ES2123501T3 - Microscopio de barrido laser de dos fotones. - Google Patents

Microscopio de barrido laser de dos fotones.

Info

Publication number
ES2123501T3
ES2123501T3 ES90917225T ES90917225T ES2123501T3 ES 2123501 T3 ES2123501 T3 ES 2123501T3 ES 90917225 T ES90917225 T ES 90917225T ES 90917225 T ES90917225 T ES 90917225T ES 2123501 T3 ES2123501 T3 ES 2123501T3
Authority
ES
Spain
Prior art keywords
laser
exploration
microscope
excitation
fluorosphores
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.)
Expired - Lifetime
Application number
ES90917225T
Other languages
English (en)
Other versions
ES2123501T5 (es
Inventor
Winfried Denk
James P Strickler
Watt W Webb
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.)
Cornell Research Foundation Inc
Original Assignee
Cornell Research Foundation Inc
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
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=23730872&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=ES2123501(T3) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Cornell Research Foundation Inc filed Critical Cornell Research Foundation Inc
Publication of ES2123501T3 publication Critical patent/ES2123501T3/es
Application granted granted Critical
Publication of ES2123501T5 publication Critical patent/ES2123501T5/es
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6402—Atomic fluorescence; Laser induced fluorescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • G01N21/6458—Fluorescence microscopy
    • 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
    • G02B21/0052—Optical details of the image generation
    • G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
    • G—PHYSICS
    • G11—INFORMATION STORAGE
    • G11B—INFORMATION STORAGE BASED ON RELATIVE MOVEMENT BETWEEN RECORD CARRIER AND TRANSDUCER
    • G11B7/00—Recording or reproducing by optical means, e.g. recording using a thermal beam of optical radiation by modifying optical properties or the physical structure, reproducing using an optical beam at lower power by sensing optical properties; Record carriers therefor
    • G11B7/004—Recording, reproducing or erasing methods; Read, write or erase circuits therefor
    • G—PHYSICS
    • G11—INFORMATION STORAGE
    • G11B—INFORMATION STORAGE BASED ON RELATIVE MOVEMENT BETWEEN RECORD CARRIER AND TRANSDUCER
    • G11B7/00—Recording or reproducing by optical means, e.g. recording using a thermal beam of optical radiation by modifying optical properties or the physical structure, reproducing using an optical beam at lower power by sensing optical properties; Record carriers therefor
    • G11B7/12—Heads, e.g. forming of the optical beam spot or modulation of the optical beam
    • G—PHYSICS
    • G11—INFORMATION STORAGE
    • G11B—INFORMATION STORAGE BASED ON RELATIVE MOVEMENT BETWEEN RECORD CARRIER AND TRANSDUCER
    • G11B7/00—Recording or reproducing by optical means, e.g. recording using a thermal beam of optical radiation by modifying optical properties or the physical structure, reproducing using an optical beam at lower power by sensing optical properties; Record carriers therefor
    • G11B7/12—Heads, e.g. forming of the optical beam spot or modulation of the optical beam
    • G11B7/135—Means for guiding the beam from the source to the record carrier or from the record carrier to the detector
    • G11B7/1362—Mirrors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6408—Fluorescence; Phosphorescence with measurement of decay time, time resolved fluorescence
    • G01N2021/6415—Fluorescence; Phosphorescence with measurement of decay time, time resolved fluorescence with two excitations, e.g. strong pump/probe flash
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/631—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited using photolysis and investigating photolysed fragments
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Optics & Photonics (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microscoopes, Condenser (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Lasers (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

