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WO2012164811A1 - 共焦点スキャナユニットおよび共焦点顕微鏡 - Google Patents

共焦点スキャナユニットおよび共焦点顕微鏡 Download PDF

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Publication number
WO2012164811A1
WO2012164811A1 PCT/JP2012/002598 JP2012002598W WO2012164811A1 WO 2012164811 A1 WO2012164811 A1 WO 2012164811A1 JP 2012002598 W JP2012002598 W JP 2012002598W WO 2012164811 A1 WO2012164811 A1 WO 2012164811A1
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Prior art keywords
confocal
collimator lens
disk
light source
scanner unit
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English (en)
French (fr)
Japanese (ja)
Inventor
健治 永井
健太 齊藤
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Hokkaido University NUC
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Hokkaido University NUC
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Priority to JP2013517824A priority Critical patent/JP5943432B2/ja
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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0036—Scanning details, e.g. scanning stages
    • G02B21/0044—Scanning details, e.g. scanning stages moving apertures, e.g. Nipkow disks, rotating lens arrays
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers

Definitions

  • the present invention relates to a multi-beam type confocal scanner unit using a microlens array disk and a nipou disk, and a confocal microscope having the confocal scanner unit.
  • Fluorescence microscopes have been used in life science research as an indispensable tool for observing proteins and molecules in living organisms.
  • a confocal (fluorescence) microscope is an indispensable tool for elucidating biological functions because it can obtain a series of tomographic images of a thick biological sample.
  • a laser is used as a light source (confocal laser microscope).
  • the confocal microscope scanning method is roughly classified into a galvanomirror method and a Nipkow disk method.
  • a galvanomirror method a beam is condensed at one point in a sample, and this beam is raster scanned (single beam scan). In this method, it takes about 1 second to obtain one confocal image (1000 ⁇ 1000 pixels).
  • the Nipkow disk system uses a disk called a Nipkow disk, which is formed with a plurality of pinholes.
  • Nipkow disk method a rotating Nipkow disk is irradiated with a surface of excitation light to generate a large number (for example, 1000) of beams. Then, the sample is scanned in parallel with the generated many beams (multi-beam scan). In this method, for example, one confocal image (1000 ⁇ 1000 pixels) can be obtained in 1/2000 seconds.
  • FIG. 1 is a schematic diagram showing a configuration of a conventional confocal microscope.
  • a conventional confocal microscope 10 includes a laser light source 20, a single mode optical fiber 30, a conventional confocal scanner unit 40, a first imaging lens 50, an objective lens 60, and a CCD camera 70.
  • the confocal scanner unit 40 includes a collimator lens 41, a mirror 42, a microlens array disk 43, a dichroic mirror 44, a nipou disk 45, and a second imaging lens 46.
  • the excitation light emitted from the laser light source 20 propagates through the single mode optical fiber 30 and is collimated by the collimator lens 41.
  • the collimated excitation light is collected by the microlens of the microlens array disk 43 and passes through the pinhole of the nipou disk 45.
  • the excitation light that has passed through the pinhole passes through the first imaging lens 50 and the objective lens 60 and is irradiated on the focal plane of the sample 80.
  • the fluorescence emitted from the sample 80 passes through the pinhole of the Nipkow disk 45 again and is reflected by the dichroic mirror 44.
  • the reflected fluorescence passes through the second imaging lens 46 and is detected by the CCD camera 70.
  • the conventional confocal microscope as shown in FIG. 1 uses a laser as a light source, the wavelength of excitation light that can be selected is limited. For this reason, the conventional confocal microscope has a problem that the fluorescent probe must be selected in accordance with the wavelength of the laser to be used.
  • a conventional confocal microscope uses a laser as a light source, there is a problem that the time, labor, and money required for setup and maintenance are very heavy.
  • a white light source for example, a mercury arc lamp or a halogen lamp
  • Non-Patent Document 1 describes a confocal microscope equipped with both a laser and a white light source (mercury arc lamp or halogen lamp) as light sources.
  • a laser when used as a light source, a sample is irradiated with excitation light using a Nipo disk (confocal observation).
