EP3175277A1 - Microscope avec dispositif séparateur de faisceau - Google Patents

Microscope avec dispositif séparateur de faisceau

Info

Publication number
EP3175277A1
EP3175277A1 EP15741575.3A EP15741575A EP3175277A1 EP 3175277 A1 EP3175277 A1 EP 3175277A1 EP 15741575 A EP15741575 A EP 15741575A EP 3175277 A1 EP3175277 A1 EP 3175277A1
Authority
EP
European Patent Office
Prior art keywords
beam path
reflection
mirror surface
microscope
dichroic mirror
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.)
Withdrawn
Application number
EP15741575.3A
Other languages
German (de)
English (en)
Inventor
Peter Euteneuer
Ralf KRÜGER
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 CMS 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 CMS GmbH filed Critical Leica Microsystems CMS GmbH
Publication of EP3175277A1 publication Critical patent/EP3175277A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • 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
    • G02B21/06—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
    • G02B21/06—Means for illuminating specimens
    • G02B21/08—Condensers
    • G02B21/14—Condensers affording illumination for phase-contrast observation
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/24—Base structure
    • G02B21/241—Devices for focusing
    • G02B21/245—Devices for focusing using auxiliary sources, detectors
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/10—Beam splitting or combining systems
    • G02B27/14—Beam splitting or combining systems operating by reflection only
    • G02B27/141—Beam splitting or combining systems operating by reflection only using dichroic mirrors

