US20030206341A1 - Microscope, especially laser scanning microscope - Google Patents

Microscope, especially laser scanning microscope Download PDF

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Publication number
US20030206341A1
US20030206341A1 US10/446,215 US44621503A US2003206341A1 US 20030206341 A1 US20030206341 A1 US 20030206341A1 US 44621503 A US44621503 A US 44621503A US 2003206341 A1 US2003206341 A1 US 2003206341A1
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Prior art keywords
microscope
wavelength
beam path
wavelengths
illumination
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US10/446,215
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Ralf Wolleschensky
Ulrich Simon
Michael Stock
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Individual
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Individual
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Priority to US10/446,215 priority Critical patent/US20030206341A1/en
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Priority to US10/819,438 priority patent/US20040190130A1/en
Abandoned legal-status Critical Current

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    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/0004Microscopes specially adapted for specific applications
    • G02B21/002Scanning microscopes
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/06Means for illuminating specimens

Definitions

  • the invention relates to laser scanning microscopes and, in particular, an improvement in such microscopes for controlling the intensity of one wavelength of illumination.
  • the primary object of the invention is to provide an improvement in laser scanning microscopes where the intensity of one wavelength of illumination is controlled.
  • a microscope especially a laser scanning microscope, comprises means for providing illumination over at least one of a single wavelength and a plurality of wavelengths and means for controlling the intensity of at least one wavelength being carried out by at least one rotatable interference filter which is arranged in an illumination beam path.
  • the at least one wavelength is at least partially reflected out of the illumination beam path.
  • FIG. 1 illustrates a microscope unit and scan head of a laser scanning microscope
  • FIGS. 2 a and 2 b illustrate rotation of dichroics for influencing the wavelengths in accordance with the invention.
  • FIG. 3 illustrates in schematic and pictorial the effect of the invention on chosen wavelengths.
  • FIG. 1 A microscope unit M and a scan head S connected thereto are shown schematically in FIG. 1.
  • a light source LQ 1 with illumination optics illuminating the object on the microscope stage T via a beam splitter ST 1 in a conventional manner is provided in the microscope.
  • a swivelable mirror S 3 serves to switch to transmitted-light illumination by means of a light source LQ 2 via the condensor KO.
  • Observation through an eyepiece OK is carried out via a tube lens TL and a mirror S 1 .
  • the scanning beam path is coupled in via the scanning lens SL and the scanner SC.
  • the light of a laser module LM is coupled in in the direction of the scanner SC via light conductor F, collimating optics KO, mirror S 2 and beam splitter ST 2 .
  • the light coming from the object travels through the scanner SC and dichroic beam splitter ST 2 in the direction of detection, represented herein by way of example by another beam splitter ST 3 for splitting into detection beam paths with pinholes PH 1 , 2 , filters Fl 1 , 2 and detectors DT 1 , 2 .
  • a plurality of lasers L 1 , L 2 , L 3 with different wavelengths are provided in the laser module; these lasers L 1 , L 2 , L 3 can also be multiline lasers. They are combined via mirrors and beam splitter S 4 , respectively, and coupled into a coupling-in unit FC in the light conductor F.
  • Dichroics DC 1 , 2 , 3 , 4 , 5 , 6 which have a wavelength-dependent and angle-dependent reflectivity are arranged in the filter unit FE. This is shown by way of example in FIG. 3 with reference to the angle-dependent reflectivity for the three wavelengths in which the mirror coating is optimized for 45 degrees, i.e., the greatest reflectivity for a determined wavelength occurs at 45 degrees. Therefore, the transmission is adjusted in a continuous manner for the respective wavelength by rotation. The rest of the wavelengths are not affected.
  • the light component that is reflected out is suppressed in a suitable manner, for example, by light traps. Since the rotation of the dichroics generates a beam offset, these dichroics are arranged in pairs for compensation.
  • pairs of dichroics DC 1 , 2 for a wavelength ⁇ 1, DC 3 , 4 for a wavelength ⁇ 2, and DC 5 , 6 for a wavelength ⁇ 3 are arranged in a continuous beam path so as to be selectively reflecting and can accordingly influence these wavelengths and compensate the beam offset by means of the paired arrangement.
  • FIG. 2 b shows another arrangement as in FIG. 1, wherein the beam offset is compensated by passing twice through the dichroics DC 1 , DC 3 , DC 5 for the three wavelengths.
  • the driving means for the rotation of the dichroics are carried out in a manner familiar to the person skilled in the art, in this case, as is shown schematically in FIGS. 2 a and b , by toothed wheels to which the dichroics are fastened, wherein the toothed wheels of the pairs of dichroics in FIG. 2 a mesh with one another and accordingly ensure a coupled movement of the pairs of toothed wheels.
  • a pinion R which is driven by a motor M is provided for driving the toothed wheels.
  • the driving of the motors can be carried out via a central control unit AE which, for example, controls a predetermined illumination and detection mode which includes the attenuation/adjustment of determined laser wavelengths.
  • one or more wavelengths can advantageously be adjusted, i.e., attenuated, continuously with respect to intensity. If the lasers should be exchangeable, a plurality of such interchangeable filter units with different wavelengths can also be provided.
  • the dichroics utilized herein are interference filters such as those supplied, for example, by Laseroptik GmbH, also in pairs for compensating beam offset. Further, dichroics of the above-mentioned type can also be used advantageously for wavelength-dependent influencing of the detection beam path, for example, in FIG. 1, in a detection beam path following the beam splitter ST 2 for the suppression of especially unwanted wavelengths, for example, of the excitation wavelength in fluorescence detection.

