MICROSCOPE AUXILIARY APPARATUS

A microscope auxiliary apparatus attachable to a microscope includes an object movable portion for moving an object in an optical axis direction of the microscope, first and second operating unit movable portions for respectively moving, in the optical axis direction, first and second operating units for operating the object, a movement instructing unit for instructing the object movable portion or the first or second operating unit movable portions to move, and a switching unit for switching a mode to first and second modes. The first mode moves one of the movable portions in the optical axis direction according to an instruction from the movement instructing unit. The second mode links movements of at least two of the movable portions and moves the at least two in the optical axis direction according to the instruction from the movement instructing unit.

BACKGROUND OF THE INVENTION

Field of the Invention

An aspect of embodiments of the present disclosure relates to a microscope auxiliary apparatus used while being attached to a microscope in order that an object such as a microscopic cell and a semiconductor element are observed or that a mechanical manipulation such as sorting, cutting, and moving an object is performed.

Description of the Related Art

When focus of a microscope is to be adjusted, manual focusing is usually performed by manually rotating a knob provided on the microscope and moving an objective lens in an optical axis direction while a position of an object is fixed in the optical axis direction. On the other hand, there is a method of using an object movable portion, generally called a “hollow stage”, on which an object can be placed and that can be electrically moved in the optical axis direction while an observation light path or illumination light path of the microscope is not blocked. This method makes it possible to electrically move an object in an optical axis direction without moving an objective lens in the optical axis direction, and thus enables autofocusing.

When autofocusing is to be performed, an image pickup unit is attached to the microscope and an image is acquired. Conventional techniques commonly used in digital cameras can be applied to autofocusing. For example, there are a contrast method with which autofocusing is performed by evaluating a contrast of an acquired image, an image pickup plane phase difference method with which autofocusing is performed by comparing two images of which an image pickup unit can detect a phase difference by pupil division in a predetermined direction, and the like.

In order that an operating unit such as a pipette, a probe, and tweezers for mechanically manipulating an object is moved in a plurality of axial directions, an operating unit movable portion, generally called a “micromanipulator”, is used while being attached to the microscope. The operating unit movable portion includes a combination of stages capable of moving an operating unit in a plurality of axial directions. Movements in the plurality of axial directions include at least a movement in an optical axis direction for focusing, and usually include movements in three axial directions of XYZ including movements in a planar direction orthogonal to the optical axis direction.

Japanese Patent Application Laid-Open No. 2008-233545 discloses a method of controlling a micromanipulator based on image information acquired by an image pickup unit. Yasuhisa Araki, “Technical Textbook for Assisted Reproductive Technology”, Ishiyaku Publishers, Inc. (hereinafter, referred to as “Araki”) discloses cell manipulation using micromanipulators.

However, in conventional configurations, an electric operation that moves the object placed on the object movable portion in the optical axis direction and an electric operation that moves the operating unit attached to the operating unit movable portion in the optical axis direction are not linked to each other. Therefore, in a case where the focus is to be readjusted after the object and the operating unit are focused on, the object movable portion and the operating unit movable portion are moved individually, which takes time to readjust the focus.

SUMMARY OF THE INVENTION

The present disclosure provides a microscope auxiliary apparatus that can shorten an operation time by linking movements, in the optical axis direction, of at least two movable portions including an object movable portion or an operating unit movable portion.

A microscope auxiliary apparatus according to one aspect of the embodiments of the present disclosure is attachable to a microscope. The microscope auxiliary apparatus includes an object movable portion, a first operating unit, a second operating unit movable portion, a movement instructing unit, and a switching unit. The object movable portion is configured to move an object in an optical axis direction of the microscope. The first operating unit movable portion is configured to move, in the optical axis direction, a first operating unit for operating the object. The second operating unit movable portion is configured to move, in the optical axis direction, a second operating unit for operating the object. The movement instructing unit is configured to instruct the object movable portion, the first operating unit movable portion, or the second operating unit movable portion to move in the optical axis direction. The switching unit is configured to switch a mode to a first mode and a second mode. The first mode is a mode that moves one of the object movable portion, the first operating unit movable portion, and the second operating unit movable portion in the optical axis direction according to an instruction from the movement instructing unit. The second mode is a mode that links movements of at least two of the object movable portion, the first operating unit movable portion, and the second operating unit movable portion and moves the at least two in the optical axis direction according to the instruction from the movement instructing unit.

A microscope auxiliary apparatus according to one aspect of the embodiments of the present disclosure is attachable to a microscope. The microscope auxiliary apparatus includes an operating unit and an image pickup unit. The operating unit movable portion is configured to move, in an optical axis direction of the microscope, an operating unit for operating an object. The image pickup unit is configured to acquire an observation image of the microscope. Based on the observation image, the operating unit movable portion moves in the optical axis direction by a same distance as a moving distance of an objective lens of the microscope in the optical axis direction while a movement of the operating unit movable portion is linked to a movement of the objective lens.

A microscope auxiliary apparatus according to one aspect of the embodiments of the present disclosure is attachable to a microscope. The microscope auxiliary apparatus includes an operating unit movable portion, an image pickup unit, an image pickup unit rotating unit, and an image rotating unit. The operating unit movable portion is configured to move, at least in an optical axis direction of the microscope, an operating unit for operating an object. The image pickup unit is configured to acquire two images by pupil division in a predetermined direction so that a phase difference is detected. The image pickup unit rotating unit is configured to rotate the image pickup unit relatively to the microscope. The image rotating unit is configured to generate a rotated image by rotating, by a predetermined angle, an image acquired by the image pickup unit.

DESCRIPTION OF THE EMBODIMENTS

Referring now to the drawings, a detailed description is given of embodiments according to the present disclosure.

