EP2208101A1 - Dispositif optique de photomanipulation - Google Patents

Dispositif optique de photomanipulation

Info

Publication number
EP2208101A1
EP2208101A1 EP08802224A EP08802224A EP2208101A1 EP 2208101 A1 EP2208101 A1 EP 2208101A1 EP 08802224 A EP08802224 A EP 08802224A EP 08802224 A EP08802224 A EP 08802224A EP 2208101 A1 EP2208101 A1 EP 2208101A1
Authority
EP
European Patent Office
Prior art keywords
manipulation
sample
light
optical arrangement
arrangement according
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Withdrawn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
EP08802224A
Other languages
German (de)
English (en)
Inventor
Christopher Power
Helmut Lippert
Benno Radt
Christian Dietrich
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.)
Carl Zeiss Microscopy GmbH
Original Assignee
Carl Zeiss MicroImaging 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 Carl Zeiss MicroImaging GmbH filed Critical Carl Zeiss MicroImaging GmbH
Publication of EP2208101A1 publication Critical patent/EP2208101A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/32—Micromanipulators structurally combined with microscopes
    • 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 object of the invention is therefore to develop an arrangement with which the manipulation in restricted sample areas can be carried out in a simple manner substantially in the focal plane of the microscope.
  • confocal sample manipulation should also be possible without the use of multiphoton effects.
  • the arrangement should also be able to enable a spatial, isotropic imaging of the sample before, after or during the manipulation.
  • This object is achieved in an optical arrangement of the type described above in that the means for photomanipulation comprise a first manipulation optics, is coupled by means of the light of a first manipulation light source in the illumination beam path for forming a substantially planar manipulation light sheet.
  • the manipulation light source can be a laser that emits light of a different wavelength or a different wavelength range than the actual illumination light source.
  • the illumination light source itself can also be used as a manipulation light source, in which case the manipulation optics and the coupling-in elements can be dispensed with. It is also conceivable to combine several lasers in one light source module, whereby, depending on the selected method, illumination or Pulation, one or more of the light sources are selected and coupled into the illumination beam path.
  • the first manipulation optics may also comprise means for structuring the light sheet.
  • the light sheet itself has a substantially perpendicular to the optical axis of the microscope objective in the coordinates X and Y, adapted to the sample field to be examined matched length and width and extending in the direction of the optical axis of the imaging lens thickness, which is in the range of a few micrometers .
  • This light sheet can be spatially structured, for example, by means of a slit, wherein the slit comprises one or more slots for spatial structuring.
  • the structuring of the light sheets by means of grids, screens or the like makes it possible, for example, to trigger photobleaching processes limited to the bright areas and then to observe FRAP processes since no bleaching processes were triggered in the dark areas.
  • a significant advantage of the SPIM technology is the ability to generate spatial images of the sample, and in the present arrangement, therefore, the sample and / or the sample holder is expediently movable, preferably mounted rotatable and displaceable. In this way, all areas of the sample can be made accessible to manipulation, in particular also to spatially strictly localized manipulation.
  • the control unit is expediently designed to control both manipulation optics.
  • the arrangement expediently has an evaluation unit which converts the light detected pixel-wise, for example on a planar CCD detector, into data, ie digital signals, and also evaluates the signal, wherein the signal conversion is often also carried out in the detection means itself.
  • FIG. 1 shows an optical arrangement for photomanipulation of a sample 1.
  • the sample 1 is picked up by a sample holder 2.
  • the sample 1 may be embedded in a gel cylinder made of agarose which is fixed in the sample holder 2.
  • the sample holder 2 is rotatably mounted, which here by the Arrow is indicated.
  • the sample holder 2 is also slidably mounted, that is movable in all three spatial directions, so that all areas of the sample 1 can be illuminated and detected.
  • the sample 1 can also be movably mounted and the sample holder 2 can be firmly constructed, so that the movements of sample 1 and sample holder 2 are decoupled.
  • the sample 1 or light which comes from the sample 1 is imaged at least partially on the detection device, ie the CCD camera 6, by imaging optics with an imaging objective 7 which is located in an imaging beam path.
  • the light sheet for illumination is essentially flat in the focus of the imaging objective 7.
  • the optical axis of the imaging lens 7 also intersects the plane of the sheet of light at a non-zero angle, preferably perpendicular, as shown in FIG.
  • the optical arrangement also has a control unit 8, which in the example has an optional nal evaluation unit 9 is combined. In the evaluation unit 9, the detected light is converted into data and evaluated. The direction in which light is detected is indicated by the arrow marked "D" between imaging objective 7 and CCD camera 6.
  • the optical arrangement furthermore has means for photomanipulation of the sample 1.
  • These means for photomanipulation comprise a first manipulation optics, by means of which light of a first manipulation light source 10 is coupled into the illumination beam path for forming a substantially planar manipulation light sheet.
  • the direction in which manipulation light is directed to the sample is indicated by the arrow marked "M" between the lens 5 and the sample holder 2.
  • the second manipulation optics can be designed as a laser scanning microscope. This allows the confocal illumination of the sample via the imaging beam path. If the deflecting mirrors 12 are configured not as fully reflective, but as partially transmissive mirrors, light from the first manipulation light source 10 can also be coupled into the imaging beam path, as indicated by the dashed line. With a corresponding configuration of the coupling element 13, even light from the illumination source 3 can be coupled into the imaging beam path and thus enables normal observation in incident light.
  • a combination of the second Manupulationsoptik as shown in Figure 3, with an arrangement according to Figure 2 is possible. Another variant is shown in FIG. 4, here the first manipulation light source 10 and the second manipulation light source 16 are identical.
  • Another application example is a sample that shows no response at all below a power density threshold, but only when it is excited above that threshold. If the power density of the light of the two manipulation light sources when impinging on the sample is in each case less than the threshold value, in the sum but above this, so a locally very narrow area can be selected in this way. If the sample has an absolute melting point in the range of the added power densities of the manipulation light sources, the sample 1 can be broken up at this point, for example by the superimposition of the two beams. This can be used, for example, for microdissection. The light source can be the same in the two cases just described.
  • irradiating a sample labeled with Dronpa 3 dyes does not cause exposure to light of 405nm wavelength nor to irradiation of 488nm wavelength light. Only with a simultaneous combination of both excitation wavelengths, bright emissions become apparent. However, in the sample 1, these emissions occur only in the region in which the two beams overlap or intersect, that is to say in a spatially very restricted area, even if the manipulation optics in each case illuminate a larger area in each case.