Definitions

  • the invention relates to a microscope according to the preamble of claim 1.
  • microscopes e.g. Inverse research microscopes
  • additional light sources for different experiments for lighting, manipulation and measurement purposes in the imaging beam path, preferably in the infinite beam path between the lens and tube lens, coupled.
  • such microscopes are usually provided with means which serve to decouple certain spectral components of the image light originating from the sample.
  • lasers are usually used whose laser light has a longer or a shorter wavelength than the actual detection light.
  • the coupling or decoupling of the light is usually carried out in a rectangular arrangement in which the optical axes of the beam paths, in which propagate on the one hand, the input or outcoupled light and the other the detection light, at an angle of 90 ° to each other are arranged.
  • This microscope contains an autofocus module with a laser light source which emits a first light beam, hereinafter referred to as a reflection sub-beam, whose wavelength lies in the infrared range of the wavelength spectrum.
  • This reflection sub-beam falls on a mirror, which is provided with a dichroic mirror surface. This is designed so that it reflects the first reflection partial beam on the sample to be examined. With reflection at the dichroic mirror surface, the reflection sub-beam changes its propagation direction by a deflection angle of 90 °.
  • the autofocus module detects a signal on the basis of which is reflected on the sample reflection partial beam, on the basis of which the imaging optics of the microscope is focused. For this purpose, the partial reflection beam coming from the sample is again reflected at the dichroic mirror surface in order to guide it into the autofocus module.
  • the dichroic mirror surface used in this prior art microscope reflects infrared light, it passes detection light in the visible range.
  • a second light beam in the form of the detection light emanating from the sample is provided, which transmits the dichroic mirror surface. This second light beam is referred to below as the transmission partial beam.
  • the dichroic mirror surface thus merges the two beam paths in which the reflection sub-beam and the transmission sub-beam propagate.
  • the two beam paths have a common beam path section which lies between the dichroic mirror surface and the sample.
  • the dichroic mirror surface is arranged so that its surface normal is arranged at an angle of 45 ° with respect to the optical axis of this common beam path section.
  • An example of such an application which is negatively influenced by the 45 ° arrangement is the differential difference contrast or, for short, DIC method, in which differences in the optical path length in the object under consideration are converted into differences in image brightness and, as a result, transparent phase objects are made visible.
  • the conventionally used 45 ° arrangement of the dichroic mirror surface also has a disadvantageous effect if, in order to increase the application flexibility of the microscope, it should be designed such that it can be introduced into or removed from the microscope beam path depending on the application. In this case occurs at a 45 ° arrangement between the two operating states in which the dichroic mirror surface is arranged in the beam path of the microscope or not, a comparatively large offset of the optical axis of the beam path, which may affect the quality of the optical image or through other components must be compensated.
  • the state of the art is additionally referred to DE 42 31 267 B4, which describes a mirror arrangement known as the "Smith divider.”
  • This mirror arrangement consists of two mirrors, one of which is formed by a divider surface which is not less than 45 °
  • this splitter surface is designed as a neutral splitter with wavelength-independent reflection or transmission effect., Therefore, it is not suitable to separate different spectral components of each other.
  • the object of the invention is a microscope refinement of the type described above so that with reduced technical complexity, a precise separation of the spectral ranges while avoiding the disadvantages described above is possible.
  • the invention achieves this object by the microscope having the features of claim 1.
  • Advantageous developments are specified in the subclaims.
  • Such a microscope comprises one or more light sources, which individually or collectively produce an illumination beam path with light in a plurality of wavelength ranges, a dichroic beam splitter arrangement with a dichroic mirror surface which is arranged between an objective optics and a tube lens in a beam path section which comprises several wavelength ranges and which by reflection generates a reflection partial beam in the direction of a reflection beam path and generated by transmission a transmission partial beam in the direction of a transmission beam whose transmitted wavelength range is different from the reflected wavelength range of the reflection beam part, wherein the beam splitter arrangement, the propagation direction of the reflection beam relative to the illumination beam path around a changes predetermined deflection angle.
  • the dichroic mirror surface is arranged at an angle of 22.5 ⁇ 7.5 ° in the beam path section.
  • the beam splitter arrangement has at least one further mirror arranged in the reflection beam path. The propagation direction of the reflection sub-beam is changed by the sum of all reflections on the dichroic mirror surface and the at least one further mirror by the predetermined deflection angle.
  • the microscope thus has a dichroic beam splitter arrangement with a dichroic mirror surface, which reflects a reflection partial beam propagating along a reflection beam path and transmits a transmission partial beam propagating along a transmission beam path whose wavelength is different from the wavelength of the reflection partial beam. Furthermore, the microscope has at least one further mirror, which is arranged in the reflection beam path outside the common beam path section which the reflection beam path and the transmission beam path have. This further mirror forms with the dichroic mirror surface a reflecting arrangement which deflects the reflection partial beam in successive reflections in such a way that the desired deflection angle is achieved. Under Deflection angle is the angle between the propagation direction of the light beam entering the beam splitter arrangement and the propagation direction of the light beam leaving the beam splitter arrangement.
