EP0706671A1 - Verfahren zur lumineszenz-rastermikroskopie und ein lumineszenzrastermikroskop - Google Patents
Verfahren zur lumineszenz-rastermikroskopie und ein lumineszenzrastermikroskopInfo
- Publication number
- EP0706671A1 EP0706671A1 EP95916571A EP95916571A EP0706671A1 EP 0706671 A1 EP0706671 A1 EP 0706671A1 EP 95916571 A EP95916571 A EP 95916571A EP 95916571 A EP95916571 A EP 95916571A EP 0706671 A1 EP0706671 A1 EP 0706671A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- laser
- luminescence
- luminescent
- scanning microscope
- light emitted
- 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
- 238000004020 luminiscence type Methods 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims abstract description 24
- 238000004621 scanning probe microscopy Methods 0.000 title claims abstract description 5
- 230000005284 excitation Effects 0.000 claims abstract description 13
- 230000005855 radiation Effects 0.000 claims abstract description 5
- 238000011156 evaluation Methods 0.000 claims description 3
- 238000000386 microscopy Methods 0.000 claims description 2
- 238000012216 screening Methods 0.000 claims description 2
- 239000004065 semiconductor Substances 0.000 claims description 2
- 238000011835 investigation Methods 0.000 claims 1
- 230000004936 stimulating effect Effects 0.000 claims 1
- 238000005259 measurement Methods 0.000 description 7
- 239000000975 dye Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000004880 explosion Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 230000009916 joint effect Effects 0.000 description 1
- 229910052743 krypton Inorganic materials 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
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
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
Definitions
- the invention relates to a method for luminescence scanning microscopy according to the preamble of the main claim and a luminescence scanning microscope for performing this method.
- REPLACEMENT LEAF luminescent, especially fluorescent dye The dye in the object is excited by the joint action of two photons and the light emitted by the dye is registered and evaluated by means of a detector. In order to capture the entire sample, the laser beam is scanned over the object. The values of all points of the object are registered, which enables a three-dimensional representation of the evaluation results.
- the two-photon excitation in connection with the scanning leads to a direct three-dimensional image of the object.
- laser pulses are used, the duration of which is in the sub-pico second range. Lasers that generate such extremely short pulses are very expensive.
- biological objects are often destroyed by the pulses of high power. Excitation of the object with laser pulses in the sub-pico-second range often leads to the luminescence signal "breaking down". This "collapse” can be clearly illustrated as a "micro-explosion” that destroys the object material.
- the object of the invention is to develop a generic method in such a way that the disadvantages mentioned above do not occur.
- Measuring times are usually applied, the sensitivity of the detector used being adapted to the measuring conditions. It has been found that if the
- lasers with continuous or pulsed radiation can be used as the light source for carrying out the measuring method according to the invention. These are not expensive.
- Conventional semiconductor or gas lasers can be used. HeNe lasers and krypton ion lasers are particularly suitable as gas lasers.
- the laser arrangement can also be designed in the form of an array. The use of two or three lasers for irradiating the same object point has proven to be particularly advantageous.
- a detector with a high signal-to-noise ratio is used to record and evaluate the luminescent light. It is particularly important that the detector used has a correspondingly high sensitivity and a has low self-noise.
- the array-like arrangement of the lasers and detectors and the resulting irradiation and measurement of several raster points makes it possible to shorten the duration of the method.
- a beam screening device is arranged upstream of the object, the beam being scanned over the object.
- Filters are also provided in the luminescence scanning microscope according to the invention for separating the light emitted by the sample from the laser light.
- a detector can also be used, which filters out the corresponding wavelength.
- FIG. 1 A schematic representation of a luminescence scanning microscope for carrying out the method according to the invention is shown in FIG. 1.
- the luminescence scanning microscope has a laser 1 for generating the laser light, a photo detector 2 for evaluating the measurement results and a lens 3 for focusing the laser beam 4 emitted by the laser light source 1 onto the object 5 to be examined.
- the laser beam 4 is controlled with a beam raster device 6 for scanning the object 5.
- the luminescence scanning microscope has filters 7 and 8 for separating the laser light 4 from the luminescent light.
- the laser light source 1 is designed as a conventional laser with continuous or pulsed radiation.
- the laser light source 1 can also be formed by an array-like arrangement of lasers.
