We present the implementation of a fast wide-field optical sectioning technique called HiLo microscopy on a fluorescence lifetime imaging microscope. HiLo microscopy is based on the fusion of two images, one with structured illumination and another with uniform illumination. Optically sectioned images are then digitally generated thanks to a fusion algorithm. HiLo images are comparable in quality with confocal images but they can be acquired faster over larger fields of view. We obtain 4D imaging by combining HiLo optical sectioning, time-gated detection, and z-displacement. We characterize the performances of this set-up in terms of 3D spatial resolution and time-resolved capabilities in both fixed- and live-cell imaging modes.

Date Published: 22 February 2013
PDF: 8 pages
Proc. SPIE 8589, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XX, 85890H (22 February 2013); doi: 10.1117/12.2003966

Published in SPIE Proceedings Vol. 8589:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XX
Carol J. Cogswell; Thomas G. Brown; Jose-Angel Conchello; Tony Wilson, Editor(s)