JP2005352372A - Optical scanning microscope system - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a clear picture in which blurring caused by pulsation is suppressed when the viable tissue or various internal organs of mammals including experimental small animals is observed in their living state. <P>SOLUTION: The optical scanning microscope system 1 is provided with: a light source; a light transmission member 17 for transmitting light from the light source; a collimating optical system 30 for converting the transmitted light into parallel light; a light scanning part 44 for scanning the parallel light in an examinee; a condensing optical system 21 for condensing the scanned light on the examinee; a pupil projection optical system 45; a light detector for detecting returned light returned from the examinee A via the condensing optical system 21, the pupil projection optical system 45, the light scanning part 44, the collimating optical system 30, and the light transmission member 17; an actuator 37 for displacing the collimating optical system 30 along the direction of the optical axis; a controller for controlling its driving; and a deflection means 44 for deflecting the light emitted from the light transmission member to a direction crossed with the optical axis thereof, and in which an actuator 37 is arranged at a space along a plane including the optical axis before and after the deflection by the deflection means 44. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an optical scanning microscope apparatus.

Conventionally, optical scanning microscope apparatuses for magnifying and observing a subject have the structures shown in Patent Document 1 and Patent Document 2. These optical scanning microscope apparatuses disclosed in Patent Documents 1 and 2 are provided with a lens driving mechanism in an objective lens in order to quickly match a focal position with respect to a subject to an observation target site.
Japanese Patent Laying-Open No. 2003-172878 ([0042], FIG. 2 etc.) JP 2001-356256 A ([0020], FIG. 2 etc.)

  However, the conventional optical scanning microscope apparatus is arranged for observation with an objective lens arranged at a predetermined interval with respect to the subject, and various organs of experimental small animals such as rats and mice remain alive. It is not assumed to be observed in vivo. That is, when observing a subject such as an experimental small animal alive, it is effective to press the tip of the objective lens against the subject in order to prevent blurring of the image due to the pulsation, The conventional optical scanning microscope apparatus has a structure in which the tip surface of the objective lens is moved in order to focus on the observation target portion by a lens driving mechanism provided in the objective lens. For this reason, the conventional optical scanning microscope apparatus cannot perform observation by bringing the tip surface of the objective lens into contact with the living body, and if other methods are not used, the image is blurred due to pulsation and a clear image is obtained. There is an inconvenience that can not be.

  Further, when magnifying and observing the state of the internal tissue of the experimental small animal, it may be necessary to observe the external state of the experimental small animal in parallel. In this case, the optical scanning type It is conceivable to perform observation by combining a microscope apparatus and a stereoscopic microscope apparatus that can observe a wide range at a lower magnification than that. Therefore, the optical scanning microscope apparatus needs to be configured so as not to block the visual field of the stereomicroscope apparatus as much as possible.

  The present invention has been made in view of the above-described circumstances, and observes living cells of mammalian cells including small experimental animals, living tissues such as muscles, or various organs such as heart and liver in a living state. An object of the present invention is to provide an optical scanning microscope apparatus that can obtain a clear image in which blurring due to pulsation is suppressed and can be miniaturized so as not to obstruct the visual field of the stereoscopic microscope apparatus as much as possible.

In order to achieve the above object, the present invention provides the following means.
The present invention includes a light source, a light transmission member that transmits light from the light source, a collimating optical system that collimates light transmitted by the light transmission member, and parallel light emitted from the collimating optical system. An optical scanning unit that scans the specimen, a condensing optical system that condenses the light scanned by the optical scanning unit on the subject, and a pupil projection that is disposed between the condensing optical system and the optical scanning unit An optical system; a light detector that detects return light from the subject through a condensing optical system, a pupil projection optical system, a light scanning unit, a collimating optical system, and a light transmission member; and a tip of the light transmission member, a collimator An actuator for displacing the optical system or pupil projection optical system along the optical axis direction, a control device for controlling the driving of the actuator, and a deflection for deflecting light emitted from the light transmission member in a direction intersecting the optical axis Means. Chueta provides a light-scanning microscope apparatus which is arranged in a space along a plane including the optical axis before and after the deflection by the deflection means.

  According to the present invention, the light emitted from the light source is transmitted through the light transmission member, and is collimated by the collimating optical system. The parallel light from the collimating optical system is scanned by the optical scanning unit and irradiated onto the subject via the pupil projection optical system and the condensing optical system. Return light from the subject returns through the same path via the condensing optical system, pupil projection optical system, optical scanning unit, collimating optical system, and light transmission member, and is detected by the photodetector.

  The light emitted from the light transmission member is deflected in the intersecting direction by the deflecting means. Therefore, light can be guided from the lateral direction with respect to the optical axis direction of the condensing optical system toward the subject. As a result, the height dimension from the subject of the optical system from the collimating optical system to the condensing optical system can be suppressed, and it can be prevented from interfering with the stereomicroscope that is disposed above it.

  And since the actuator which displaces either the tip part of these light transmission members, a collimating optical system, or the pupil projection optical system is arranged in the space along the plane including the optical axis before and after deflection by the deflection means, the actuator Without projecting in the lateral direction, the projected area onto the subject can be kept small so that the stereomicroscope is not obstructed.

  Further, the actuator is operated by the operation of the control device, and any one of the distal end portion of the light transmission member, the collimating optical system, or the pupil projection optical system is displaced along the optical axis direction thereof, thereby condensing optics. The focal position can be changed while the tip position of the system is fixed. That is, observation can be performed with the tip of the condensing optical system pressed against a subject such as an experimental small animal. As a result, it is possible to prevent image blur due to the pulsation of the subject and obtain a clear image.

  According to the present invention, since the focal position can be adjusted without displacing the light irradiation end face of the condensing optical system, observation is performed while adjusting the focal position while pressing the light irradiation end face of the condensing optical system against the subject. It can be performed. In this case, by arranging an actuator for displacing the tip of the light transmission member, the collimating optical system or the pupil projection optical system along the optical axis direction along the plane including the optical axis before and after the deflection, the actuator is moved in the width direction. The projection area on the subject is reduced, and the subject is observed from the opposite side of the subject with the condensing optical system interposed therebetween, for example, so as not to interfere with the stereoscopic microscope observation. There is an effect that can be.

An optical scanning microscope observation apparatus according to an embodiment of the present invention will be described below with reference to FIGS.
The optical scanning microscope apparatus 1 according to the present embodiment is used in the microscope observation system 2 shown in FIG. As shown in FIG. 1, the microscope observation system 2 includes a stage 3 on which a subject A such as an experimental small animal is placed, and an optical scanning microscope observation apparatus according to the present embodiment disposed above the stage 3. 1 and a stereomicroscope observation device 4 disposed further above.

  The stereomicroscope observation device 4 is provided on a support column 6 rising from a base 5 provided with a stage 3 so as to be movable in the vertical direction, and includes an eyepiece unit 7 and a CCD camera 8 for observing the subject A at a relatively small magnification. I have. The CCD camera 8 is connected to a first monitor 10 by a cable 9 so that a captured image can be observed on the first monitor 10.

As shown in FIG. 1, the optical scanning microscope observation apparatus 1 according to the present embodiment includes a microscope observation apparatus body 11 and a light source 12, a photodetector 13, a control apparatus 14, and a second monitor 15 connected thereto. It has.
The microscope observation apparatus main body 11 is also provided on a column 16 rising from the base 5 so as to be movable in the vertical direction and tiltable in an arbitrary angle direction. The microscope observation apparatus body 11 and the light source 12 are connected by an optical fiber (light transmission member) 17. Further, the microscope observation apparatus main body 11 and the control apparatus 14 are connected to the photodetector 13 and the control apparatus 14 by cables 18 and 19, respectively. On the second monitor 15, an image subjected to image processing in the control device 14 is displayed.

  As shown in FIG. 2, the microscope observation apparatus body 11 includes a housing 20 attached to one end of the optical fiber 17 and an objective lens unit (condensing optical system) 21 attached to the housing 20. Yes. The optical fiber 17 is covered with a sheath 22. The distal end opening of the sheath 22 and the housing 20 are connected in a sealed state by a watertight seal 23. Further, in the sheath 22, a cable 18 that connects each device in the microscope observation apparatus body 11, for example, a galvano mirror unit, a stepping motor, an identification code reader, and the like, which will be described later, and the control device 14 is wired.

