JP2015084060A - Microscope apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a microscope apparatus capable of performing highly reliable fluorescence observations without the use of a darkened room and being made compact.SOLUTION: A microscope apparatus 500 comprises a measurement unit 100, a PC 200, a measurement light supplying unit 300, and a display 400. The measurement unit 100 includes a housing 101. The housing 101 houses a pattern providing part 110, a light receiving part 120, a transmitted light supplying part 130, a stage 140, a stage drive device 141, a filter unit 150, a lens unit 160, and a control board 170. A measured object S is placed on the stage 140. The inside of the housing 101 is kept dark, and a fluorescence observation is performed for the measured object S. The measurement light supplying unit 300 includes a power supply device 310 and a light projection part 320. The power supply device 310 supplies electric power to each component in the measurement unit 100 from the outside of the housing 101. The light projection part 320 generates light for fluorescence observations of the measured object S and applies it to the measurement unit 100.

Description

  The present invention relates to a microscope apparatus capable of performing fluorescence observation.

  A fluorescence microscope capable of performing fluorescence observation without using a dark room has been proposed. For example, an inverted fluorescence microscope described in Patent Document 1 includes a main body case, a specimen cover, and a lower cover. The specimen cover and the lower cover are provided on the front surface of the main body case. A stage, a bright-field transmission illumination system, an epi-illumination system, an imaging system, a power supply unit, a power supply board, a controller, and the like are mounted inside the main body case.

  The specimen cover is provided so as to be movable in the vertical direction between an open position at the same height as the lower cover and a closed position above the lower cover. The bright field transmission illumination system includes a transmission illumination optical unit. The transmitted illumination optical unit is provided so as to be movable between a regular position located above the stage and a jump position that is farther from the stage than the regular position.

  With the specimen cover in the open position and the transmitted illumination optical unit in the flipped-up position, the stage provided in the main body case is exposed to the space outside the main body case. In this state, the user can place the object on the stage. Further, the user can take out the object from the stage.

  On the other hand, in the state where the specimen cover is in the closed position and the transmission illumination optical unit is in the normal position, the periphery of the stage is in a dark room state. Thereby, at the time of fluorescence observation, weak fluorescence emitted from the object is guided to the imaging system.

JP 2006-162664 A

  As described above, in the inverted fluorescent microscope of Patent Document 1, the epi-illumination system and the power supply unit are provided inside the main body case. The epi-illumination system includes a mercury lamp for generating light used for fluorescence observation. When the power supply unit is activated during the fluorescence observation and the mercury lamp is driven, a large amount of heat is generated in the main body case. A large amount of heat generated in the main body case causes a temperature drift in each component in the main body case.

  Therefore, the inverted fluorescent microscope of Patent Document 1 is provided with an electric fan unit that discharges heat generated in the main body case to the outside of the main body case. When the size of the main body case is large, heat generated by the power supply unit and the mercury lamp is easily diffused into the space inside the main body case. Therefore, in order to sufficiently discharge the heat in the main body case, a large electric fan unit having a high heat exhaust capability is required, and the exhaust air cooling passage formed in the main body case is also increased in size. In this case, the main body case becomes larger and the entire inverted fluorescence microscope becomes larger.

  An object of the present invention is to provide a microscope apparatus that can perform highly reliable fluorescence observation without using a darkroom and is miniaturized.

  (1) A microscope apparatus according to the present invention includes a stage on which an object is placed, a first light source device that generates first light, and an object placed on the stage by the first light source device. A first optical device that irradiates the generated first light, a second light source device that generates the second light, and a second light source device that is generated on the object placed on the stage by the second light source device. The second optical device that irradiates the second light and the first light that passes through the object when the object is irradiated with the first light is used to image the object, and the object is An imaging device configured to receive an image of the object by receiving fluorescence emitted from the object when the second light is irradiated, and a second light directed from the second optical device toward the object. Condenses and directs the first light transmitted through the object and the fluorescence emitted from the object to the imaging device. An optical system, an imaging visual field position moving unit that moves the imaging visual field position of the object by the imaging device, and at least one of the object and the optical system by moving along the optical axis of the optical system. A focal position moving unit that moves the focal position of the optical system with respect to the object by changing a relative distance between the first light source device, the first optical device, and the second light source. Device, second optical device, imaging device, operation device for instructing operations of imaging visual field position moving unit and focus position moving unit, first light source device in response to command given from operating device, first An optical device, a second light source device, a second optical device, an imaging device, a control device that controls the imaging visual field position moving unit and the focal position moving unit, a stage, a second optical device, an imaging device, an optical system, Moving the field of view And a first housing that houses the focal position moving unit, a first light source device, a first optical device, a second light source device, a second optical device, an imaging device, an imaging visual field position moving unit, a focal position A power supply device that supplies power to the moving unit and the control device, and the first housing is configured to bring a space including the object mounted on the stage and the imaging device into a darkroom state, and the second housing The light source device, the power supply device, and the operation device are provided outside the first housing.

  In the microscope apparatus, an object is placed on a stage. The object placed on the stage is irradiated with the first light generated by the first light source device. In this case, the first light that passes through the object is guided to the imaging device by the optical system, and the object is imaged.

  Further, the object placed on the stage is irradiated with the second light generated by the second light source device while being condensed by the optical system. In this case, the fluorescence emitted from the object is guided to the imaging device by the optical system, and the object is imaged.

  The stage, the second optical device, the imaging device, the optical system, the imaging visual field position moving unit, and the focal position moving unit are accommodated in the first casing. In the first housing, the space including the object placed on the stage and the imaging device is in a dark room state. Thereby, by irradiating the object with the second light, highly reliable fluorescence observation can be performed without installing the microscope apparatus in the darkroom.

  The second light source device and the power supply device are provided outside the first housing. Accordingly, the first housing can be downsized, and the heat exhaust device and the air cooling passage for discharging the heat generated in the first housing to the outside of the housing can be downsized. As a result, the first housing can be further reduced in size, and the entire microscope apparatus can be reduced in size.

  The operating device is provided outside the first housing. When the control device responds to a command from the operation device, the operations of the second optical device, the imaging device, the imaging visual field position moving unit, and the focal position moving unit are controlled inside the first housing. Thereby, the user can move the imaging visual field position of the object by the imaging apparatus without moving the first housing, and can move the focal position of the optical system with respect to the object.

  (2) The first housing may further accommodate a control device.

  In this case, the first light source device, the first optical device, the second optical device, the imaging device, the imaging visual field position moving unit, the focal position moving unit, and the wiring for electrically connecting the control device are shortened. be able to. Thereby, since generation | occurrence | production of noise can be suppressed, it becomes possible to perform observation with higher reliability.

  (3) The microscope apparatus may further include a second casing provided separately from the first casing, and the second casing may house the second light source device and the power supply device.

  In this case, the second light source device and the power supply device can be handled integrally.

  (4) The second light source device may include at least one of a metal halide lamp, a mercury lamp, a xenon lamp, and a light emitting diode.

  Metal halide lamps, mercury lamps, and xenon lamps are high-intensity discharge lamps, and generally generate a large amount of heat during light emission. On the other hand, light emitting diodes are said to generate relatively little heat during light emission. Even in such a light emitting diode, high luminance is required to realize fluorescence observation. Therefore, when the light emitting diode is used for fluorescence observation, a large amount of heat is generated from the light emitting diode during the light emission. Also in these cases, since the second light source device is provided outside the first housing, the stage, the second optical device, the imaging device, the optical system, and the imaging visual field position moving unit are separated from the second light source device. Unaffected by the heat generated. Therefore, a decrease in observation reliability due to temperature drift is suppressed.

  (5) The microscope apparatus further includes an installation member, and a support that supports the stage, the second optical device, the imaging device, the optical system, the imaging visual field position moving unit, and the focal position moving unit. The first casing may be attached to the support body so as not to contact the installation member.

  In this case, the vibration generated on the installation surface on which the installation member is provided is absorbed by the elastic member. Further, since the first casing does not contact the installation member, vibration transmitted from the installation surface to the installation member is not transmitted to the first casing. Therefore, a decrease in observation reliability caused by vibration generated on the installation surface is suppressed.

  (6) The support body is disposed so as to face each other, and extends between the first and second plate-like members extending in one direction, and between the first plate-like member and the second plate-like member. 3rd, 4th and 5th plate-shaped members provided so as to connect the plate-shaped member and the second plate-shaped member, and the third, 4th and 5th plate-shaped members are The fourth plate member is provided at different positions so that the fourth plate member is positioned between the third and fifth plate members in one direction and the stage is integrated with the fourth plate member. May be provided.

  In this case, in one direction, the third and fifth plate members are provided so as to sandwich the fourth plate member that supports the stage. Thereby, compared with the case where the 1st plate-like member and the 2nd plate-like member are connected only by the 3rd and 5th plate-like members, the rigidity of the whole support body supports the stage. It is significantly improved by the plate-like member. Therefore, in order to ensure high rigidity, it is not necessary to increase the thickness of each plate-like member constituting the support. As a result, the support can be reduced in size and weight.

