WO2018051515A1 - Appareil d'observation - Google Patents

Appareil d'observation Download PDF

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Publication number
WO2018051515A1
WO2018051515A1 PCT/JP2016/077572 JP2016077572W WO2018051515A1 WO 2018051515 A1 WO2018051515 A1 WO 2018051515A1 JP 2016077572 W JP2016077572 W JP 2016077572W WO 2018051515 A1 WO2018051515 A1 WO 2018051515A1
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WO
WIPO (PCT)
Prior art keywords
unit
light source
imaging
sample
photographing
Prior art date
Application number
PCT/JP2016/077572
Other languages
English (en)
Japanese (ja)
Inventor
高橋　晋太郎
Original Assignee
オリンパス株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by オリンパス株式会社 filed Critical オリンパス株式会社
Priority to JP2018539490A priority Critical patent/JPWO2018051515A1/ja
Priority to PCT/JP2016/077572 priority patent/WO2018051515A1/fr
Publication of WO2018051515A1 publication Critical patent/WO2018051515A1/fr

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes

Definitions

  • the present invention relates to an observation apparatus.
  • Patent Document 1 an observation apparatus for observing cells adhered to the bottom surface of a culture vessel while culturing them in an incubator is known (for example, see Patent Document 1).
  • Patent Document 1 both a light source unit that irradiates cells with illumination light and an imaging unit that images cells are arranged below the sample, and the illumination position and A configuration for changing the shooting position is disclosed.
  • the present invention has been made in view of the above-described circumstances, and an object thereof is to provide an observation apparatus capable of observing a sample such as a cell without increasing the size of the apparatus.
  • One embodiment of the present invention includes a stage that supports a sample, a light source unit that is disposed below the sample supported by the stage, and emits illumination light that illuminates the sample upward, and is supported by the stage.
  • An imaging unit that is disposed below the sample and that images the sample, and an imaging position in which the imaging position by the imaging unit is changed within a predetermined imaging range by moving the light source unit and the imaging unit in the horizontal direction.
  • An observation apparatus includes a moving unit and a light source moving unit that changes a position of the light source unit with respect to the imaging unit in accordance with the imaging position within the predetermined imaging range.
  • the sample supported by the stage and illuminated by the illumination light from the light source unit is photographed by the photographing unit.
  • the imaging position moving unit to change the illumination position and the imaging position, samples at different positions are imaged. Therefore, it is possible to observe a sample in a photographing range wider than the field of view of the photographing unit.
  • the position of the light source unit with respect to the imaging unit is changed by the light source moving unit according to the imaging position within the imaging range. That is, the position of the light source unit is changed so that the light source unit is positioned inside the moving range of the imaging unit corresponding to the imaging range when the imaging position is at or near the end in the imaging range. Accordingly, it is not necessary to secure a moving space for moving the photographing unit and the moving unit in excess of the moving range of the photographing unit in consideration of the space for the light source unit. The size can be minimized and the apparatus can be miniaturized.
  • the light source unit and the photographing unit are held, the holding unit is capable of translationally moving in the horizontal direction and rotatable about a vertical rotation axis, and the photographing position moving unit includes the holding unit.
  • the light source moving unit may cause the light source moving unit to rotate the holding unit around the rotation axis. In this way, when the position of the light source unit relative to the imaging unit is changed by the rotation of the holding unit, the relative positional relationship between the imaging unit and the light source unit is not converted. Thereby, the illumination conditions can be made the same in the images acquired at any photographing position.
  • the imaging unit is held, the first holding unit capable of translational movement in the horizontal direction, the light source unit is held, and the periphery of the imaging unit can be rotated around a vertical rotation axis.
  • a second holding unit, the photographing position moving unit translates the first holding unit in the horizontal direction, and the light source moving unit rotates the second holding unit around the rotation axis. You may let them.
  • the position of the light source unit relative to the image capturing unit is changed by rotating the light source unit around the image capturing unit, the position of the image capturing unit with respect to the sample does not change. There is no rotation of the sample image in the accompanying image. Thereby, it is possible to acquire an image with the same orientation of the sample image at any photographing position.
  • the imaging unit includes a two-dimensional imaging element having a rectangular imaging surface having a short side and a long side longer than the short side, and the light source moving unit is provided on the long side of the imaging surface.
