Two-Photon Laser Scanning Fluorescence Microscopy

Winfried Denk, James H. Strickler, Watt W. Webb

DSEID: DSEID-005-9324453
DOI: 10.1126/science.2321027
Journal: Science
Publisher: American Association for the Advancement of Science (AAAS)
Published: 1990-4-6
Status: temporarily_unreachable

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 photobleaching 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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Metadata

Title: Two-Photon Laser Scanning Fluorescence Microscopy
Delta ID: DSEID-005-9324453
Authors: Winfried Denk, James H. Strickler, Watt W. Webb
Abstract source: crossref
Source URL: https://doi.org/10.1126/science.2321027
Access: closed_or_uncertain
Licence: unknown
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