Abstract

A microscope, especially a laser scanning microscope, with illumination over one wavelength and/or a plurality of wavelengths, wherein a controlling of the intensity of at least one wavelength is carried out by at least one rotatable interference filter which is arranged in the illumination beam path, wherein the at least one wavelength is at least partially reflected out of the illumination beam path and a plurality of filters for different wavelengths can be arranged one behind the other in the illumination beam path.

Description

    BACKGROUND OF THE INVENTION cl a) FIELD OF THE INVENTION
  • The invention relates to laser scanning microscopes and, in particular, an improvement in such microscopes for controlling the intensity of one wavelength of illumination. [0001]
  • OBJECT AND SUMMARY OF THE INVENTION
  • The primary object of the invention is to provide an improvement in laser scanning microscopes where the intensity of one wavelength of illumination is controlled. [0002]
  • In accordance with the invention, a microscope, especially a laser scanning microscope, comprises means for providing illumination over at least one of a single wavelength and a plurality of wavelengths and means for controlling the intensity of at least one wavelength being carried out by at least one rotatable interference filter which is arranged in an illumination beam path. The at least one wavelength is at least partially reflected out of the illumination beam path.[0003]
  • BRIEF DESCRIPTION OF THE DRAWINGS
  • In the drawings: [0004]
  • FIG. 1 illustrates a microscope unit and scan head of a laser scanning microscope; [0005]
  • FIGS. 2[0006] a and 2 b illustrate rotation of dichroics for influencing the wavelengths in accordance with the invention; and
  • FIG. 3 illustrates in schematic and pictorial the effect of the invention on chosen wavelengths. [0007]
  • DESCRIPTION OF THE PREFERRED EMBODIMENTS
  • A microscope unit M and a scan head S connected thereto are shown schematically in FIG. 1. [0008]
  • A light source LQ[0009] 1 with illumination optics illuminating the object on the microscope stage T via a beam splitter ST1 in a conventional manner is provided in the microscope.
  • A swivelable mirror S[0010] 3 serves to switch to transmitted-light illumination by means of a light source LQ2 via the condensor KO.
  • Observation through an eyepiece OK is carried out via a tube lens TL and a mirror S[0011] 1. In addition, by means of this mirror or beam splitter S1, the scanning beam path is coupled in via the scanning lens SL and the scanner SC.
  • The light of a laser module LM is coupled in in the direction of the scanner SC via light conductor F, collimating optics KO, mirror S[0012] 2 and beam splitter ST2.
  • The light coming from the object travels through the scanner SC and dichroic beam splitter ST[0013] 2 in the direction of detection, represented herein by way of example by another beam splitter ST3 for splitting into detection beam paths with pinholes PH1, 2, filters Fl1, 2 and detectors DT1, 2.
  • A plurality of lasers L[0014] 1, L2, L3 with different wavelengths are provided in the laser module; these lasers L1, L2, L3 can also be multiline lasers. They are combined via mirrors and beam splitter S4, respectively, and coupled into a coupling-in unit FC in the light conductor F.
  • Before being coupled in, they pass a mirror S[0015] 5 and a filter unit FE, as is shown in FIG. 2b, and are deflected, again via a filter unit FE, in the direction of the coupling-in unit by a reflector R.