Abstract
Description
Claims
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE4414940 | 1994-04-28 | ||
DE4414940A DE4414940C2 (de) | 1994-04-28 | 1994-04-28 | Lumineszenz-Rastermikroskop mit zwei Photonen Anregung |
PCT/DE1995/000566 WO1995030166A1 (de) | 1994-04-28 | 1995-04-27 | Verfahren zur lumineszenz-rastermikroskopie und ein lumineszenzrastermikroskop |
Publications (1)
Publication Number | Publication Date |
---|---|
EP0706671A1 true EP0706671A1 (de) | 1996-04-17 |
Family
ID=6516737
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP95916571A Ceased EP0706671A1 (de) | 1994-04-28 | 1995-04-27 | Verfahren zur lumineszenz-rastermikroskopie und ein lumineszenzrastermikroskop |
Country Status (4)
Country | Link |
---|---|
US (1) | US5777732A (de) |
EP (1) | EP0706671A1 (de) |
DE (1) | DE4414940C2 (de) |
WO (1) | WO1995030166A1 (de) |
Families Citing this family (59)
Publication number | Priority date | Publication date | Assignee | Title |
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WO1997011355A1 (en) * | 1995-09-19 | 1997-03-27 | Cornell Research Foundation, Inc. | Multi-photon laser microscopy |
DE19629141A1 (de) * | 1996-07-19 | 1998-04-16 | Bayer Ag | Verfahren und Vorrichtung zum Screening von Molekülen bezüglich ihres individuellen Bindungsverhaltens zu mindestens einem vorgegebenen Ligand |
DE19653413C2 (de) | 1996-12-22 | 2002-02-07 | Stefan Hell | Rastermikroskop, bei dem eine Probe in mehreren Probenpunkten gleichzeitig optisch angeregt wird |
CA2279574C (en) | 1997-01-31 | 2007-07-24 | The Horticulture & Food Research Institute Of New Zealand Ltd. | Optical apparatus |
JP3816632B2 (ja) * | 1997-05-14 | 2006-08-30 | オリンパス株式会社 | 走査型顕微鏡 |
US6020591A (en) * | 1997-07-11 | 2000-02-01 | Imra America, Inc. | Two-photon microscopy with plane wave illumination |
DE19733193B4 (de) * | 1997-08-01 | 2005-09-08 | Carl Zeiss Jena Gmbh | Mikroskop mit adaptiver Optik |
DE19733195B4 (de) * | 1997-08-01 | 2006-04-06 | Carl Zeiss Jena Gmbh | Hoch-Kompaktes Laser Scanning Mikroskop mit integriertem Kurzpuls Laser |
DE19733194B4 (de) * | 1997-08-01 | 2005-06-16 | Carl Zeiss Jena Gmbh | Laser-Scanning-Mikroskop |
US6149867A (en) | 1997-12-31 | 2000-11-21 | Xy, Inc. | Sheath fluids and collection systems for sex-specific cytometer sorting of sperm |
US6169289B1 (en) | 1998-01-27 | 2001-01-02 | Wisconsin Alumni Research Foundation | Signal enhancement for fluorescence microscopy |
US6342397B1 (en) * | 1998-06-04 | 2002-01-29 | Erkki Soini | Homogeneous biospecific assay using a solid phase, two-photon excitation and confocal fluorescence detection |
PT1917974E (pt) | 1998-07-30 | 2011-02-22 | Xy Llc | Sistema de inseminação artificial não cirúrgica em equinos |
DE19851240C1 (de) * | 1998-11-06 | 2000-03-02 | Europ Lab Molekularbiolog | Fluoreszenzmikroskop mit Mehrphotonenanregung |
DE19901381A1 (de) * | 1999-01-15 | 2000-07-20 | Joerg Enderlein | Verfahren und Vorrichtung zur optischen Detektion eines Partikels |
DE19908883A1 (de) | 1999-03-02 | 2000-09-07 | Rainer Heintzmann | Verfahren zur Erhöhung der Auflösung optischer Abbildung |
US6403332B1 (en) | 1999-07-30 | 2002-06-11 | California Institute Of Technology | System and method for monitoring cellular activity |
US7024316B1 (en) * | 1999-10-21 | 2006-04-04 | Dakocytomation Colorado, Inc. | Transiently dynamic flow cytometer analysis system |
EP1102101A1 (de) * | 1999-11-22 | 2001-05-23 | Leica Microsystems Heidelberg GmbH | Laserscanmikroskop |