  • a mercury arc lamp or a halogen lamp when used as a light source, the sample is irradiated with excitation light without using a Nipkow disc (non-confocal observation) in order to avoid darkening the fluorescent image. Therefore, in the confocal microscope described in Non-Patent Document 1, when a white light source is used, high-speed observation using a nipou disk cannot be performed.
  • Non-Patent Document 2 describes a confocal microscope using a mercury arc lamp as a light source.
  • FIG. 2 is a schematic diagram showing a configuration of a conventional confocal microscope described in Non-Patent Document 2.
  • the confocal microscope 10 'described in Non-Patent Document 2 has a white light source 20' (mercury arc lamp) instead of the laser light source 20 (see FIG. 1).
  • the confocal microscope 10 ′ includes a multimode optical fiber 30 ′ instead of the single mode optical fiber 30 (see FIG. 1) in order to eliminate unevenness in the light intensity of white light.
  • the confocal microscope 10 ' has a filter wheel 90 for detecting only fluorescence of a specific wavelength.
  • a mercury arc lamp is used as a light source, a sample is irradiated with excitation light using a nipou disk (confocal observation).
  • Non-Patent Document 2 fluorescence observation can be performed at high speed without limiting the types of fluorescent probes that can be used.
  • the excitation efficiency is low, and a sufficient signal may not be obtained in high-speed observation (described later).
  • An object of the present invention is to provide a Niipou disc type confocal scanner unit capable of obtaining a sufficient signal in high-speed observation even when a white light source is used as a light source, and a confocal microscope having the confocal scanner unit. That is.
  • the inventor has determined that the core radius of the multimode optical fiber, the focal length of the collimator lens, the distance between the centers of the microlens array disk and the nipou disk, and the distance between the centers of the pinholes in the nipou disk satisfy a predetermined condition. It has been found that the above-mentioned problems can be solved by selecting a collimator lens, and further studies are made to complete the present invention.
  • the present invention relates to the following confocal scanner unit.
  • a collimator lens that collimates light emitted from a multimode optical fiber; a microlens array disk having a plurality of microlenses that collect light collimated by the collimator lens; and And a Nipkow disk having a plurality of pinholes at positions corresponding to the condensing points of the plurality of microlenses; and the collimator lens includes a central portion of an output end of the multimode optical fiber;
  • the collimator lens is arranged such that the distance from the center of the collimator lens is the focal length f of the collimator lens; the focal length of the collimator lens is f (mm), and the radius of the core of the multimode optical fiber is y and 1 (mm), the said micro-lens array disc Nipo
  • the center-to-center distance between the disk and D1 (mm), the center distance between the pin hole in the Nipkow disk is taken as D2 (mm), the following equation
  • the present invention also relates to the following confocal microscope.
  • a confocal microscope comprising: the confocal scanner unit according to [1]; a light source that emits excitation light; and a multimode optical fiber that propagates the excitation light to the confocal scanner unit.
  • the light source is a white light source.
  • the light source is an LED light source, a mercury arc lamp, a xenon arc lamp, or a halogen lamp.
  • the confocal scanner unit of the present invention can obtain a sufficient signal in high-speed observation even when a white light source is used as the light source.
  • FIGS. 10A to 10F are the results of photographing confocal fluorescence images of HeLa cells.
  • FIG. 3 is a schematic diagram showing a configuration of a confocal microscope according to an embodiment of the present invention.
  • the confocal microscope 100 includes a light source 110, a multimode optical fiber 120, a confocal scanner unit 130 of the present invention, a first imaging lens 140, an objective lens 150, A filter wheel 160 and a CCD camera 170 are included.
  • the light source 110 emits excitation light to the multimode optical fiber 120.
  • the type of the light source 110 is not particularly limited, and a white light source or the like can be used as well as a conventionally used laser light source.
  • white light sources include LED light sources, mercury arc lamps, xenon arc lamps, and halogen lamps.
  • the multimode optical fiber 120 propagates the excitation light emitted from the light source 110 to the confocal scanner unit 130.
  • a multimode optical fiber is used instead of a single mode optical fiber in order to eliminate unevenness in the light intensity of the excitation light (see Non-Patent Document 2).
  • the confocal scanner unit 130 includes a collimator lens 131, a microlens array disk 132, a dichroic mirror 133, a nipou disk 134, and a second imaging lens 135.