  (7) A light guide member that guides the second light generated by the second light source device to the second optical device is provided between the second light source device and the first housing. The housing has a back surface portion, and the second optical device includes an optical connection portion, a reflective member, a reflective light modulation element, and a filter device, and the optical connection portion can connect the light guide member through the back surface portion. The light guide member is supported so that the second light emitted from the light guide member is directed forward, and the reflection member is provided at a position in front of the optical connection portion and emitted from the light guide member. The second light is configured to reflect backward, and the reflective light modulation element is provided at a position behind the reflective member, and receives the second light reflected from the reflective member and receives the second light. The filter device is configured to reflect light toward the front while modulating the light. Provided in a position, it may be configured to direct the object mounted on the stage of light in a wavelength region predetermined in the second light reflected from the reflective optical modulation element.

  In this case, in the first housing, the second light emitted from the light guide member is reflected by the reflecting member and enters the reflective light modulation element. In the reflection type light modulation element, the second light reflected from the reflection member is reflected while being modulated. Of the reflected second light, light in a predetermined wavelength region is guided to the object by the filter device.

  In this way, the modulated second light can be applied to the object. Thereby, various observation images of the object can be generated based on the fluorescence emitted from the object.

  According to said structure, in order to make 2nd light inject into a reflective light modulation element, it is not necessary to bend a part of light guide member within a 1st housing | casing. Therefore, the enlargement of the first housing is suppressed. In addition, since the light guide member is connected through the back surface of the first housing, the user can use the space in front of the first housing as the work space.

  (8) The first housing is provided with an opening for maintenance of the second optical device and the optical system. The microscope device includes a lid member for opening and closing the opening of the first housing, and opening and closing of the lid member. An open / close detector for detecting a state, a light guiding state for guiding the second light generated by the second light source device to the second optical device, and a second light generated by the second light source device; A light-shielding device configured to be able to shift to a light-shielding state that is not guided to the optical device, and the control device includes the light-shielding device when the lid member switches from the closed state to the open state based on the detection of the open / close detector. The light guiding state may be shifted to the light shielding state.

  In this case, at the time of maintenance of the second optical device and the optical system, the second light is prevented from leaking from the inside of the first housing through the opening to the outside of the first housing. As a result, the second light is prevented from entering the eyes of the user.

  According to the present invention, it is possible to perform highly reliable fluorescence observation without using a darkroom, and the miniaturization of the microscope apparatus is realized.

1 is an external perspective view of a microscope apparatus according to an embodiment of the present invention. It is a figure for demonstrating the hardware constitutions of the microscope apparatus of FIG. It is a schematic diagram which shows the optical path in a measurement part and a measurement light supply part. It is a block diagram which shows the control system of the microscope apparatus of FIG. It is a block diagram which shows the structure of CPU. It is a schematic diagram showing fluorescence emitted from a measuring object when measuring light is irradiated to a measuring object. It is a figure which shows an example of the fluorescence observation image obtained by irradiating uniform measurement light to the predetermined observation range in a measurement object. It is a figure which shows an example of the sectioning observation method. It is an external appearance perspective view which shows the state of the measurement part at the time of replacement | exchange work of a measuring object, an objective lens, and a filter cube. It is a disassembled perspective view of the support body provided in the inside of a 1st housing | casing. It is a disassembled perspective view of the some component supported by the support body of FIG. It is a disassembled perspective view which shows the some support | pillar and base which support the support body of FIG. It is an external appearance perspective view which shows the attachment method of the housing | casing to the support body of FIG. It is a schematic diagram which shows the 1st structural example of the microscope apparatus which concerns on other embodiment. It is a schematic diagram which shows the 2nd structural example of the microscope apparatus which concerns on other embodiment. It is a schematic diagram which shows the 3rd structural example of the microscope apparatus which concerns on other embodiment. It is a schematic diagram which shows the 4th structural example of the microscope apparatus which concerns on other embodiment.

(1) Configuration of Microscope Device A microscope device according to an embodiment of the present invention will be described. In the microscope apparatus according to the present embodiment, it is possible to perform fluorescence observation of the measurement object. Further, in the microscope apparatus, bright field observation, dark field observation, phase difference observation, differential interference observation, declination observation, and polarization observation using light transmitted through the measurement object can be performed. In the following description, bright field observation, dark field observation, phase difference observation, differential interference observation, declination observation, and polarization observation using light transmitted through a measurement object are collectively referred to as transmission observation.

  FIG. 1 is an external perspective view of a microscope apparatus according to an embodiment of the present invention. As shown in FIG. 1, the microscope apparatus 500 of this example mainly includes a measurement unit 100, a PC (personal computer) 200, a measurement light supply unit 300, and a display unit 400. Measuring unit 100 and PC 200 are connected by wiring WR1. The measurement unit 100 and the measurement light supply unit 300 are connected by the wiring WR2 and the light guide member 330. In the example of FIG. 1, a single notebook personal computer is used as a configuration in which the PC 200 and the display unit 400 are integrated. As the wiring WR1, a LAN (local area network) cable, a USB (universal serial bus) cable, or the like is used. As the wiring WR2, a cable in which a power supply line and a signal line are integrated is used.

  FIG. 2 is a diagram for explaining a hardware configuration of the microscope apparatus 500 of FIG. In FIG. 2, longitudinal sections of the measurement unit 100 and the measurement light supply unit 300 are schematically shown.

  As shown in FIG. 2, the measurement light supply unit 300 includes a housing 301, a power supply device 310, a light projecting unit 320, and a heat exhaust device 340. The power supply device 310, the light projecting unit 320, and the heat exhaust device 340 are accommodated in the housing 301. The power supply device 310 supplies power to the light projecting unit 320 and also supplies power to the measuring unit 100 via the wiring WR2.

  An opening 309 for exhaust heat is formed in the housing 301 of the measurement light supply unit 300. Exhaust heat device 340 includes an exhaust heat fan and a motor that drives the exhaust heat fan, and exhausts heat generated by power supply device 310 and light projecting unit 320 to the outside of housing 301 through opening 309.

  The measuring unit 100 includes a housing 101, a plurality of support columns 106, a pedestal 107, a pattern applying unit 110, a light receiving unit 120, a transmitted light supply unit 130, a stage 140, a stage driving device 141, a filter unit 150, a lens unit 160, and a control board. 170 is included.

  The housing 101 accommodates a pattern applying unit 110, a light receiving unit 120, a transmitted light supply unit 130, a stage 140, a stage driving device 141, a filter unit 150, a lens unit 160, a control board 170, and driving circuits (not shown) of each component. To do. The housing 101 includes a front surface portion 101a, a back surface portion 101b, one side surface portion 101c (FIG. 1), another side surface portion 101d (FIG. 1), and an upper surface portion 101e. An upper surface lid 102 is attached to the upper surface portion 101e. A front cover 103 is attached to the center of the front part 101a. With the top cover 102 and the front cover 103 closed, the space including the stage 140 and the light receiving unit 120 is in a dark room state.

  In the housing 101, the stage frame 11, the base frame 12, and the upper frame 16 are provided so as to be lined up and down and parallel to the horizontal plane. The stage frame 11, the base frame 12, and the upper frame 16 constitute a part of a support body to be described later, and the support body is supported by a plurality of support columns 106 and a pedestal 107. The stage frame 11 is located above the base frame 12, and the upper frame 16 is located above the stage frame 11. The stage frame 11 and the base frame 12 are formed with openings for allowing light used for observing the measurement object S to pass therethrough.

  An opening is also formed at the center of the stage 140. The stage 140 is integrally attached to the upper surface of the stage frame 11 so that the openings of the stage frame 11 and the stage 140 overlap each other. Further, a stage driving device 141 is attached to the upper surface of the stage frame 11 together with the stage 140.

  On the stage 140, the measuring object S is placed using a petri dish or a preparation. A plane on the stage 140 on which the measurement object S is placed is called a placement surface. In this example, the placement surface is maintained parallel to the horizontal plane.

  In the following description, as indicated by arrows in FIG. 2, two directions orthogonal to each other on the placement surface are defined as an X direction and a Y direction, and a direction perpendicular to the placement surface is defined as a Z direction. The X direction corresponds to the horizontal direction of the measurement unit 100, the Y direction corresponds to the front-rear direction of the measurement unit 100, and the Z direction corresponds to the vertical direction of the measurement unit 100. The stage 140 is an XY stage and is configured to be movable in the X direction and the Y direction. The stage driving device 141 moves the stage 140 in the X direction and the Y direction in response to a signal given from the control board 170.