First Embodiment

First, a description is given of a microscope system according to a first embodiment of the present disclosure.FIGS.1A and1Bare entire views of a microscope system10including a microscope (inverted microscope)100and a microscope auxiliary apparatus and illustrate a state in which the microscope auxiliary apparatus is attached to the microscope100. The microscope auxiliary apparatus includes an object movable portion1, an operating unit movable portions2L and2R, a controller3, and a console4.FIG.1Aillustrates a front view of the microscope system10, andFIG.1Billustrates a right view of the microscope system10. InFIGS.1A and1B, for clarity, some dimensions are exaggerated, some components are omitted, and some internal components are drawn with solid lines instead of dotted lines.

The microscope100is configured to cause an illumination optical system101to illuminate an object103placed on a transmissive observation tray102and to allow observation of the object103through an observation optical system104. The observation optical system104allows an enlarged image obtained through an objective lens104ato be observed with a naked eye at an eyepiece lens104bvia unillustrated other lenses or an unillustrated refractive optical system. Focus can be adjusted by finely moving the objective lens104ain an optical axis direction (vertical direction inFIGS.1A and1B). This fine movement is performed by rotating a knob105provided on the microscope100, and the rotation amount of the knob105is converted into a fine moving amount of the objective lens104avia a deceleration transmission mechanism (not illustrated).

The object movable portion1, which is a part of the microscope auxiliary apparatus, is generally called a “hollow stage”, etc., and an observation tray102on which an object103is placed on the object movable portion1. The object movable portion1includes a driving mechanism that can move the object103in the optical axis direction of the microscope100.

The operating unit movable portions2L and2R, which are part of the microscope auxiliary apparatus, are generally called “micromanipulators”, etc., and are attached to left and right sides of the microscope100. The operating unit movable portions2L and2R include driving mechanisms that can move the operating unit2La and2Ra for manipulating (operating) the object103at least in the optical axis direction of the microscope100. Normally, the operating unit movable portions2L and2R can move in three axial directions of XYZ including a planar direction orthogonal to the optical axis direction. The operating unit movable portions2L and2R may also move in directions of rotational axes of yaw, pitch, and roll. Further, a high-speed driving mechanism for coarse movement and a high-resolution driving mechanism for fine movement may be separately provided.

The controller3, which is part of the microscope auxiliary apparatus, controls the object movable portion1and the operating unit movable portions2L and2R and includes a CPU3afor controlling the entire system, and other peripheral circuits. The other peripheral circuits include a driving circuit C3bfor driving the object movable portion1and driving circuits R3cand R3dfor respectively driving the driving mechanisms of the operating unit movable portions2L and2R.

The driving circuit C3bhas a function of controlling a movement of the observation tray102, which is placed on the object movable portion1, in the optical axis direction. By moving the observation tray102in the optical axis direction, the driving circuit C3bcan align the object103placed on the observation tray102with a focus position of the objective lens104a. The driving circuits R3cand R3dhave functions of controlling movements of the operating unit movable portions2L and2R in the optical axis direction. By moving the operating units2La and2Ra in the optical axis direction, positions of tips of the operating units2La and2Ra can be aligned with the focus position of the objective lens104a.

The console4for giving instructions to the controller3is provided with several inputting units for inputting necessary instructions to the object movable portion1and the operating unit movable portions2L and2R.

With reference toFIGS.2A to2C, a description is given of functions of the console4.FIG.2Ais an explanatory diagram of functions of the console4. Dials4aL,4aC, and4aR are inputting units for adjusting the positions of the object movable portion1and the operating unit movable portions2L and2R in the optical axis direction. The dial4aL is for the left operating unit movable portion2L, the dial4aC is for the object movable portion1, and the dial4aR is for the right operating unit movable portion2R. According to rotation amounts of the dials4aL,4aC, and4aR, the respective positions of the movable portions in the optical axis direction can be moved in directions of arrows DL, DC, and DR. Thus, the dials4aL,4aC, and4aR function as a movement instructing unit that instructs the object movable portion1or the operating unit movable portion2L or2R to move in the optical axis direction of the microscope100. The dials4aL,4aC, and4aR are an example of a manual movement instructing unit for inputting moving amounts and moving directions according to the rotation amount. In this embodiment, as a manual movement instructing unit via which a moving amount and a moving direction are input, a sliding-type inputting unit, a lever-tilting-type inputting unit, a touch panel, and the like can also be applied.

Inputting units similar to the dials4aL,4aC, and4aR are to be also provided for movements of the operating unit movable portions2L and2R in directions other than the optical axis, such as XY directions.FIG.2Billustrates an example in which dials LX, LY, RX and RY are added for movements of the operating unit movable portions2L and2R in the XY directions. Alternatively, as illustrated inFIG.2C, two sets of button switches X, Y, and Z may be provided, and the button switches X, Y, and Z may be used to switch directions in which the operating unit movable portions2L and2R are moved by operations on the dials4aL and4aR. For example, when a button switch X as a changeover switch is pressed, operating the dial4aL moves the operating unit movable portion2L in the X direction. In this embodiment, an input unit such as a stick-shaped lever may be used instead of the dial. Further, a function may be provided with which magnifications of moving amounts of the object movable portion1and the operating unit movable portions2L and2R with respect to the respective rotation amounts of the dials can be freely changed. A description is omitted of the inputting units for a movement in a direction other than the optical axis direction and the unit for changing the magnifications.

InFIG.2A, motion modes can be switched by operating left and right changeover switches4bLC and4bRC. In this embodiment, the changeover switches4bLC and4bRC are a changeover switch capable of reciprocating in two directions including “ON” for enabling a “linked mode” described below and “OFF” for disabling the “linked mode”.