  • the light source unit may be moved to a position aligned in a direction substantially perpendicular to the center and the long side.
  • a difference in brightness may occur in an image due to a difference in the incident condition of light on each position on the imaging surface according to the relative positional relationship between each position on the imaging surface and the light source unit.
  • the light source unit includes a light source body that emits the illumination light, a lens that converges the illumination light emitted from the light source body, and a diffusion plate that diffuses the illumination light converged by the lens. It may be. By doing so, the illumination light emitted from the light source as a divergent light beam is converged by the lens, whereby the illumination efficiency of the sample by the illumination light can be improved and a brighter image can be obtained. Further, by diffusing the illumination light with the diffusion plate, it is possible to reduce brightness unevenness included in the illumination light and obtain an image with reduced illumination unevenness.
  • the illumination light reflected by the reflection part is irradiated onto the sample from above, and the illumination light transmitted through the sample is incident on the imaging part, whereby the sample is imaged.
  • the illumination efficiency is improved by increasing the proportion of the illumination light emitted from the light source portion that contributes to the illumination of the sample, compared to the configuration in which the illumination light is reflected by the upper plate of the container that accommodates the sample. Can be improved.
  • the distance between the light source unit and the reflection unit and the distance between the imaging unit and the reflection unit are constant regardless of the type of the container, the illumination characteristics can be stabilized.
  • the observation apparatus 100 is arranged with a stage 2 supporting the container 1 containing the sample A and a horizontal space below the stage 2 in the horizontal direction.
  • the light source unit 3 and the imaging unit 4 that holds the light source unit 3 and the imaging unit 4, and the imaging position at which the imaging unit 4 changes the imaging position by moving the movable table 5 in the horizontal direction.
  • a moving unit 6 and a light source moving unit 7 that changes the position of the light source unit 3 with respect to the photographing unit 4 are provided.
  • the container 1 is a closed container formed of an entirely optically transparent material, such as a cell culture flask or dish, and has a top plate 1a and a bottom plate 1b facing each other.
  • the upper plate 1a is provided with a reflecting surface for reflecting the illumination light from the light source unit 3 downward.
  • the sample A is, for example, a cell that adheres to the bottom plate 1b and is cultured in the medium B.
  • the stage 2 is made of a flat plate-like member arranged horizontally, and the container 1 is placed on the stage 2.
  • the stage 2 is made of an optically transparent material such as glass so as to transmit illumination light.
  • the stage 2 is composed of a top plate of a casing 8 formed of a rectangular parallelepiped sealed container, and the light source unit 3, the imaging unit 4, the movable base 5, and the imaging position moving unit are provided in the casing 8. 6 and the light source moving part 7 are accommodated.
  • the light source unit 3 includes a light source body 3a such as an LED, and emits illumination light obliquely upward from the light source body 3a.
  • the illumination light emitted from the light source unit 3 passes through the stage 2 and the bottom plate 1b of the container 1 and is reflected downward on the reflection surface of the upper plate 1a of the container 1. As a result, the sample A is irradiated with illumination light from above to below.
  • the imaging unit 4 is an imaging device that acquires an image signal by capturing an image of the sample A connected by the objective lens 41 and the objective lens 41 that is arranged so that the optical axis is substantially along the vertical direction and collects the illumination light. And an element 42.
  • the objective lens 41 is disposed at a position where illumination light reflected by the upper plate 1a and transmitted through the sample A, the bottom plate 1b, and the stage 2 is incident.
  • the focus of the objective lens 41 can be moved in the optical axis direction by a focus adjustment mechanism (not shown).
  • the objective lens 41 having a large depth of field may be used so that the adjustment of the focal position is not necessary.
  • the objective lens 41 has a narrower field of view than the bottom plate 1 b of the container 1. Therefore, the range observed at a time by the imaging unit 4 is narrower than that of the bottom plate 1b. However, by moving the imaging position of the imaging unit 4 by the imaging position moving unit 6 as described later, a wide range of samples A can be obtained. Can be observed.
  • An image signal acquired by the image sensor 42 is wirelessly transmitted to a device arranged outside the housing 8 by a transmission / reception unit 9 provided in the housing 8 and displayed on a display.
