Whole-field five-dimensional fluorescence microscopy combining lifetime and spectral resolution with optical sectioning

OPT LETT 26 (17): 1338-1340 SEP 1 2001 Siegel J, Elson DS, Webb SED, et al.

We report a novel whole-field three-dimensional f luorescence lifetime imaging microscope that incoporatesmultispectral imaging to provide five-dimensional (5-D) f luorescence microscopy. This instrument, which canacquire a 5-D data set in less than a minute, is based on potentially compact and inexpensive diode-pumpedsolid-state laser technology. We demonstrate that spectral discrimination as well as optical sectioning minimizeartifacts in lifetime determination and illustrate how spectral discrimination improves the lifetime contrastof biological tissue.
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