The housing 20 includes a cover member 24 and a reference member 25 that is fixedly disposed therein. The reference member 25 functions as a base for fixing or slidably supporting various members described later.
The reference member 25 is a substantially rectangular parallelepiped block-shaped member, and includes a first through hole 26 penetrating in the longitudinal direction and a second through hole 27 orthogonal to the first through hole 26. A ferrule 28 to which the tip of the optical fiber 17 is attached is fixed to one end opening of the first through hole 26 by a bracket 29. The ferrule 28 cuts the front end face obliquely to form the outgoing end face of the optical fiber 17 obliquely with respect to the length direction, and the reflected light in the optical fiber 17 on the outgoing end face is a photodetector 13 which will be described later. It is comprised so that it may prevent returning to.

A collimating lens unit 30 is supported in the first through hole 26 of the reference member 25 so as to be movable along the length direction of the first through hole 26. The collimating lens unit 30 houses one or more collimating lenses 31 in a lens frame 32.
The reference member 25 has a lead screw 34 extending in parallel to the first through hole 26 in a hole 33 provided in parallel to the first through hole 26, and a stepping motor for rotating the lead screw 34 about the longitudinal axis. An actuator 37 comprising a nut 35 and a nut 36 screwed into the lead screw 34 and linearly moved in the direction along the first through hole 26 by the rotation of the lead screw 34 is accommodated.
Note that other driving means such as a DC motor with an encoder may be used instead of the stepping motor.

  The hole 33 and the first through hole 26 are connected to each other by a communication portion 38, and the nut 36 and the lens frame 32 are connected by a connection portion including a spherical bearing 39. That is, the lens frame 32 and the nut 36 are connected by a spherical bearing 39 with a degree of freedom of rotation about an axis perpendicular to the longitudinal direction of the first through hole 26. That is, even if a blur occurs between the first through hole 26 and the lead screw 34, an excessive internal force does not act on the first through hole 26 and the lens frame 32. A rotation stop of the spherical bearing 39 is constituted by a groove (not shown) provided in the reference member 25 along the longitudinal direction of the first through hole 26.

  In the first through-hole 26, operating range detectors 40 for detecting the lens frame 32 are provided at both ends of the moving range of the lens frame 32. Further, in the hole 33 in which the lead screw 34 is accommodated, operation range detectors 41 for detecting the nut 36 are provided at both ends of the movement range of the nut 36. Any one of these operation range detectors 40 and 41 may be used.

  Further, an abutting surface 42 that abuts the end surface of the lens frame 32 is provided at one end of the first through hole 26. Since the lead screw 34 is continuously rotated in one direction by the operation of the stepping motor 35 and the end surface of the lens frame 32 is abutted against the abutting surface 42, the stepping motor 35 is stepped out by further rotating. The origin position can be accurately reproduced by rotating the lead screw 34 in a direction to return by a predetermined rotation angle.

  Further, a bracket 29 for attaching the ferrule 28 to the reference member 25 is provided with a transparent glass 43 that separates the ferrule 28 and the collimating lens unit 30 and allows light to pass therethrough. It is conceivable that dust is generated by sliding in the first through hole 26 of the collimator lens unit 30, but this is to prevent the generated dust from adhering to the ferrule 28.

  In the present embodiment, the actuator 37 that linearly moves the collimating lens unit 30 along the length direction of the first through hole 26 includes the central axis of the first through hole 26 and the central axis of the second through hole 27. It is arrange | positioned along the plane containing. That is, when the housing 20 is disposed above the objective lens unit 21, the lead screw 34, the nut 36, and the stepping motor 35 are disposed below the first through hole 26 so as not to protrude in the lateral direction. . As a result, the width dimension of the housing 20 can be kept small, the projected area onto the subject A from the upper side is reduced, and the field of view of the stereomicroscope observation device 4 disposed thereabove is not blocked. ing.

Further, the reference member 25 is formed with an inclined surface 25 a that forms an angle of 45 ° with respect to the axis of the through holes 26 and 27 in the vicinity of the intersection of the first through hole 26 and the second through hole 27. . A galvano mirror unit (light scanning unit, deflecting means) 44 capable of two-dimensional scanning is fixed to the inclined surface 25a so as to close the through holes 26 and 27. As a result, the laser beam that has passed through the first through hole 26 is deflected by approximately 90 ° by the galvano mirror unit 44 and guided into the second through hole 27.
In place of the galvanometer mirror unit, a scanning unit driven by a piezo element, a polygon mirror or the like may be used.

A pupil projection lens unit 45 having a pupil projection lens 45 a is fixed to one end opening of the second through hole 27 by a bracket 46. The pupil projection lens unit 45 focuses the laser beam deflected by the galvanometer mirror unit 44 on the intermediate image position B at one end. As a result, the laser beam is prevented from divergence of off-axis light, and the beam diameter is reduced, so that the laser beam can pass through the elongated objective lens unit 21.
In addition, a circumferential groove 46a is formed over the entire circumference of the outer peripheral surface of the bracket 46 of the pupil projection lens unit 45, and an O-ring 47 is disposed in the circumferential groove 46a.

  The objective lens unit 21 includes a substantially cylindrical lens frame 48 fitted to the outer surface of the bracket 46 of the pupil projection lens unit 45, and houses one or more objective lenses 49 therein. The objective lens 49 is configured to re-image the intermediate image formed by the pupil projection lens unit 45 on the subject A.

  The lens frame 48 and the bracket 46 are sealed in a watertight state by the O-ring 47. The lens frame 48 and the cover member 24 of the housing 20 are also sealed with a watertight seal 50.

An identification code portion 51 formed by cutting out a part of the end face of the lens frame 48 on the side to be fitted to the bracket 46 is provided. This identification code | cord | chord part 51 has a shape intrinsic | native to each objective lens unit 21, when there are multiple objective lens units 21. As shown in FIG. In a state where the objective lens unit 21 is fitted to the bracket 46, an identification code detector 52 for reading the identification code unit 51 is disposed on the reference member 25 at a position facing the identification code unit 51.
An attachment / detachment sensor (not shown) may be provided in the housing 20 to detect the attachment / detachment state of the objective lens unit 21.

  Further, by deflecting the light by the galvanometer mirror unit 44 in this way, when the objective lens unit 21 is arranged in the vertical direction, the first through hole 26 is arranged in a substantially horizontal direction, and thus the objective lens unit 21 is arranged. The distance from the tip to the upper end of the housing 20 can be suppressed. As a result, the distance from the stereomicroscope observation device 4 disposed above the housing 20 to the subject A can be shortened.

  Reference numeral 53 in the figure denotes an attachment portion for fixing the microscope observation apparatus main body 11 to an arm 54 extending from the support column 16, and this attachment portion 53 is also fixed to the reference member 25, and is watertight between the cover member 24. Sealed by a seal 55.

  For example, the light source 12 is a laser light source capable of selectively emitting excitation light having a plurality of wavelengths. Selection means for selecting the wavelength of the light source 12 is included in the control device 14. The photodetector 13 is, for example, a photomultiplier tube (PMT), detects fluorescence separated from the return light by the dichroic mirror 56 in the light source 12, and inputs it to the control device 14 as an image signal. It is configured as follows.

  As shown in FIG. 3, the control device 14 outputs an instruction for changing the wavelength of the emitted light to the light source 12. In addition, a control unit 57 that outputs an operation command to the stepping motor 35 of the microscope observation apparatus body 11 and outputs a deflection angle command to the galvanometer mirror unit 44 is provided. Further, output signals from the operation range detectors 40 and 41 and the identification code detector 52 are input from the microscope observation apparatus body 11. Furthermore, captured image information is input from the photodetector 13.

  For example, an input unit 58 such as a keyboard, a mouse, or a joystick is connected to the control device 14, and image information of an image to be acquired, for example, an image magnification, an image depth position, an image viewing range, or a frame rate is set. It can be input. And the control apparatus 14 calculates the movement command to the said actuator 37 based on the input image information.

  Further, a database 59 is provided in the control device 14. In the database 59, the identification codes indicated by the identification code units 51 of all the objective lens units 21 to be used and the information unique to the objective lens unit 21, for example, the nominal magnification and the unique magnification of the objective lens unit 21 are stored. , Magnification error, magnification characteristic function indicating the relationship between light wavelength and magnification, aberration characteristic function indicating the relationship between light wavelength and aberration for each type of aberration, serial number of the objective lens unit 21, and the like. Is remembered.

