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

We report a novel whole-field three-dimensional fluorescence lifetime imagi ng microscope that incoporates multispectral imaging to provide five-dimens ional (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 inex pensive diode-pumped solid-state laser technology. We demonstrate that spec tral discrimination as well as optical sectioning minimize artifacts in lif etime determination and illustrate how spectral discrimination improves the lifetime contrast of biological tissue. (C) 2001 Optical Society of Americ a.