By disabling the linked mode, the mode is switched to a non-linked mode in which the object movable portion1and the operating unit movable portions2L and2R are independently moved according to the instructions to the dials4aL,4aC, and4aR, respectively. By enabling the linked mode, the mode is switched to the linked mode in which at least two movable portions of the object movable portion1and the operating unit movable portions2L and2R are simultaneously moved by the same moving amount according to the instructions to the dials4aL,4aC, and4aR. Thus, the changeover switches4bLC and4bRC function as a switching unit that switches the mode to the non-linked mode (first mode) and the linked mode (second mode).

FIG.3Ais an explanatory diagram of processes in this embodiment. First, with reference toFIG.3A, a description is given of the non-linked mode during manual focusing. First, the changeover switches4bLC and4bRC are all switched to a linked mode OFF side. When the dial4aC is operated, only the object movable portion1moves in the optical axis direction (DC direction inFIG.3A). Therefore, while an image is viewed, it is possible to focus on a desired portion of the object103placed on the observation tray102. At this time, the focus on the operating units2La and2Ra attached to the operating unit movable portions2L and2R remains unchanged.

Similarly, when the dials4aL and4aR are operated, only the operating units2La and2Ra respectively attached to the left and right operating unit movable portions2L and2R move in the optical axis direction (DL and DR directions inFIG.3A). Therefore, while an image is viewed, it is possible to focus on the operating units2La and2Ra. At this time, the focus on the object103remains unchanged.

Next, with reference toFIG.3B, a description is given of the linked mode during manual focusing. A description is given of an example in which movements of the left operating unit movable portion2L and the object movable portion1are linked. First, the changeover switch4bLC is switched to a linked mode ON side, and the changeover switch4bRC is switched to the linked mode OFF side. As in a state where the linked mode is disabled, when the dial4aC is operated, the object movable portion1moves in the optical axis direction, and thereby, while an image is viewed, it is possible to focus on a desired portion of the object103placed on the observation tray102. Here, in a state where the linked mode is enabled, the left operating unit movable portion2L for which the linked mode is selected also moves in the optical axis direction by the same moving amount and at the same speed as the moving amount and the speed of the object movable portion1. As a result, the focus on the operating unit2La attached to the left operating unit movable portion2L also changes similarly to the focus on the object103. On the other hand, since the operating unit2Ra attached to the right operating unit movable portion2R, for which the linked mode is not selected, does not move in the optical axis direction, the focus thereon does not change.

Similarly, by switching the changeover switch4bLC to the linked mode OFF side and the changeover switch4bRC to the linked mode ON side, movements of the right operating unit movable portion2R and object movable portion1can be linked. By switching both the changeover switches4bLC and4bRC to the linked mode ON side, movements of the three movable portions of the operating unit movable portions2L and2R and the object movable portion1can be linked.

FIG.4is a flow chart illustrating the flow of a response to the dial4aC as an example of the processes illustrated inFIGS.3A and3B. First, in step S101, the controller3determines whether or not it is in a reception state for receiving an operation on the dial4aC. If it is in the reception state, the process proceeds to step S102. On the other hand, if it is not in the reception state, this flow ends.

In step S102, the controller3determines whether or not there is an input to the dial4aC. If there is an input, the process proceeds to step S103. On the other hand, if there is no input, the process returns to step S101.

In step S103, the controller3determines whether or not the changeover switch4bLC is OFF. If the changeover switch4bLC is OFF, the process proceeds to step S104. On the other hand, if the changeover switch4bLC is ON, the process proceeds to step S105.

In step S104, since the changeover switch4bLC is OFF, the microscope auxiliary apparatus acts in the non-linked mode. At this time, the controller3moves only the object movable portion1in the optical axis direction according to the amount of input to the dial4aL, and the process returns to step S101.

In step S105, since the changeover switch4bLC is ON, the microscope auxiliary apparatus acts in the linked mode. At this time, the controller3moves the object movable portion1and the operating unit movable portion2L in the optical axis direction according to the amount of input to the dial4aL, and the process returns to step S101.

A description is given of effects of the linked mode in cell manipulation as an example. In cell manipulation, a holding pipette HP is attached to the left operating unit movable portion2L, and an injection pipette IP is attached to the right operating unit movable portion2R. The holding pipette HP sucks and holds an egg or a fertilized egg, which is the object103. The injection pipette IP injects sperm into the egg or a special cell, etc. into the fertilized egg. A detailed description thereof is given in Araki.

FIGS.5A to5Care explanatory diagrams of the effects in this embodiment.FIG.5Ais a diagram of a state immediately after the holding pipette HP and the injection pipette IP are attached. The positions of the holding pipette HP and the injection pipette IP in the optical axis direction are different, and thus the moving distances DL and DR to the focus position are different. Therefore, the linked mode is disabled, and the dials4aL and4aR are individually operated so that the holding pipette HP and the injection pipette IP individually move and their respective tips are aligned with the focus position. After that, the positions in the planar direction and tilts of the holding pipette HP and the injection pipette IP are properly adjusted.

FIG.5Bis a diagram illustrating a state after the tips of the holding pipette HP and the injection pipette IP are aligned with the focus position, and the positions in the plane direction, the tilts, and the like have been properly adjusted. Then, a petri dish, which is the observation tray102, on which a cell, which is the object103, is placed is set within a field of view of the microscope. The petri dish is set after the positions, tilts, etc. of the holding pipette HP and the injection pipette IP are adjusted because the cell has been stored in a culture chamber and the time the cell are kept out of the culture chamber is to be minimized. Here, in order that the petri dish is set, the holding pipette HP and the injection pipette IP are to be retracted. At this time, rather than moving the holding pipette HP and the injection pipette IP individually, enabling the linked mode and retracting them by a same distance at a same time can shorten the operation time. After the holding pipette HP and the injection pipette IP are retracted, the petri dish is to be set, and then the holding pipette HP and the injection pipette IP are to be returned to the focus position again. At this time as well, the operation time can be shortened by enabling the linked mode and simultaneously moving the holding pipette HP and the injection pipette IP by a same distance. This is because if both of them retract by the same distance and return by the same distance, the positions thereof are aligned with the focus position.