The aberration characteristic function indicating the relationship between the wavelength of light and the aberration for each type of aberration is information for each wavelength of the aberration characteristic of the objective lens unit 21. For example, since there are various types of distortion such as “pincushion type” and “barrel type” for each objective lens unit, each objective lens unit 21 has database 59 as information on the type of distortion and its correction value. To have. The correction value is variably set for each wavelength.
Based on the identification code of the objective lens unit 21 read by the identification code detector 52, various corresponding unique information can be read out and used together with the image information to calculate a movement command to the actuator 37. It has become.

  The control device 14 has a function of correcting the image signal obtained by the photodetector 13 based on the read aberration characteristic function. The corrected image signal thus corrected is output to the second monitor 15.

The operation of the thus configured scanning microscope observation apparatus 1 according to this embodiment will be described below.
In order to observe the subject A with the microscope observation system 2 described above, first, the subject A such as a mouse or rat, a small experimental animal, or the like is fixed to the stage 3, and the stereomicroscope observation apparatus 4 disposed above the subject A. While displaying the image of the site to be observed on the first monitor 10 using, the incision is made in the epidermis to expose the internal tissue. At this time, the microscope observation apparatus main body 11 of the scanning microscope observation apparatus 1 is disposed at a position retracted from the visual field of the stereoscopic microscope observation apparatus 4.

  In the scanning microscope observation apparatus 1, an objective lens unit 21 suitable for achieving a desired observation magnification is attached to the housing 20. The objective lens unit 21 and the housing 20 are sealed by an O-ring 47 and a watertight seal 50 so that moisture does not enter the inside. When the objective lens unit 21 is attached to the housing 20, the identification code indicated by the identification code unit 51 provided in the objective lens unit 21 is read by the identification code detector 52 and sent to the control device 14. In the control device 14, the database 59 is searched using the sent identification code as a key, and information unique to the objective lens unit 21 is read out.

When the preparation is completed, the microscope observation apparatus main body 11 of the scanning microscope observation apparatus 1 is inserted between the stereomicroscope observation apparatus 4 and the subject A.
According to the scanning microscope observation apparatus 1 according to the present embodiment, the actuator 37 for displacing the collimating lens unit 30 is disposed along a plane including the first through hole 26 and the second through hole 27, and its width. Since the dimension is set to be sufficiently small, the scanning microscope observation device 1 does not obstruct the visual field of the stereoscopic microscope observation device 4 so much and the observation of the observation target portion by the stereoscopic microscope observation device 4 is continued. Can be observed.

  In addition, since the first through hole 26 and the second through hole 27 that guide light are arranged at an angle of 90 ° with each other by the galvano mirror unit 44, the second through hole 27 and the objective that follows the second through hole 27 are disposed. When the lens unit 21 is arranged in the vertical direction, the first through hole 26 can be arranged substantially horizontally. Therefore, since the laser beam applied to the subject A can be guided from the side without being guided from directly above, the dimension in the height direction can be reduced. As a result, the distance between the stereoscopic microscope observation device 4 and the subject A can be reduced.

  And the front-end | tip of the objective lens unit 21 provided in the microscope observation apparatus main body 11 is pressed against the observation object site | part of the subject A exposed. Thereby, even if the subject A generates pulsation, the tip of the objective lens unit 21 can suppress the pulsation of the site to be observed and prevent the image from blurring.

In this state, the control device 14, the light source 12, the photodetector 13, and the like are operated. Based on the image information input from the input unit 58, the control device 14 performs the wavelength command for the light source 12, the operation command for the actuator 37, A deflection angle command for the mirror unit 44 is output.
When a wavelength command to the light source 12 is sent to the light source 12, the light source 12 is set to output laser light having a designated wavelength by a wavelength adjusting unit (not shown).

  The laser light emitted from the light source 12 is propagated through the optical fiber 17 into the housing 20 of the microscope observation apparatus body 11. The tip of the optical fiber 17 is fixed to the reference member 25 in the housing 20 by a ferrule 28, and laser light is emitted from the tip surface toward the first through hole 26. Since the tip surface of the optical fiber 17 is cut obliquely, the reflected light from the tip surface is prevented from returning through the optical fiber 17 and being detected by the photodetector 13.

  The laser light emitted from the tip of the optical fiber 17 is converted into parallel light by passing through the collimating lens unit 30 and is incident on the galvanometer mirror unit 44. Since the galvanometer mirror unit 44 is disposed at an angle of 45 ° with respect to the optical axis of the collimating lens unit 30, the light incident from the collimating lens unit 30 is deflected by 90 ° and output. Yes. The galvanometer mirror unit 44 scans the laser beam in a two-dimensional direction in response to a deflection angle command from the control device 14. Thereby, when the deflected laser light passes through the pupil projection lens unit 45 and the objective lens unit 21 and is irradiated with the subject A, the field of view is scanned two-dimensionally and input by the input means 58. The object A is irradiated over the entire area.

  The laser light deflected in the galvanometer mirror unit 44 forms an intermediate image by the pupil projection lens unit 45 and then is re-imaged on the subject A by the objective lens unit 21. When the subject A is irradiated with the laser light, the subject A emits fluorescence, and the emitted fluorescence is emitted from the objective lens unit 21, the pupil projection lens unit 45, the galvanometer mirror unit 44, the collimating lens unit 30, and the optical fiber. 17 returns to the same optical path through 17, is separated from the laser beam by the dichroic mirror 56, and is detected by the photodetector 13.

  When an operation command for the actuator 37 is sent to the stepping motor 35, the nut 36 is moved in the direction along the first through hole 26 by rotating the lead screw 34 by the commanded rotation angle. When the nut 36 is moved, the lens frame 32 connected to the nut 36 is displaced, and the internal collimating lens 31 is displaced together with the lens frame 32, whereby the laser at the tip of the objective lens unit 21 is displaced. The focal position of the light will be adjusted.

  Further, when changing the observation magnification by the scanning microscope observation apparatus 1, the magnification is inputted from the input means 58. In the control device 14, the movement amount of the actuator 37 for achieving the input magnification is calculated again, and an operation command is sent to the stepping motor 35. Thereby, the collimating lens unit 30 is displaced along the optical axis direction, and the focal position of the laser light at the tip of the objective lens unit 21 is changed.

  In this case, according to the scanning microscope observation apparatus 1 of the present embodiment, the unique information of the objective lens unit 21 read from the database 59 is used for calculating the operation command of the actuator 37. That is, the magnification of the objective lens unit 21 is deviated from the nominal magnification due to individual differences of optical components such as the lenses 49 constituting the objective lens unit 21, and the information is read from the database 59, whereby the actuator This is reflected in the generation of 37 operation commands. Thereby, even if the objective lens unit 21 is replaced, an operation command reflecting the individual difference of the objective lens unit 21 can always be generated, and the specified magnification can be achieved with high accuracy.

  Further, when setting and changing the focal position, it is only necessary to displace the collimating lens unit 30 in the housing 20 without displacing the tip of the objective lens unit 21, so that the pulsation of the subject A is continuously suppressed. Can be observed. Therefore, a clear image without blur can be obtained.

  Further, since the nut 36 of the actuator 37 for displacing the collimating lens unit 30 is connected to the lens frame 32 of the collimating lens unit 30 by the spherical bearing 39, both of them can be relatively rotated around an axis perpendicular to the optical axis. . As a result, even if an attachment error of the lens frame 32 to the housing 20 or an attachment error between the lens frame 32 and the nut 36 occurs, the attachment error can be absorbed by rotating both of them relatively.

In the scanning microscope observation apparatus 1 according to the present embodiment, an identification code unique to the objective lens unit 21 is displayed by the notch 51 provided in the objective lens unit 21. These identification codes may be adopted. Further, instead of the database 59 connected to the control device 14, the objective lens unit 21 itself may be provided with an IC chip storing unique information.
According to the identification code, there is an advantage that it is possible to correct an error inherent to each component accompanying the miniaturization of the mechanical component and the optical component, and to obtain a desired image and display accurate image information. .

  The arrangement of the actuator 37 is not limited to the above embodiment. It has a light source, light transmission member, collimating optical system, optical scanning unit, condensing optical system, pupil projection optical system, photodetector, actuator, and control device, and uses a condensing optical system that has been reduced in size and diameter. This is effective for general optical scanning microscopes.

  Further, an identification code (intra-housing optical system identification code) indicating unique information such as magnification and aberration of the pupil projection lens unit 45 and the collimating lens unit 30 provided in the microscope observation apparatus main body 11 is connected to the control device 14, for example. The reader 60 (not shown) may be provided in the control device 14. Further, the control device 14 may be provided with another database (second database) 59 ′ capable of searching for the corresponding unique information using the read identification code as a key.