  • the collimator lens 131 collimates the excitation light emitted from the multimode optical fiber 120.
  • the collimator lens 131 is arranged such that the distance between the center of the emission end of the multimode optical fiber 120 and the center of the collimator lens is the focal length f of the collimator lens (see FIG. 6).
  • the microlens array disk 132 is a circular substrate on which a plurality of microlenses are arranged.
  • the plurality of microlenses are arranged in the same pattern as the pinhole of the Niipou disk 134.
  • the microlens array disk 132 and the nipou disk 134 are connected by a connecting drum so as to be parallel to each other, and can rotate integrally around a rotation axis.
  • the Nipou disk 134 is a circular light shielding substrate (pinhole array disk) having a plurality of pinholes.
  • the Nipkow disk 134 has a pinhole at a position corresponding to the condensing point of the microlens of the microlens array disk 132.
  • the kind of light shielding substrate is not particularly limited.
  • the light shielding substrate is a glass substrate having a light shielding film formed on the surface thereof.
  • the pinhole arrangement method is not particularly limited, but an equal pitch spiral arrangement is preferable from the viewpoint of preventing uneven illumination and uneven scan. When pinholes are arranged in an equi-pitch spiral arrangement, the center-to-center distance between adjacent pinholes is the same distance ("center-to-center distance D2" described later).
  • the dichroic mirror 133 is disposed between the microlens array disk 132 and the nipou disk 134.
  • the dichroic mirror 133 passes the excitation light incident from the microlens array disk 132 side to the Nipkow disk 134 side.
  • the dichroic mirror 133 reflects fluorescence incident from the side of the Nipkow disk 134 to the second imaging lens 135 side.
  • the second imaging lens 135 causes the CCD camera 170 to image the fluorescence reflected by the dichroic mirror 133.
  • the first imaging lens 140 and the objective lens 150 constitute an infinite correction optical system.
  • the first imaging lens 140 and the objective lens 150 collect the excitation light that has passed through the pinhole of the Niipou disc 134 on the focal plane of the sample 180.
  • the first imaging lens 140 and the objective lens 150 collect the fluorescence emitted from the sample 180 in the pinhole of the nipou disk 134.
  • the filter wheel 160 has various filters, and allows only the fluorescence having a specific wavelength among the fluorescence from the sample 180 to pass through.
  • the CCD camera 170 detects the fluorescence that has passed through the filter of the filter wheel 160.
  • the excitation light emitted from the light source 110 propagates through the multimode optical fiber 120 and is emitted from the emission end of the multimode optical fiber 120.
  • the excitation light is collimated by the collimator lens 131 and irradiated to the microlens array disk 132.
  • the excitation light is focused on the corresponding pinhole of the Niipou disk 134 by the action of each microlens of the microlens array disk 132.
  • the excitation light that has passed through the pinhole passes through the first imaging lens 140 and the objective lens 150 and is focused on the focal plane of the sample 180.
  • the sample 180 that has received the excitation light emits fluorescence.
  • the fluorescence emitted from the sample 180 passes through the first imaging lens 140 and the objective lens 150 and returns to the confocal scanner unit 130.
  • the fluorescent light again passes through the pinhole of the Nipkow disk 134 and is reflected by the dichroic mirror 133.
  • the reflected fluorescence passes through the second imaging lens 135 and the filter wheel 160 and is detected by the CCD camera 170.
  • the microlens array disk 132 and the Nipo disk 134 are rotated, a plurality of beams (excitation light) that have passed through the pinholes scan the focal plane of the sample 180 in parallel (multi-beam scan). Further, the fluorescence emitted from the sample 180 scans the imaging surface of the CCD camera 170 after passing through the same pinhole. Thereby, the fluorescence of the focal plane of the sample 180 is detected by the CCD camera 170. Light other than the focal plane cannot pass through the pinhole and therefore cannot reach the CCD camera 170. Therefore, the CCD camera 170 can capture a confocal image consisting only of the fluorescence on the focal plane of the sample 180.
