US20010021063A1 - Microscope and segmenting device for a microscope - Google Patents

Microscope and segmenting device for a microscope Download PDF

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Publication number
US20010021063A1
US20010021063A1 US09/769,215 US76921501A US2001021063A1 US 20010021063 A1 US20010021063 A1 US 20010021063A1 US 76921501 A US76921501 A US 76921501A US 2001021063 A1 US2001021063 A1 US 2001021063A1
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US
United States
Prior art keywords
light
segmenting device
microscope according
segmenting
microscope
Prior art date
Legal status (The legal status 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 status listed.)
Abandoned
Application number
US09/769,215
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English (en)
Inventor
Werner Knebel
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Leica Microsystems CMS GmbH
Original Assignee
Leica Microsystems Heidelberg GmbH
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 Leica Microsystems Heidelberg GmbH filed Critical Leica Microsystems Heidelberg GmbH
Assigned to LEICA MICROSYSTEMS HEIDELBERG GMBH reassignment LEICA MICROSYSTEMS HEIDELBERG GMBH ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: KNEBEL, WERNER
Publication of US20010021063A1 publication Critical patent/US20010021063A1/en
Priority to US10/266,005 priority Critical patent/US6836359B2/en
Abandoned legal-status Critical Current

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Classifications

    • 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
    • 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
    • G02B21/0052—Optical details of the image generation
    • 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
    • G02B21/0052—Optical details of the image generation
    • G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G02B21/08—Condensers
    • G02B21/082—Condensers for incident illumination only
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/18—Arrangements with more than one light path, e.g. for comparing two specimens