  In this case, as shown in FIG. 4, the corresponding unique information includes image characteristics indicating the relationship between the movement amount and magnification of the collimating lens unit 30 or the pupil projection lens unit 45 for each wavelength. Then, if the operation command to the actuator 37 is calculated based on the searched unique information, the input image information is considered in consideration of individual differences between the pupil projection lens unit 45 and the collimating lens unit 30. It can be achieved with high accuracy.

  In the above embodiment, the image information desired to be acquired is input by an input device such as a keyboard (not shown) provided in the control device 14, but instead of this, as shown in FIG. A dial 61 and a scale 62 for inputting image information, for example, a magnification, may be provided on the side surface. The operator can set the image information with the dial 61 while operating the microscope observation apparatus body 11, and the setting is simple. Further, the dial 61 and the scale 62 may be provided on the controller 63 connected to the control device 14 as shown in FIG. Reference numeral 64 in the figure denotes a liquid crystal screen that displays a set value.

Further, as shown in FIG. 7, a window portion 65 that exposes the nut 36 of the internal actuator 37 is provided on the side surface of the housing 20, and a scale 66 is provided on the edge of the window portion 65, so that the position of the nut 36 is provided. The current magnification may be displayed.
Further, as shown in FIG. 8, a liquid crystal screen 64 and a scale 66 arranged at the edge thereof are provided on the side surface of the housing 20, and a magnification that varies depending on the position of the internal collimating lens unit 30 is displayed on the liquid crystal screen 64. You may decide to do it. In this case, an encoder that detects the position of the nut 36 of the actuator 37 may be incorporated, and the detection result may be displayed on the liquid crystal screen 64.
Further, as shown in FIG. 9, instead of the liquid crystal display, magnification display may be performed by an array of LEDs 67.

  Further, instead of providing the objective lens unit 21 with an identification code or an IC chip, as shown in FIGS. 14A and 14B, storage containers 110A and 110B for storing the objective lens units 21A and 21B, etc. And the identification codes 112A and 112B may be attached to the storage containers 110A and 110B. The identification codes 112A and 112B can store information on the magnification, numerical aperture, corresponding wavelength range, and observation atmosphere of the objective lens units 21A and 21B. By doing in this way, even when the objective lens units 21A, 21B, etc. are extremely downsized and cannot be given an identification code, the storage containers 110A, 110B, etc. can secure a sufficient installation space. There is an advantage. In addition, even when the objective lens units 21A and 21B are attached to the living body in advance, the identification codes 112A and 112B can be read by the storage containers 110A and 110B separate from the objective lens units 21A and 21B. There is also.

  Further, in this case, it is desirable to arrange protective members 111A and 111B that can accommodate only the corresponding objective lens units 21A and 21B in the storage containers 110A and 110B. In the example shown in FIG. 14, the protection members 111A and 111B are configured by cylindrical members having insertion holes that match the outer diameters of the objective lens units 21A and 21B.

Next, a scanning microscope observation apparatus 70 according to the second embodiment of the present invention will be described below with reference to FIG.
In the description of the present embodiment, the same reference numerals are given to portions having the same configuration as the scanning microscope observation apparatus 1 according to the first embodiment described above, and the description will be simplified.

  The scanning microscope observation apparatus 70 according to the present embodiment includes a ferrule 28 in which one end of the optical fiber 17 is fixed, while the scanning microscope observation apparatus 1 according to the first embodiment displaces the collimating lens unit 30. This is different from the first embodiment in that the (tip portion of the light transmission member) is displaced in the optical axis direction. .

The ferrule 28 is slidably disposed in the first through hole 26. In addition, an actuator 74 including a plunger 72 for causing the rod 71 to protrude and retract in the optical axis direction and a connecting member 73 attached to the rod 71 of the plunger 72 is provided, and the connecting member 73 is engaged with a groove 28 a provided in the ferrule 28. As a result, the linear displacement of the rod 71 is transmitted as the linear displacement of the ferrule 28. In the vicinity of the connecting member 73, an encoder 75 arranged along the moving direction is provided, and the linear position of the ferrule 28 is detected by the encoder 75.
In addition, the collimating lens unit 30 provided so as to be displaceable in the first embodiment is fixed in the reference member 25 of the housing 20.

According to the scanning microscope observation apparatus 70 according to the present embodiment configured as described above, the distal end surface of the objective lens unit 21 is abutted against the subject A without being displaced as in the first embodiment. The focal position can be moved.
Since the ferrule 28 is lighter than the collimating lens unit 30 and can be easily displaced by the actuator 74, there is an advantage that the focal position adjustment is easy.

Next, a scanning microscope observation apparatus 80 according to a third embodiment of the present invention will be described below with reference to FIGS.
In the description of the present embodiment, the same reference numerals are given to portions having the same configuration as those of the scanning microscope observation apparatus 80 according to the first and second embodiments described above, and the description will be simplified.

  In the scanning microscope observation apparatus 80 according to the present embodiment, the scanning microscope observation apparatuses 1 and 70 according to the first and second embodiments have moved the collimating lens unit 30 or the ferrule 28 in the optical axis direction. On the other hand, the collimating lens unit 30 and the ferrule 28 are fixed to the housing 20, and the pupil projection lens unit 81 is displaced along the optical axis direction.

  In the pupil projection lens unit 81, a lens frame 81b provided with a pupil projection lens 81a is slidably disposed in the housing 20, and is always urged upward by a compression spring 82 disposed below the lens frame 81b. The lens frame 81b is placed in contact with the upper surface of the lens frame 81b and rotated around the horizontal axis, thereby applying a force in a direction to push down the lens frame 81b from above according to the rotation angle. A cam 83 is provided. The cam 83 is rotated by a rotating body 85 rotatably supported by a sliding bearing 84 and a flexible shaft 86 that rotates the rotating body 85 about a horizontal axis.

The flexible shaft 86 is rotated around its axis by a motor or handle (not shown) provided on the control device 14 side, so that the rotational force is transmitted to the rotating body 84 and further to the cam 83. .
In the figure, reference numeral 87 denotes a distance measuring sensor that detects the position of the lens frame 81b.
According to the scanning microscope observation apparatus 80 according to the present embodiment configured as described above, the displacement is performed while the front end surface of the objective lens unit 21 is abutted against the subject A as in the first and second embodiments. It is possible to move the focal position without making it.
Instead of the flexible shaft 86 and the rotating body 85, the cam 83 may be directly rotated by a motor.

Next, a scanning microscope observation apparatus 90 according to a fourth embodiment of the present invention will be described below with reference to FIG.
A scanning microscope observation apparatus 90 according to the present embodiment is an example in which a light source 91 and a control device 92 are arranged in a microscope observation apparatus main body 93 as shown in FIG.

  The scanning microscope observation apparatus 90 according to the present embodiment includes a light source 91, a collimator 94 that collimates light emitted from the light source 91, a fixed mirror 95 that reflects parallel light emitted from the collimator 94, and a fixed A galvano mirror unit 96 that scans light reflected by the mirror 95 in a two-dimensional direction, a pupil projection lens 97, an objective lens 98, and light that has passed through the objective lens 98 is deflected by 90 ° to irradiate the subject A. The beam splitter 100 that branches the fluorescence returning from the deflection unit 99, the subject A, the deflection unit 99, the objective lens 98, the pupil projection lens 97, the galvano mirror unit 96, the fixed mirror 95, and the collimator 94. And a photodetector 101 for detecting fluorescence.

The light source 91 and the photodetector 101 are provided with linear movement mechanisms 102 and 103 for displacing the light source 91 and the photodetector 101 along the respective optical axis directions. Is controlled by. The control device 92 operates the linear movement mechanisms 102 and 103 based on the image information input by the input means 58. At this time, the linear movement mechanism 102 of the light source 91 and the linear movement mechanism 103 of the photodetector 101 are operated. These are configured to move by the same distance in the direction along each optical axis.
The collimator 94, the fixed mirror 95, and the galvanometer mirror unit 96 are arranged in a cylindrical housing 104 fixed to the microscope observation apparatus main body 93, and have a cylindrical shape including a pupil projection lens 97, an objective lens 98, and deflecting means 99. The lens unit 105 is fixed to the housing 104 by a watertight seal 106.
In the figure, reference numerals 107 and 108 denote pinholes for narrowing the light emitted from the light source 91.