  In this example, the measuring object S is a biological specimen containing various proteins. When performing fluorescence observation, the measurement object S is coated with a fluorescent reagent that is fused to a specific protein. Fluorescent reagents include, for example, GFP (green fluorescent protein), TexasRed (Texas Red) and DAPI (diamidinophenylindole). GFP absorbs light near a wavelength of 490 nm and emits light near a wavelength of 510 nm. TexasRed absorbs light near a wavelength of 596 nm and emits light near a wavelength of 620 nm. DAPI absorbs light near a wavelength of 345 nm and emits light near a wavelength of 455 nm.

  The pattern applying unit 110, the filter unit 150, and the lens unit 160 are attached to the upper surface of the base frame 12, and the light receiving unit 120 is provided below the base frame 12.

  On the base frame 12, the filter unit 150 is disposed at a position in front of the pattern applying unit 110. A transmitted light supply unit 130 is attached to the upper surface of the upper frame 16.

  FIG. 3 is a schematic diagram illustrating optical paths in the measurement unit 100 and the measurement light supply unit 300. In FIG. 3, the details of each component of the measurement unit 100 and the measurement light supply unit 300 are shown along with the optical path. In FIG. 3, as in FIG. 2, the X direction, the Y direction, and the Z direction are indicated by arrows.

  As shown in FIG. 3, the light projecting unit 320 of the measurement light supply unit 300 includes a measurement light source 321, a light reduction mechanism 322, a light shielding mechanism 323, and an optical connector 324. One end of the light guide member 330 is connected to the optical connector 324. In this example, the light guide member 330 is a liquid light guide. The light guide member 330 may be, for example, a glass fiber or a quartz fiber.

  The measurement light source 321 is, for example, a metal halide lamp. The measurement light source 321 may be another light source such as a mercury lamp, a xenon lamp, or a white LED (light emitting diode). The measurement light source 321 is used as a light source for fluorescence observation of the measurement object S. Hereinafter, the light emitted from the measurement light source 321 is referred to as measurement light.

  The light shielding mechanism 323 is, for example, a mechanical shutter. The light shielding mechanism 323 is disposed so as to be positioned on the optical path of the measurement light emitted from the measurement light source 321. When the light shielding mechanism 323 is in the light guide state, the measurement light passes through the light shielding mechanism 323 and enters one end of the light guide member 330. On the other hand, when the light blocking mechanism 323 is in the light blocking state, the measurement light is blocked and does not enter one end of the light guide member 330. The light blocking mechanism 323 may include a light modulation element that can switch between passing and blocking of the measurement light. The light shielding mechanism 323 is in a light guiding state when fluorescence observation of the measurement object S is performed, and is in a light shielding state when fluorescence observation of the measurement object S is not performed.

  The dimming mechanism 322 includes a plurality of ND (Neutral Density) filters having different transmittances. The dimming mechanism 322 is arranged so that any one of the plurality of ND filters is positioned on the optical path of the measurement light that has passed through the light shielding mechanism 323. By selectively switching the ND filter located on the optical path of the measurement light, the intensity of the measurement light passing through the dimming mechanism 322 can be adjusted. The dimming mechanism 322 may include a light modulation element capable of adjusting the intensity of the measurement light instead of the plurality of ND filters.

  The pattern providing unit 110 of the measurement unit 100 includes an optical connector 111, a light modulation element 112, and a plurality (two in this example) of mirrors 113. The other end of the light guide member 330 is connected to the optical connector 111 from the rear of the measurement unit 100 through the back surface 101b. The optical connector 111 supports the other end of the light guide member 330 so that measurement light guided by the light guide member 330 during fluorescence observation is emitted from the rear of the measurement unit 100 toward the front.

  One mirror 113 is disposed in front of the optical connector 111 so as to be inclined with respect to the XY plane. Further, the other mirror 113 is disposed above the one mirror 113 so as to be inclined with respect to the XY plane. Further, the light modulation element 112 is disposed at a position behind the two mirrors 113 and above the two mirrors 113. The measurement light emitted from the other end of the optical connector 111 is reflected by the one mirror 113 so as to be directed upward from below the measurement unit 100. Further, the measurement light reflected by one mirror 113 is reflected by the other mirror 113 so as to go from diagonally forward to diagonally upward and enter the light modulation element 112.

  The light modulation element 112 is, for example, a DMD (digital micromirror device). The light modulation element 112 may be an LCOS (Liquid Crystal on Silicon). The light incident on the light modulation element 112 is converted into a preset pattern and a preset intensity (brightness) by a pattern generation unit 212 described later, and is reflected so as to go from the rear to the front of the measurement unit 100. The The measurement light reflected by the light modulation element 112 of the pattern application unit 110 passes through a light projection lens (not shown) and enters the filter unit 150.

  As described above, in this example, a reflection type device such as DMD or LCOS is used as the light modulation element 112. However, the light modulation element 112 may be a transmissive device such as an LCD (liquid crystal display) instead of the reflective device.

  The filter unit 150 includes a plurality of filter cubes 151, a filter turret 152, and a filter turret driver 153. Each filter cube 151 includes a frame 151a, an excitation filter 151b, a dichroic mirror 151c, and an absorption filter 151d. The frame 151a is a cubic member that supports the excitation filter 151b, the dichroic mirror 151c, and the absorption filter 151d.

  The plurality of filter cubes 151 respectively correspond to a plurality of types of fluorescent reagents used for fluorescence observation of the measurement object S. For example, when the above GFP, TexasRed, and DAPI are used as the fluorescent reagent, three types of filter cubes 151 corresponding to GFP, TexasRed, and DAPI are provided in the filter unit 150.

  In this case, the excitation filter 151b of the filter cube 151 corresponding to GFP passes light near the wavelength of 490 nm, and the absorption filter 151d passes light near the wavelength of 510 nm. The dichroic mirror 151c reflects light having a wavelength of about 490 nm and transmits light having a wavelength of about 510 nm.

  Further, the excitation filter 151b of the filter cube 151 corresponding to TexasRed passes light near 596 nm, and the absorption filter 151d passes light near 620 nm. The dichroic mirror 151c reflects light having a wavelength of about 596 nm and transmits light having a wavelength of about 620 nm.

  Further, the excitation filter 151b of the filter cube 151 corresponding to DAPI passes light near 345 nm, and the absorption filter 151d passes light near 455 nm. The dichroic mirror 151c reflects light having a wavelength of about 345 nm and allows light having a wavelength of about 455 nm to pass therethrough.

  The filter turret 152 has a disk shape. The filter turret 152 is provided with four through holes at an angular interval of 90 ° with respect to the central axis. The filter turret 152 is rotatably supported around the central axis of the filter turret 152 on the base frame 12 of FIG. When the filter turret driving unit 153 drives the filter turret 152, the filter turret 152 rotates.

  Each of the plurality of filter cubes 151 is provided on the filter turret 152 such that the excitation filter 151b is directed in a direction opposite to the central axis of the filter turret 152 and overlaps one of the four through holes.

  In the present embodiment, three filter cubes 151 corresponding to the three types of fluorescent reagents are attached on the filter turret 152 so as to overlap the three through holes of the filter turret 152.

  The user can select a desired filter cube 151 by rotating the filter turret 152 during fluorescence observation. The selected filter cube 151 is arranged on the observation axis OA parallel to the Z direction passing through the measurement object S on the stage 140 so that the measurement light is incident on the excitation filter 151b. The observation axis OA coincides with the optical axis of the light receiving unit 120 extending in the Z direction. When the measurement light is incident on the excitation filter 151b, a part of the wavelength region of the measurement light passes through the excitation filter 151b. The light that has passed through the excitation filter 151b is reflected upward by the dichroic mirror 151c.

  As described above, in this embodiment, three filter cubes 151 are attached to the three through holes of the filter turret 152, and the filter cube 151 is not attached to the remaining one through hole. Therefore, it is possible to perform bright field observation without using the filter cube 151 by arranging a through hole to which the filter cube 151 is not attached on the observation axis OA (optical path of measurement light).

  The lens unit 160 includes a plurality of objective lenses 161, a lens turret 162, a focal length adjustment mechanism 163, a lens support plate 163p, a lens turret drive unit 164, and a focal length adjustment drive unit 165. The plurality of objective lenses 161 have different magnifications.

  A focal length adjustment mechanism 163 is mounted on the base frame 12 of FIG. The focal length adjustment mechanism 163 supports the lens support plate 163p so as to be parallel to the XY plane and movable in the Z direction.

  A lens turret 162 and a lens turret driving unit 164 are provided on the lens support plate 163p. The lens turret 162 has a disk shape. The lens turret 162 is formed with six through holes at an angular interval of 60 ° with respect to the central axis. The lens turret 162 is supported so as to be rotatable around the central axis of the lens turret 162. When the lens turret driving unit 164 drives the lens turret 162, the lens turret 162 rotates.