Definitions

  • the invention relates to a microscope.
  • the invention relates to a confocal scanning microscopy having a segmenting device.
  • the invention relates to a segmenting device which is used, in particular, in the abovenamed microscope or confocal microscope.
  • a microscope generally comprises a light source and a focussing optical system with the aid of which the light from the source is focussed onto a pinhole stop.
  • a beam splitter a scanning device for beam control, a microscope optical system, a detection stop and detectors for detecting detection and/or fluorescent light are provided.
  • the illuminating light is mostly coupled in via the beam splitter.
  • the focus of the light beam is moved with the aid of the scanning device in a sample plane.
  • a transmitted-light arrangement for the fluorescent light or the transmitted light—the transmission of the exciting light—, for example, to be detected on the condenser side, that is to say on the side of a condenser arranged downstream of the object.
  • the detection-light beam then does not pass via the scanning mirrors to the detector.
  • Such an arrangement is denoted as a non-descanning arrangement.
  • a microscope for simultaneously detecting fluorescent and transmitted light is already known to the applicant from an earlier patent application.
  • the known microscope comprises downstream of a condenser a light-splitting device in the form of at least one colour beam splitter which spatially separates or splits the fluorescent light from the transmitted light.
  • the segmenting device required for contrast is arranged in the form of a segment stop in a Fourier plane of the transmitted light downstream of the light-splitting device or downstream of the colour beam splitter. This requires a long detection-light path downstream of the light-splitting device, which path also has to be set up in addition to the standard microscope equipment.
  • a microscope which comprises: a light source for illuminating an object to be investigated, a an optical device for splitting transmitted light passing through the object and fluorescent light generated in the object, a segmenting device acting on the transmitted light, wherein the segmenting device is arranged between the object and the optical device.
  • a confocal microscope which comprises: a laser light source for illuminating an object to be investigated, a an optical device for splitting transmitted light passing through the object and fluorescent light generated in the object, a segmenting device acting on the transmitted light, wherein the segmenting device is arranged between the object and the optical device.
  • a segmenting device which comprises: a transparent substrate and a colour-selective coating formed on said transparent substrate.
  • the advantage of the invention is, that an optimized arrangement of the segmenting device solves the above object in a surprisingly simple way.
  • the segmenting device is no longer arranged downstream of the light-splitting device, but between the object and the light-splitting device.
  • a detection-light path, already prescribed by the arrangement of the light-splitting device, between the object and the light-splitting device is employed in this case, in addition, by virtue of the arrangement of the segmenting device on this detection-light path. The creation of an additional detection-light path is thereby avoided.
  • the microscope according to the invention has a detection-light path which is as short as possible with the aid of structurally simple means.
  • a condenser for the transmitted light and the fluorescent light could be arranged on the side of the object averted from the light source.
  • the segmenting device could then be arranged in a simple way between the condenser and the optical device, in which case with regard to segmenting which is as effective as possible, it is advantageous to arrange the segmenting device in a Fourier plane between the condenser and the optical device.
  • the segmenting device could act exclusively on the transmitted light, in which case the fluorescent light can pass through the segmenting device without being influenced.
  • the device for inserting optical components into a beam path could have a revolver mechanism or a sliding magazine in order to ensure a particularly reliable insertion of the optical components into the beam path. Not only a very reliable, but also a very simple insertion of a segmenting device into the beam path is achieved thereby.
  • segmenting device could have a segment stop, a segment phase stop or a segment phase filter.
  • the segmenting device could have a transparent substrate with a colour-selective coating.
  • This implements not only a particularly simple segmenting device, but also an exceptionally thin segmenting device which can be inserted without difficulty into a microscope of the type mentioned at the beginning.
  • the coating could in this case be opaque to the transmitted light and transparent to the fluorescent light.
  • the coating could advantageously be a dielectric coating.
  • the coating could be vapour-deposited onto the substrate. Reliable adhesion of the coating on the substrate can thereby be achieved.
  • the segmenting device could have a suitably configured colour filter.
  • the colour filter could, in particular, be tailored in accordance with a known segment stop.
  • Various materials come into consideration as colour filter material.
  • a configuration of the colour filter from plastic, preferably plastic film, is particularly easy to handle.
  • the colour filter it is also conceivable for the colour filter to be configured from glass.
  • the segmenting device could have a colour LCD matrix.
  • a colour LCD matrix permits variation of the segment or stop shape and variation of the colour properties without further redesigning.
  • the optically active part of the segmenting device could be arranged perpendicular to an optical axis of the microscope.
  • the optically active part is formed by an essentially flat region.
  • the coating would be the optically active part of the segmenting device.
  • the optically active part of the segmenting device could be arranged in a fashion deviating from the perpendicular to an optical axis of the microscope.
  • the aim is, in particular, a slight deviation which does not influence the optical action of the segmenting device.
  • the segmenting device or the optically active part of the segmenting device can have different geometrical shapes, depending on the application.
  • the optical device is configured as a light-splitting device and could have at least one colour beam splitter.
  • a plurality of colour beam splitters could, specifically, be arranged in series to enable different wavelengths or wavelength regions to be split.
  • the light-splitting device could have at least one partially transparent mirror.
  • a bandpass or blocking filter could be arranged downstream of this mirror or these mirrors.
  • a plurality of such mirrors could be arranged in series, if appropriate with a downstream bandpass or blocking filter. It is also possible thereby to split the fluorescent light into a plurality of spectral regions.
  • the fluorescent light and the transmitted light could be detected in the same detector.
  • the fluorescent light and the transmitted light could, however, also be detected in different detectors.
  • a laser could be used in a particularly advantageous way as light source. However, it is also conceivable to use other suitable light sources.
  • the segmenting device could, for example, operate using the Dodt principle or using the Hoffinan principle.
  • the segmenting device does not —as is usual—comprise a solid opaque segment stop.
  • the coating is transparent to the fluorescent light, while it is opaque to the transmitted light—two—or multiphoton exciting light.
  • FIG. 1 shows a diagrammatic illustration of an exemplary embodiment of a microscope according to the invention
  • FIG. 2 shows a diagrammatic illustration of an exemplary embodiment of a segmenting device according to the invention with a suitably coated substrate.
  • FIG. 1 shows an exemplary embodiment of a microscope according to the invention in a diagrammatic illustration.
  • the microscope is a confocal laser scanning microscope.
  • the microscope has a light source 1 designed as a laser.
  • the light source 1 emits an illuminating light beam 2 which is reflected to a scanning device 4 via a main beam splitter 3 .
  • the scanning device 4 leads the illuminating light beam 2 through a microscope optical system or an objective 5 via an object 6 .
  • Both the transmitted light passing through the object 6 and the fluorescent light produced in the object 6 reach via a condenser 7 and a deflecting mirror 8 a first colour beam splitter 9 which splits the spectrally lower-wave region 10 of the fluorescent light and reflects it to a fluorescent-light detector 11 .
  • a colour beam splitter 12 the spectrally higher-wave region 13 of the fluorescent light is reflected to a further fluorescent-light detector 14 .
  • the transmitted light 15 reaches a transmitted-light detector 16 arranged in the straight
  • the microscope consequently has a light source 1 for illuminating an object 6 to be investigated. Furthermore, the microscope has an optical device which is configures as a light-splitting device with two colour beam splitters 9 and 12 for splitting transmitted light 15 passing through the object 6 and fluorescent light 10 and 13 produced in the object. Finally, the microscope comprises a segmenting device, in the form of a colour segment stop 17 , which acts on the transmitted light 15 . With regard to a detection-light path which is as short as possible, the segmenting device is arranged between the object 6 or the condenser 7 and the light-splitting device. The microscope is suitable for transmitted-light contrast microscopy on account of the segmenting device.
  • the microscope shown in FIG. 1 further has a detector 18 which is arranged on the objective side and is normally not used in transmitted-light contrast microscopy.
  • Both fluorescent-light detectors 11 and 14 are arranged on the side of the object 6 averted from the light source 1 . Furthermore, the transmitted-light detector 16 is arranged on the side of the object 6 averted from the light source 1 .
  • the segmenting device in the form of the colour segment stop 17 is arranged in a Fourier plane between the condenser 7 and the light-splitting device, more precisely the deflecting mirror 8 .
  • a revolver mechanism or a sliding magazine can be used to insert the colour segment stop 17 .
  • FIG. 2 shows a diagrammatic representation of an exemplary embodiment of a segmenting device according to the invention.
  • the segmenting device has a flat substrate 19 with a coating 20 .
  • the coating 20 is preferably dielectric and vapour-deposited onto the substrate 19 .
  • the coating 20 vapour-deposited onto the transparent substrate is opaque to the transmitted light and transparent to the fluorescent light.
  • the dielectric coating 20 is transparent to light of wavelengths which are less than 790 nm. Light of higher wavelengths is reflected.
  • the wavelength limit with regard to the transmission and reflection of light can, however, also be at another suitable wavelength.

