EP1141761A1 - System for introducing optical tweezers and/or a treatment beam into a microscope - Google Patents
System for introducing optical tweezers and/or a treatment beam into a microscopeInfo
- Publication number
- EP1141761A1 EP1141761A1 EP00971409A EP00971409A EP1141761A1 EP 1141761 A1 EP1141761 A1 EP 1141761A1 EP 00971409 A EP00971409 A EP 00971409A EP 00971409 A EP00971409 A EP 00971409A EP 1141761 A1 EP1141761 A1 EP 1141761A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- optical tweezers
- microscope
- optical
- focus
- 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.)
- Ceased
Links
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
Definitions
- the invention allows the spatial fixation of microscopic objects in the laser scanning microscope, even while moving the object plane, for example during image acquisition. This means that even moving objects can be imaged sharply.
- Optical tweezers have proven to be an interesting tool for a number of biological work techniques.
- the combination of laser scanning microscopes with laser microtechnologies suggests an expansion of the experimental possibilities.
- LSM images of moving objects especially inside unopened cells, often do not produce satisfactory images because many subcellular structures move during the scan time.
- the optical tweezers are ideal for gentle (vital) fixation.
- a spatially defined displacement of fixed objects is possible with the optical tweezers.
- Application examples for the use of compensated optical tweezers in the laser scanning microscope are the examination of organisms, for example chloroplasts, or the holding of objects moved by motor proteins. In the latter case, force measurements are even possible under suitable conditions.
- moving objects for example particles in suspension or certain organelles, cannot be imaged sharply without being fixed by compensated optical tweezers.
- Optical tweezers coupled through the lens have their focus in the object plane. If the object plane is shifted in parallel by the three-dimensional image acquisition process ("scanning"), the focus of the optical tweezers is also displaced. This means that objects which are held by the optical tweezers are also displaced. However, this is undesirable during image acquisition Therefore the Displacement of the object plane can be compensated for by a suitable device in the beam path of the optical tweezers.
- the invention is always necessary in laser scanning microscopes from different manufacturers when the optical tweezers are coupled in through the lens and the third dimension in the image acquisition by moving the lens or the stage or another method that focuses the optical tweezers relative to the sample postponed, developed.
- the sample is no longer freely accessible from above, which, for example, makes applications with microinjection or temperature control devices very difficult, if not impossible.
- the latter also applies to structures in which the optical fixing of particles is carried out by glass fibers provided with microlenses, which are guided directly onto the sample.
- there are problems with the sterility of the sample since the glass fibers have to be immersed in thicker layers of liquid if particles are to be fixed on the underside of the liquid.
- FIG. 1 shows a schematic representation of the effect of the invention
- Figure 2 the application in a microscope like a laser scanning -
- the optical tweezers are
- Microscope objective guided into the object plane It is adjusted so that microscopic particles located in the object plane are held, that is, the focus of the optical tweezers is in the object plane.
- the object plane In the three-dimensional image acquisition by a laser scanning microscope, however, the object plane must be shifted in parallel in order to open up the third dimension protruding from the object plane. This also shifts the focus of the optical tweezers, which leads to an undesired shift of the fixed particles. Without compensation for this shift, three-dimensional objects held by the optical tweezers cannot be recorded in three-dimensional resolution.
- z-compensation Compensation for the displacement of the object plane, hereinafter referred to as z-compensation, consists of variable optical elements which are inserted in the beam path of the optical tweezers and which compensate for the movement of the object plane.
- the z-compensation effects a compensating movement of the optical tweezers which takes place simultaneously with the movement of the object plane, so that the position of the fixed object in the sample is retained.
- the compensation is implemented via an electromechanically displaceable optical element in the coupling system of the optical tweezers.
- the exact position of the object plane is taken from the control electronics of the laser scanning microscope during the image acquisition process. Accordingly, the displaceable optical element in the coupling system of the optical tweezers is moved under computer control, so that the position of the fixed object relative to the sample is retained. In principle, it is not necessary to take the position of the object plane from the control electronics of the laser scanning microscope, since the position of the relevant optical elements can also be detected electromechanically or optically. However, this is associated with a greater effort.
- the z-compensation can be combined with the microscope holder on the optical fiber.
- the result is a compact unit with a minimum of optical elements.
- the object to be examined is scanned in various xy sections.
- the object plane is between the cuts by the laser Scanning microscope moved.
- the position of the focus of the optical tweezers is brought back to the starting point by manually shifting the additional optical element located in the beam path of the optical tweezers. This process is repeated for each xy section.
- the z-compensation described here also allows the coupling of a so-called multitrap, an optical tweezer, in which one or more laser beams are directed onto several objects for fixation. This can also be done by using a scanner mirror to alternate a beam at a high frequency is directed onto several objects in such a way that they remain fixed, even if the laser beam does not permanently irradiate the corresponding object.
- a laser microbeam can be coupled in compensated (a laser microbeam is a short-pulsed laser beam that is coupled into a microscope to perform micromaterial processing).
- the same optics as for the optical tweezers can be used for the coupling of the laser micro beam.
- a z-compensated laser micro-beam enables precise material processing during image acquisition, for example to examine the light-material interaction in detail.
- Figure 2 shows a microscopic beam path with a sample P, an objective O and a tube lens TL.
- a laser beam L1 which scans the sample P in the x / y direction, is coupled in via a deflection mirror US via a scan lens SL and an x / y scanner SC as well as a deflection mirror US1 and a dichroic beam splitter ST1.
- the beam focus in the sample is here shifted by the objective O in Z-
- Height is adjusted via a control unit AS so that the sample can be scanned at different Z positions.
- the radiation coming from the sample passes in the opposite way via the
- an HBO - is connected via a further steel divider ST2 and a lens L.
- Lighting can be coupled.
- corresponding pulsing optics 01, 02 are used to continue pulsing via the beam splitter ST2 and a further beam splitter ST3
- Laser beam L2 for optical cutting and another laser beam L3 coupled as optical tweezers (Optical Tweezer).
- the light coupling can be an indirect coupling via light guides, to which collimation optics are arranged.
- Axis changes the beam focus position of the respective laser in sample P.
- correction optics 01, 02 are in the variant V1 along the optical
- the position of the focus within the sample can be changed in a defined manner in the Z direction.
- an object held with the optical tweezers can always be advantageous at the same point in the lens when the lens shifts in the Z direction
- Cutting laser L2 can be moved accordingly and thus the position of the cut can be chosen as desired and also decoupled from the position of laser L3.
- a common displaceable correction lens 03 is provided for the lasers L2, L3.
- decoupling of the movement of L2 and L3 can be achieved by means of different optics which can additionally be inserted into the beam path of the laser L2.
- multibeam tweezer i.e. tweezers that use one or more laser beams to hold several
- Objects can be used, possible.
- Scanner mirror is directed in high frequency onto several objects in such a way that they can be held simultaneously (C.Hoyer, S.Monajembashi, K.O. Greulich:
- Organelles can often not be focused because they move during image acquisition. Sharp, three-dimensional images are only possible through the use of compensated optical tweezers that enable the fixation of the organelles during image acquisition. In this way, cell organelles such as chloroplasts or mitochondria can be fixed in living cells and depicted sharply in three dimensions. Organelles that normally do not move, such as secretory vesicles or the gravesceptor, can be deflected from the original position with the optical tweezers and the response of the cell (reorganization) can be examined three-dimensionally. The cytoskeleton dynamics in living cells can also be examined by deflection from the resting position.
- Spheroids can be manipulated and examined three-dimensionally as an in-vivo model for tissue using z-compensated optical tweezers in a laser scanning microscope.
