CA2231222A1 - Multi-photon laser microscopy - Google Patents

Multi-photon laser microscopy

Info

Publication number
CA2231222A1
CA2231222A1 CA002231222A CA2231222A CA2231222A1 CA 2231222 A1 CA2231222 A1 CA 2231222A1 CA 002231222 A CA002231222 A CA 002231222A CA 2231222 A CA2231222 A CA 2231222A CA 2231222 A1 CA2231222 A1 CA 2231222A1
Authority
CA
Canada
Prior art keywords
fluorophores
fluoroscent
laser
wavelength range
photobleaching
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.)
Granted
Application number
CA002231222A
Other languages
French (fr)
Other versions
CA2231222C (en
Inventor
Watt W. Webb
Chris Xu
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.
Watt W. Webb
Chris Xu
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=21708403&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=CA2231222(A1) "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., Watt W. Webb, Chris Xu filed Critical Cornell Research Foundation, Inc.
Publication of CA2231222A1 publication Critical patent/CA2231222A1/en
Application granted granted Critical
Publication of CA2231222C publication Critical patent/CA2231222C/en
Anticipated expiration legal-status Critical
Expired - Fee Related 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
    • G02B21/002—Scanning 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
    • G02B21/0052—Optical details of the image generation
    • G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
    • 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
    • G02B21/008—Details of detection or image processing, including general computer control
    • G02B21/0084—Details of detection or image processing, including general computer control time-scale detection, e.g. strobed, ultra-fast, heterodyne detection
    • 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

Abstract

A laser scanning micrsocope (10) produces molecular excitation in a target material (14) by simultaneous absorption of three or more photons to thereby provide intrinsic three-dimensional resolution.
Fluorophores having single photon absorption in the short (ultraviolet or visible) wavelength range are excited by a beam (16) of strongly focused subpicosecond pulses of laser light of relatively long (red or infrared) wavelength range. The fluorophores absorb at about one third, one fourth or even smaller fraction of the laser wavelength to produce fluoroscent images of living cells and other microscopic objects. The fluoroscent emission from the fluorophores increases cubicly, quarticly or even higher power law with the excitation intensity so that by focusing the laser light, fluorescence as well as photobleaching are confined to the vicinity of the focal plane. This feature provides depth of field resolution comparable to that produced by confocal laser scanning microscopes, and in addition reduces photobleaching and phototoxicity.
CA002231222A 1995-09-19 1996-09-18 Multi-photon laser microscopy Expired - Fee Related CA2231222C (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US395795P 1995-09-19 1995-09-19
US60/003,957 1995-09-19
PCT/US1996/014519 WO1997011355A1 (en) 1995-09-19 1996-09-18 Multi-photon laser microscopy

Publications (2)

Publication Number Publication Date
CA2231222A1 true CA2231222A1 (en) 1997-03-27
CA2231222C CA2231222C (en) 2001-12-11

Family

ID=21708403

Family Applications (1)

Application Number Title Priority Date Filing Date
CA002231222A Expired - Fee Related CA2231222C (en) 1995-09-19 1996-09-18 Multi-photon laser microscopy

Country Status (9)

Country Link
US (2) US6166385A (en)
EP (1) EP0852716B1 (en)
JP (3) JPH10512959A (en)
AU (1) AU700560B2 (en)
CA (1) CA2231222C (en)
DE (2) DE852716T1 (en)
NO (1) NO981032L (en)
NZ (1) NZ318277A (en)
WO (1) WO1997011355A1 (en)

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Also Published As

Publication number Publication date
EP0852716A1 (en) 1998-07-15
DE69635521D1 (en) 2006-01-05
WO1997011355A1 (en) 1997-03-27
DE852716T1 (en) 2001-07-19
NZ318277A (en) 1999-02-25
EP0852716B1 (en) 2005-11-30
JP2002139436A (en) 2002-05-17
CA2231222C (en) 2001-12-11
NO981032L (en) 1998-05-04
JPH10512959A (en) 1998-12-08
DE69635521T2 (en) 2006-08-17
EP0852716A4 (en) 1999-01-13
AU6972496A (en) 1997-04-09
AU700560B2 (en) 1999-01-07
NO981032D0 (en) 1998-03-09
US6166385A (en) 2000-12-26
JP2006106004A (en) 2006-04-20
US6344653B1 (en) 2002-02-05

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