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Lightsheet fluorescence lifetime imaging microscopy with wide‐field time‐correlated single photon counting
Author(s) -
Hirvonen Liisa M.,
Nedbal Jakub,
Almutairi Norah,
Phillips Thomas A.,
Becker Wolfgang,
Conneely Thomas,
Milnes James,
Cox Susan,
Stürzenbaum Stephen,
Suhling Klaus
Publication year - 2020
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201960099
Subject(s) - optics , microscopy , picosecond , photon counting , fluorescence lifetime imaging microscopy , fluorescence , detector , materials science , photon , image intensifier , temporal resolution , fluorescence microscope , resolution (logic) , two photon excitation microscopy , optoelectronics , physics , laser , computer science , artificial intelligence
We report on wide‐field time‐correlated single photon counting (TCSPC)‐based fluorescence lifetime imaging microscopy (FLIM) with lightsheet illumination. A pulsed diode laser is used for excitation, and a crossed delay line anode image intensifier, effectively a single‐photon sensitive camera, is used to record the position and arrival time of the photons with picosecond time resolution, combining low illumination intensity of microwatts with wide‐field data collection. We pair this detector with the lightsheet illumination technique, and apply it to 3D FLIM imaging of dye gradients in human cancer cell spheroids, and C. elegans .