  • Dichroics DC[0016] 1, 2, 3, 4, 5, 6 which have a wavelength-dependent and angle-dependent reflectivity are arranged in the filter unit FE. This is shown by way of example in FIG. 3 with reference to the angle-dependent reflectivity for the three wavelengths in which the mirror coating is optimized for 45 degrees, i.e., the greatest reflectivity for a determined wavelength occurs at 45 degrees. Therefore, the transmission is adjusted in a continuous manner for the respective wavelength by rotation. The rest of the wavelengths are not affected.
  • The optimization at 45 degrees is given by way of example; other angles could also be selected for the greatest reflectivity. [0017]
  • The light component that is reflected out is suppressed in a suitable manner, for example, by light traps. Since the rotation of the dichroics generates a beam offset, these dichroics are arranged in pairs for compensation. [0018]
  • In FIGS. 2[0019] a and 3, pairs of dichroics DC1, 2 for a wavelength λ1, DC3, 4 for a wavelength λ2, and DC5, 6 for a wavelength λ3 are arranged in a continuous beam path so as to be selectively reflecting and can accordingly influence these wavelengths and compensate the beam offset by means of the paired arrangement.
  • FIG. 2[0020] b shows another arrangement as in FIG. 1, wherein the beam offset is compensated by passing twice through the dichroics DC1, DC3, DC5 for the three wavelengths.
  • The driving means for the rotation of the dichroics are carried out in a manner familiar to the person skilled in the art, in this case, as is shown schematically in FIGS. 2[0021] a and b, by toothed wheels to which the dichroics are fastened, wherein the toothed wheels of the pairs of dichroics in FIG. 2a mesh with one another and accordingly ensure a coupled movement of the pairs of toothed wheels.
  • A pinion R which is driven by a motor M is provided for driving the toothed wheels. The driving of the motors can be carried out via a central control unit AE which, for example, controls a predetermined illumination and detection mode which includes the attenuation/adjustment of determined laser wavelengths. [0022]
  • In a multi-wavelength laser, or when a plurality of laser wavelengths are coupled into a microscope jointly, especially in a laser scanning microscope, one or more wavelengths can advantageously be adjusted, i.e., attenuated, continuously with respect to intensity. If the lasers should be exchangeable, a plurality of such interchangeable filter units with different wavelengths can also be provided. [0023]
  • The dichroics utilized herein are interference filters such as those supplied, for example, by Laseroptik GmbH, also in pairs for compensating beam offset. Further, dichroics of the above-mentioned type can also be used advantageously for wavelength-dependent influencing of the detection beam path, for example, in FIG. 1, in a detection beam path following the beam splitter ST[0024] 2 for the suppression of especially unwanted wavelengths, for example, of the excitation wavelength in fluorescence detection.
  • While the foregoing description and drawings represent the preferred embodiments of the present invention, it will be obvious to those skilled in the art that various changes and modifications may be made therein without departing from the true spirit and scope of the present invention. [0025]

Claims (4)

