and the sample or the sample holder come into contact.

4. The microscope system according to claim 3, wherein the control device determines the size of the predetermined range according to the gel immersion objective lens.

5. The microscope system according to claim 4, wherein the microscope further comprises: a plurality of objective lenses including the gel immersion objective lens; a switching unit having a plurality of mounting holes to which the plurality of objective lenses are attached, and which switches between the objective lenses arranged on the optical path from among the plurality of objective lenses, and the control device determines the size of the predetermined range based on the value of a parameter stored in association with the gel immersion objective lens when the switching unit determines that the gel immersion objective lens is arranged on the optical path.

6. A microscope system according to claim 1 or claim 2, wherein the control device displays the predetermined range on a map image displayed on the display unit, and the map image is an image of the sample that captures a wider area than the field of view of the microscope.

Images