Open AccessOptically sectioned wide-field fluorescence lifetime imaging microscopy enabled by structured illumination
Author(s) -
Taylor Hinsdale,
Cory Olsovsky,
Jose J. Rico-Jimenez,
Kristen C. Maitland,
Javier A. Jo,
Bilal H. Malik
Publication year - 2017
Publication title -
biomedical optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.8.001455
Subject(s) - fluorophore , fluorescence , microscopy , fluorescence lifetime imaging microscopy , fluorescence microscope , optics , materials science , cheek pouch , microscope , physics , hamster , biology , microbiology and biotechnology
In this paper, we demonstrate the ability of structured illumination microscopy to enhance the ability of fluorescence lifetime imaging to resolve fluorescence lifetimes in relatively thick samples that possess distinct but spectrally overlapping fluorescent layers. Structured illumination fluorescent lifetime imaging microscopy (SI-FLIM) is shown to be able to accurately reconstruct lifetime values in homogenous fluorophore samples (POPOP, NADH, and FAD) as well as accurately measure fluorescent lifetime in two layer models that are layered with NADH/FAD over POPOP, where NADH/FAD and POPOP have spectral overlap. Finally, the ability of SI-FLIM was demonstrated in a hamster cheek pouch ex vivo to show that more accurate lifetimes could be measured for each layer of interest in the oral mucosa (epithelium and submucosa).