What is claimed is:
1. A microscope, especially a laser scanning microscope, comprising:
means for providing illumination over at least one of a single wavelength and a plurality of wavelengths; and
means for controlling the intensity of at least one wavelength being carried out by at least one rotatable interference filter which is arranged in an illumination beam path, said at least one wavelength being at least partially reflected out of the illumination beam path.
2. The microscope according to claim 1, wherein a plurality of filters for different wavelengths are arranged one behind the other in the illumination beam path.
3. The microscope according to claim 1, wherein pairs of filters with identical wavelength characteristics are provided for at least one wavelength for compensating the beam offset.
4. The microscope according to claim 1, wherein at least one filter is traversed twice in that a reflector returns the illumination light via the filter.
US10/446,215 1998-08-04 2003-05-27 Microscope, especially laser scanning microscope Abandoned US20030206341A1 (en)

Priority Applications (2)

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US10/446,215 US20030206341A1 (en) 1998-08-04 2003-05-27 Microscope, especially laser scanning microscope
US10/819,438 US20040190130A1 (en) 1998-08-04 2004-04-06 Microscope, especially laser scanning microscope

Applications Claiming Priority (4)

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DE19835068A DE19835068A1 (en) 1998-08-04 1998-08-04 Microscope, esp. laser-scanning microscope, has illumination with intensity of wavelength(s) controlled via rotatable interference filter(s) in illumination beam path
DE19835068.6 1998-08-04
US09/366,883 US6594074B1 (en) 1998-08-04 1999-08-04 Microscope, especially laser scanning microscope with rotatable interference filters
US10/446,215 US20030206341A1 (en) 1998-08-04 2003-05-27 Microscope, especially laser scanning microscope

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US09/366,883 Continuation US6594074B1 (en) 1998-08-04 1999-08-04 Microscope, especially laser scanning microscope with rotatable interference filters

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US10/819,438 Continuation US20040190130A1 (en) 1998-08-04 2004-04-06 Microscope, especially laser scanning microscope

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US10/446,215 Abandoned US20030206341A1 (en) 1998-08-04 2003-05-27 Microscope, especially laser scanning microscope
US10/819,438 Abandoned US20040190130A1 (en) 1998-08-04 2004-04-06 Microscope, especially laser scanning microscope

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Cited By (3)

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US20100091287A1 (en) * 2008-10-10 2010-04-15 Christopher Power Method for Imaging a Sample Using a Microscope, and Microscope and Data Storage Center
WO2018089864A1 (en) * 2016-11-12 2018-05-17 Caliber Imaging & Diagnostics, Inc. Confocal microscope with positionable imaging head
US10459211B2 (en) 2016-10-11 2019-10-29 Caliber Imaging & Diagnostics, Inc. Resonant scanner interoperation with movable stage

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DE20012378U1 (en) * 2000-07-17 2000-10-19 Leica Microsystems Arrangement for spectrally sensitive incident and transmitted light illumination
DE10102033C5 (en) * 2001-01-18 2011-01-13 Leica Microsystems Cms Gmbh Device and scanning microscope for the simultaneous detection of multiple spectral regions of a light beam
DE10233074B4 (en) 2002-07-19 2005-05-19 Leica Microsystems Heidelberg Gmbh Optical device for combining light beams and scanning microscope
US20050017197A1 (en) * 2003-07-26 2005-01-27 Leica Microsystems Heidelberg Gmbh Scanning microscope and method for scanning microscopy
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JP4445796B2 (en) * 2004-05-17 2010-04-07 オリンパス株式会社 microscope
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US10459211B2 (en) 2016-10-11 2019-10-29 Caliber Imaging & Diagnostics, Inc. Resonant scanner interoperation with movable stage
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WO2018089864A1 (en) * 2016-11-12 2018-05-17 Caliber Imaging & Diagnostics, Inc. Confocal microscope with positionable imaging head
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US11796786B2 (en) 2016-11-12 2023-10-24 Caliber Imaging & Diagnostics, Inc. Confocal microscope with positionable imaging head

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JP2000056227A (en) 2000-02-25
US6594074B1 (en) 2003-07-15
DE19835068A1 (en) 2000-02-10
US20040190130A1 (en) 2004-09-30

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