Abstract

L'invention concerne un dispositif optique pour la photomanipulation d'un échantillon (1). Ce dispositif comprend un porte-échantillon (2) recevant l'échantillon (1) et un système d'éclairage comprenant une source d'éclairage (3) et une trajectoire de faisceau d'éclairage, permettant d'éclairer l'échantillon (1) au moyen d'un feuillet lumineux. Ce dispositif comprend également un système de détection qui détecte la lumière émise par l'échantillon (1) ainsi qu'une optique d'imagerie, qui reproduit une image au moins partielle de l'échantillon (1) dans le dispositif de détection, par l'intermédiaire d'un objectif (7) de capture d'image situé sur une trajectoire de formation d'image, le feuillet lumineux étant sensiblement plan dans le foyer de l'objectif (7) de capture d'image, et l'objectif (7) de capture d'image présentant un axe optique formant une intersection avec le plan du feuillet lumineux, avec un angle différent de zéro, de préférence verticalement. Le dispositif comprend en outre une unité de commande (8) et des moyens de photomanipulation de l'échantillon (1). Dans un tel dispositif optique, les moyens de photomanipulation comprennent une première optique de manipulation, qui permet l'injection de lumière provenant d'une première source lumineuse de manipulation (10) dans la trajectoire du faisceau d'éclairage afin de former un feuillet lumineux de manipulation sensiblement plan.
EP08802224A 2007-09-28 2008-09-16 Dispositif optique de photomanipulation Withdrawn EP2208101A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102007047464.6A DE102007047464B4 (de) 2007-09-28 2007-09-28 Optische Anordnung zur Photomanipulation
PCT/EP2008/007690 WO2009043473A1 (fr) 2007-09-28 2008-09-16 Dispositif optique de photomanipulation

Publications (1)

Publication Number Publication Date
EP2208101A1 true EP2208101A1 (fr) 2010-07-21

Family

ID=40206477

Family Applications (1)

Application Number Title Priority Date Filing Date
EP08802224A Withdrawn EP2208101A1 (fr) 2007-09-28 2008-09-16 Dispositif optique de photomanipulation

Country Status (5)

Country Link
US (1) US8547634B2 (fr)
EP (1) EP2208101A1 (fr)
JP (1) JP5636280B2 (fr)
DE (1) DE102007047464B4 (fr)
WO (1) WO2009043473A1 (fr)