  In the present embodiment, six objective lenses 161 having different magnifications are attached on the lens turret 162 so as to overlap the six through holes of the lens turret 162. In this state, the six objective lenses 161 are located below the stage 140 and above the filter unit 150.

  The user can select one objective lens 161 used for observation of the measuring object S by rotating the lens turret 162. The selected objective lens 161 is disposed on the observation axis OA.

  The focal length adjustment driving unit 165 drives the focal length adjustment mechanism 163 to move the lens support plate 163p in the Z direction. Thereby, the relative distance in the Z direction between the measuring object S on the stage 140 and the selected objective lens 161 is adjusted.

  During fluorescence observation of the measurement object S, the measurement light reflected by the dichroic mirror 151c of the filter cube 151 passes through the opening of the stage 140 while being condensed by the selected objective lens 161, and is irradiated on the measurement object S. The When the measurement object S is irradiated with measurement light, fluorescence is emitted from the fluorescent reagent applied to the measurement object S. The fluorescence emitted below the measuring object S passes through the selected objective lens 161, the filter cube 151 located below the objective lens 161 and the opening formed in the base frame 12, and enters the light receiving unit 120. Incident.

  The transmitted light supply unit 130 attached on the upper frame 16 of FIG. 2 includes a fixed unit 130A and a swinging unit 130B. The fixing unit 130A includes a transmission light source 131, a diaphragm adjustment unit 132, and a transmission optical system 133, and is fixed on the upper surface of the upper frame 16 (FIG. 2).

  The transmissive light source 131 is, for example, a white LED. The transmissive light source 131 may be another light source such as a halogen lamp. The transmission light source 131 is used as a light source for transmission observation of the measurement object S. Hereinafter, the light emitted from the transmissive light source 131 is referred to as transmitted light. The aperture adjustment unit 132 includes an aperture and a phase difference slit, and is used for adjusting the brightness of the transmitted light irradiated on the measuring object S and for performing phase difference observation using the transmitted light. The transmissive optical system 133 includes a relay lens and guides the transmitted light emitted from the transmissive light source 131 to the front of the fixed portion 130A. The relay lens is arranged such that a conjugate relationship is maintained between the diaphragm and phase difference slit included in the diaphragm adjustment unit 132 and the front focal position of the condenser lens 135.

  The swing part 130B includes a mirror 134, a condenser lens 135, and a swing mechanism (not shown). The swinging portion 130B is configured to be swingable about an axis parallel to the X direction in a state where the upper surface lid 102 of FIG. 2 is opened. As a result, the swinging unit 130B has a normal position where the condenser lens 135 faces the measurement object S on the stage 140 in the Z direction and a separation position where the condenser lens 135 does not face the measurement object S on the stage 140 in the Z direction. Move between.

  At the time of transmission observation of the measuring object S, the rocking part 130B is manually moved to the normal position by the user. In a state where the swinging part 130B is in the normal position, the transmitted light guided forward from the fixed part 130A is reflected downward by the mirror 134 and passes through the measuring object S on the stage 140 through the condenser lens 135.

  As described above, in the filter unit 150, the through hole of the filter turret 152 that is not provided with the filter cube 151 is selected during transmission observation. In this case, the transmitted light that has passed through the measuring object S enters the light receiving unit 120 through the objective lens 161 selected by the user, the through hole of the filter turret 152, and the opening formed in the base frame 12.

  The light receiving unit 120 includes a camera 121, a color filter 122, and an imaging lens 123. The camera 121 is, for example, a CCD (charge coupled device) camera including an image sensor. The image sensor is, for example, a monochrome CCD. The imaging device may be another imaging device such as a CMOS (complementary metal oxide semiconductor) image sensor.

  The fluorescence or transmitted light incident on the light receiving unit 120 is collected and imaged by the imaging lens 123 and then received by the camera 121. Thereby, the image of the measuring object S is obtained. From each pixel of the image sensor of the camera 121, an analog electric signal (hereinafter referred to as a light reception signal) corresponding to the amount of received light is output to the control board 170.

  Unlike a color CCD, a monochrome CCD does not need to be provided with pixels that receive red wavelength light, pixels that receive green wavelength light, and pixels that receive blue wavelength light. Therefore, the measurement resolution of the monochrome CCD is higher than the resolution of the color CCD. Further, unlike a color CCD, a monochrome CCD does not require a color filter for each pixel. Therefore, the sensitivity of the monochrome CCD is higher than that of the color CCD. For these reasons, the camera 121 in this example is provided with a monochrome CCD. On the other hand, when the color CCD has sufficient resolution and sensitivity, the image sensor may be a color CCD.

  On the control board 170, an A / D converter (analog / digital converter) and a FIFO (First In First Out) memory (not shown) are mounted. The light reception signal output from the camera 121 is sampled at a constant sampling period by the A / D converter and converted into a digital signal based on control by the PC 200. Digital signals output from the A / D converter are sequentially stored in the FIFO memory. The digital signals stored in the FIFO memory are sequentially transferred to the PC 200 as pixel data.

(2) Control System of Microscope Device FIG. 4 is a block diagram showing a control system of the microscope device 500 of FIG. In the measurement unit 100, the control board 170 performs operations of the pattern applying unit 110, the light receiving unit 120, the transmitted light supply unit 130, the stage driving device 141, the filter unit 150, and the lens unit 160 in response to a command given from the PC 200. Control. Further, the control board 170 transfers a digital signal based on the light reception signal output from the camera 121 (FIG. 3) to the PC 200 as described above. Further, the control board 170 controls the operations of the power supply device 310 and the light projecting unit 320 of the measurement light supply unit 300 in response to a command from the PC 200.

  The PC 200 includes a CPU (Central Processing Unit) 210, a ROM (Read Only Memory) 220, a RAM (Random Access Memory) 230, a storage device 240, and an operation unit 250. The operation unit 250 is configured to be operated by a user to give a command to the control board 170 of the measurement unit 100, and includes a keyboard and a pointing device. A mouse or a joystick is used as the pointing device.

  The display unit 400 is configured by, for example, an LCD panel or an organic EL (electroluminescence) panel. The ROM 220 stores a system program. The RAM 230 stores various data and functions as a work area for the CPU 210. The storage device 240 is composed of a hard disk or the like. The storage device 240 stores a control program for the microscope apparatus 500 and an image processing program. The storage device 240 is used for storing various data such as pixel data provided from the measurement unit 100.

  FIG. 5 is a block diagram showing the configuration of the CPU 210. As illustrated in FIG. 5, the CPU 210 includes an image data generation unit 211, a pattern generation unit 212, and a control unit 213. The image data generation unit 211 generates image data based on the pixel data provided from the measurement unit 100 by executing the control program and the image processing program. Image data is a set of a plurality of pixel data.

  The pattern generation unit 212 generates a pattern to be irradiated on the measurement object as the pattern of measurement light emitted from the light modulation element 112 in FIG. The control unit 213 irradiates the measurement object S with measurement light having a predetermined pattern by controlling the light modulation element 112 via the control board 170 in FIG. 2 based on the pattern generated by the pattern generation unit 212. While shifting the phase of the pattern.

  Further, the control unit 213 gives instructions to the control board 170 to control the operations of the light receiving unit 120, the transmitted light supply unit 130, the stage 140, the filter unit 150, the lens unit 160, and the light projecting unit 320 via the control board 170. Control. Further, the control unit 213 performs various processes on the generated image data using the RAM 230 and causes the display unit 400 to display an image based on the image data.

(3) Function of pattern imparting unit FIG. 6 is a schematic diagram illustrating fluorescence emitted from the measurement object S when the measurement object S is irradiated with measurement light. As shown in FIG. 6, when the observation target portion sp1 of the measurement target S is arranged at the focal point of the objective lens 161, the measurement light passing through the objective lens 161 is the observation target portion of the measurement target S as indicated by a dotted line. Condensed toward sp1. Thereby, when a fluorescent reagent is present in the observation target portion sp1 of the measurement object S, fluorescence is emitted from the observation target portion sp1. A part of the fluorescence enters the light receiving unit 120 (FIG. 3) through the objective lens 161 as indicated by a thick arrow.

  When the measurement light is focused on the observation target portion sp1 of the measurement target S by the objective lens 161, the fluorescent reagent existing at a position out of the focus of the objective lens 161 also receives the measurement light and emits fluorescence. . In the example of FIG. 6, fluorescence is also emitted from portions sp2 and sp3 other than the observation target portion sp1. When the portions sp2 and sp3 are located at positions away from the depth of field DF of the objective lens 161, the fluorescence emitted from the portions sp2 and sp3 enters the light receiving unit 120 as stray light.

  FIG. 7 is a diagram illustrating an example of a fluorescence observation image obtained by irradiating a predetermined observation range on the measurement object S with uniform measurement light. For the above reason, when the measurement light is uniformly irradiated to the predetermined observation range in the measurement object S, as shown in FIG. 7, the fluorescence emitted from the position outside the depth of field DF of the objective lens 161 is stray light. Is incident on the light receiving unit 120 to reduce the overall contrast of the fluorescence observation image.