  According to the scanning microscope observation apparatus 90 according to the present embodiment configured as described above, the scanning microscope observation apparatus 1 according to the first to third embodiments is achieved by displacing the light source 91 along the optical axis. , 70, 80, the focal position can be moved without displacing the tip position of the lens unit 105, and the focus position is adjusted while the tip position of the lens unit 105 is pressed against the subject A. Can do. Further, since the light guided from the lateral direction is deflected by the deflecting means 99 and irradiated onto the subject A, there is an effect that the field of view of the stereomicroscope observation device 4 is not further blocked.

  In the example shown in FIG. 3, the image information of the image to be acquired, for example, the image magnification, the image depth position, the image viewing range or the frame rate is input by the input unit 58. 3, a position detector for detecting the position of the actuator is provided, and image information of an image to be acquired by the photodetector 13 is calculated by the control device 57 based on the detected position of the actuator. Then, after correction based on error information unique to the objective lens unit 21 and the internal optical system of the housing 20, it may be displayed on the image information display means.

  In this case, the actuator position is detected in the first embodiment by a rotation angle detector of the stepping motor 35, a position detector such as an encoder (not shown), the encoder 75 in the second embodiment, and the distance sensor in the third embodiment. 88, the fourth embodiment is an encoder (not shown) that is provided to the linear movement mechanisms 102 and 103.

[Appendix]
In addition, the invention of the following structure is guide | induced from these embodiments.
1. A light source, a light transmission member that transmits light from the light source, a collimating optical system that collimates the light transmitted by the light transmission member, and parallel light emitted from the collimating optical system is scanned on the subject. An optical scanning unit, a condensing optical system that condenses light scanned by the optical scanning unit on a subject, and a pupil projection optical system disposed between the condensing optical system and the optical scanning unit, Photodetector that detects return light returning from the subject through the condensing optical system, pupil projection optical system, optical scanning unit, collimating optical system, and light transmission member, and the tip of the light transmission member, collimating optical system or pupil An actuator for displacing the projection optical system along the optical axis direction, a control device for controlling the driving of the actuator, and a deflection means for deflecting light emitted from the light transmission member in a direction intersecting the optical axis , Said actuator But optical-scanning microscope apparatus which is arranged in a space along a plane including the optical axis before and after the deflection by the deflection means.

According to the present invention, the light emitted from the light source is transmitted through the light transmission member, and is collimated by the collimating optical system. The parallel light from the collimating optical system is scanned by the optical scanning unit and irradiated onto the subject via the pupil projection optical system and the condensing optical system. Return light from the subject returns through the same path via the condensing optical system, pupil projection optical system, optical scanning unit, collimating optical system, and light transmission member, and is detected by the photodetector.
The light emitted from the light transmission member is deflected in the intersecting direction by the deflecting means. Therefore, light can be guided from the lateral direction with respect to the optical axis direction of the condensing optical system toward the subject. As a result, the height dimension from the subject of the optical system from the collimating optical system to the condensing optical system can be suppressed, and it can be prevented from interfering with the stereomicroscope that is disposed above it.

And since the actuator which displaces either the tip part of these light transmission members, a collimating optical system, or the pupil projection optical system is arranged in the space along the plane including the optical axis before and after deflection by the deflection means, the actuator Without projecting in the lateral direction, the projected area onto the subject can be kept small so that the stereomicroscope is not obstructed.
Further, the actuator is operated by the operation of the control device, and any one of the distal end portion of the light transmission member, the collimating optical system, or the pupil projection optical system is displaced along the optical axis direction thereof, thereby condensing optics. The focal position can be changed while the tip position of the system is fixed. That is, observation can be performed with the tip of the condensing optical system pressed against a subject such as an experimental small animal. As a result, it is possible to prevent image blur due to the pulsation of the subject and obtain a clear image.

2. A connecting portion between the actuator and the tip of the light transmission member displaced by the actuator, a collimating optical system, or a pupil projection optical system can be rotated about an axis perpendicular to the optical axis before and after the deflection by the deflecting unit. Item 2. The optical scanning microscope apparatus according to Item 1, which is connected.
According to this invention, when the tip of the light transmission member, the collimating optical system, or the pupil projection optical system is displaced by the operation of the actuator, a force is applied from the actuator to the connecting portion. When the tip of the light transmission member, collimating optical system, or pupil projection optical system is downsized, a force that is distorted by the frictional force with the surroundings acts when the displacement occurs, but the connecting part is before and after being deflected by the deflecting means. It is possible to smoothly displace by rotating and displacing around an axis perpendicular to the optical axis of the optical axis to release the force.

3. The optical scanning microscope apparatus according to Item 2, wherein the connection portion is connected by a spherical bearing.
According to the spherical bearing, the above action can be achieved with a simple configuration.

4). The optical scanning microscope observation apparatus according to any one of additional items 1 to 3, wherein the deflecting unit includes the optical scanning unit.
By combining the deflecting means with the optical scanning unit, the number of parts can be reduced and the apparatus can be downsized.

5). 5. The optical scanning according to claim 1, wherein the actuator includes a lead screw, a driving unit that rotationally drives the lead screw about its axis, and a nut that is screwed into the lead screw and has the connection portion. Type microscope equipment.
Due to the operation of the driving means, the lead screw is rotated around its axis, and the nut screwed to the lead screw is linearly moved along the length direction of the lead screw. Since the connection portion is provided on the nut, the focal position is adjusted by simply displacing the tip of the light transmission member, the collimating optical system, or the pupil projection optical system by linear movement of the nut. It becomes possible.

6). The optical scanning microscope apparatus according to any one of appendix 1 to appendix 4, wherein the actuator includes a plunger that displaces a shaft having the connecting portion attached to a tip in a longitudinal direction thereof.
7). The actuator is attached to any one of a cam having a cam surface that is displaced in the optical axis direction, a tip of the light transmission member that is displaced by the actuator, a collimating optical system, or a pupil projection optical system. The optical scanning microscope apparatus according to any one of additional items 1 to 4, further comprising a cam follower that is moved in an axial direction.
Similarly, the actuator can be configured simply by these.

8). Item 8. The optical scanning microscope apparatus according to Item 7, further comprising an angle detector that detects a rotation angle of the cam.
It is possible to detect the rotation angle of the cam and control the position of the tip of the light transmission member, the collimating optical system, or the pupil projection optical system with higher accuracy.

9. The optical scanning according to any one of appendices 1 to 8, further comprising an operation range detector that detects both ends of a moving portion of the light transmission member displaced by the actuator and a collimating optical system or a pupil projection optical system. Type microscope equipment.
By operating the motion range detector, the tip of the light transmission member that is displaced by the actuator and both ends of the movement range of the collimating optical system or pupil projection optical system are detected, so that displacement beyond the movement range is prevented. The

10. The optical scanning microscope observation apparatus according to any one of appendix 1 to appendix 9, further comprising a position detector that detects a position of the tip of the light transmission member displaced by the actuator, a collimating optical system, or a pupil projection optical system. .
The position detector can detect the position of the tip of the light transmission member, the collimating optical system, or the pupil projection optical system, and control the position more accurately.

11. The optical scanning microscope according to any one of appendix 1 to appendix 10, wherein the collimating optical system is movably provided and includes a transparent member that seals between a front end surface of the light transmission member and the collimating optical system. apparatus.
Although it is conceivable that dust is generated from the sliding portion or the like by the movement of the collimating optical system, it is possible to protect the dust generated by the operation of the transparent member from adhering to the tip surface of the light transmission member.

12 The optical scanning microscope apparatus according to any one of appendices 1 to 11, wherein the optical scanning unit includes a proximity galvanometer mirror.
According to the proximity galvanometer mirror, the light scanning unit can be easily configured, and the parallel light from the collimating optical system can be scanned with high accuracy.

13. The light source is capable of emitting light of a plurality of wavelengths, and includes a selection unit that selects the wavelength of light emitted from the light source, and the control device controls the actuator according to the wavelength selected by the selection unit The optical scanning microscope apparatus according to any one of Additional Item 1 to Additional Item 12.
When the wavelength of the light source is switched, the focal position changes according to the wavelength. Therefore, the control device controls the actuator according to the wavelength, thereby reducing the occurrence of chromatic aberration and reducing the number of lenses. .