  • the focal length of the collimator lens 131 is f (mm)
  • the core radius of the multimode optical fiber 120 is y 1 (mm)
  • the microlens array disk 132 and the Niipou disk 134 When the center distance is D1 (mm) and the center distance between pinholes in the Niipou disc 134 is D2 (mm), the following expression (1) is satisfied.
  • y 1 (mm) / f (mm) on the left side means the diffusion angle ⁇ 1 (rad) of the excitation light collimated by the collimator lens 131.
  • the diffusion angle ⁇ 1 is y 1 / f (rad).
  • “D2 (mm) / D1 (mm)” on the right side means the angle ⁇ 2 (rad) shown in FIG. 4B.
  • the angle ⁇ 2 (rad) is a pin adjacent to the pinhole corresponding to the center of the microlens and the line (optical axis) connecting the center of the microlens and the center of the corresponding pinhole. The angle of the line connecting the center of the hole.
  • ⁇ 1 is the diffusion angle ⁇ 1 (rad) of the collimated excitation light.
  • ⁇ 2 is an angle for the excitation light that has passed through the microlens to pass through a pinhole adjacent to the corresponding pinhole.
  • FIG. 5 is a schematic diagram showing the configuration of the optical system of the conventional confocal microscope 10 'shown in FIG.
  • the light when pump light is introduced into a conventional confocal scanner unit 40 using a multimode optical fiber 30 ′, the light is emitted from the end (near the outer periphery) of the core of the multimode optical fiber 30 ′. Since the light A passes through the collimator lens 41 and proceeds obliquely with respect to the optical axis, it cannot pass through the pinhole of the Niipou disc 45. That is, the light A emitted from the end of the core cannot be used as excitation light.
  • the optical system of the conventional confocal scanner unit 40 is designed for a laser that is an ideal point light source.
  • a lens having a long focal length (for example, 100 mm) and a small numerical aperture (for example, 0.1 or less) is used as the collimator lens 41.
  • a collimator lens having a long focal length is used, it is difficult to satisfy the above formula (1).
  • the present inventor thought that the pumping efficiency could be increased if the light emitted from the end of the core of the multimode optical fiber could also be used as the pumping light. Therefore, as a result of intensive studies, the present inventors have found that if the optical system is constructed so as to satisfy the above formula (1), the light emitted from the end of the core of the multimode optical fiber can also be used as the excitation light. It was.
  • FIG. 6 is a schematic diagram showing the configuration of the optical system of the confocal microscope 100 of the present invention shown in FIG.
  • the optical system is constructed so as to satisfy the above formula (1) (for example, if a collimator lens 131 with a short focal length is selected), from the end of the core of the multimode optical fiber 120.
  • the emitted light A can also pass through the pinhole of the Nipkow disk 134 after passing through the collimator lens 131.
  • the light A emitted from the end portion of the core passes through the microlens and then obliquely passes through the pinhole adjacent to the pinhole corresponding to the microlens.
  • the light that has passed through the pinhole obliquely is focused on the focal plane of the sample 180 in the same manner as the light that has passed straight through the pinhole. Therefore, the light A emitted from the end portion of the core can also contribute as excitation light.
  • the confocal scanner unit of the present invention can also use the light emitted from the end of the core of the multimode optical fiber as the excitation light, so that the excitation efficiency is higher than that of the conventional confocal scanner unit. Are better.
  • the confocal scanner unit of the present invention is applied to the confocal microscope.
  • the application of the confocal scanner unit of the present invention is not limited to the confocal microscope.
  • the confocal scanner unit of the present invention can be applied to a confocal endoscope, an optical coherence tomography (OCT) apparatus, and the like.
  • This example shows the results of evaluating the performance of a confocal microscope having the confocal scanner unit of the present invention.
  • optical system White light source multi-mode optical fiber (APCH1000; core diameter 1 mm, numerical aperture 0.39, length 2 m; Fiberguide industries Inc.), collimator lens, confocal scanner unit (CSU10; Yokogawa Electric Corporation) ), An electric inverted microscope (Eclipse Ti-E; Nikon Corporation), a filter wheel (Ludl Electronic Products Ltd.) and an EM-CCD camera (ImagEM; Hamamatsu Photonics Corporation) were used to construct an optical system.