US7208265B1 (en) | 1999-11-24 | 2007-04-24 | Xy, Inc. | Method of cryopreserving selected sperm cells |
BR0110731A (pt) | 2000-05-09 | 2004-04-27 | Xy Inc | Populações de espermatózoides contendo cromossomo x e cromossomo y de alta pureza |
DE20122783U1 (de) * | 2000-06-17 | 2007-11-15 | Leica Microsystems Cms Gmbh | Anordnung zum Untersuchen mikroskopischer Präparate mit einem Scanmikroskop und Beleuchtungseinrichtung für ein Scanmikroskop |
DE10115486A1 (de) | 2000-06-17 | 2001-12-20 | Leica Microsystems | Verschränkte-Photonen-Mikroskop |
US6898367B2 (en) | 2000-06-17 | 2005-05-24 | Leica Microsystems Heidelberg Gmbh | Method and instrument for microscopy |
EP1164401B1 (de) | 2000-06-17 | 2005-03-09 | Leica Microsystems Heidelberg GmbH | Verschränkte-Photonen-Mikroskop |
US6687000B1 (en) | 2000-06-26 | 2004-02-03 | Wisconsin Alumni Research Foundation | Photon-sorting spectroscopic microscope system |
DE10039520A1 (de) | 2000-08-08 | 2002-02-21 | Leica Microsystems | Vorrichtung zur Untersuchung und Manipulation von mikroskopischen Objekten |
DE10044308A1 (de) * | 2000-09-07 | 2002-03-21 | Leica Microsystems | Verfahren und Vorrichtung zur Detektion von Fluoreszenzlicht bei der konfokalen Rastermikroskopie |
DE10045245A1 (de) * | 2000-09-13 | 2002-03-28 | Siemens Ag | Einrichtung für optische Inspektion einer auf Defekte hin zu prüfenden Oberfläche eines Objekts |
EP1207387A1 (de) * | 2000-11-20 | 2002-05-22 | Institut Curie | Multiphotonabbildungsvorrichtung |
US7713687B2 (en) | 2000-11-29 | 2010-05-11 | Xy, Inc. | System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations |
AU3768902A (en) | 2000-11-29 | 2002-06-11 | Xy Inc | System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations |
US6552794B2 (en) * | 2001-04-04 | 2003-04-22 | Applied Spectral Imaging Ltd. | Optical detection method for improved sensitivity |
DE10120425C2 (de) * | 2001-04-26 | 2003-12-18 | Leica Microsystems | Scanmikroskop |
US20030211009A1 (en) * | 2001-05-18 | 2003-11-13 | Buchanan Kris S. | Rapid multi-material sample input system |
WO2003060610A1 (de) * | 2002-01-16 | 2003-07-24 | Carl Zeiss Jena Gmbh | Verfahren und anordnungen zur mikroskopischen abbildung |
DE10206980A1 (de) | 2002-02-20 | 2003-08-21 | Leica Microsystems | Mikroskop, Detektor und Verfahren zur Mikroskopie |
DE10228374A1 (de) | 2002-06-25 | 2004-01-15 | Leica Microsystems Heidelberg Gmbh | Verfahren zur Mikroskopie und Mikroskop |
AU2003265362B2 (en) | 2002-08-01 | 2009-11-05 | Xy, Llc. | Low pressure sperm cell separation system |
US8486618B2 (en) | 2002-08-01 | 2013-07-16 | Xy, Llc | Heterogeneous inseminate system |
AU2003265471B2 (en) | 2002-08-15 | 2009-08-06 | Xy, Llc. | High resolution flow cytometer |
US7169548B2 (en) | 2002-09-13 | 2007-01-30 | Xy, Inc. | Sperm cell processing and preservation systems |
DK2308417T3 (da) | 2003-03-28 | 2016-07-04 | Inguran Llc | Apparat og fremgangsmåder til tilvejebringelse af sorterede partikler |
DE10314750A1 (de) * | 2003-03-31 | 2004-11-04 | Leica Microsystems Heidelberg Gmbh | Rastermikroskop zur Detektion eines Objekts |
US7151270B2 (en) * | 2003-05-02 | 2006-12-19 | Leica Microsystems Cms Gmbh | Method for classifying object image regions of an object to be detected using a scanning microscope |
AU2004242121B2 (en) | 2003-05-15 | 2010-06-24 | Xy, Llc. | Efficient haploid cell sorting for flow cytometer systems |