  • the movable table 5 is made of a disk-shaped member, and includes a fixed portion 51 and a rotating portion 52 that are arranged in the thickness direction.
  • the rotating unit 52 is disposed on the fixed unit 51, and the light source unit 3 and the photographing unit 4 are fixed on the rotating unit 52.
  • the rotating part 52 is rotatably supported by a rotating bearing (not shown) around a rotating shaft 53 disposed along the central axis of the fixed part 51.
  • the photographing position moving unit 6 includes two linear motion mechanisms 61 and 62 that are orthogonally arranged and a control unit 63 that controls the linear motion mechanisms 61 and 62, and moves the movable base 5 in a direction along the stage 2. It is configured to be movable.
  • the first linear motion mechanism 61 includes a guide rail 61a fixed to the housing 8, a slider 61b supported so as to be movable in the first horizontal direction X along the guide rail 61a, and a drive for moving the slider 61b. And a mechanism 61c.
  • the drive mechanism 61c includes an X-axis motor 61d and a ball screw 61e.
  • the second linear motion mechanism 62 includes a guide rail 62a fixed to the slider 61b of the first linear motion mechanism 61, and a slider 62b supported so as to be movable in the second horizontal direction Y along the guide rail 62a. And a drive mechanism 62c for moving the slider 62b.
  • the drive mechanism 62c includes a Y-axis motor 62d and a ball screw 62e.
  • the movable base 5 is fixed on the slider 62 b, translated in the first direction X by the first linear motion mechanism 61, and translated in the second direction Y by the second linear motion mechanism 62.
  • the movable table 5 is movable within a predetermined movement range defined by the lengths of the guide rails 61a and 62a, and the photographing position by the photographing unit 4 is also a predetermined value defined by the lengths of the guide rails 61a and 62a. It can be changed within the shooting range.
  • guide rails 61 a and 62 a are provided so that the movable base 5 can move to the vicinity of each side wall of the housing 8, and the entire inside of the housing 8 is moved by the movable base 5. It is a range.
  • the control unit 63 controls the motors 61 d and 62 d of the linear motion mechanisms 61 and 62 based on the command signal received by the transmission / reception unit 9, and determines the shooting position by the shooting unit 4 as a command signal. Move to the position specified by.
  • a command signal for moving the shooting position in a desired direction is transmitted to the transmission / reception unit 9 using the shooting position input means 20. By doing so, the photographing position can be changed.
  • the light source moving unit 7 is provided on the movable table 5 and includes a rotation motor 7 a and a gear 7 b for rotating the rotation unit 52 around the rotation axis 53 with respect to the fixed unit 51.
  • the gear 7b rotates around a horizontal axis by the operation of the rotary motor 7a, the rotating part 52 that contacts the gear 7b is sent in the circumferential direction around the rotating shaft 53.
  • the light source moving unit 7 includes a control unit 7c that controls the rotation motor 7a according to the shooting position within the shooting range.
  • the control unit 7c holds a rotation angle table in which the photographing position and the rotation angle of the movable table 5 are associated with each other.
  • the control unit 7c reads the rotation angle corresponding to the imaging position specified by the command signal from the rotation angle table, and the rotation angle of the movable base 5 coincides with the read rotation angle.
  • the rotation of the movable table 5 is controlled.
  • the rotation angle corresponding to the photographing position at or near the end of the photographing range is such that the photographing unit 4 is closest to the end of the photographing range (that is, the photographing unit 4 is closest to the side wall of the housing 8).
  • the angle is set so that it approaches. Therefore, as shown in FIG. 4, when the shooting position is at or near the end of the shooting range, the rotation angle of the rotation unit 52 is such that the light source unit 3 is positioned inside the shooting range relative to the shooting unit 4. Is adjusted. In FIG. 4, some components are not shown in order to simplify the drawing.
  • the operation of the observation apparatus 100 according to the present embodiment configured as described above will be described below.
  • the observation apparatus 100 is arranged in the incubator with the container 1 placed on the stage 2, performs imaging in the incubator in accordance with a command signal transmitted by the operator, and outputs an image signal to the apparatus arranged outside the incubator. Send.
  • the illumination light emitted from the light source unit 3 passes through the stage 2 and the bottom plate 1b and is reflected by the top plate 1a, thereby illuminating the cell A from above.
  • the illumination light transmitted through the cell A enters the housing 8 via the stage 2 and is collected by the objective lens 41.
  • the image formed by the objective lens 41 is captured by the image sensor 42, and an image signal is acquired.
  • the image signal is wirelessly transmitted to a device arranged outside the incubator by the transmission / reception unit 9 and displayed on the display. Thereby, the operator can observe the cells A being cultured in the incubator without taking them out of the incubator.