  Therefore, in the measurement unit 100 of this example, in order to obtain a fluorescence observation image with high contrast, sectioning observation using patterned measurement light described below is performed.

  In sectioning observation, the measurement object S is irradiated with measurement light having a predetermined pattern, and the spatial phase of the predetermined pattern is moved by a certain amount. The measurement light having a predetermined pattern has an intensity that periodically changes in one direction (for example, the Y direction) or two directions (for example, the X direction and the Y direction) on the XY plane, for example. Hereinafter, measurement light having a pattern is referred to as pattern measurement light.

  The pattern of the pattern measurement light and its phase are controlled by the light modulation element 112. Here, the portion of the pattern measurement light having an intensity equal to or higher than a predetermined value is called a bright portion, and the portion of the pattern measurement light having an intensity smaller than the predetermined value is called a dark portion.

  The pattern measurement light includes, for example, a plurality of linear bright portions that are parallel to one direction (for example, the X direction) and arranged at substantially equal intervals in another direction (for example, the Y direction) orthogonal to the one direction. Striped measurement light having a cross section including a plurality of linear dark portions between the portions can be used.

  FIG. 8 is a diagram illustrating an example of the sectioning observation method. In sectioning observation, as shown in FIGS. 8A to 8D, the bright part of the pattern measurement light (in this example, the striped measurement light) is irradiated at least once on all parts of the measurement object S. The phase of the pattern measurement light pattern is shifted by a certain amount. Further, every time the phase of the pattern is moved by a certain amount, the fluorescence emitted from the measuring object S is detected. Thereby, a plurality of image data of the measuring object S is generated.

  Hereinafter, the image data obtained when the measurement object S is irradiated with the pattern measurement light is referred to as pattern image data. An image based on the pattern image data is called a pattern image.

  In each pattern image data, pixel data corresponding to a bright portion of the pattern measurement light has a high value (luminance value), and pixel data corresponding to a dark portion of the pattern measurement light has a low value (luminance value). Therefore, as indicated by arrows in FIGS. 8A to 8D, in each pattern image, the pixel portion corresponding to the bright portion of the pattern measurement light is bright and the pixel portion corresponding to the dark portion of the pattern measurement light. Is dark.

  Here, each pattern image is affected by stray light. In order to obtain image data from which the influence of stray light has been removed, the following processing is executed.

  First, a component (hereinafter referred to as a focusing component) representing the degree of contrast is calculated from the plurality of pattern image data using the values of the plurality of pixel data for each pixel. Image data is generated by stitching together pixels having in-focus components. The image data generated in this way is called sectioning image data. An image based on the sectioning image data is called a sectioning image.

  In the pattern image data generated using the above-described striped measurement light, the focus component is, for example, the difference between the maximum value (maximum luminance value) and the minimum value (minimum luminance value) of the pixel data, or the pixel data The standard deviation of values.

  In this example, for each pixel, the difference between the pixel data of the pattern image data when the bright part and the dark part of the pattern measurement light are irradiated is calculated as a focusing component. Sectioning image data is generated by connecting the focus components for all the calculated pixels. For each pixel, the value of the pixel data of the pattern image data when the dark portion of the pattern measurement light is irradiated corresponds to the stray light component. Therefore, sectioning image data from which the influence of stray light is removed can be obtained. As a result, as shown in FIG. 8E, it is possible to obtain a fluorescence observation image (sectioning image) with high contrast from which the influence of stray light is removed.

  As the pattern measurement light, for example, sine wave measurement light having a cross section including a pattern that is parallel to the X direction and whose intensity changes in a sine wave shape in the Y direction can be used instead of the striped measurement light. In pattern image data generated using sinusoidal measurement light, the focus component is, for example, the amplitude (peak to peak) of pixel data. Further, as the pattern measurement light, instead of the striped measurement light, dot-shaped measurement light having a cross section including a plurality of dot-like bright portions arranged at substantially equal intervals in the X direction and the Y direction can be used. In the pattern image data generated using the dot-shaped measurement light, the focus component is, for example, the difference between the maximum value (maximum luminance value) and the minimum value (minimum luminance value) of the pixel data, or the value of the pixel data. Standard deviation.

(4) Replacement Operation of Measurement Object, Objective Lens, and Filter Cube FIG. 9 is an external perspective view showing the state of the measurement unit 100 during the replacement operation of the measurement object S, the objective lens 161, and the filter cube 151. As shown in FIG. 9A, an upper opening OP <b> 1 is formed in the housing 101 of the measurement unit 100. An upper surface lid 102 capable of opening and closing the upper opening OP1 is attached to a part of the upper surface portion 101e of the housing 101 via a hinge 102H (FIG. 2). When the measurement object S placed on the stage 140 is exchanged, the upper surface lid 102 is opened as shown by a thick arrow in FIG. Thereafter, as indicated by a thick arrow in FIG. 9B, the swinging part 130B of the transmitted light supply part 130 is manually swung from the normal position np1 to the separation position cp1. Thereby, the user can easily perform the exchange operation of the measuring object S from the front of the measuring unit 100 through the upper opening OP1.

  A swing part detector S1 is provided in the vicinity of the fixed part 130A. The swing part detector S1 is a magnetic sensor including, for example, a magnet and a Hall element. The swing part detector S1 detects whether or not the swing part 130B is at the normal position np1, and gives a detection result to the control board 170 (FIG. 2). When the swinging part 130B is not in the normal position np1, the control board 170 puts the light shielding mechanism 323 (FIG. 3) of the light projecting part 320 (FIG. 3) into a light shielding state, and transmits the light source 131 (FIG. ) Stop supplying power to. Accordingly, it is possible to prevent the measurement light from being irradiated onto the stage 140 and the transmitted light from being emitted to the front of the measurement unit 100 from the fixed unit 130A.

  The control board 170 controls the light modulation element 112 of the pattern applying unit 110 instead of putting the light shielding mechanism 323 (FIG. 3) in a light shielding state when the swinging unit 130B is not in the normal position np1. The measurement light may be prevented from being irradiated toward the stage 140.

  The housing 101 further has a front opening OP2. A front lid 103 capable of opening and closing the front opening OP2 is attached to a part of the front surface portion 101a of the housing 101 via a hinge 103H (FIG. 2).

  When the objective lens 161 attached to the lens turret 162 in FIG. 3 is exchanged, the front cover 103 is manually opened as shown by a thick dotted line arrow in FIG. In addition, the front lid 103 is also manually opened when the filter cube 151 attached to the filter turret 152 of FIG. 3 is replaced. In these cases, the user can easily replace the objective lens 161 and the filter cube 151 from the front of the measurement unit 100 through the front opening OP2.

  In the vicinity of the front cover 103, a front cover detector S2 is provided. The front lid detector S2 is a magnetic sensor including, for example, a magnet and a Hall element, detects the open / closed state of the front lid 103, and gives the detection result to the control board 170. The control board 170 switches the light shielding mechanism 323 (FIG. 3) of the light projecting unit 320 (FIG. 3) from the light guiding state to the light shielding state when the front cover 103 is switched from the closed state to the open state based on the detection result. . Further, the control board 170 stops supplying power from the power supply device 310 to the transmissive light source 131 (FIG. 3). This prevents the measurement light from being emitted from the pattern applying unit 110 (FIG. 3) toward the filter unit 150 and the transmitted light from being emitted from the transmitted light supply unit 130 (FIG. 3) to the filter unit 150. The

  The control board 170 controls the light modulation element 112 of the pattern applying unit 110 instead of putting the light shielding mechanism 323 (FIG. 3) in the light shielding state when the front cover 103 is switched from the closed state to the open state. Thus, the measurement light may be prevented from being emitted toward the filter unit 150.

  In this way, the measurement light and the transmitted light are prevented from leaking out of the housing 101. Therefore, the measurement light and the transmitted light are prevented from entering the eyes of the user.

  The top cover 102 and the front cover 103 are manually closed by the user. In this case, light is prevented from entering the measurement object S and the light receiving unit 120 on the stage 140 from the outside of the housing 101. Thereby, as described above, the space including the measurement object S and the light receiving unit 120 placed on the placement surface of the stage 140 is in a dark room state. Therefore, only fluorescence emitted from the measuring object S can be incident on the light receiving unit 120 during fluorescence observation.

(5) Details of Structure of Measuring Unit (5-1) Frame Structure Inside the first casing 101 of FIG. 2, the pattern applying unit 110, the light receiving unit 120, the transmitted light supply unit 130, the stage 140, and the stage The driving device 141, the filter unit 150, and the lens unit 160 are supported by a support including the stage frame 11, the base frame 12, and the upper frame 16.