FIG.5Cis a diagram illustrating a state just before the injection pipette IP punctures the vicinity of the center of the cell held by the holding pipette HP. In a case where the position of the injection pipette IP in the optical axis direction is to be readjusted, only the injection pipette IP is to be moved and the positions of the others are to be maintained, so the linked mode is disabled and the dial4aR is operated.

On the other hand, in a case where the position of the cell in the optical axis direction is to be readjusted, the linked mode is enabled for the operating unit movable portion2L and the object movable portion1. The operation time can be shortened by operating the dial4aL or the dial4aC and simultaneously moving the operating unit movable portion2L and the object movable portion1. Here, in a case where only the object movable portion1is moved with the linked mode disabled, the relative positions of the cells and the holding pipette HP are changed and the holding of the cells may become unstable.

In a case where a conventional microscope auxiliary apparatus not provided with the linked mode is used, the objective lens104ais first adjusted so that the focus on the cell is readjusted. Thereafter, the position of the injection pipette IP is adjusted so that the focus on the injection pipette IP is readjusted, which takes time for the operation. On the other hand, in the case where the microscope auxiliary apparatus according to this embodiment is used, the linked mode is enabled so that movements of at least two movable portions of the object movable portion1and the operating unit movable portions2L and2R in the optical axis direction are linked. Thereby, the operation time can be shortened. The linked mode is not to be enabled all the time since there is a case where the linked mode is to be disabled, such as a case where the focus is to be adjusted immediately after the pipette is attached as illustrated inFIG.5A. For this reason, a switch for enabling and disabling the linked mode is practically required.

In this embodiment, a description is given of a switch that mechanically reciprocates as an example of switching unit, but the switching unit may be a unit such as a touch switch and a foot pedal or may be, instead of a physical switch, a software switch that switches the mode in response to sounds or the like. This point is also similarly applied to each of the following embodiments.

In this embodiment, an example is described in which two sets of switches for enabling and disabling the linked mode are provided and the object movable portion1and the left and right operating unit movable portions2L and2R are objects to be moved. However, this embodiment is not limited to this, and a switch may be further provided with which movements are linked of the left and right operating unit movable portions2L and2R. Alternatively, one switch and a movable portion selecting unit (console4) may be provided, and the movable portion selecting unit may be used for selecting portions to be simultaneously moved by the same moving amount in the linked mode from the object movable portion1and the operating unit movable portions2L and2R. As long as there are a plurality of objects to be moved, a similar configuration is possible even when the number of the objects to be moved is not three. In this embodiment, an example is described of an inverted microscope as the microscope100, but a similar configuration is possible in a case where the microscope100is a real-image microscope with which an object is observed from an upper part or is a microscope of another type. These points are similarly applied to each of the following examples.

As described above, the microscope auxiliary apparatus attachable to the microscope100according to this embodiment includes the object movable portion1, the first operating unit movable portion (operating unit movable portion2L), and the second operating unit movable portion (operating unit movable portion2R). The microscope auxiliary apparatus includes the movement instructing unit (dials4aL,4aC, and4aR) and the switching unit (changeover switches4bLC and4bRC). The object movable portion moves the object103in the optical axis direction of the microscope. The first operating unit movable portion moves, in the optical axis direction, the first operating unit (operating unit2La) for manipulating the object. The second operating unit movable portion moves, in the optical axis direction, the second operating unit (operating unit2Ra) for manipulating the object. The movement instructing unit instructs the object movable portion, the first operating unit movable portion, or the second operating unit movable portion to move in the optical axis direction. The switching unit switches the mode to the first mode and to the second mode. The first mode is a mode (non-linked mode) that moves one of the object movable portion, the first operating unit movable portion, and the second operating unit movable portion in the optical axis direction according to the instruction from the movement instructing unit. The second mode is a mode (linked mode) that links movements of at least two of the object movable portion, the first operating unit movable portion, and the second operating unit movable portion in the optical axis direction and moves the at least two according to the instruction from the movement instructing unit. That is, the second mode is a mode that simultaneously moves at least two of the object movable portion, the first operating unit movable portion, and the second operating unit movable portion by a same moving amount in the optical axis direction according to the instruction from the movement instructing unit.

According to this embodiment, a microscope auxiliary apparatus can be provided that can shorten an operation time by linking movements of at least two movable portions including an object movable portion or an operating unit movable portion in an optical axis direction.

Second Embodiment

Next, a description is given of a microscope system according to a second embodiment of the present disclosure.FIGS.6A and6Bare entire views of a microscope system10aincluding a microscope (inverted microscope)100and a microscope auxiliary apparatus and illustrate a state in which the microscope auxiliary apparatus is attached to the microscope100. The microscope auxiliary apparatus includes an object movable portion1, an operating unit movable portions2L and2R, a display unit (display part)12, a controller13, and a console14.FIG.6Aillustrates a front view of the microscope system10a, andFIG.6Billustrates a right view of the microscope system10a. InFIGS.6A and6B, for clarity, some dimensions are exaggerated, some components are omitted, and some internal components are drawn with solid lines instead of dotted lines.

With reference toFIGS.6A and6B, a description is given of functions added in this embodiment to the functions described in the first embodiment. An unillustrated prism is added to the observation optical system, and an enlarged image can be formed on the image pickup unit11. The image pickup unit11may be a dedicated device or may have a configuration such that a general digital camera is attached via a predetermined mount adapter. The image pickup unit11can acquire an image for performing autofocus of a “contrast method” or an “image pickup plane phase difference method” like a general digital camera.