  • APCH1000 core diameter 1 mm, numerical aperture 0.39, length 2 m; Fiberguide industries Inc.
  • collimator lens CSU10; Yokogawa Electric Corporation
  • An electric inverted microscope Eclipse Ti-E; Nikon Corporation
  • a filter wheel Lidl Electronic Products Ltd.
  • EM-CCD camera ImagEM; Hamamatsu Photonics Corporation
  • FIG. 7 is a schematic diagram showing the configuration of the constructed optical system.
  • reference numeral 110 denotes a light source (white light source)
  • reference numeral 120 denotes a multimode optical fiber
  • reference numeral 131 denotes a collimator lens
  • reference numeral 40 denotes a commercially available confocal scanner unit (see FIG. 1)
  • reference numeral 140 denotes a first imaging.
  • Reference numeral 150 denotes an objective lens
  • reference numeral 160 denotes a filter wheel
  • reference numeral 170 denotes a CCD camera
  • reference numeral 180 denotes a sample.
  • a lens having a magnification of 4 to 20 times (focal length of 10 to 50 mm).
  • the excitation light that has passed through the collimator lens 131 installed separately after removing the mirror 42 of the confocal scanner unit 40 is applied to the microlens array disk 43 of the confocal scanner unit 40.
  • the optical system was adjusted to irradiate. Whichever lens is used, the exit end of the multimode optical fiber 120 is placed at the focal position of the collimator lens 131 (or the collimator lens 41).
  • the collimator lens 131 is a lens with a magnification of 4 times (Plan Apo 4x; focal length 50 mm, numerical aperture 0.20; Nikon Corporation), and a lens with a magnification of 10 times (Plan Apo 10x; focal length 20 mm, numerical aperture 0.45; Nikon Corporation) or a lens with a magnification of 20 times (Plan-Fluor 20x; focal length 10 mm, numerical aperture 0.50; Nikon Corporation) was used.
  • As the objective lens 150 of the microscope an oil immersion lens (Plan Fluor 40x; numerical aperture 1.30; Nikon Corporation) having a magnification of 40 times was used.
  • a multimode optical fiber having a core diameter of 1 mm is used. Therefore, when a lens with a magnification of 4 times (focal length 50 mm) is used, the diffusion angle ⁇ 1 is 0.010 rad. Similarly, when using 10X magnification of the lens (focal length 20 mm), the diffusion angle theta 1 is 0.025rad, and when using a magnification 20x of the lens (focal length 10 mm), the diffusion angle theta 1 0. 050 rad. Further, when the collimator lens 41 (focal length of 100 mm or more) included in the confocal scanner unit 40 is used, the diffusion angle ⁇ 1 is 0.005 rad or less.
  • the center distance D1 between the microlens array disk 43 and the tip disk 45 included in the confocal scanner unit 40 is 10 mm.
  • the intensity of excitation light at the tip of the objective lens 150 was measured.
  • a white light source (SPECTRA7 Light Engine; Lumencor, Inc.) was used as the light source.
  • Excitation light having a wavelength of 475/28 nm (transmission center wavelength / half width) from the light source 110 was introduced into the multimode optical fiber 120.
  • the excitation light propagated through the multimode optical fiber 120 is applied to a collimator lens 131 having a magnification of 4 to 20 times (focal length 10 to 50 mm) or a fiber port for laser of a confocal scanner unit (indicated by “B” in FIG. 7). Introduced.
  • the intensity of the excitation light was measured using an optical power meter (3664; Hioki Electric Co., Ltd.).
  • FIG. 8 is a graph showing measurement results of the intensity of excitation light.
  • the magnification is 10 times (focal length 20 mm) or 20 times magnification (focal length 10 mm), compared to the case where excitation light is introduced into the laser fiber port (collimator lens focal length 100 mm or more).
  • the intensity of the excitation light was remarkably improved to 3.7 times and 3.6 times, respectively.
  • a collimator lens having a magnification of 4 times was used, the intensity of the excitation light was hardly improved (1.3 times). From this result, it is understood that the intensity of the excitation light is remarkably increased by using a collimator lens satisfying ⁇ 1 ⁇ ⁇ 2 .
  • the fluorescence intensity from the fluorescent beads was measured.