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  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Microscoopes, Condenser (AREA)
US09/769,215 2000-01-28 2001-01-24 Microscope and segmenting device for a microscope Abandoned US20010021063A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US10/266,005 US6836359B2 (en) 2000-01-28 2002-10-07 Microscope and segmenting device for a microscope

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10003762 2000-01-28
DE10003762.3 2000-01-28

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US10/266,005 Continuation US6836359B2 (en) 2000-01-28 2002-10-07 Microscope and segmenting device for a microscope

Publications (1)

Publication Number Publication Date
US20010021063A1 true US20010021063A1 (en) 2001-09-13

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US09/769,215 Abandoned US20010021063A1 (en) 2000-01-28 2001-01-24 Microscope and segmenting device for a microscope
US10/266,005 Expired - Lifetime US6836359B2 (en) 2000-01-28 2002-10-07 Microscope and segmenting device for a microscope

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Country Status (3)

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US (2) US20010021063A1 (ja)
JP (1) JP2001242383A (ja)
DE (1) DE10024135B4 (ja)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030035104A1 (en) * 2001-07-06 2003-02-20 Jurgen Beuthan Optical diagnosis system for small animal imaging
US11636627B2 (en) * 2016-08-28 2023-04-25 Augmentiqs Medical Ltd. System for histological examination of tissue specimens

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6898367B2 (en) * 2000-06-17 2005-05-24 Leica Microsystems Heidelberg Gmbh Method and instrument for microscopy
DE10314750A1 (de) * 2003-03-31 2004-11-04 Leica Microsystems Heidelberg Gmbh Rastermikroskop zur Detektion eines Objekts
US6993230B2 (en) * 2003-08-01 2006-01-31 The United States Of America As Represented By The Secretary Of The Navy Hollow core photonic band gap infrared fibers
DE10351414A1 (de) * 2003-10-30 2005-06-23 Carl Zeiss Jena Gmbh Laser-Scanning-Mikroskop mit einem non-descannten Detektions- und/oder Beobachtungsstrahlengang
DE102004044629B4 (de) 2004-09-13 2023-01-12 Leica Microsystems Cms Gmbh Verfahren zum simultanen Nachweis mehrerer Spektralbereiche eines Lichtstrahls
JP5112430B2 (ja) * 2006-06-29 2013-01-09 エージェンシー フォー サイエンス，テクノロジー アンド リサーチ マトリックス関連の組織動態及び疾患を定量するシステム及び方法
DE102007047461A1 (de) * 2007-09-28 2009-04-02 Carl Zeiss Microimaging Gmbh Verfahren und optische Anordnung zur Untersuchung einer Probe
CN104122662B (zh) * 2014-08-15 2017-01-18 北京大学 一种超高密度超分辨光学闪烁显微成像系统及方法

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US4109999A (en) * 1976-11-01 1978-08-29 Mamiya Camera Co., Ltd. Illuminating device for slit lamp microscopes
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JP3708246B2 (ja) * 1996-09-19 2005-10-19 オリンパス株式会社 光制御部材を有する光学顕微鏡
DE19644662C2 (de) * 1996-10-25 2000-04-13 Leica Microsystems Beleuchtungseinrichtung für ein Mikroskop
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030035104A1 (en) * 2001-07-06 2003-02-20 Jurgen Beuthan Optical diagnosis system for small animal imaging
US6833915B2 (en) * 2001-07-06 2004-12-21 Siemens Aktiengesellschaft Optical diagnosis system for small animal imaging
US11636627B2 (en) * 2016-08-28 2023-04-25 Augmentiqs Medical Ltd. System for histological examination of tissue specimens

Also Published As

Publication number Publication date
US6836359B2 (en) 2004-12-28
DE10024135A1 (de) 2001-08-16
JP2001242383A (ja) 2001-09-07
US20030030900A1 (en) 2003-02-13
DE10024135B4 (de) 2004-07-08

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AS Assignment

Owner name: LEICA MICROSYSTEMS HEIDELBERG GMBH, GERMANY

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:KNEBEL, WERNER;REEL/FRAME:011743/0600

Effective date: 20010222

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION