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Whole-field five-dimensional fluorescence microscopy combining lifetime and spectral resolution with optical sectioning
Authors:Siegel J  Elson D S  Webb S E  Parsons-Karavassilis D  Lévêque-Fort S  Cole M J  Lever M J  French P M  Neil M A  Juskaitis R  Sucharov L O  Wilson T
Abstract:We report a novel whole-field three-dimensional fluorescence lifetime imaging microscope that incoporates multispectral imaging to provide five-dimensional (5-D) fluorescence microscopy. This instrument, which can acquire a 5-D data set in less than a minute, is based on potentially compact and inexpensive diode-pumped solid-state laser technology. We demonstrate that spectral discrimination as well as optical sectioning minimize artifacts in lifetime determination and illustrate how spectral discrimination improves the lifetime contrast of biological tissue.
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