US7706863B2 (en) | 2004-01-21 | 2010-04-27 | University Of Washington | Methods for assessing a physiological state of a mammalian retina |
EP2801363B1 (de) | 2004-03-29 | 2018-02-21 | Inguran, LLC | Verfahren zur lagerung von sortierten spermatozoen |
CA2574499C (en) | 2004-07-22 | 2016-11-29 | Monsanto Technology Llc | Process for enriching a population of sperm cells |
PT1771729E (pt) | 2004-07-27 | 2015-12-31 | Beckman Coulter Inc | Acentuação da discriminação da citometria de fluxo com transformação geométrica |
EP1889039B1 (de) * | 2005-05-31 | 2015-04-22 | W.O.M. World of Medicine AG | Verfahren und vorrichtung zur optischen charakterisierung von gewebe |
DE102006029809B3 (de) | 2006-06-28 | 2007-11-08 | Ltb Lasertechnik Berlin Gmbh | Ortsaufgelöstes Messverfahren für die Detektion von Melanin in Fluorophorgemischen in einer Festkörperprobe |
US8217992B2 (en) | 2007-01-11 | 2012-07-10 | The Jackson Laboratory | Microscopic imaging techniques |
US7772569B2 (en) * | 2008-04-01 | 2010-08-10 | The Jackson Laboratory | 3D biplane microscopy |
DE102009029831A1 (de) | 2009-06-17 | 2011-01-13 | W.O.M. World Of Medicine Ag | Vorrichtung und Verfahren für die Mehr-Photonen-Fluoreszenzmikroskopie zur Gewinnung von Informationen aus biologischem Gewebe |
DE102011007751B4 (de) | 2011-04-20 | 2023-10-19 | Carl Zeiss Microscopy Gmbh | Weitfeldmikroskop und Verfahren zur Weitfeldmikroskopie |
JP2020502558A (ja) | 2016-11-10 | 2020-01-23 | ザ トラスティーズ オブ コロンビア ユニバーシティ イン ザ シティ オブ ニューヨーク | 大型試料のための高速・高解像度イメージング方法 |
CN109718476A (zh) * | 2018-12-28 | 2019-05-07 | 中国科学院苏州生物医学工程技术研究所 | 三维扫描双光子刺激系统及其刺激方法 |
CN110146473B (zh) * | 2019-04-16 | 2020-10-13 | 浙江大学 | 一种轴向超分辨的双光子荧光显微装置及方法 |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
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AU3849685A (en) * | 1984-02-16 | 1985-08-22 | Molecular Biophysics Technology, Inc. | Pusled light selective photcysis for sterilization of biological fluid |
US5022757A (en) * | 1989-01-23 | 1991-06-11 | Modell Mark D | Heterodyne system and method for sensing a target substance |
US5034613A (en) * | 1989-11-14 | 1991-07-23 | Cornell Research Foundation, Inc. | Two-photon laser microscopy |
DE4035799C2 (de) * | 1990-11-10 | 1995-10-12 | Groskopf Rudolf Dr Ing | Vorrichtung zur dreidimensionalen optischen Untersuchung eines Objektes |
DE4113279C2 (de) * | 1991-04-24 | 1996-08-29 | Miodrag Dipl Phys Milicev | Konfokales optisches Rastermikroskop |
US5196709A (en) * | 1991-05-03 | 1993-03-23 | University Of Maryland Systems | Fluorometry method and apparatus using a semiconductor laser diode as a light source |
US5462879A (en) * | 1993-10-14 | 1995-10-31 | Minnesota Mining And Manufacturing Company | Method of sensing with emission quenching sensors |
-
1994
- 1994-04-28 DE DE4414940A patent/DE4414940C2/de not_active Revoked
-
1995
- 1995-04-27 US US08/571,839 patent/US5777732A/en not_active Expired - Lifetime
- 1995-04-27 WO PCT/DE1995/000566 patent/WO1995030166A1/de not_active Application Discontinuation
- 1995-04-27 EP EP95916571A patent/EP0706671A1/de not_active Ceased
Non-Patent Citations (1)
Title |
---|
See references of WO9530166A1 * |
Also Published As
Publication number | Publication date |
---|---|
WO1995030166A1 (de) | 1995-11-09 |
DE4414940C2 (de) | 1998-07-02 |
DE4414940A1 (de) | 1995-11-02 |
US5777732A (en) | 1998-07-07 |
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Legal Events
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