  • the invention makes it possible, in departure from the conventional 45 ° arrangement to align the dichroic mirror surface at a smaller angle than 45 ° relative to the optical axis of the common beam path section.
  • the smaller this angle the easier it is to achieve the desired sharp separation of the spectral regions with a layer structure such as is commonly used to realize a dichroic mirror surface.
  • the arrangement according to the invention enables the sharp separation of the spectral ranges, without having to provide further optical components such as an infrared cut filter. This sharp separation is especially needed for fluorescence applications on living cells where highly sensitive cameras are used. Rest light from other spectral regions would be perceived here as very disturbing background structures.
  • the microscope according to the invention is particularly profitable in all polarization-optical applications, e.g. in the D IC method.
  • the orientation of the dichroic mirror surface according to the invention also causes the extent of the mirror surface along the optical axis of the beam path to be smaller than is the case in the conventional 45 ° arrangement.
  • the dichroic mirror surface along the optical axis requires less space than in the 45 ° arrangement. This favors a particularly compact microscope structure, which is particularly important in the dimensioning of the distance-sensitive area between the objective lens and the tube lens, in which the dichroic beam splitter assembly is usually advantageous.
  • the alignment of the mirror surface reduces the optical axis offset of the beam path that occurs between the two operating states in which the mirror surface is arranged in the beam path or removed therefrom.
  • the smaller installation space along the optical axis also has a favorable effect here.
  • the surface normal of the dichroic mirror surface with respect to the optical axis of the common beam path section is inclined only half as much compared to the conventionally used 45 ° arrangement.
  • Such an arrangement of less than 22.5 ° has proved to be favorable, in particular with regard to the desired sharp separation of the spectral ranges and with regard to the desired polarization neutrality.
  • the solution according to the invention is not strictly limited to the aforementioned 22.5 ° arrangement. Accordingly, the angle at which the dichroic mirror surface is arranged opposite the optical axis of the common beam path section can be varied within a tolerance range of ⁇ 7.5 °. This still ensures that it is smaller than the commonly provided angle of 45 °.
  • the intended technical effects namely a reduced beam offset, a sharp separation of the spectral regions and / or a broad polarization neutrality are achieved.
  • the beam splitter assembly according to the invention consists in this case only of two mirror surfaces, whereby the optical structure can be kept simple.
  • the microscope according to the invention has an adjusting device for removing and introducing the dichroic mirror surface from or into the microscope Beam path section on. This increases the application flexibility of the microscope.
  • the dichroic mirror surface is formed in a particularly preferred embodiment, as it were as a switchable splitter surface, i. Depending on the application, it can be introduced into and removed from the beam path section, it is advantageous to mount the further mirror firmly in the microscope, e.g. in his tripod. This is possible because the further mirror is outside the common beam path section. In an operating state in which the dichroic mirror surface is removed from the beam path section, therefore, the further mirror is also inoperative.
  • an autofocus module with an autofocus light source which emits autofocus light onto the beam splitter arrangement.
  • This embodiment can be realized for example in such a way that the dichroic mirror surface, which is otherwise removed from the beam path section, is introduced into the beam path section for starting up the autofocus module.
  • the autofocus light source of the autofocus module preferably emits the partial reflection beam in the form of light whose wavelength lies in the infrared range.
  • the dichroic mirror surface is arranged in an infinite beam path. This is particularly advantageous when the dichroic mirror surface is designed as a switchable element, i. Depending on the application is introduced into the beam path section or removed therefrom.
  • Figure 1 is a schematic representation with components of a microscope, which represents a first embodiment of the invention
  • FIG. 2 shows a schematic representation of the dichroic beam splitter arrangement provided in the microscope according to FIG. 1;
  • Figure 3 is a schematic representation of a microscope as a comparative example;
  • Figure 4 is a schematic representation of the dichroic beam splitter arrangement used in the microscope of Figure 3;
  • Figure 5 is a graph showing the wavelength-dependent transmission of the dichroic mirror surface used in the microscope of Figure 1;
  • FIG. 6a is a graph showing the wavelength-dependent transmission of an im
  • FIG. 6b shows a graph showing the wavelength-dependent transmission of an im
  • Figure 7 is a graph illustrating the angular dependence of the polarization neutrality of the example of reflection on a glass plate.
  • FIG. 1 shows a microscope 10, which represents a first embodiment of the invention. Only components of the microscope 10 that are necessary for the understanding of the invention are shown in FIG.
  • the microscope 10 according to FIG. 1 comprises an objective optics 12 which is formed from a plurality of lens groups 14, 16, 18 and 20.
  • the objective optics 12 serves to image a sample 22 via a tube lens 24 into an intermediate image plane 26.
  • the microscope 10 further comprises an autofocus module 28, which has the function of focusing the objective optics 12 in a manner known per se on the sample 22.