UN MICROSCOPIO DE EXPLORACION LASERICO (10) QUE PRODUCE UNA EXCITACION MOLECULAR EN UNA MATERIA DE BLANCO MEDIANTE LA ABSORCION SIMULTANEA DE DOS FOTONES PARA ASI PROPORCIONAR UNA RESOLUCION TRIDIMENSIONAL INTRINSECA. LOS FLUOROFOROS QUE TIENEN UNA ABSORCION DE FOTONES SENCILLA EN LA GAMA DE LONGITUDES DE ONDAS CORTAS (ULTRAVIOLETA O VISIBLE) SON EXCITADOS POR UNA SUCESION DE IMPULSOS DE SUBPICOSEGUNDOS FUERTEMENTE ENFOCADOS DE LUZ LASER (14) DE GAMA DE LONGITUDES DE ONDAS RELATIVAMENTE LARGAS (ROJAS O INFRAROJAS). LOS FLUOROSFOROS ABSORBEN ALREDEDOR DE LA MITAD DE LA LONGITUD DE ONDA LASERICA PARA PRODUCIR IMAGENES FLUORESCENTES DE CELULAS VIVAS Y OTROS OBJETOS MICROSCOPICOS. LA EMISION FLUORESCENTE DE LOS FLUOROSFOROS AUMENTA CUADRATICAMENTE CON LA INTENSIDAD DE EXCITACION DE MODO QUE A BASE DE ENFOCAR FUERTEMENTE LA LUZ LASERICA (14), LA FLUORESCENCIA ASI COMO EL FOTOBLANQUEO SON CONFINADOS CERCA DEL PLANO FOCAL (18). ESTA CARACTERISTICA PROPORCIONA UNA PROFUNDIDAD DE RESOLUCION DE CAMPOCOMPARABLE A LA PRODUCIDA POR LOS MICROSCOPICOS DE EXPLORACION LASERICOS CONFOCALES Y ADEMAS REDUCE EL FOTOBLANQUEO. LA EXPLORACION DEL RAYO LASER (14), POR UN MICROSCOPIO DE EXPLORACION LASERICO (10), PERMITE LA FORMACION DE IMAGENES MEDIANTE LA RECOGIDA DE UNA FLUORESCENCIA EXCITADA POR DOS FOTONES DESDE CADA PUNTO EN EL OBJETO EXPLORADO AL TIEMPO QUE TODAVIA SATISFACE EL REQUISITO PARA UNA INTENSIDAD DE EXCITACION MUY ALTA OBTENIDA A BASE DE ENFOCAR EL RAYO LASER (14) Y DE COMPRIMIR EL HAZ POR TIEMPO DE IMPULSOS. LOS IMPULSOS ENFOCADOS PROPORCIONAN TAMBIEN UNA FOTOQUIMICA TRIDIMENSIONAL RESUELTA ESPACIALMENTE QUE ES PARTICULARMENTE UTIL EN LA LIBERACION FOTOLITICA DE MOLECULAS EFECTORAS ENJAULADAS.
ES90917225T 1989-11-14 1990-11-13 Microscopio de barrido laser de dos fotones. Expired - Lifetime ES2123501T5 (es)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US436045 1989-11-14
US07/436,045 US5034613A (en) 1989-11-14 1989-11-14 Two-photon laser microscopy

Publications (2)

Publication Number Publication Date
ES2123501T3 true ES2123501T3 (es) 1999-01-16
ES2123501T5 ES2123501T5 (es) 2004-07-16

Family

ID=23730872

Family Applications (2)

Application Number Title Priority Date Filing Date
ES97108581T Expired - Lifetime ES2210414T3 (es) 1989-11-14 1990-11-13 Excitacion molecular de dos fotones en un microscopio de exploracion laser.
ES90917225T Expired - Lifetime ES2123501T5 (es) 1989-11-14 1990-11-13 Microscopio de barrido laser de dos fotones.

Family Applications Before (1)

Application Number Title Priority Date Filing Date
ES97108581T Expired - Lifetime ES2210414T3 (es) 1989-11-14 1990-11-13 Excitacion molecular de dos fotones en un microscopio de exploracion laser.

Country Status (9)

Country Link
US (1) US5034613A (es)
EP (2) EP0500717B2 (es)
JP (1) JP2848952B2 (es)
AT (2) ATE170636T1 (es)
DE (2) DE69034117T2 (es)
DK (1) DK0807814T3 (es)
ES (2) ES2210414T3 (es)
HK (1) HK1010411A1 (es)
WO (1) WO1991007651A1 (es)

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