  FIG. 10 is an exploded perspective view of a support provided inside the first housing 101. In FIG. 10 and FIGS. 11 to 13 described later, the X direction, the Y direction, and the Z direction are indicated by arrows as in FIG.

  As shown in FIG. 10, the support 10 includes a stage frame 11, a base frame 12, two side frames 13, two reinforcing frames 14, a back frame 15, and an upper frame 16. Each frame is a plate-like member made of a metal material such as an aluminum alloy. Each frame may be made of resin.

  The base frame 12 is a plate-like member having one side and the other side, and is arranged in parallel to the XY plane. One side and the other side of the base frame 12 are orthogonal to the X direction. A rectangular cutout is formed from the rear at the center of the base frame 12 in the X direction.

  The two side frames 13 are plate-like members having the same shape, and are attached to the vicinity of one side and the other side of the upper surface of the base frame 12 so as to be parallel to the YZ plane and face each other. The front upper end portion of each side frame 13 is lower than the rear upper end portion.

  The two reinforcing frames 14 are attached between the two side frames 13 so as to be parallel to the XZ plane and face each other. The back frame 15 is attached to the rear ends of the two side frames 13 so as to be parallel to the XZ plane.

  The stage frame 11 is a plate-like member, and is attached to the front upper end portions of the two side frames 13 so as to be parallel to the XY plane. The upper frame 16 is a plate-like member and is attached to upper rear portions of the two side frames 13 so as to be parallel to the XY plane. Thus, the support body 10 is produced by connecting the plurality of frames 11 to 16. A control board 170 is attached to the back frame 15 from the back of the support 10.

  In the support body 10 of this example, two side frames 13 parallel to the YZ plane have three frames (stage frame 11, base frame 12 and upper frame 16 parallel to the XY plane at three different positions in the Z direction. ). Accordingly, the two side frames 13 are connected by two frames (any two of the stage frame 11, the base frame 12, and the upper frame 16) parallel to the XY plane at two different positions in the Z direction. Compared to the case, the rigidity of the support 10 is remarkably improved. Therefore, in order to ensure high rigidity, it is not necessary to increase the thickness of each frame 11-16 which comprises the support body 10. FIG. As a result, the support 10 can be reduced in size and weight.

(5-2) A plurality of components supported by the support body Fig. 11 is an exploded perspective view of a plurality of components supported by the support body 10 of Fig. 10. As shown in FIG. 11, a storage box 120 </ b> B whose upper surface is open is attached to the lower surface of the base frame 12. The light receiving unit 120 is housed in the housing box 120B. In addition, in the storage box 120B, a drive circuit and a heat exhaust fan (not shown) are stored together with the light receiving unit 120.

  The pattern applying unit 110 is attached to a portion of the base frame 12 where the notch is formed. A filter unit 150 is attached to the upper surface of the base frame 12 at a position in front of the pattern applying unit 110. Further, a focal length adjustment mechanism 163 of the lens unit 160 is attached to one side of the filter unit 150 on the upper surface of the base frame 12. In this state, the lens turret 162 of the lens unit 160 is supported so as to be movable in the Z direction by the focal length adjustment mechanism 163 at a position above the filter turret 152. A transmitted light supply unit 130 is attached on the upper surface of the upper frame 16.

(5-3) A plurality of columns, pedestals, and housings FIG. 12 is an exploded perspective view showing a plurality of columns 106 and pedestals 107 that support the support 10 of FIG. As shown in FIG. 12, the base 107 is a plate-like member having a substantially rectangular shape. Four support columns 106 are attached to the four corners of the base 107 so as to extend upward. An elastic member 106 d is attached to the upper end of each column 106.

  Each elastic member 106d has a configuration in which a coiled spring is covered with a gel material. As the gel material, polyurethane gel can be used. The gel material preferably has a low hardness.

  The support 10 is placed on the four elastic members 106d. In this state, the two front elastic members 106 d and the support column 106 support the front portion of the lower surface of the stage frame 11. Further, the rear two elastic members 106 d and the support column 106 support the lower end portion of the back frame 15. A connecting member (not shown) having a lower surface with which the two elastic members 106d abut is attached to the lower end portion of the back frame 15 in advance.

  As described above, the housing 101 is further attached to the support body 10 in a state where the support body 10 is supported by the four support columns 106, the four elastic members 106 d, and the pedestal 107.

  FIG. 13 is an external perspective view showing a method of attaching the housing 101 to the support 10 of FIG. As shown in FIG. 13, the front surface portion 101 a of the housing 101 is attached to the front portion of the support 10 using screws or the like, and the back surface portion 101 b of the housing 101 is attached to the rear portion of the support 10 using screws or the like. It is attached. Further, one side surface portion 101c of the housing 101 is attached to one side portion of the support body 10 using a screw or the like, and the other side surface portion 101d of the housing 101 is attached to the other side portion of the support body 10 using a screw or the like. Attached. Further, the upper surface portion 101e of the housing 101 is attached to the upper portion of the support 10 using screws or the like.

  In this way, the housing 101 is formed by attaching the front surface portion 101a, the back surface portion 101b, the one side surface portion 101c, the other side surface portion 101d, and the upper surface portion 101e to the support body 10. In this state, the casing 101 is supported by the support body 10 without contacting any of the four columns 106, the four elastic members 106d, and the pedestal 107.

  As described above, in the measurement unit 100 of this example, the support 10 (FIG. 2), the casing 101 (FIG. 2), the pattern application unit 110 (FIG. 2), the light receiving unit 120 (FIG. 2), and the transmitted light supply The unit 130 (FIG. 2), the stage 140 (FIG. 2), the stage driving device 141 (FIG. 2), the filter unit 150 (FIG. 2), and the lens unit 160 (FIG. 2) are divided into four via four elastic members 106d. It is supported by the column 106 and the base 107.

  Thereby, even when vibration occurs on the installation surface on which the measurement unit 100 is installed, the vibration transmitted from the installation surface to the base 107 and the four columns 106 is absorbed by the four elastic members 106d, and the support 10 Not transmitted to. Therefore, a decrease in observation reliability due to vibrations generated outside the measurement unit 100 is suppressed.

  As shown in FIGS. 2 and 3, the housing 101 includes a light modulation element 112 of the pattern applying unit 110, a camera 121 of the light receiving unit 120, a transmissive light source 131 of the transmitted light supply unit 130, and a control board 170 as heat sources. Exists. Therefore, the support 10 of FIG. 11 has a heat exhaust device (not shown) for exhausting heat generated from the light modulation element 112, the camera 121, the transmissive light source 131, and the control board 170 to the outside of the housing 101. Provided. Also in this case, the high-frequency vibration generated in the support 10 by the operation of the heat exhausting device is absorbed by the four elastic members 106d in FIG. Therefore, a decrease in observation reliability due to vibration generated inside the measurement unit 100 is suppressed.

(6) Effect In the measurement unit 100, the space including the measurement object S and the light receiving unit 120 placed on the placement surface of the stage 140 is in a dark room state by closing the top cover 102 and the front cover 103. . Thereby, only the fluorescence emitted from the measuring object S during the fluorescence observation can be incident on the light receiving unit 120. Therefore, highly reliable fluorescence observation can be performed without installing the measuring unit 100 in a dark room.

  In the microscope apparatus 500, a power supply device 310 that generates a large amount of heat during operation and a measurement light source 321 that generates a large amount of heat when driven are provided outside the casing 101 of the measurement unit 100. Therefore, the housing 101 can be downsized, and the heat exhaust device and the air cooling passage for discharging the heat generated in the housing 101 to the outside of the housing 101 can also be downsized. As a result, the housing 101 can be further downsized, and the entire microscope apparatus 500 can be downsized.

  In response to a command given from the PC 200 when the user operates the operation unit 250, the control board 170 of the measurement unit 100 responds to a command imparting unit 110, a light receiving unit 120, a transmitted light supply unit 130 in the housing 101, The operations of the stage driving device 141, the filter unit 150, and the lens unit 160 are controlled. Thereby, the user can move the imaging visual field position of the measuring object S without touching the housing 101 and can adjust the focus position of the objective lens 161.

  In the present embodiment, a control board 170 is provided inside the housing 101 as shown in FIG. In this case, the wiring connecting the control board 170, the pattern applying unit 110, the light receiving unit 120, the transmitted light supply unit 130, the stage driving device 141, the filter unit 150, and the lens unit 160 can be shortened. Thereby, since the generation of noise is suppressed, it becomes possible to perform observation with higher reliability.

  As described above, in the measurement unit 100, the other end of the light guide member 330 is connected to the optical connector 111 of the pattern applying unit 110 from the back of the measurement unit 100 through the back surface portion 101b. In this case, it is not necessary to bend a part of the light guide member 330 in the housing 101 in order to make the measurement light incident on the light modulation element 112 of the pattern applying unit 110. Therefore, the enlargement of the housing 101 is further suppressed. Further, the user can use the space in front of the housing 101 as a work space.