  • the sample used was a 0.2 ⁇ m diameter fluorescent bead (Tetraspeck; Invitrogen Corporation).
  • the excitation light is 475/28 nm light from a white light source (SPECTRA7 Light Engine; Lumencor, Inc.), 470/40 nm light from a 100 W mercury arc lamp (OSRAM GmbH), or an argon ion laser (Sapphire 488 LP). Light with a wavelength of 488 nm from Coherent, Inc) was used.
  • Light having a wavelength of 475/28 nm from a white light source was introduced into a collimator lens (focal length 20 mm) having a magnification of 10 times.
  • a collimator lens having a magnification of 10 times.
  • light with a wavelength of 470/40 nm from a mercury arc lamp or a wavelength of 488 nm from an argon ion laser was introduced into the laser fiber port of the confocal scanner unit (focal length of collimator lens of 100 mm or more).
  • the distribution of fluorescence intensity from the fluorescent beads irradiated with excitation light under each condition was measured, and the average value of 10 beads was obtained.
  • FIG. 9 is a graph showing the distribution of fluorescence intensity under each condition.
  • the full width at half maximum in the x-axis direction was 0.31 ⁇ m
  • the full width at half maximum in the y-axis direction was 0.31 ⁇ m
  • the full width at half maximum in the z-axis direction was 0.63 ⁇ m.
  • the full width at half maximum in the x axis direction was 0.32 ⁇ m
  • the full width at half maximum in the y axis direction was 0.33 ⁇ m
  • the full width at half maximum in the z axis direction was 0.84 ⁇ m.
  • the full width at half maximum in the x-axis direction was 0.28 ⁇ m
  • the full width at half maximum in the y-axis direction was 0.29 ⁇ m
  • the full width at half maximum in the z-axis direction was 0.61 ⁇ m.
  • the confocal microscope of the present invention has a spatial resolution comparable to that of the conventional confocal microscope.
  • FIG. 10A is a ratio image (YFP / CFP; 1-3 seconds after histamine stimulation) taken using the confocal microscope of the present invention.
  • FIG. 10D is a ratio image (YFP / CFP; 1-3 seconds after histamine stimulation) taken using a conventional confocal microscope.
  • 10A and 10D are images taken of the same cell. The ratio image was created by dividing the YFP fluorescence intensity by the CFP fluorescence intensity for each pixel using the acquired YFP image (535 nm) and CFP image (480 nm).
  • FIG. 10B is a graph showing the change over time of the average ratio value in the region of interest (region surrounded by a square) shown in FIG. 10A.
  • FIG. 10E is a graph showing the change over time of the average ratio value in the region of interest shown in FIG. 10D.
  • FIG. 10C is a graph showing the change over time in the average fluorescence intensity of YFP and CFP in the region of interest shown in FIG. 10A.
  • FIG. 10F is a graph showing the time-dependent change in the average fluorescence intensity of YFP and CFP in the region of interest shown in FIG. 10D.
  • the interpolated view of FIG. 10F is a graph in which the vertical axis of the graph of FIG. 10F is enlarged.
  • the confocal scanner unit of the present invention can be applied to, for example, a confocal microscope, a confocal endoscope, and an optical coherence tomography apparatus.

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  • Optics & Photonics (AREA)
  • Microscoopes, Condenser (AREA)
PCT/JP2012/002598 2011-06-01 2012-04-13 共焦点スキャナユニットおよび共焦点顕微鏡 Ceased WO2012164811A1 (ja)

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JP2000126116A (ja) * 1998-10-28 2000-05-09 Olympus Optical Co Ltd 光診断システム
JP2005283659A (ja) * 2004-03-26 2005-10-13 Olympus Corp 走査型共焦点顕微鏡
JP2007121590A (ja) * 2005-10-27 2007-05-17 Yokogawa Electric Corp 共焦点スキャナ

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000126116A (ja) * 1998-10-28 2000-05-09 Olympus Optical Co Ltd 光診断システム
JP2005283659A (ja) * 2004-03-26 2005-10-13 Olympus Corp 走査型共焦点顕微鏡
JP2007121590A (ja) * 2005-10-27 2007-05-17 Yokogawa Electric Corp 共焦点スキャナ

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