14 14. The optical scanning microscope apparatus according to any one of additional items 1 to 13, wherein a tip portion of the light transmission member, a collimating optical system, an optical scanning unit, and a pupil projection optical system are accommodated in a casing.
15. Item 15. The light according to Item 14, wherein the housing is provided with a window that allows a distal end portion of the light transmission member displaced by the actuator, a collimating optical system, or a part of the pupil projection optical system to be observed from the outside. Scanning microscope observation device.
By doing in this way, the operator can visually recognize the current magnification by the position of the tip of the light transmission member that can be observed through the window, the position of the collimating optical system or the pupil projection optical system.

16. Item 15. The optical scanning microscope apparatus according to Item 14 or Item 15, wherein the condensing optical system is detachably attached to the housing.
By making the condensing optical system detachable from the housing, the condensing optical system can be placed in a state of being set on the subject. Therefore, it is possible to fix the condensing optical system to the subject and observe the same position of the subject for a long time.

17. The condensing optical system is provided with an identification code indicating information unique to the condensing optical system, the housing is provided with a reading device that reads the identification code, and the control device is provided with the read identification code. Item 17. The optical scanning microscope apparatus according to Item 16, which controls the actuator based on the first item.
Because there are individual differences in the condensing optical system, when the one condensing optical system is switched to another condensing optical system, etc., the control device reads each condensing optical system by reading the identification code with the reading device. Thus, it is possible to perform proper focus position adjustment by the actuator.

18. A database for storing the identification code and information specific to the condensing optical system in association with each other, and the control device searches the database using the identification code read by the reading device as a key, and the condensing optical system obtained Item 18. The optical scanning microscope apparatus according to Item 17, wherein the actuator is controlled based on the unique information.
By doing so, information unique to the condensing optical system can be stored in the database, and the identification code can be simplified. Further, it is possible to acquire a lot of information such as lens data and aberrations using a simple identification code, and the actuator can be controlled with higher accuracy.

19. In the database, the identification code and aberration information of the condensing optical system are stored in association with each other, the database is searched using the identification code read by the reader as a key, and the obtained aberration information of the condensing optical system is obtained. Item 19. The optical scanning microscope observation device according to Item 18, further comprising image correction means for correcting an image of the subject detected by the photodetector based on the above.
According to this invention, since the aberration information of the condensing optical system is acquired from the database using the identification code read by the reading device as a key, the image is corrected by the image correcting means based on the acquired aberration information. A clear image can be obtained.

20. The optical scanning microscope apparatus according to any one of supplementary items 16 to 19, wherein the condensing optical system and the housing are connected in a watertight state.
When both are connected in a watertight state, these liquids can enter between the condensing optical system and the housing even in an environment where body fluids or cleaning fluids exist, for example, when observing the inside of a small experimental animal. Can be prevented.

21. Item 21. The optical scanning microscope apparatus according to any one of items 16 to 20, further comprising an attachment / detachment sensor that detects attachment / detachment of the condensing optical system to the housing.
Since the attachment / detachment state of the condensing optical system to the housing is detected by the operation of the attachment / detachment sensor, it is possible to prevent waste of work due to observation in an incomplete attachment state and intrusion of liquid.

22. The housing is detachably connected to the control device by a connector, and the housing includes an in-housing optical system identification code indicating information unique to the collimating optical system and the pupil projection optical system in the housing. The connector includes an optical system information reading device in the housing that reads the optical system identification code in the housing, and the control device controls the actuator based on the read optical system identification code in the housing. Item 22. The optical scanning microscope apparatus according to any one of Items 14 to 21.
When the housing and the control device are connected by a connector, the optical system identification code in the housing provided in the housing is read by the optical system information reading device in the housing provided in the connector. Since there are optical systems with unique information such as collimating optical system and pupil projection optical system in the housing, by reading this and controlling the actuator based on the read optical system identification code in the housing, more The focal position can be adjusted with high accuracy.

23. A second database for storing the internal optical system identification code and information unique to the collimating optical system and the pupil projection optical system in association with each other, and the control device reads the internal optical system information reading device; Item 23. The optical scanning microscope apparatus according to Item 22, wherein the second database is searched using the in-vivo optical system identification code as a key, and the actuator is controlled based on the obtained information.
By doing so, it is possible to simplify the optical system identification code in the housing, and it is possible to acquire a lot of information such as lens data and aberration from the second database using a simple identification code, The actuator can be controlled with higher accuracy.

24. In the second database, the optical system identification code and the aberration information of the collimating optical system and the pupil projection optical system are stored in association with each other, and the image correction unit is read by the optical system information reading device in the casing. A supplementary note that searches the second database using the in-vivo optical system identification code as a key, and corrects the image of the subject detected by the photodetector based on the obtained aberration information of the collimating optical system and the pupil projection optical system Item 20. An optical scanning microscope observation apparatus according to Item 19.
According to this invention, the aberration information of the collimating optical system and the pupil projection optical system is acquired from the second database using the in-casing optical system identification code read by the in-casing optical system information reading device as a key. A clear image can be obtained by correcting the image by the image correcting means based on the aberration information.

25. Item 1. An image information input unit that inputs image information of an image to be acquired by the photodetector, wherein the control device controls the actuator based on the image information input by the image information input unit. 25. An optical scanning microscope apparatus according to any one of items 24 to 24.
When the operator inputs image information of an image to be acquired by the image information input means, the actuator is controlled by the control means based on the input image information. Since the actuator is automatically controlled, adjustment work can be saved.

26. Image information input means for inputting image information of an image to be acquired by the photodetector is provided on the outer surface of the housing, and the control device is configured to control the actuator based on the image information input by the image information input means. 25. The optical scanning microscope apparatus according to any one of appendices 14 to 24, which controls
When the operator inputs image information on the outer surface of the casing, the actuator is controlled based on the input image information. The operability can be improved by providing the image information input means on the outer surface of the casing.

27. Image information of an image to be acquired by the photodetector is calculated based on the position of the tip of the light transmission member, the collimating optical system, or the pupil projection optical system that is detected by the position detection unit and is displaced by the actuator. Item 11. The optical scanning microscope apparatus according to Item 10, further comprising an image information calculation unit and image information display means for displaying the calculated image information.
When the actuator is actuated, the position of the light transmission member, the collimating optical system, or the pupil projection optical system is detected by the operation of the position detecting means, and the image information calculation unit should acquire based on the detected position. Image information of the image is calculated. Since the calculated image information is displayed on the image information display means, the operator can know the image information of the currently captured image by looking at the display on the image information display means.

28. Providing unique information input means for inputting unique information of the condensing optical system, the tip of the light transmission member detected by the position detecting means and displaced by the actuator, a collimating optical system, or a pupil projection optical system An image information calculation unit that calculates image information of an image to be acquired by the photodetector based on the position of the light collecting optical system and the input information unique to the condensing optical system, and image information display means for displaying the calculated image information An optical scanning microscope observation apparatus according to Additional Item 10, comprising:
By doing in this way, the image information calculation unit should acquire the image information of the image to be acquired based on the tip of the light transmission member, the position of the collimating optical system or the pupil projection optical system, and the information specific to the condensing optical system. Therefore, more accurate image information is displayed on the image information display means.

29. 29. The optical scanning microscope observation device according to additional item 28, wherein the information unique to the condensing optical system is any one of a magnification, a depth, and a visual field range of the condensing optical system.
By inputting a magnification specific to the condensing optical system, it is possible to display image information with high accuracy regardless of any variation in magnification, depth, or field of view due to individual differences.

30. An optical system information input means for inputting information unique to the collimating optical system and the pupil projection optical system, the tip of the light transmission member detected by the position detecting means and displaced by the actuator; a collimating optical system; Alternatively, an image information calculation unit that calculates image information of an image to be acquired by the photodetector based on the position of the pupil projection optical system and the input information specific to the collimating optical system and the pupil projection optical system is calculated. The optical scanning microscope apparatus according to appendix 10, comprising image information display means for displaying the image information.
Since the image information calculation unit calculates the image information of the image to be acquired by the photodetector using the information specific to the collimating optical system and the pupil projection optical system input by the optical system information input means, the collimating optical system In addition, more accurate image information can be displayed regardless of individual differences in pupil projection optical systems.

31. The optical scanning microscope apparatus according to item 30, wherein the information unique to the collimating optical system and the pupil projection optical system is a magnification of the collimating optical system and the pupil projection optical system.
In particular, when the size is reduced and sufficient processing accuracy cannot be obtained, the magnification of the collimating optical system and pupil projection optical system will have individual differences. Since it is input by the information input means, it is possible to calculate image information with high accuracy based on this and display it on the image information display means.