  • the autofocus module 28 has a laser light source 30 which emits a partial reflection beam 32 whose wavelength is in the infrared range.
  • the reflection sub-beam 32 emitted by the laser light source 30 passes through a notch filter 34, that of those Spectral components of the reflection sub-beam 32 filters out, which are in the visible wavelength spectrum.
  • the reflection sub-beam 32 then passes through a collector lens 31 and a field lens 36 and is reflected at a mirror element 38 in an optical transmission system 40, which is formed of a plurality of lens groups 42, 44, 46 and 48 and a diaphragm 50.
  • the reflection sub-beam 32 is incident on a dichroic beam splitter array 52 formed of a dichroic mirror surface 54 and a mirror 56.
  • the dichroic beam splitter arrangement 52 is shown in more detail in FIG.
  • the dichroic mirror surface 54 is applied as a layer structure on a transparent, plane-parallel carrier plate 58.
  • the total reflecting mirror 56 is provided on another plane-parallel support plate 60.
  • the reflection sub-beam 32 is first reflected at the mirror 56 and then at the dichroic first mirror surface 54.
  • the dichroic mirror surface 54 is designed to reflect the partial reflection beam 32 whose wavelength is in the infrared range while transmitting light whose wavelength is in the visible range.
  • the dichroic mirror surface 54 of the partial reflection beam 32 After reflection at the dichroic mirror surface 54 of the partial reflection beam 32 passes into the objective lens 12 and is focused by this on the sample 22.
  • the part of the reflection partial beam 32 reflected by the specimen 22 returns via the objective optics 12 to the dichroic mirror surface 54, which deflects the partial reflection beam 32 onto the second mirror surface 56. This reflects the reflection partial beam 32 back into the transmission system 40.
  • the reflection partial beam 32 is then reflected at the mirror element 38 by means of pupil division and imaged by a filter 62 onto the detector 64.
  • the filter 62 and the detector 64 are part of the autofocus module 28. The latter evaluates a signal which the reflection sub-beam 32 generates on the detector 64 and controls a focusing device via this signal in order to focus the objective lens 12 on the sample 22.
  • the components provided for this control are omitted in FIG.
  • the reflection sub-beam 32 propagates in the microscope 10 along a reflection beam path, which is generally designated 66 in FIG.
  • the detection light coming from the sample 22, which is conducted via the objective optics 12 and the tube lens 24 to the intermediate image plane 26, propagates in the form of a transmission partial beam 68 along a transmission beam path denoted by 70 in FIG.
  • the wavelength of this transmission partial beam should be in the visible wavelength spectrum.
  • the dichroic mirror surface 54 is formed to transmit light whose wavelength is in the visible range.
  • the reflection beam path 66 and the transmission beam path 70 have, between the dichroic mirror surface 54 and the sample 22, a common beam path section with an optical axis O designated by 72 in FIG.
  • the optical axis O of this common beam path section 72 is positioned on the dichroic surface 54 such that it has an angle ⁇ of 22.5 ° with its surface normal N-i.
  • the surface normal of the second mirror surface 56 denoted by N2 in FIG. 2 with the direction of incidence of the reflection partial beam 32 incident on it also includes an angle of 22.5 °, which is denoted by ⁇ in FIG.
  • FIGS. 3 and 4 show a comparative example not according to the invention, as conventionally used in the prior art.
  • the exemplary embodiment illustrated in FIGS. 1 and 2 essentially differs from the comparative example according to FIGS. 3 and 4 by the dichroic beam splitter arrangement 52.
  • the dichroic mirror surface designated by 54 ' is provided in the comparative example.
  • the 22.5 ° arrangement used in the exemplary embodiment according to the invention has the advantage of a smaller beam offset x which occurs between the two operating states in which the carrier plate 58 or 58 'in the reflection beam path 66 is arranged or removed from this.
  • the respective carrier plate 58 or 58 ' is a 2 mm thick plate made of BK7.
  • the beam offset decreases from 0.67 mm in the 45 ° arrangement to 0.28 mm in the 22.5 ° arrangement according to the invention.
  • FIGS. 5, 6a and 6b A further advantage of the solution according to the invention.
  • FIG. 5 shows an example of the wavelength-dependent transmission of the dichroic mirror surface 54 in the 22.5 ° arrangement according to the invention, which is to be contrasted with the corresponding graph of FIG. 6a, which shows the transmission of the dichroic mirror surface 54 'in the usual 45 ° arrangement (see Figure 4).
  • the arrangement according to the invention enables a clearly sharper separation of the spectral ranges under consideration, in this embodiment the visible range and the infrared range.
  • the usual 45 ° arrangement of the dichroic mirror surface 54 ' is often followed by an infrared cut filter whose wavelength-dependent transmission is shown in Figure 6b.
  • Such a filter is no longer required in the embodiment shown in Figures 1 and 2.
  • FIG. 7 illustrates how the arrangement of the dichroic mirror surface 54 in the reflection beam path 66 affects the polarization neutrality.
  • the reflectivities R s and R P for the reflection of perpendicular or parallel polarized light on a glass plate, which has a refractive index of 1.5 are shown in FIG.
  • the greater the deflection angle ⁇ the more the two reflectivities R s and R P deviate from one another.
  • the inventive 22.5 arrangement of the dichroic mirror surface 54 is significantly better in polarization neutrality than the conventional 45 ° arrangement.
  • the solution according to the invention is not limited to the 22.5 ° arrangement provided in the specifically described exemplary embodiment.
  • the angle at which the surface normal N-1 of the dichroic mirror surface 54 is arranged opposite to the optical axis O of the common beam path section 72 may also be different within a range of ⁇ 7.5 °, as long as it is sufficiently small to reach the desired one technical effects, namely to achieve a reduced beam offset, a sharp separation of the spectral ranges and / or extensive polarization neutrality.