A controller13includes an image processing circuit13ethat performs processing on an image acquired by the image pickup unit11, in addition to a CPU13a, a driving circuit C13b, a driving circuit L13c, and a driving circuit R13d. The image processing circuit13ecan perform various image processing on the image acquired by the image pickup unit11and output the processed image to an external display unit12as an image. Further, using the image acquired by the image pickup unit11, the image processing circuit13ecan output, to the CPU13a, image information for performing autofocus of the “contrast method” or the “image pickup plane phase difference method”.

Button switches14cL,14cC, and14cR of the console14are inputting units capable of detecting that they are pressed and of triggering a predetermined process. In this embodiment, the button switches14cL,14cC, and14cR are assigned a function of triggering an autofocusing process. When the button switch14cC is pressed, the object movable portion1is moved in the optical axis direction and the autofocusing process is started. Moving the object movable portion1adjusts focus on the object103placed on the observation tray102. As described above, conventional techniques of various methods can be used for automatically determining that the object is focused on. A method of recognizing an area in the vicinity of the object103may be a method of automatically determining the area by image recognition, or a method of manually specifying the area in advance by using the display unit12.

Similarly, the button switches14cL and14cR are assigned functions of triggering autofocusing processes for the operating units2La and2Ra attached to the operating unit movable portions2L and2R. The autofocusing method and the method of recognizing the areas in the vicinity of the operating units2La and2Ra are similar to the methods used when the button switch14cC is used.

In this embodiment, as in the first embodiment, dials14aL,14aC, and14aR can be used when an instruction for manual focusing is given, and button switches14cL,14cC, and14cR added in this embodiment can be used when an instruction for autofocusing is given. Thus, in this embodiment, the dials4aL,4aC, and4aR and the button switches14cL,14cC, and14cR function as a movement instructing unit that instructs the object movable portion1or the operating unit movable portion2L or2R to move in the optical axis direction of the microscope100. The button switches14cL,14cC, and14cR are an example of an automatic movement instructing unit that, when being pressed, starts a movement based on a predetermined input signal (with a predetermined input signal as a trigger) and automatically stops when a predetermined condition is satisfied. Alternatively, a touch panel can also be applied as an automatic movement instructing unit that automatically stops when a predetermined condition is satisfied.

FIGS.7A and7Bare explanatory diagrams of processes in this embodiment. First, with reference toFIG.7A, a description is given of the non-linked mode during autofocusing. First, the changeover switches4bLC and4bRC are all switched to the linked mode OFF side. When the button switch14cC is pressed, only the object movable portion1moves in the optical axis direction (DC direction inFIG.7A). Thereby, it is possible to autofocus onto the object103placed on the observation tray102. At this time, the focus on the operating units2La and2Ra attached to the operating unit movable portions2L and2R remains unchanged. Similarly, when the button switches14cL and14cR are pressed, only the operating units2La and2Ra attached to the operating unit movable portions2L and2R move in the optical axis direction (DL direction and DR direction inFIG.7A), and it is possible to autofocus onto the operating units2La and2Ra. At this time, the focus on the object103remains unchanged.

Next, with reference toFIG.7B, a description is given of the linked mode during autofocusing. A description is given of an example in which a movements of the left operating unit movable portion2L and the object movable portion1are linked. First, the changeover switch4bLC is switched to the linked mode ON side, and the changeover switch4bRC is switched to the linked mode OFF side. As in a case where the linked mode is disabled, when the button switch14cC is pressed, the object movable portion1moves in the optical axis direction so that the object103placed on the observation tray102is focused on. Here, in a state where the linked mode is enabled, the operating unit movable portion2L for which the linked mode is selected also moves in the optical axis direction by the same moving amount at the same speed as the moving amount and the speed of the object movable portion1. As a result, the focus on the operating unit2La attached to the operating unit movable portion2L changes as in the focus on the object103. On the other hand, since the operating unit2Ra attached to the operating unit movable portion2R for which the linked mode is not selected does not move in the optical axis direction, the focus does not change.

Similarly, by switching the changeover switch4bLC to the linked mode OFF side and the changeover switch44bRC to the linked mode ON side, movements of the operating unit movable portion2R and the object movable portion1can be linked. Also, by switching both the changeover switches4bLC and4bRC to the linked mode ON side, movements of three of the operating unit movable portions2L and2R and the object movable portion1can be linked.

FIG.8is a flow chart illustrating the flow of a response to the button switch14cC as an example of the processes illustrated inFIGS.7A and7B. First, in step S201, the controller13determines whether or not it is in a reception state for receiving an operation on the button switch14cC. If it is in the reception state, the process proceeds to step S202. On the other hand, if it is not in the reception state, this flow ends.

In step S202, the controller13determines whether or not there is an input to the button switch14cC. If there is an input, the process proceeds to step S203. On the other hand, if there is no input, the process returns to step S201.

In step S203, the controller13determines whether or not the changeover switch4bLC is OFF. If the changeover switch4bLC is OFF, the process proceeds to step S204. On the other hand, if the changeover switch4bLC is ON, the process proceeds to step S205.

In step S204, since the changeover switch4bLC is OFF, the microscope auxiliary apparatus acts in the non-linked mode. At this time, the controller13autofocuses onto the object by moving only the object movable portion1in the optical axis direction, and the process ends.

In step S205, since the changeover switch4bLC is ON, the microscope auxiliary apparatus acts in the linked mode. At this time, the controller13autofocuses onto the object by moving only the object movable portion1in the optical axis direction and, at the same time, moves the operating unit movable portion2L in the optical axis direction by the same amount as the moving amount of the object movable portion1, and this flow ends.

A description is given of effects of the linked mode in cell manipulation as an example.

