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.
© 2001 Optical Society of America
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.6920) Medical optics and biotechnology : Time-resolved imaging
(180.0180) Microscopy : Microscopy
(180.2520) Microscopy : Fluorescence microscopy
J. Siegel, D. S. Elson, S. E. D. Webb, D. Parsons-Karavassilis, S. Lévêque-Fort, M. J. Cole, M. J. Lever, P. M. W. French, M. A. A. Neil, R. Juskaitis, L. O. Sucharov, and T. Wilson, "Whole-field five-dimensional fluorescence microscopy combining lifetime and spectral resolution with optical sectioning," Opt. Lett. 26, 1338-1340 (2001)