Two-photon laser scanning fluorescence microscopy

Science
W DenkW W Webb

Abstract

Molecular excitation by the simultaneous absorption of two photons provides intrinsic three-dimensional resolution in laser scanning fluorescence microscopy. The excitation of fluorophores having single-photon absorption in the ultraviolet with a stream of strongly focused subpicosecond pulses of red laser light has made possible fluorescence images of living cells and other microscopic objects. The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation. This technique also provides unprecedented capabilities for three-dimensional, spatially resolved photochemistry, particularly photolytic release of caged effector molecules.

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Jun 27, 2009·Annual Review of Neuroscience·Brian A WiltMark J Schnitzer
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Sep 2, 2004·Photochemistry and Photobiology·Brett A King, Dennis H Oh
Dec 14, 2004·Photochemistry and Photobiology·Changhuei Yang
Apr 6, 2005·Photochemistry and Photobiology·Kevin W Eliceiri, Curtis Rueden
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Nov 28, 2006·Frontiers in Bioscience : a Journal and Virtual Library·Peter G BushAndrew C Hall
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May 1, 2009·HFSP Journal·Julien VermotMichael Liebling
Feb 2, 2010·HFSP Journal·Alexandre SpechtMaurice Goeldner
May 19, 2010·International Journal of Molecular Sciences·Michael WagnerHerbert Schneckenburger
Sep 12, 2013·International Journal of Molecular Sciences·Masood SamimVirginijus Barzda
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Mar 2, 2010·Journal of Visualized Experiments : JoVE·Kelley D Sullivan, Edward B Brown
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Jul 4, 2012·Journal of Pathology Informatics·Yu ChenLiron Pantanowitz
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Dec 1, 2010·Chemical Communications : Chem Comm·Ryuhei NishiyabuJohn S Fossey
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Sep 26, 2009·The Journal of Chemical Physics·Masayoshi NakanoBenoît Champagne
Oct 2, 2009·The Review of Scientific Instruments·Partha Pratim Mondal
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Apr 10, 2009·The Journal of Chemical Physics·Jonathan A N FisherArjun G Yodh
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Oct 7, 2011·The Review of Scientific Instruments·David T ClarkeMarisa L Martin-Fernandez
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Jun 15, 2011·The Journal of Chemical Physics·Kasper KristensenAntonio Rizzo
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Jan 2, 2014·Analytical Chemistry·Rajan AroraMarlan O Scully
Dec 2, 2009·The Review of Scientific Instruments·Jiun-Yann YuShi-Wei Chu
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May 6, 2010·The Review of Scientific Instruments·Partha Pratim MondalAlberto Diaspro
Jan 6, 2006·The Journal of Chemical Physics·Ion CohanoschiFlorencio E Hernández
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