FIGS.9A and9Bare explanatory diagrams of the effects in this embodiment.FIG.9Ais a diagram of a state immediately after the holding pipette HP and the injection pipette IP are attached. Since the positions of the holding pipette HP and the injection pipette IP in the optical axis direction are different, moving distances DL and DR to the focus position are different. Therefore, the linked mode is disabled, and the button switches14cL and14cR are individually pressed so that autofocusing is individually performed onto the holding pipette HP and the injection pipette IP. After that, the positions in the planar direction, tilts, etc. of the holding pipette HP and the injection pipette IP are appropriately adjusted.

FIG.9Bis a diagram illustrating a state just before the injection pipette IP punctures the vicinity of the center of the cell held by the holding pipette HP. In a case where the position of the injection pipette IP in the optical axis direction is to be readjusted, only the injection pipette IP is to be moved and the positions of the others are to be maintained, so the linked mode is disabled and the button switch14cR is pressed.

On the other hand, in a case where the position of the cell in the optical axis direction is to be readjusted, the linked mode is enabled for the operating unit movable portion2L and the object movable portion1. The operation time can be shortened by operating the button switch14cL or the button switch14cC and simultaneously moving the operating unit movable portion2L and the object movable portion1. Here, in a case where only the object movable portion1is moved with the linked mode disabled, the relative positions of the cells and the holding pipette HP are changed and the holding of the cells may become unstable.

In a case where a conventional microscope auxiliary apparatus not provided with the linked mode is used, the objective lens104ais first adjusted so that the focus on the cell is readjusted. Thereafter, the position of the injection pipette IP is adjusted so that the focus on the injection pipette IP is readjusted, which takes time for the operation. On the other hand, in the case where the microscope auxiliary apparatus according to this embodiment is used, the linked mode is enabled so that movements of at least two movable portions of the object movable portion1and the operating unit movable portions2L and2R in the optical axis direction are linked. Thereby, the operation time can be shortened. The linked mode is not to be enabled all the time since there is a case where the linked mode is to be disabled, such as a case where the focus is to be adjusted immediately after the pipette is attached as illustrated inFIG.9A. For this reason, a switch for enabling and disabling the linked mode is practically required.

According to this embodiment, a microscope auxiliary apparatus can be provided that can shorten an operation time by linking movements of at least two movable portions including an object movable portion or an operating unit movable portion in an optical axis direction.

Third Embodiment

Next, a description is given of a microscope system according to a third embodiment of the present disclosure.FIGS.10A and10Bare entire views of a microscope system10bincluding a microscope (inverted microscope)100and a microscope auxiliary apparatus and illustrate a state in which the microscope auxiliary apparatus is attached to the microscope100. The microscope auxiliary apparatus includes operating unit movable portions2L and2R, a display unit (display part)12, a controller23, and a console24.FIG.10Aillustrates a front view of the microscope system10b, andFIG.10Billustrates a right view of the microscope system10b. InFIGS.10A and10B, for clarity, some dimensions are exaggerated, some components are omitted, and some internal components are drawn with solid lines instead of dotted lines.

With reference toFIGS.10A and10B, a description is given of functions removed or added in this embodiment from or to the functions described in the second embodiment. In this embodiment, the object movable portion1and the driving circuit C in the second embodiment are not provided. Therefore, in this embodiment, focus on an object is adjusted by rotating the knob105of the microscope and moving the objective lens104ain the optical axis direction. The controller23includes a CPU23a, a driving circuit L23c, a driving circuit R23d, and an image processing circuit23e.

Changeover switches24dL and24dR can be operated left and right so that the operation mode is switched. The changeover switches24dL and24dR are changeover switches capable of reciprocating in two directions including “ON” for enabling a “linked mode” described below and “OFF” for disabling the “linked mode”.

In a case where the linked mode is disabled, a non-linked mode, which is conventional, is enabled in which even when the knob105is operated and the objective lens104ais moved in the optical axis direction, the operating unit movable portions2L and2R do not move in the optical axis direction. On the other hand, in a case where the “linked mode” is enabled, when the knob105is operated and the objective lens104amoves in the optical axis direction, the operating unit movable portions2L and2R are simultaneously moved by the same moving amount as the moving amount of the objective lens104a. Thus, the changeover switches24dL and24dR function as a switching unit that switches the mode to the linked mode (second mode) and to a non-linked mode (first mode).

A description is given of a method of simultaneously moving the operating unit movable portions2L and2R by the same moving amount as the objective lens104awhen the objective lens104ais moved in the optical axis direction in the linked mode. When the objective lens104ais moved, the focus position moves in the optical axis direction, and thus the focus on the operating units2La and2Ra attached to the operating unit movable portions2L and2R changes. A change in the focus can be detected as temporal changes acquired as a result of calculation of a predetermined in-focus evaluation value in the image pickup unit11. The predetermined in-focus evaluation value is acquired by calculating a contrast of the acquired image in a case of the contrast method, and by comparing two images from which a phase difference can be detected by pupil division in a predetermined direction in a case of the image pickup plane phase difference method. These are conventional techniques commonly used in digital cameras. Then, by performing feedback control such that the in-focus evaluation value is maintained of the observation image of the microscope100acquired by the image pickup unit11, the position of the objective lens104ain the optical axis direction can be maintained relatively to the focus position. As a result, movements of the operating unit movable portions2L and2R in the optical axis direction can be linked and they are moved by the same distance as the moving distance of the objective lens104aof the microscope100in the optical axis direction.

FIG.11is a block diagram of the feedback control performed in the linked mode. The controller23stores in advance an initial value f0 of an in-focus evaluation value f as a target value. The CPU23a(comparing unit231) calculates a difference df between the target value f0 and a current in-focus evaluation value f. The CPU23a(controlling unit)232performs calculation such that the difference f0−f=df is converted from the difference in the in-focus evaluation values to a shift amount in the optical axis direction, and sets the calculated value to a target value dZ of a relative moving amount of the operating unit movable portion, which is an object to be controlled. The controller23(driving circuit233) drives the operating unit movable portion so as to relatively move it by dZ, and a current position Z of the operating unit movable portion is output. The controller23(detector234) also calculates the in-focus evaluation value f at the current position Z and performs feedback control. In this manner, the feedback control is performed such that the in-focus evaluation value of the observation image of the microscope100acquired by the image pickup unit11is maintained at a constant value. As a result, the movement of the operating unit movable portion in the optical axis direction can be linked to the movement of the objective lens104aof the microscope so that the operating unit movable portion is moved by the same distance as the moving distance in the optical axis direction of the objective lens104a.