  • the operator When the operator wants to observe the cells A at different positions on the bottom plate 1b, the operator transmits a command signal for moving the imaging position in either direction.
  • the imaging position moving unit 6 moves the movable base 5 to the linear motion mechanisms 61 and 62 so that the imaging position by the imaging unit 4 is arranged at the position specified by the command signal. To translate horizontally.
  • the shooting unit 4 is positioned on the side wall side of the housing 8, and the light source unit 3 is the shooting unit 4.
  • the rotation angle of the rotating unit 52 of the movable table 5 is adjusted by the light source moving unit 7 so as to be located on the inner side.
  • the light source unit 3 when the photographing position is at or near the end of the photographing range, the light source unit 3 is sandwiched between the photographing unit 4 and the side wall of the housing 8 to thereby provide the light source unit 3. Since the position of the light source unit 3 with respect to the image capturing unit 4 is adjusted so as not to interfere with the side wall of the housing 8, the image capturing unit 4 can be moved to a position closer to the side wall of the housing 8. Thereby, there is an advantage that the moving space of the movable base 5 can be minimized with respect to the photographing range by the photographing unit 4 and the housing 8 can be downsized in the horizontal direction. In particular, in the present embodiment, the horizontal dimension of the stage 2 can be suppressed substantially equal to the shooting range. Further, by disposing both the light source unit 3 and the photographing unit 4 below the stage 2, there is an advantage that the entire apparatus can be downsized in the height direction.
  • the position of the light source unit 3 with respect to the photographing unit 4 is changed while maintaining the relative positional relationship between the light source unit 3 and the photographing unit 4. If the relative positional relationship between the light source unit 3 and the imaging unit 4 changes, the incident condition of the illumination light on the objective lens 41 and the image sensor 42 changes, and the brightness of the image changes.
  • the imaging surface 42a of the imaging element 42 has a rectangular shape having a short side and a long side, the brightness at each position in the image is determined by the light source unit. 3 and the relative positional relationship between the image sensor 42. According to the present embodiment, the position of the light source unit 3 relative to the photographing unit 4 can be changed without causing a change in brightness in the image.
  • the light source unit 3 and the imaging unit 4 are provided at substantially symmetrical positions with respect to the rotation axis 53 of the movable table 5, but the arrangement of the light source unit 3 and the imaging unit 4 in the movable table 5 is as follows. It is not limited to this.
  • the center of the imaging unit 4 may be provided on the rotation shaft 53 and the light source unit 3 may be provided around the imaging unit 4.
  • observation apparatus 200 Refers to the second embodiment of the present invention with reference to the drawings.
  • a configuration different from that of the first embodiment will be described, and a configuration common to the first embodiment will be denoted by the same reference numeral and description thereof will be omitted.
  • the observation apparatus 200 according to this embodiment replaces the light source unit 3 and the image capturing unit 4 integrally with each other, and rotates and revolves (revolves) the light source unit 3 around the image capturing unit 4 with respect to the image capturing unit 4.
  • the second embodiment is different from the first embodiment in that the position of the light source unit 3 is changed.
  • the imaging unit 4 is fixed to a movable table (first holding unit) 501 fixed on the slider 62 b, and the imaging unit 4 is translated by the translational movement of the movable table 501 by the imaging position moving unit 6.
  • the shooting position can be changed.
  • the light source unit 3 is held by a cylindrical rotating cylinder (second holding unit) 10 disposed outside the photographing unit 4.
  • the rotating cylinder 10 is disposed coaxially with the optical axis of the photographing unit 4 and is supported on the movable base 501 so as to be rotatable around the optical axis.
  • the light source unit 3 rotates around the photographing unit 4 so that the position of the light source unit 3 with respect to the photographing unit 4 is changed.
  • the light source moving unit 71 is provided on the movable base 501 and includes a rotation motor 71a and a gear 71b for rotating the rotating cylinder 10 around the optical axis.
  • a rotation motor 71a rotates around an axis parallel to the optical axis by the operation of the rotary motor 71a
  • the rotary cylinder 10 in contact with the gear 71b is sent in the circumferential direction around the optical axis.
  • the control unit 71c of the light source moving unit 71 controls the rotation motor 71a according to the shooting position within the shooting range.
  • the control unit 71c holds a rotation angle table in which the photographing position and the rotation angle of the rotary cylinder 10 are associated with each other.