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

Abstract

L'invention concerne un microscope (10), comprenant une ou plusieurs sources de lumière (30) qui génèrent de manière individuelle ou collective un trajet de faisceau d'éclairage avec une lumière dans plusieurs plages de longueur d'onde, un dispositif séparateur de faisceau dichroïque (52) avec une surface de miroir dichroïque (54) qui est disposée entre un optique d'objectif (12) et une lentille de barillet (24) dans une portion (72) du trajet de faisceau comprenant plusieurs plages de longueur d'onde, et qui génère, par réflexion, un faisceau partiel de réflexion (32) en direction d'un trajet de faisceau de réflexion (66) et génère, par transmission, un faisceau partiel de transmission (68) en direction d'un trajet de faisceau de transmission (70) dont la plage de longueur d'onde transmise est différente de la plage de longueur d'onde réfléchie du faisceau partiel de réflexion (32). Selon l'invention, le dispositif séparateur de faisceau (52) modifie la direction de propagation du faisceau partiel de réflexion (32) selon un angle de déviation (a) prédéfini vis-à-vis du trajet de faisceau d'éclairage. La surface de miroir dichroïque (54) est disposée à un angle de 22,5 ± 7,5° dans la portion de trajet de faisceau (72), et le dispositif séparateur de faisceau (52) comporte au moins un autre miroir (56) disposé dans le trajet de faisceau de réflexion (66), la direction de propagation du faisceau partiel de réflexion (32) étant modifiée par la somme de toutes les réflexions au niveau de la surface de miroir dichroïque (54) et d'au moins un autre miroir (56) selon l'angle de déviation (a) prédéfini.
EP15741575.3A 2014-07-28 2015-07-28 Microscope avec dispositif séparateur de faisceau Withdrawn EP3175277A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102014110606.7A DE102014110606B4 (de) 2014-07-28 2014-07-28 Mikroskop mit einer Strahlteileranordnung
PCT/EP2015/067315 WO2016016270A1 (fr) 2014-07-28 2015-07-28 Microscope avec dispositif séparateur de faisceau

Publications (1)

Publication Number Publication Date
EP3175277A1 true EP3175277A1 (fr) 2017-06-07

Family

ID=53719786

Family Applications (1)

Application Number Title Priority Date Filing Date
EP15741575.3A Withdrawn EP3175277A1 (fr) 2014-07-28 2015-07-28 Microscope avec dispositif séparateur de faisceau

Country Status (6)

Country Link
US (1) US11002978B2 (fr)
EP (1) EP3175277A1 (fr)
JP (1) JP6742984B2 (fr)
CN (1) CN106575030A (fr)
DE (1) DE102014110606B4 (fr)
WO (1) WO2016016270A1 (fr)

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Publication number Priority date Publication date Assignee Title
JP6608249B2 (ja) * 2015-11-09 2019-11-20 株式会社ミツトヨ 光ピックアップ装置
CN106873079A (zh) * 2017-04-27 2017-06-20 苏州高新区建金建智能科技有限公司 一种光纤分束装置
DE102017218449B3 (de) 2017-10-16 2019-02-21 Leica Microsystems Cms Gmbh Mikroskopieverfahren mit Fokusstabilisierung, Recheneinheit, Mikroskopsystem und Computerprogrammprodukt
DE102020216541A1 (de) 2020-12-23 2022-06-23 Robert Bosch Gesellschaft mit beschränkter Haftung Vorrichtung und Verfahren für eine Fluoreszenzmessung für eine Analyse einer biochemischen Probe

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JP3699762B2 (ja) 1996-01-08 2005-09-28 オリンパス株式会社 走査型光学顕微鏡
JP3999479B2 (ja) * 2001-07-10 2007-10-31 オリンパス株式会社 光学装置
JP4270884B2 (ja) 2002-01-10 2009-06-03 オリンパス株式会社 顕微鏡システム
JP4370554B2 (ja) 2002-06-14 2009-11-25 株式会社ニコン オートフォーカス装置およびオートフォーカス付き顕微鏡
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Also Published As

Publication number Publication date
DE102014110606B4 (de) 2017-10-19
WO2016016270A1 (fr) 2016-02-04
JP6742984B2 (ja) 2020-08-19
JP2017522605A (ja) 2017-08-10
DE102014110606A1 (de) 2016-01-28
US11002978B2 (en) 2021-05-11
US20170212356A1 (en) 2017-07-27
CN106575030A (zh) 2017-04-19

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