(7) Other Embodiments (7-1) In the above embodiment, the control board 170 is accommodated in the housing 101. The control board 170 generates a relatively large amount of heat during operation. Therefore, the control board 170 may be housed in the housing 301 of the measurement light supply unit 300 instead of being housed in the housing 101.

  FIG. 14 is a schematic diagram illustrating a first configuration example of a microscope apparatus 500 according to another embodiment. As shown in FIG. 14, in this example, the control board 170 is accommodated in the housing 301 of the measurement light supply unit 300. In addition, the control board 170 in the housing 301 and the PC 200 are connected by the wiring WR1. Further, the control board 170 is connected to the pattern applying unit 110, the light receiving unit 120, the transmitted light supply unit 130, the stage driving device 141, the filter unit 150, and the lens unit 160 in the housing 101 through the wiring WR <b> 11.

  In this case, the number of heat sources provided in the housing 101 is reduced as compared with the microscope apparatus 500 of FIGS. This prevents temperature drift due to heat generated from the control board 170 in each component in the housing 101. In this case, the configuration for discharging the heat generated in the housing 101 to the outside can be further downsized, and the housing 101 can be further downsized. As a result, further downsizing of the entire microscope apparatus 500 is realized.

  (7-2) In the above embodiment, the transmissive light source 131 of the transmissive light supply unit 130 is accommodated in the housing 101. Not limited to this, only the transmissive light source 131 of the transmissive light supply unit 130 may be housed in the housing 301 of the measurement light supply unit 300 instead of being housed in the housing 101.

  FIG. 15 is a schematic diagram illustrating a second configuration example of a microscope apparatus 500 according to another embodiment. In FIG. 15, illustration of wirings between the measurement unit 100, the PC 200, the measurement light supply unit 300, and the display unit 400 is omitted.

  As shown in FIG. 15, in this example, the transmissive light source 131 is accommodated in the housing 301 of the measurement light supply unit 300. A transmitted light projector 350 is provided in the housing 301. The transmitted light projector 350 includes an optical connector 351 and a transmitted light source 131. Further, an optical connector 353 is provided in the fixed portion 130 </ b> A in the housing 101 instead of the transmissive light source 131. Further, the optical connector 351 of the measurement light supply unit 300 and the optical connector 353 of the measurement unit 100 are connected by the light guide member 352. The light guide member 352 is a liquid light guide. The light guide member 352 may be, for example, a glass fiber, a quartz fiber, or a plastic fiber.

  Even in this case, the number of heat sources provided in the housing 101 is reduced as compared with the microscope apparatus 500 of FIGS. This prevents temperature drift due to heat generated from the transmissive light source 131 from occurring in each component in the housing 101. In addition, the configuration for exhausting heat generated in the housing 101 to the outside can be further reduced, and the housing 101 can be further reduced in size. As a result, further downsizing of the entire microscope apparatus 500 is realized.

  (7-3) In the above embodiment, the transmissive light source 131 of the transmitted light supply unit 130 is accommodated in the housing 101. However, the measurement light emitted from the measurement light source 321 may be used as transmitted light during transmission observation instead of providing the transmission light source 131 in the housing 101.

  FIG. 16 is a schematic diagram illustrating a third configuration example of a microscope apparatus 500 according to another embodiment. In FIG. 16, illustration of wirings between the measurement unit 100, the PC 200, the measurement light supply unit 300, and the display unit 400 is omitted.

  In this example, measurement light emitted from the measurement light source 321 is used as transmitted light during transmission observation. In this case, as shown in FIG. 16, the light projecting unit 320 is provided with an optical connector 351 and an optical path switching device 360 in addition to the dimming mechanism 322, the light shielding mechanism 323, and the optical connector 324. Further, an optical connector 353 is provided in the fixed portion 130 </ b> A in the housing 101 in place of the transmissive light source 131. Further, the optical connector 351 of the measurement light supply unit 300 and the optical connector 353 of the measurement unit 100 are connected by the light guide member 352.

  The optical path switching device 360 includes a first mirror 361, a second mirror 362, and a mirror driving unit 363. The first mirror 361 is provided between the light reduction mechanism 322 and the optical connector 324, and is supported so as to be swingable about an axis parallel to the horizontal direction. The second mirror 362 is provided in the vicinity of the optical connector 351 so as to be inclined by 45 ° with respect to the horizontal direction.

  The mirror driving unit 363 holds the first mirror 361 in parallel with the horizontal direction during fluorescence observation in the measurement unit 100. As a result, light emitted from the measurement light source 321 enters one end of the light guide member 330 as measurement light. On the other hand, the mirror driving unit 363 swings the first mirror 361 so as to be inclined by 45 ° with respect to the horizontal direction during transmission observation in the measurement unit 100. Thereby, the light emitted from the measurement light source 321 is reflected by the first mirror 361 so as to be directed to the second mirror 362. The light reflected by the first mirror 361 is further reflected by the second mirror 362 and enters one end of the light guide member 352 as transmitted light.

  In this case, the number of light sources used is reduced as compared with the microscope apparatus 500 of FIGS. Therefore, further downsizing of the entire microscope apparatus 500 is realized. In this example, instead of the first mirror 361 and the mirror drive unit 363, an optical modulation element capable of switching the optical path of the measurement light and its drive circuit may be used.

  (7-4) In the above embodiment, the pattern applying unit 110, the light receiving unit 120, the transmitted light supply unit 130, the stage driving device 141, the filter unit 150, and the lens unit 160 are driven in the housing 101. The The drive circuit for the pattern applying unit 110, the light receiving unit 120, the transmitted light supply unit 130, the stage drive device 141, the filter unit 150, and the lens unit 160 is housed in the housing 301 of the measurement light supply unit 300. Also good.

  FIG. 17 is a schematic diagram illustrating a fourth configuration example of a microscope apparatus 500 according to another embodiment. The microscope apparatus 500 of this example has a configuration different from the microscope apparatus 500 of FIG. 14 in the following points.

  In the microscope apparatus 500 of the present example, the pattern applying unit 110, the light receiving unit 120, the transmitted light source 131 of the transmitted light supply unit 130, the aperture adjusting unit 132 of the transmitted light supply unit 130, in the housing 301 of the measurement light supply unit 300, A drive circuit 370 for driving the stage drive device 141, the filter unit 150, and the lens unit 160 is provided. In this case, between the measurement unit 100 and the measurement light supply unit 300, as indicated by a thick solid line, the pattern applying unit 110, the light receiving unit 120, the transmission light source 131, the aperture adjustment unit 132, the stage driving device 141, the filter unit 150, and The lens unit 160 and the drive circuit 370 are connected by a plurality of wires.

  The drive circuit generates heat during operation. Therefore, also in this case, the number of heat sources provided in the housing 101 is reduced as compared with the microscope apparatus 500 of FIGS. This prevents temperature drift caused by heat generated from the drive circuit in each component in the housing 101. In addition, the configuration for exhausting heat generated in the housing 101 to the outside can be further reduced, and the housing 101 can be further reduced in size. As a result, further downsizing of the entire microscope apparatus 500 is realized.

  (7-5) In the above embodiment, the other end of the light guide member 330 is connected to the optical connector 111 from the rear of the measurement unit 100 through the back surface 101b. Not limited to this, the other end of the light guide member 330 may be connected to the optical connector 111 from the side of the measurement unit 100 through the one side surface portion 101c (FIG. 1) or the other side surface portion 101d (FIG. 1). Even in this case, the user can use the space in front of the housing 101 as a work space.

  (7-6) In the above embodiment, one objective lens 161 used for observation is selected from the plurality of objective lenses 161 in the lens unit 160. As the lens turret 162 rotates, the selected objective lens 161 is positioned on the observation axis OA. Not limited to this, the lens unit 160 may be provided with a zoom lens and its driving device instead of the plurality of objective lenses 161, the lens turret 162 and the lens turret driving unit 164. In this case, for example, the user operates the operation unit 250 in FIG. 4 to specify the magnification. Thereby, the control board 170 adjusts the magnification of the zoom lens so that the designated magnification is obtained.

  (7-7) In the above embodiment, the focal length adjustment driving unit 165 adjusts the distance between the measuring object S and the objective lens 161 by moving the objective lens 161 in the Z direction. Not limited to this, the focal distance adjustment driving unit 165 may adjust the distance between the measurement object S and the objective lens 161 by moving the stage 140 in the Z direction.

  (7-8) In the above embodiment, the measurement object S can be observed with a plurality of different magnifications by switching the plurality of objective lenses 161. However, the measurement unit 100 may be provided with only one objective lens 161. In this case, the objective lens 161 may be configured to be detachable from the measuring unit 100. Thereby, size reduction and simplification of the configuration of the measurement unit 100 are realized.