  • the control unit 71c reads the rotation angle corresponding to the imaging position specified by the command signal from the rotation angle table, and the rotation angle of the rotary cylinder 10 matches the read rotation angle.
  • the rotation of the rotary cylinder 10 is controlled.
  • the rotation angle corresponding to the photographing position at or near the end of the photographing range is such that the photographing unit 4 is closest to the end of the photographing range (that is, the photographing unit 4 is closest to the side wall of the housing 8).
  • the angle is set so that it approaches. Therefore, as shown in FIG. 7, when the shooting position is at or near the end of the shooting range, the rotation angle of the rotary cylinder 10 is set so that the light source unit 3 is positioned inside the shooting range with respect to the shooting unit 4. Is adjusted.
  • illustration of a part of the configuration is omitted to simplify the drawing.
  • the shooting position moving unit 6 specifies the shooting position by the shooting unit 4 in response to the command signal.
  • the movable base 501 is translated in the horizontal direction by the linear motion mechanism so as to be arranged at the position.
  • the shooting unit 4 is positioned on the side wall side of the housing 8, and the light source unit 3 is the shooting unit 4.
  • the rotation angle of the rotating cylinder 10 is adjusted by the light source moving unit 71 so as to be located on the inner side.
  • the light source unit 3 when the photographing position is at or near the end of the photographing range, the light source unit 3 is sandwiched between the photographing unit 4 and the side wall of the housing 8 to thereby provide the light source unit 3. Since the position of the light source unit 3 with respect to the image capturing unit 4 is adjusted so as not to interfere with the side wall of the housing 8, the image capturing unit 4 can be moved to a position closer to the side wall of the housing 8. Thereby, there is an advantage that the moving space of the movable base 501 can be minimized with respect to the photographing range by the photographing unit 4 and the housing 8 can be downsized in the horizontal direction. In particular, in the present embodiment, the horizontal dimension of the stage 2 can be suppressed substantially equal to the shooting range. Further, by disposing both the light source unit 3 and the photographing unit 4 below the stage 2, there is an advantage that the entire apparatus can be downsized in the height direction.
  • the imaging unit 4 does not rotate, so the orientation of the imaging unit 4 with respect to the sample A is unchanged. Therefore, unlike the first embodiment, the image does not rotate in the image with the change of the position of the light source unit 3 with respect to the imaging unit 4, and the orientation of the image of the sample A is the same at all imaging positions. There is an advantage that a certain image can be acquired.
  • the imaging element 42 is a two-dimensional imaging element having a rectangular imaging surface 42a having a short side and a long side longer than the short side.
  • the light source unit 3 is disposed at a position where the light source unit 3 is arranged in a position substantially aligned with the substantially center of the long side of the image pickup surface 42 a of the image pickup device 42 and the long side. It is preferable that the moving unit 7 rotationally moves the light source unit 3. In this case, the light source moving unit 7 rotates the rotary cylinder 10 in units of 180 ° to move the light source unit 3 between two positions sandwiching the imaging element 42 in the short side direction.
  • the ratio of the light transmitted through the objective lens 41 to the image sensor 42 is reduced, and the image becomes dark.
  • the farther from the light source unit 3 in the imaging surface 42 a the light having a larger incident angle of the sample A arrives, so the image becomes darker.
  • the bottom plate 1b to which the sample A is bonded is in an optically conjugate position with the imaging surface 42a, and the imaging range of the sample A is a rectangle with the same orientation as the imaging surface 42a.
  • the light source unit 31 converges the illumination light emitted from the light source body 3a and the illumination light converged by the lens 3b.
  • a diffusion plate 3c for diffusing may be further provided. According to this modification, the illumination efficiency can be improved and the brightness of the image can be improved by converging the illumination light emitted as a divergent light beam from the light source body 3a. Further, by diffusing the illumination light by the diffusion plate 3c, it is possible to obtain an image in which the brightness of the illumination light is made uniform and the brightness unevenness is reduced.
  • a reflection unit 12 that reflects illumination light may be provided above the stage 2.
  • the reflection part 12 is arrange
  • the illumination efficiency can be improved by using a material having a higher reflectance than the upper plate 1a for the reflecting surface of the reflecting portion 12.
  • the height with respect to the sample A of the upper plate 1a changes with kinds of the container 1
  • the height with respect to the sample A can be made constant by arrange