Landscapes
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Microscoopes, Condenser (AREA)
- Lens Barrels (AREA)
Abstract
Description
Claims
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19954933A DE19954933A1 (en) | 1999-11-10 | 1999-11-10 | Arrangement for coupling optical tweezers and / or a processing beam into a microscope |
DE19954933 | 1999-11-10 | ||
PCT/EP2000/010808 WO2001035150A1 (en) | 1999-11-10 | 2000-11-02 | System for introducing optical tweezers and/or a treatment beam into a microscope |
Publications (1)
Publication Number | Publication Date |
---|---|
EP1141761A1 true EP1141761A1 (en) | 2001-10-10 |
Family
ID=7929118
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP00971409A Ceased EP1141761A1 (en) | 1999-11-10 | 2000-11-02 | System for introducing optical tweezers and/or a treatment beam into a microscope |
Country Status (5)
Country | Link |
---|---|
US (1) | US6850363B1 (en) |
EP (1) | EP1141761A1 (en) |
JP (1) | JP2003514252A (en) |
DE (1) | DE19954933A1 (en) |
WO (1) | WO2001035150A1 (en) |
Families Citing this family (22)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE10233549B4 (en) | 2002-07-23 | 2021-10-14 | Leica Microsystems Cms Gmbh | Scanning microscope with manipulation light beam and method for scanning microscopy |
DE10247249A1 (en) | 2002-10-10 | 2004-04-22 | Leica Microsystems Heidelberg Gmbh | Scanning microscope with a mirror for coupling a manipulation light beam |
US7800750B2 (en) * | 2003-09-19 | 2010-09-21 | The Regents Of The University Of California | Optical trap utilizing a reflecting mirror for alignment |
WO2005029139A2 (en) * | 2003-09-19 | 2005-03-31 | The Regents Of The University Of California | A light-force sensor and method for measuring axial optical-trap forces from changes in light momentum along an optic axis |
US7586684B2 (en) * | 2005-01-21 | 2009-09-08 | New York University | Solute characterization by optoelectronkinetic potentiometry in an inclined array of optical traps |
DE102005046638C5 (en) | 2005-09-29 | 2024-02-15 | Leica Microsystems Cms Gmbh | Scanning microscope and method for sample manipulation with a manipulation light beam in a scanning microscope |
KR101157176B1 (en) * | 2005-12-20 | 2012-06-20 | 삼성전자주식회사 | Microfluidic device and method for concentration or purification of cells or viruses |
US7718953B2 (en) * | 2006-04-12 | 2010-05-18 | University Of Delaware | Electromagnetic/optical tweezers using a full 3D negative-refraction flat lens |
WO2008127410A2 (en) * | 2006-11-07 | 2008-10-23 | New York University | Holographic microfabrication and characterization system for soft matter and biological systems |
DK2100127T3 (en) * | 2006-11-20 | 2017-01-30 | Nanotemper Tech Gmbh | Rapid thermo-optic particle characterization |
US8741570B2 (en) * | 2008-02-06 | 2014-06-03 | Ludwig-Maximilians-Universitat Munchen | Thermo-optical characterisation of nucleic acid molecules |
US8174742B2 (en) | 2008-03-14 | 2012-05-08 | New York University | System for applying optical forces from phase gradients |
WO2009141390A1 (en) * | 2008-05-20 | 2009-11-26 | Ludwig-Maximilians-Universität München | Method and device for particle analysis using thermophoresis |
US9316578B2 (en) | 2008-10-30 | 2016-04-19 | New York University | Automated real-time particle characterization and three-dimensional velocimetry with holographic video microscopy |
DE102010027720A1 (en) * | 2010-04-14 | 2011-10-20 | Carl Zeiss Microlmaging Gmbh | Methods and devices for position and force detection |
WO2016077472A1 (en) | 2014-11-12 | 2016-05-19 | New York University | Colloidal fingerprints for soft materials using total holographic characterization |
US11385157B2 (en) | 2016-02-08 | 2022-07-12 | New York University | Holographic characterization of protein aggregates |
EP3538941A4 (en) | 2016-11-10 | 2020-06-17 | The Trustees of Columbia University in the City of New York | Rapid high-resolution imaging methods for large samples |
DE102018213965A1 (en) * | 2018-08-20 | 2020-02-20 | Universität Ulm | Device and method for the detection of a spatial extension of at least one adherent biological cell |
US11543338B2 (en) | 2019-10-25 | 2023-01-03 | New York University | Holographic characterization of irregular particles |
