Open AccessParallel excitation-emission multiplexed fluorescence lifetime confocal microscopy for live cell imaging
Author(s) -
Ming Zhao,
Yu Liu,
Leilei Peng
Publication year - 2014
Publication title -
optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.22.010221
Subject(s) - multiplexing , fluorescence lifetime imaging microscopy , optics , confocal , confocal microscopy , microscope , microscopy , materials science , fluorescence , fluorescence microscope , spectral imaging , biological imaging , optoelectronics , physics , computer science , telecommunications
We present a novel excitation-emission multiplexed fluorescence lifetime microscopy (FLIM) method that surpasses current FLIM techniques in multiplexing capability. The method employs Fourier multiplexing to simultaneously acquire confocal fluorescence lifetime images of multiple excitation wavelength and emission color combinations at 44,000 pixels/sec. The system is built with low-cost CW laser sources and standard PMTs with versatile spectral configuration, which can be implemented as an add-on to commercial confocal microscopes. The Fourier lifetime confocal method allows fast multiplexed FLIM imaging, which makes it possible to monitor multiple biological processes in live cells. The low cost and compatibility with commercial systems could also make multiplexed FLIM more accessible to biological research community.