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  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

La présente invention concerne un appareil d'observation (100) qui comporte : un étage (2) qui supporte un échantillon (A) ; une unité de source de lumière (3) qui est disposée sous l'échantillon (A) et qui émet de la lumière d'éclairage vers le haut ; une unité imageuse (4) qui est disposée sous l'échantillon et qui capture des images de l'échantillon (A) ; une unité de déplacement de position (6) d'imagerie qui déplace l'unité de source de lumière (3) et l'unité imageuse (4) dans une direction horizontale pour changer, dans une plage d'imagerie préétablie, la position à imager au moyen de l'unité imageuse (4) ; et une unité de déplacement de source de lumière (7) pour le changement de la position de l'unité de source de lumière (3) par rapport à l'unité imageuse (4) en fonction de la position d'imagerie dans la plage d'imagerie préétablie.
PCT/JP2016/077572 2016-09-16 2016-09-16 Appareil d'observation WO2018051515A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2018539490A JPWO2018051515A1 (ja) 2016-09-16 2016-09-16 観察装置
PCT/JP2016/077572 WO2018051515A1 (fr) 2016-09-16 2016-09-16 Appareil d'observation

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP2016/077572 WO2018051515A1 (fr) 2016-09-16 2016-09-16 Appareil d'observation

Publications (1)

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WO2018051515A1 true WO2018051515A1 (fr) 2018-03-22

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PCT/JP2016/077572 WO2018051515A1 (fr) 2016-09-16 2016-09-16 Appareil d'observation

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JP (1) JPWO2018051515A1 (fr)
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004361485A (ja) * 2003-06-02 2004-12-24 Nikon Corp 顕微鏡装置
JP2010257585A (ja) * 2009-04-21 2010-11-11 Nikon Corp 照明装置及びこの照明装置を備えた光学装置
WO2011132586A1 (fr) * 2010-04-23 2011-10-27 浜松ホトニクス株式会社 Procédé et appareil pour l'observation de cellules

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004361485A (ja) * 2003-06-02 2004-12-24 Nikon Corp 顕微鏡装置
JP2010257585A (ja) * 2009-04-21 2010-11-11 Nikon Corp 照明装置及びこの照明装置を備えた光学装置
WO2011132586A1 (fr) * 2010-04-23 2011-10-27 浜松ホトニクス株式会社 Procédé et appareil pour l'observation de cellules

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