CN113136324A (en) * | 2020-01-19 | 2021-07-20 | 中国科学院青岛生物能源与过程研究所 | Coupling device, microscope-optical tweezers single cell sorting system and sorting method thereof |
US11948302B2 (en) | 2020-03-09 | 2024-04-02 | New York University | Automated holographic video microscopy assay |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH10206742A (en) * | 1996-11-21 | 1998-08-07 | Olympus Optical Co Ltd | Laser scanning microscope |
Family Cites Families (14)
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US4289378A (en) | 1978-06-21 | 1981-09-15 | Ernst Remy | Apparatus for adjusting the focal point of an operating laser beam focused by an objective |
DE3214268A1 (en) * | 1982-04-17 | 1983-10-20 | Fa. Carl Zeiss, 7920 Heidenheim | OPTICAL ADJUSTMENT |
US5022743A (en) * | 1987-03-27 | 1991-06-11 | The Board Of Trustees Of The Leland Stanford Junior University | Scanning confocal optical microscope |
KR900002716B1 (en) * | 1987-11-26 | 1990-04-23 | 재단법인한국전자 통신연구소 | Beam focusing device of laser-grapher |
GB2231681B (en) * | 1989-05-05 | 1993-04-21 | Hatfield Polytechnic | Optical microscopes |
JP3129471B2 (en) * | 1991-06-01 | 2001-01-29 | 科学技術振興事業団 | Multi-beam particle operation method |
JPH052136A (en) * | 1991-06-24 | 1993-01-08 | Nikon Corp | Particulate observing method and microscope used for the same |
DE4300698A1 (en) * | 1993-01-13 | 1994-07-14 | Raimund Schuetze | Device and method for handling, processing and observing small particles, in particular biological particles |
JP3339244B2 (en) * | 1995-02-23 | 2002-10-28 | 株式会社ニコン | Epi-fluorescence microscope |
DE19616216A1 (en) * | 1996-04-23 | 1997-10-30 | P A L M Gmbh | Laser beam process and assembly separates individual cells from tissue mass |
US5952651A (en) * | 1996-06-10 | 1999-09-14 | Moritex Corporation | Laser manipulation apparatus and cell plate used therefor |
US6075643A (en) * | 1997-10-24 | 2000-06-13 | Olympus Optical Co., Ltd. | Reflected fluorescence microscope with multiple laser and excitation light sources |
DE19801139B4 (en) | 1998-01-14 | 2016-05-12 | Till Photonics Gmbh | Point Scanning Luminescence Microscope |
WO2000057231A1 (en) * | 1999-03-19 | 2000-09-28 | Olympus Optical Co., Ltd. | Scanning confocal microscope |
-
1999
- 1999-11-10 DE DE19954933A patent/DE19954933A1/en not_active Withdrawn
-
2000
- 2000-11-02 JP JP2001536624A patent/JP2003514252A/en active Pending
- 2000-11-02 US US09/869,951 patent/US6850363B1/en not_active Expired - Lifetime
- 2000-11-02 EP EP00971409A patent/EP1141761A1/en not_active Ceased
- 2000-11-02 WO PCT/EP2000/010808 patent/WO2001035150A1/en active Application Filing
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH10206742A (en) * | 1996-11-21 | 1998-08-07 | Olympus Optical Co Ltd | Laser scanning microscope |
US6094300A (en) * | 1996-11-21 | 2000-07-25 | Olympus Optical Co., Ltd. | Laser scanning microscope |
Non-Patent Citations (1)
Title |
---|
See also references of WO0135150A1 * |
Also Published As
Publication number | Publication date |
---|---|
US6850363B1 (en) | 2005-02-01 |
DE19954933A1 (en) | 2001-05-17 |
WO2001035150A1 (en) | 2001-05-17 |
JP2003514252A (en) | 2003-04-15 |
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Legal Events
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PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
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17P | Request for examination filed |
Effective date: 20010704 |
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Kind code of ref document: A1 Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE TR |
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RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: MICHELMANN, ANJA Inventor name: WENDENBURG, RONALD Inventor name: GREULICH, KARL, OTTO Inventor name: UHL, VOLKER Inventor name: MONAJEMBASHI, SHAMCI |
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17Q | First examination report despatched |
Effective date: 20070920 |
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STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION HAS BEEN REFUSED |
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18R | Application refused |
Effective date: 20090702 |