The feedback control in this embodiment maintains the in-focus evaluation value, and does not raise the in-focus evaluation value to bring it closer to in focus. This is because the purpose of the linked mode is not to bring the state into an in-focus state, but to move the operating unit movable portions2L and2R by the same amount as the moving amount of the objective lens104ain the optical axis direction.

A description is given of effects of the linked mode in cell manipulation as an example.FIGS.12A to12Care explanatory diagrams of the effects in this embodiment.FIG.12Ais a diagram of a state immediately after the holding pipette HP and the injection pipette IP are attached. The positions of the holding pipette HP and the injection pipette IP in the optical axis direction are different, and thus the moving distances DL and DR to the focus position are different. Therefore, the linked mode is disabled, and the dials24aL and24aR are individually operated so that the holding pipette HP and the injection pipette IP individually move and their respective tips are aligned with the focus position. After that, the positions in the planar direction, tilts, etc. of the holding pipette HP and the injection pipette IP are properly adjusted.

FIG.12Bis a diagram illustrating a state in which a petri dish, which is the observation tray102, on which a cell, which is the object103, is placed is set within a field of view of the microscope after the tips of the holding pipette HP and the injection pipette IP are aligned with the focus position and adjustment is properly performed. The petri dish is set after the positions, tilts, etc. of the holding pipette HP and the injection pipette IP are adjusted because the cell has been stored under a proper environment in a culture chamber and the time the cell is kept out of the culture chamber is to be minimized. After that, the set object103is to be focused on. Here, the changeover switches24dL and24dR are switched so that the linked mode is enabled, the objective lens is adjusted, and the focus position is moved in the direction of the arrow DC. When the focus plane moves to the center of object103, since the linked mode is enabled, the holding pipette HP and the injection pipette IP are also moved similarly to the objective lens104a, and thus the holding pipette HP and the injection pipette IP can be moved by operating only the objective lens104a. Conventionally, movements of the objective lens104a, the holding pipette HP, and the injection pipette IP are not linked, and therefore three operations are performed. On the other hand, according to this embodiment, the operation time can be shortened because the operation is completed with a single operation.

FIG.12Cis a diagram illustrating a state just before the injection pipette IP punctures the vicinity of the center of the cell held by the holding pipette HP. In this state, in a case where the position of the cell in the optical axis direction is to be readjusted, the relative positions of the holding pipette HP and the cell are to be maintained, and therefore the changeover switch24dL is switched to the linked mode OFF side. On the other hand, the focus on the injection pipette IP is to be maintained, the changeover switch24dR is switched to the linked mode ON side. Adjusting the position of the objective lens104ain this state can adjust the focus on the cell while maintaining the relative positions of the cell and the holding pipette HP holding the cell, and the movement of the injection pipette IP whose focus is to be maintained is linked to the movement of the objective lens104a. As a result, the focus can be finely adjusted with a single operation, which shortens the operation time. At this time, the linked mode is not to be enabled all the time since there is a case where the linked mode is to be disabled, so a switch for enabling and disabling the linked mode is practically required.

In this embodiment, even in a case where the linked mode is selected, if the in-focus evaluation value of the observation image is equal to or smaller than a predetermined value, since there is no point in linking the movements, the movement of the operating unit movable portion in the optical axis direction may not be linked to the movement of the objective lens.

Fourth Embodiment

Next, a description is given of a microscope system according to a fourth embodiment of the present disclosure.FIGS.13A to13Care entire views of a microscope system10cincluding a microscope (inverted microscope)100and a microscope auxiliary apparatus and illustrate a state in which the microscope auxiliary apparatus is attached to the microscope100. The microscope auxiliary apparatus includes operating unit movable portions2L and2R, a display unit12, and an image processing unit20.FIG.13Aillustrates a front view of the microscope system10c, andFIGS.13B and13Cillustrate a left view of the microscope system10c. InFIGS.13A to13C, for clarity, some dimensions are exaggerated, some components are omitted, and some internal components are drawn with solid lines instead of dotted lines.

With reference toFIGS.13A to13C, a description is given of functions added in this embodiment to the functions described in the third embodiment. An image pickup unit (image pickup apparatus)11in this embodiment is capable of acquiring two images from which a phase difference can be detected by pupil division in a predetermined direction, and includes an image pickup sensor capable of acquiring an observation image (captured image) of the microscope100. The image pickup unit11is capable of so-called image pickup plane phase difference AF. The image pickup unit11is attached to the microscope100via a mount11a. The mount11ain this embodiment is rotatable about the optical axis of the image pickup unit11as indicated by arrows inFIGS.13A to13C. Therefore, the mount11afunctions as an image pickup unit rotating unit capable of rotating the image pickup unit11relatively to the microscope100.

An image processing apparatus (image processing unit)20includes an image rotating unit201that generates a rotated image acquired by rotating, by a predetermined angle, an image that has been acquired by the image pickup unit11, and an image outputting unit202capable of outputting the rotated image rotated by the predetermined angle to an external display unit12. With these configurations, even in a case where the image pickup unit11is rotated by a predetermined angle by the mount11a, which is the image pickup unit rotating unit, and the observation image is acquired in a state of being rotated by the predetermined angle, the image rotating unit201can generate an image that is reversely rotated by the predetermined angle. As a result, the image outputting unit can output, to the display unit12, an image captured in a state where the image pickup unit11is not rotated by the predetermined angle.