  Similarly, in the above-described embodiment, by switching the plurality of filter cubes 151, it is possible to perform fluorescence observation of the measurement object S with measurement light in different wavelength regions. Not only this but the measurement part 100 may be provided with only one filter cube 151. In this case, the filter cube 151 may be configured to be detachable from the measuring unit 100. Thereby, size reduction and simplification of the configuration of the measurement unit 100 are realized.

(8) Correspondence between each constituent element of claim and each part of the embodiment Hereinafter, an example of correspondence between each constituent element of the claim and each constituent element of the embodiment will be described. It is not limited to examples.

  In the above embodiment, the measuring object S is an example of the object, the stage 140 is an example of the stage, the transmitted light is an example of the first light, and the transmitted light source 131 of the fixing unit 130A is the first. The oscillating unit 130B is an example of the first optical device, the measurement light is an example of the second light, and the measurement light source 321 of the light projecting unit 320 is the second light source device. For example, the pattern applying unit 110 and the filter unit 150 are examples of the second optical device, and the light receiving unit 120 is an example of the imaging device.

  The plurality of objective lenses 161 of the lens unit 160 is an example of an optical system, the stage driving device 141 is an example of an imaging visual field position moving unit, and the focal length adjusting mechanism 163 and the focal length adjusting driving unit 165 of the lens unit 160. Is an example of the focal position moving unit, the operation unit 250 of the PC 200 is an example of the operation device, the control board 170 is an example of the control device, the case 101 is an example of the first case, and the power supply device 310 is an example of a power supply apparatus, and the microscope apparatus 500 is an example of a microscope apparatus.

  The housing 301 is an example of the second housing, the plurality of support columns 106 and the base 107 are examples of installation members, the support body 10 is an example of a support body, and the plurality of elastic members 106d are elastic members. The Z direction is an example of one direction, the two side frames 13 are examples of the first and second plate-like members, and the base frame 12, the stage frame 11, and the upper frame 16 are the first, respectively. 3, examples of fourth and fifth plate-like members.

  The light guide member 330 is an example of a light guide member, the back surface portion 101b is an example of a back surface portion, the optical connector 111 is an example of an optical connection portion, the mirror 113 is an example of a reflection member, and light modulation is performed. The element 112 is an example of a reflection type light modulation element, the filter unit 150 is an example of a filter device, the front opening OP2 of the housing 101 is an example of an opening, the front cover 103 is an example of a lid member, The front lid detector S2 is an example of an open / close detector, and the light shielding mechanism 323 of the light projecting unit 320 is an example of a light shielding device.

  As each constituent element in the claims, various other constituent elements having configurations or functions described in the claims can be used.

  The present invention can be effectively used for a microscope apparatus.

DESCRIPTION OF SYMBOLS 10 Support body 11 Stage frame 12 Base frame 13 Side frame 14 Reinforcement frame 15 Back frame 16 Upper frame 100 Measuring part 101,301 Case 101a Front part 101b Rear part 101c One side part 101d Other side part 101e Upper surface part 102 Upper surface cover 102H , 103H Hinge 103 Front cover 106 Post 106d Elastic member 107 Base 110 Pattern imparting part 111, 324, 351, 353 Optical connector 112 Light modulation element 113, 134, 361, 362 Mirror 120 Light receiving part 120B Accommodation box 121 Camera 122 Color filter 123 Imaging lens 130 Transmitted light supply unit 130A Fixed unit 130B Oscillating unit 131 Transmitted light source 132 Aperture adjustment unit 133 Transmission optical system 135 Condenser lens 14 Stage 141 Stage driving device 150 Filter unit 151 Filter cube 151a Frame 151b Excitation filter 151c Dichroic mirror 151d Absorption filter 152 Filter turret 153 Filter turret drive unit 160 Lens unit 161 Objective lens 162 Lens turret 163 Focal length adjustment mechanism 163p Lens support plate 164 Turret drive unit 165 Focal length adjustment drive unit 170 Control board 200 PC
210 CPU
220 ROM
230 RAM
DESCRIPTION OF SYMBOLS 240 Memory | storage device 250 Operation part 300 Measurement light supply part 309 Opening 310 Power supply device 320 Light projection part 321 Measurement light source 322 Light reduction mechanism 323 Light shielding mechanism 330,352 Light guide member 340 Heat exhaust apparatus 350 Transmitted light light projection part 360 363 Mirror drive unit 370 Drive circuit 400 Display unit 500 Microscope device cp1 Separation position DF Depth of field np1 Normal position OA Observation axis OP1 Upper opening OP2 Front opening S Measurement object S1 Swing unit detector S2 Front lid detector sp1 , Sp2, sp3 part WR1, WR2, WR11 wiring

Claims (8)

A stage on which the object is placed;
A first light source device for generating first light;
A first optical device that irradiates an object placed on the stage with a first light generated by the first light source device;
A second light source device for generating second light;
A second optical device that irradiates the object placed on the stage with the second light generated by the second light source device;
When the object is irradiated with the first light, the object is imaged by receiving the first light transmitted through the object, and emitted from the object when the object is irradiated with the second light. An imaging device configured to image an object by receiving received fluorescence; and
An optical system for condensing the second light from the second optical device toward the object, and guiding the first light transmitted through the object and the fluorescence emitted from the object to the imaging device;
An imaging visual field position moving unit that moves an imaging visual field position of an object by the imaging device;
A focal position of the optical system with respect to the object by changing a relative distance between the object and the optical system by moving at least one of the object and the optical system along the optical axis of the optical system. A focal position moving part for moving
Operated by a user, the first light source device, the first optical device, the second light source device, the second optical device, the imaging device, the imaging visual field position moving unit, and the focal position moving unit An operating device for instructing the operation of
In response to a command given from the operation device, the first light source device, the first optical device, the second light source device, the second optical device, the imaging device, the imaging visual field position moving unit, and A control device for controlling the focal position moving unit;
A first housing that houses the stage, the second optical device, the imaging device, the optical system, the imaging visual field position moving unit, and the focal position moving unit;
Power to the first light source device, the first optical device, the second light source device, the second optical device, the imaging device, the imaging visual field position moving unit, the focus position moving unit, and the control device And a power supply for supplying
The first housing is configured to bring a space including the object placed on the stage and the imaging device into a darkroom state,
The microscope apparatus, wherein the second light source device, the power supply device, and the operation device are provided outside the first casing. The microscope apparatus according to claim 1, wherein the first housing further accommodates the control device. A second housing provided separately from the first housing;
The microscope apparatus according to claim 1, wherein the second housing accommodates the second light source device and the power supply device. The microscope apparatus according to claim 1, wherein the second light source device includes at least one of a metal halide lamp, a mercury lamp, a xenon lamp, and a light emitting diode. An installation member;
A support for supporting the stage, the second optical device, the imaging device, the optical system, the imaging visual field position moving unit, and the focal position moving unit;
The support is supported on the installation member via an elastic member,
The microscope apparatus according to claim 1, wherein the first housing is attached to the support body so as not to contact the installation member. The support is disposed so as to face each other, and extends in one direction, the first and second plate members,
Third, fourth, and fifth portions provided to connect the first plate member and the second plate member between the first plate member and the second plate member. A plate-shaped member,
The third, fourth, and fifth plate-like members are orthogonal to the one direction, and the fourth plate-like member is located between the third and fifth plate-like members in the one direction. Provided in different positions from each other,
The microscope apparatus according to claim 5, wherein the stage is integrally provided on the fourth plate member. Between the second light source device and the first housing, a light guide member that guides the second light generated by the second light source device to the second optical device is provided,
The first housing has a back surface;
The second optical device includes an optical connection portion, a reflective member, a reflective light modulation element, and a filter device,
The optical connection portion is configured to be connectable to the light guide member through the back surface portion, and supports the light guide member so that the second light emitted from the light guide member is directed forward,
The reflection member is provided at a position in front of the optical connection portion, and is configured to reflect back the second light emitted from the light guide member,
The reflection type light modulation element is provided at a position behind the reflection member, receives the second light reflected from the reflection member, and reflects the received second light toward the front while modulating the received second light. Configured as
The filter device is provided at a position in front of the reflective light modulation element, and mounts light of a predetermined wavelength region out of the second light reflected from the reflective light modulation element on the stage. The microscope apparatus according to any one of claims 1 to 6, wherein the microscope apparatus is configured to be guided to a target object. In the first housing, an opening for maintenance of the second optical device and the optical system is formed,
A lid member for opening and closing the opening of the first housing;
An open / close detector for detecting an open / closed state of the lid member;
A light guide state in which the second light generated by the second light source device is guided to the second optical device, and the second light generated by the second light source device is guided to the second optical device. A light-shielding device configured to be capable of shifting to a non-light-shielding state,
8. The control device according to claim 1, wherein the control device shifts the light shielding device from the light guiding state to the light shielding state when the lid member is switched from a closed state to an open state based on detection by the open / close detector. The microscope apparatus according to claim 1.

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