32. Item 32. The optical scanning microscope apparatus according to any one of Item 25 to Item 31, wherein the image information is any one of a magnification, a depth, and a visual field range of an image to be acquired.
By displaying these pieces of information by the image information display means, the operator can improve the current shooting state and the observation accuracy of the obtained image.

33. A light source, an optical scanning unit that scans incident light from the light source on the subject, a condensing optical system that collects incident light scanned by the optical scanning unit on the subject, and light emitted from the light source A deflecting unit that deflects in a direction intersecting the optical axis, and a separating unit that separates the returning light from the subject that returns from the subject through the condensing optical system and the optical scanning unit from the incident light. A detector for detecting the return light, a light source moving means for displacing the light source in the optical axis direction, and a detection for displacing the photodetector in the optical axis direction by the same displacement amount as the displacement amount of the light source. An optical scanning microscope apparatus comprising a detector moving means and a control device that controls driving of the detector moving means.

  According to the present invention, the light emitted from the light source is scanned by the light scanning unit and irradiated onto the subject via the condensing optical system. Since the light from the light source is deflected in the intersecting direction by the deflecting means and irradiated on the subject, the light can be incident from the lateral direction with respect to the direction of irradiation with respect to the subject. Can be avoided. The return light from the subject returns through the same path via the condensing optical system and the optical scanning unit, is separated from the incident light by the separation means, and is detected by the photodetector. In this case, the focal position can be adjusted without displacing the tip position of the condensing optical system by changing the light source position by operating the light source moving means. Further, in this case, a clear image can be obtained by displacing the detector by the same displacement amount as the light source displacement amount by the operation of the control means.

In order to observe various organs of experimental small animals such as rats and mice alive using an optical scanning microscope device, a microscope is used so that the objective lens unit of the microscope device can be brought close to the experimental small animal that is the subject. It is important that the objective lens unit of the apparatus is reduced in diameter. When the diameter of the condensing optical system is reduced, the objective lens and the mechanical parts that hold the objective lens become smaller, and an error occurs in the magnification and the focus position in the assembly process. An error inherent to each objective lens unit needs to be corrected when the objective lens unit is mounted on the microscope body. In the conventional optical scanning microscope apparatus, it is not assumed that errors caused by individual differences between the objective lens and the mechanical parts that hold the objective lens are corrected.
The following additional items 34 to 55 are made in view of the above-described circumstances, and are intended to correct errors inherent in the objective lens unit when the objective lens unit is mounted on the microscope body.

34. A light source, a light transmission member that transmits light from the light source, a collimating optical system that collimates the light transmitted by the light transmission member, and parallel light emitted from the collimating optical system is scanned on the subject. An optical scanning unit, a condensing optical system that condenses light scanned by the optical scanning unit on a subject, and a pupil projection optical system disposed between the condensing optical system and the optical scanning unit, Photodetector that detects return light returning from the subject through the condensing optical system, pupil projection optical system, optical scanning unit, collimating optical system, and light transmission member, and the tip of the light transmission member, collimating optical system or pupil An optical scanning microscope comprising: an actuator for displacing the projection optical system along the optical axis direction; and a control device for controlling driving of the actuator, wherein the condensing optical system is detachably attached to the housing apparatus.

  According to this invention, the actuator is operated by the operation of the control device, and any one of the tip portion of the light transmission member, the collimating optical system, or the pupil projection optical system is displaced along the optical axis direction thereof. The focal position can be changed while the tip position of the condensing optical system is fixed. That is, observation can be performed with the tip of the condensing optical system pressed against a subject such as an experimental small animal. As a result, it is possible to prevent image blur due to the pulsation of the subject and obtain a clear image.

  In this case, by making the condensing optical system detachable from the housing, the condensing optical system can be placed in a state of being set on the subject. Therefore, it is possible to fix the condensing optical system to the subject and observe the same position of the subject for a long time.

35. The condensing optical system is provided with an identification code indicating information unique to the condensing optical system, the housing is provided with a reading device that reads the identification code, and the control device is provided with the read identification code. Item 34. The optical scanning microscope apparatus according to Item 34, which controls the actuator based on the first item.
According to the present invention, even if there is an error inherent in each condensing optical system, it is possible to correct the condensing optical system at the time of mounting, and display desired image settings or image information being acquired.

36. A database for storing the identification code and information specific to the condensing optical system in association with each other, and the control device searches the database using the identification code read by the reading device as a key, and the condensing optical system obtained Item 36. The optical scanning microscope apparatus according to Item 35, wherein the actuator is controlled based on the unique information.
By doing so, information unique to the condensing optical system can be stored in the database, and the identification code can be simplified. Further, it is possible to acquire a lot of information such as lens data and aberrations using a simple identification code, and the actuator can be controlled with higher accuracy.

37. In the database, the identification code and aberration information of the condensing optical system are stored in association with each other, the database is searched using the identification code read by the reader as a key, and the obtained aberration information of the condensing optical system is obtained. 37. The optical scanning microscope observation apparatus according to additional item 36, further comprising: an image correction unit that corrects the image of the subject detected by the photodetector based on the above.
According to this invention, since the aberration information of the condensing optical system is acquired from the database using the identification code read by the reading device as a key, the image is corrected by the image correcting means based on the acquired aberration information. A clear image can be obtained.

38. The optical scanning microscope apparatus according to any one of appendices 34 to 37, wherein the condensing optical system and the casing are connected in a watertight state.
When both are connected in a watertight state, these liquids can enter between the condensing optical system and the housing even in an environment where body fluids or cleaning fluids exist, for example, when observing the inside of a small experimental animal. Can be prevented.

39. 39. The optical scanning microscope apparatus according to any one of appendices 34 to 38, further comprising an attachment / detachment sensor that detects attachment / detachment of the condensing optical system to / from the housing.
Since the attachment / detachment state of the condensing optical system to the housing is detected by the operation of the attachment / detachment sensor, it is possible to prevent waste of work due to observation in an incomplete attachment state and intrusion of liquid.

40. The housing is detachably connected to the control device by a connector, and the housing includes an in-housing optical system identification code indicating information unique to the collimating optical system and the pupil projection optical system in the housing. The connector includes an optical system information reading device in the housing that reads the optical system identification code in the housing, and the control device controls the actuator based on the optical system identification code in the housing that is read. Item 34. The optical scanning microscope apparatus according to any one of Items 34 to 39.
When the housing and the control device are connected by a connector, the optical system identification code in the housing provided in the housing is read by the optical system information reading device in the housing provided in the connector. Since there are optical systems with unique information such as collimating optical system and pupil projection optical system in the housing, by reading this and controlling the actuator based on the in-housing optical system identification code, The focal position can be adjusted with high accuracy.

41. A second database for storing the internal optical system identification code and information unique to the collimating optical system and the pupil projection optical system in association with each other, and the control device reads the internal optical system information reading device; 41. The optical scanning microscope apparatus according to Additional Item 40, wherein the second database is searched using the in-vivo optical system identification code as a key, and the actuator is controlled based on the obtained information.
By doing so, it is possible to simplify the optical system identification code in the housing, and it is possible to acquire a lot of information such as lens data and aberration from the second database using a simple identification code, The actuator can be controlled with higher accuracy.

42. The in-casing optical system identification code and the aberration information of the collimating optical system and the pupil projection optical system are stored in the second database in association with each other, and the image correcting means is read by the in-casing optical system information reading device. The second database is searched using the in-housing optical system identification code as a key, and the image of the object detected by the photodetector is corrected based on the obtained aberration information of the collimating optical system and the pupil projection optical system. 42. The optical scanning microscope observation apparatus according to any one of Supplementary Item 34 to Supplementary Item 41.
According to this invention, the aberration information of the collimating optical system and the pupil projection optical system is acquired from the second database using the in-casing optical system identification code read by the in-casing optical system information reading device as a key. A clear image can be obtained by correcting the image by the image correcting means based on the aberration information.

43. Item 34. Image information input means for inputting image information of an image to be acquired by the photodetector, and the control device controls the actuator based on the image information input by the image information input means. 45. The optical scanning microscope apparatus according to any one of items 42 to 42.
When the operator inputs image information of an image to be acquired by the image information input means, the actuator is controlled by the control means based on the input image information. Since the actuator is automatically controlled, adjustment work can be saved.

44. Image information input means for inputting image information of an image to be acquired by the photodetector is provided on the outer surface of the housing, and the control device is configured to control the actuator based on the image information input by the image information input means. 44. The optical scanning microscope apparatus according to any one of appendices 34 to 43, which controls
When the operator inputs image information on the outer surface of the housing, the actuator is controlled based on the input image information. By providing the image information input means on the outer surface of the casing, the operability can be improved.