Next, with reference toFIGS.14A to14E, a description is given of effects of this embodiment in a case where the holding pipette HP and the injection pipette IP are inserted from the left and right directions as an example.FIGS.14A to14Eare explanatory diagrams of the effects in this embodiment.

FIG.14Ais an optical image observed through an eyepiece lens104bwhen a cell is manipulated, and, normally, a part of the optical image is cut out as it is and output to the display unit12as illustrated inFIG.14B. This is because an observed view may be output and recorded as it is.

Here, a pupil division direction of the image pickup plane phase difference AF is normally a horizontal direction ofFIG.14B. On the other hand, in cell manipulation work, the holding pipette HP or the injection pipette IP is normally inserted into the field of view from the left and right. For this reason, as illustrated inFIG.14B, an image is acquired that includes few straight lines in a vertical direction (direction orthogonal to the pupil division direction), and parallax in the image pickup plane phase difference AF is small, which may reduce AF accuracy. Therefore, the image pickup unit11is attached while being rotated by a predetermined angle (90 degrees in this embodiment) so that an image is acquired as illustrated inFIG.14C. As a result, the parallax in the image pickup plane phase difference AF is increased, which can improve the AF accuracy.

However, in a case where the image is output and recorded as it is, an image rotated by 90 degrees relatively to the optical image is output and recorded, as illustrated inFIG.14D. As a result, it becomes uncomfortable to view the image on the display unit12or to reproduce the recorded image later. Therefore, the image rotating unit201generate an image reversely rotated by 90 degrees and outputs and records the image as illustrated inFIG.14E, which causes missing in the image at a peripheral part but can avoid the discomfort when the image is viewed on the display unit12or the recorded image is reproduced later.

According to this embodiment, it is possible to improve the AF accuracy of the image pickup plane phase difference AF. As a result, the accuracy can be improved of the linked movement of at least two movable portions including the object movable portion or the operating unit movable portion in the optical axis direction, and the operation time can be shortened.

Fifth Embodiment

Next, a description is given of a microscope system according to a fifth embodiment of the present disclosure.FIGS.15A to15Care entire views of a microscope system10dincluding a microscope (inverted microscope)100and a microscope auxiliary apparatus and illustrate a state in which the microscope auxiliary apparatus is attached to the microscope100. The microscope auxiliary apparatus includes operating unit movable portions2L and2R, a display unit12, and an image processing unit20.FIG.15Aillustrates a front view of the microscope system10, andFIGS.15B and15Cillustrate left views of the microscope system10. InFIGS.15A and15C, for clarity, some dimensions are exaggerated, some components are omitted, and some internal components are drawn with solid lines instead of dotted lines.

With reference toFIGS.15A to15C, a description is given of functions added in this embodiment to the functions described in the fourth embodiment. An image processing apparatus (image processing unit)20in this embodiment includes a dial20aand a level display20bin addition to an image rotating unit201and an image outputting unit202. The dial20ais a rotational angle changing unit that changes an angle by which the image rotating unit201rotates an image. The level display20bis a display unit that displays a shift change between two parallax images acquired by the image pickup unit11. In the level display20b, the larger an illuminated LED frame on a right side, the greater the parallax between the two images, and thus it is indicated that the image is captured with good AF accuracy of the image pickup plane phase difference AF.

Next, with reference toFIGS.16A to16E, a description is given of effects of this embodiment, and an example is described of a case where probes are brought into contact with cell tissue from various directions.FIGS.16A to16Eare explanatory diagrams of the effects of this embodiment.FIG.16Ais an optical image observed through the eyepiece lens104bwhen three probes P are brought into contact with cell tissue203. Normally, as illustrated inFIG.16B, part of the optical image is cut out as it is and output to the display unit12. This is because an observed optical image may be output and record as it is.

A pupil division direction of the image pickup plane phase difference AF is normally a horizontal direction ofFIG.16B. In this work, the probes are brought into contact with the cell tissue203from various directions. For this reason, as illustrated inFIG.16B, the image includes few straight lines crossing at 45 degrees or less in a vertical direction (the direction orthogonal to the pupil division direction), and the parallax in the image pickup plane phase difference AF is small, which may lower the AF accuracy. Therefore, the image pickup unit11is attached while being rotated by an angle θ, and an image such as that illustrated inFIG.16Cis acquired so that the parallax in the image pickup plane phase difference AF becomes large, which can improve the AF accuracy.

However, in a case where the image is output and recorded as it is, an image rotated by the angle θ from the optical image is output and recorded, as illustrated inFIG.16D. As a result, it becomes uncomfortable to view the image on the display unit12or to reproduce the recorded image later. Therefore, the image rotating unit201generate an image reversely rotated by the angle θ and outputs and records the image as illustrated inFIG.16E, which causes missing in the image at a peripheral part but can avoid the discomfort when the image is viewed on the display unit12or the recorded image is reproduced later. The angle θ by which the image pickup unit11is rotated can be set to an angle at which the level display20bindicates the maximum level while the display on the level display20bis referred to. By using the dial20a, the angle by which an image is reversely rotated when the image rotating unit201generates an image can be adjusted to an angle such that the angles of the optical image inFIG.16Aand the output image inFIG.16Eare equivalent to each other.

In this embodiment, an example is described in which an image is manually rotated via the dial20a, but the rotation angle θ of the image pickup unit11may be detected by a sensor or the like on the mount11aand the image rotating unit may automatically determine the angle by which the image is reversely rotated when generating an image.

According to this embodiment, it is possible to improve AF accuracy of image pickup plane phase difference AF. As a result, accuracy can be improved of linked movements of at least two movable portions including the object movable portion or the operating unit movable portion in the optical axis direction, and the operation time can be shortened.