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DE102008009216A1 (de) 2008-02-13 2009-08-20 Carl Zeiss Microimaging Gmbh Vorrichtung und Verfahren zum räumlich hochauflösenden Abbilden einer Struktur einer Probe
GB0813090D0 (en) * 2008-07-17 2008-08-27 Univ St Andrews Optical trap
DE102009044986A1 (de) 2009-09-24 2011-03-31 Carl Zeiss Microimaging Gmbh Mikroskop
EP2494399A4 (fr) * 2009-10-29 2018-01-17 California Institute of Technology Microscope à feuille de lumière par illumination et excitation multiphoton
US8711211B2 (en) 2010-06-14 2014-04-29 Howard Hughes Medical Institute Bessel beam plane illumination microscope
US10051240B2 (en) 2010-06-14 2018-08-14 Howard Hughes Medical Institute Structured plane illumination microscopy
US8575570B2 (en) 2010-08-25 2013-11-05 California Institute Of Technology Simultaneous orthogonal light sheet microscopy and computed optical tomography
DE102010039950B4 (de) * 2010-08-30 2021-07-22 Leica Microsystems Cms Gmbh Mikroskop mit Mikro- und Makro-Objektiven
DE102010060121C5 (de) * 2010-10-22 2023-09-28 Leica Microsystems Cms Gmbh SPIM-Mikroskop mit sequenziellem Lightsheet
DE102012221955A1 (de) * 2012-11-30 2014-06-05 Leica Microsystems (Schweiz) Ag Beleuchtungseinrichtung für ein Operationsmikroskop
DE102013213781A1 (de) * 2013-03-20 2014-09-25 Leica Microsystems Cms Gmbh Verfahren und optische Anordnung zum Manipulieren und Abbilden einer mikroskopischen Probe
US9971136B2 (en) 2013-03-21 2018-05-15 ETH Zürich Method and device to achieve spatially confined photointeraction at the focal volume of a microscope
DE102013205115A1 (de) 2013-03-22 2014-09-25 Leica Microsystems Cms Gmbh SPIM-Anordnung
JP2015135463A (ja) * 2013-12-19 2015-07-27 オリンパス株式会社 顕微鏡装置、及び、顕微鏡システム
DE102014204994A1 (de) * 2014-03-18 2015-09-24 Carl Zeiss Microscopy Gmbh Verfahren zur Fluoreszenzmikroskopie einer Probe
US10247672B2 (en) * 2014-09-29 2019-04-02 Howard Hughes Medical Institute Non-linear structured illumination microscopy
US10795144B2 (en) 2014-12-06 2020-10-06 Howard Hughes Medical Institute Microscopy with structured plane illumination and point accumulation for imaging and nanoscale topography
WO2016125281A1 (fr) 2015-02-05 2016-08-11 株式会社ニコン Microscope à éclairage structuré, procédé d'observation, et programme de commande
JP2018004777A (ja) * 2016-06-28 2018-01-11 オリンパス株式会社 光シート顕微鏡、及び、光シート顕微鏡の制御方法
DE102016120683A1 (de) * 2016-10-28 2018-05-03 Carl Zeiss Microscopy Gmbh Lichtblattmikroskop
DE102017118691A1 (de) * 2017-08-16 2019-02-21 Georg-August-Universität Göttingen Stiftung Öffentlichen Rechts, Universitätsmedizin Verfahren zur Lichtblatt-mikroskopischen Untersuchung von insbesondere biologischen Proben und Lichtblatt-Mikroskop
CN108303374A (zh) * 2018-02-05 2018-07-20 河南师范大学 一种可改光强的非线性测量系统
US20210208170A1 (en) * 2018-05-25 2021-07-08 Spovum Technologies Private Limited [In/In] A method for regulated manipulation of a biological sample and a system thereof
JP7134839B2 (ja) * 2018-11-07 2022-09-12 株式会社エビデント 顕微鏡装置、制御方法、及び、プログラム
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EP4163692A1 (fr) * 2020-03-31 2023-04-12 Leica Microsystems CMS GmbH Microscope à feuille de lumière et procédé de manipulation d'une zone cible d'un échantillon
JP2023537536A (ja) * 2020-08-14 2023-09-01 ライカ マイクロシステムズ シーエムエス ゲゼルシャフト ミット ベシュレンクテル ハフツング 反射器を用いて光シートを生成するための光学装置、レトロフィットキットおよび方法

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Also Published As

Publication number Publication date
DE102007047464A1 (de) 2009-04-02
WO2009043473A1 (fr) 2009-04-09
JP5636280B2 (ja) 2014-12-03
DE102007047464B4 (de) 2023-03-02
US8547634B2 (en) 2013-10-01
US20100193673A1 (en) 2010-08-05
JP2010540995A (ja) 2010-12-24

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