45. Item 34. The optical scanning microscope observation device according to Item 34, further comprising a position detector that detects the position of the tip of the light transmission member displaced by the actuator, the collimating optical system, or the pupil projection optical system.
The position detector can detect the position of the tip of the light transmission member, the collimating optical system, or the pupil projection optical system, and control the position more accurately.

46. Image information of an image to be acquired by the photodetector is calculated based on the position of the tip of the light transmission member, the collimating optical system, or the pupil projection optical system that is detected by the position detection unit and is displaced by the actuator. Item 46. The optical scanning microscope observation apparatus according to Item 45, further comprising an image information calculation unit and image information display means for displaying the calculated image information.
When the actuator is actuated, the position of the light transmission member, the collimating optical system, or the pupil projection optical system is detected by the operation of the position detecting means, and the image information calculation unit should acquire based on the detected position. Image information of the image is calculated. Since the calculated image information is displayed on the image information display means, the operator can know the image information of the currently captured image by looking at the display on the image information display means.

47. Providing unique information input means for inputting unique information of the condensing optical system, the tip of the light transmission member detected by the position detecting means and displaced by the actuator, a collimating optical system, or a pupil projection optical system An image information calculation unit that calculates image information of an image to be acquired by the photodetector based on the position of the light collecting optical system and the input information unique to the condensing optical system, and image information display means for displaying the calculated image information An optical scanning microscope observation apparatus according to Additional Item 45, comprising:
By doing in this way, the image information calculation unit should acquire the image information of the image to be acquired based on the tip of the light transmission member, the position of the collimating optical system or the pupil projection optical system, and the information specific to the condensing optical system. Therefore, more accurate image information is displayed on the image information display means.

48. 48. The optical scanning microscope observation device according to additional item 47, wherein the information unique to the condensing optical system is any one of a magnification, a depth, and a visual field range of the condensing optical system.
By inputting a magnification specific to the condensing optical system, it is possible to display image information with high accuracy regardless of any variation in magnification, depth, or field of view due to individual differences.

49. An optical system information input means for inputting information unique to the collimating optical system and the pupil projection optical system, the tip of the light transmission member detected by the position detecting means and displaced by the actuator; a collimating optical system; Alternatively, an image information calculation unit that calculates image information of an image to be acquired by the photodetector based on the position of the pupil projection optical system and the input information specific to the collimating optical system and the pupil projection optical system is calculated. 46. An optical scanning microscope observation apparatus according to additional item 45, comprising image information display means for displaying the image information.
Since the image information calculation unit calculates the image information of the image to be acquired by the photodetector using the information specific to the collimating optical system and the pupil projection optical system input by the optical system information input means, the collimating optical system In addition, more accurate image information can be displayed regardless of individual differences in pupil projection optical systems.

50. Item 50. The optical scanning microscope observation apparatus according to Item 49, wherein the information unique to the collimating optical system and the pupil projection optical system is a magnification of the collimating optical system and the pupil projection optical system.
In particular, when the size is reduced and sufficient processing accuracy cannot be obtained, the magnification of the collimating optical system and pupil projection optical system will have individual differences. Since it is input by the information input means, it is possible to calculate image information with high accuracy based on this and display it on the image information display means.

51. The optical scanning microscope apparatus according to any one of appendix 43 to appendix 50, wherein the image information is any one of a magnification, a depth, and a visual field range of an image to be acquired.
By displaying these pieces of information by the image information display means, the operator can improve the current shooting state and the observation accuracy of the obtained image.

52. A laser light scanning microscope comprising: an optical scanning unit having an optical fiber, a collimating unit, and an optical scanning unit; and a condensing optical system having an elongated rigid cylindrical insertion portion and detachably attached to the optical scanning unit A storage container for storing the condensing optical system in which an identification code indicating information unique to the condensing optical system is provided.
According to this invention, even when the condensing optical system is too small to attach an identification code, the identification code of the condensing optical system can be used, and the condensing optical system is already attached to a living body. The identification code can be input even in the state in which it is displayed.

53. Item 53. The storage container according to Item 52, further comprising a protection unit that can accommodate only the condensing optical system corresponding to the identification code.
By comprising in this way, a condensing optical system and a storage container can be matched on a one-to-one basis.

54. Item 54. The storage container according to item 53 or item 53, wherein the identification code is a radio wave individual recognition (RFID) method.
With this configuration, the identification code can be easily read.

55. 55. The storage container according to any one of additional items 52 to 54, wherein the identification code includes at least one information of a magnification, a numerical aperture, a corresponding wavelength range, or an observation atmosphere.
By reading the identification code, it is possible to easily acquire such information.

  Note that other embodiments formed by partially combining the above-described embodiments and the like belong to the present invention.

1 is a schematic diagram showing a microscope observation system including a scanning microscope observation apparatus according to a first embodiment of the present invention. It is a longitudinal cross-sectional view which shows the detailed structure of the scanning microscope observation apparatus which concerns on the 1st Embodiment of this invention. It is a block diagram which shows the function of the control apparatus of the scanning microscope observation apparatus of FIG. It is a graph which shows an example of the characteristic of the collimating lens unit used in the control apparatus of the scanning microscope observation apparatus of FIG. It is a side view which shows the 1st modification of the input means of the scanning microscope observation apparatus of FIG. It is a figure which shows the 2nd modification of the input means of the scanning microscope observation apparatus of FIG. It is a side view which shows the case where it is a modification of the scanning microscope observation apparatus of FIG. 2, and has a window part which displays image information. FIG. 9 is a modification of FIG. 7 and is a side view showing a case where a liquid crystal screen for displaying image information is provided. FIG. 8 is a side view showing a modification example of FIG. 7 and having an LED array for displaying image information. It is a longitudinal cross-sectional view which shows the detailed structure of the scanning microscope observation apparatus which concerns on the 2nd Embodiment of this invention. It is a longitudinal cross-sectional view which shows the detailed structure of the scanning microscope observation apparatus which concerns on the 3rd Embodiment of this invention. 12 is a longitudinal sectional view showing a cam 83 for displacing a pupil projection lens 81a in the scanning microscope observation apparatus of FIG. It is a longitudinal cross-sectional view which shows the detailed structure of the scanning microscope observation apparatus concerning the 4th Embodiment of this invention. It is a perspective view which shows the example of the storage container provided with the identification code | symbol.

Explanation of symbols

A subject 1, 70, 80, 90 optical scanning microscope device 12, 91 light source 13, 101 photodetector 14 control device (selection means, image correction means, image information calculation section)
17 Optical fiber (light transmission member)
21 Objective lens unit (condensing optical system)
28 Ferrule (tip)
30 Collimating lens unit (collimating optical system)
34 Lead screw 35 Stepping motor (drive means)
36 Nut 37 Actuator 39 Spherical bearing (connection part)
40, 41 Operating range detector 20 Housing 43 Transparent glass (transparent member)
44,96 Galvanometer mirror unit (optical scanning unit, deflection means)
45 Pupil projection lens unit (pupil projection optical system)
51 Identification code part (identification code)
52 Identification code detector (reading device: unique information input means)
59 Database 61 Dial (image information input means)
64 LCD screen (image information display means)
65 Window (window)
67 LED (image information display means)
71 Shaft 72 Plunger 75 Encoder (position detector)
81b Lens frame (cam follower)
83 Cam 92 Control device 100 Beam splitter (separating means)
102 Linear movement mechanism (light source movement means)
103 Linear moving mechanism (detector moving means)
105 Lens unit (condensing optical system)

Claims (1)

A light source;
A light transmission member for transmitting light from the light source;
A collimating optical system that collimates the light transmitted by the light transmission member;
An optical scanning unit that scans the subject with parallel light emitted from the collimating optical system;
A condensing optical system for condensing the light scanned by the light scanning unit on the subject;
A pupil projection optical system disposed between the condensing optical system and the optical scanning unit;
A photodetector that detects return light returning from the subject through a condensing optical system, a pupil projection optical system, a light scanning unit, a collimating optical system, and a light transmission member;
An actuator for displacing the tip of the light transmission member, the collimating optical system or the pupil projection optical system along the optical axis direction;
A control device for controlling the drive of the actuator;
Deflecting means for deflecting light emitted from the light transmission member in a direction intersecting the optical axis;
An optical scanning microscope apparatus in which the actuator is disposed in a space along a plane including an optical axis before and after deflection by the deflection means.

Images