Lightsheet fluorescence lifetime imaging microscopy (FLIM) with wide-field time-correlated single photon counting (TCSPC)

Liisa M Hirvonen, Jakub Nedbal, Norah Almutairi, Thomas A Phillips, Wolfgang Becker, Thomas Conneely, James Milnes, Susan Cox, Stephen Stürzenbaum, Klaus Suhling

Research output: Contribution to journalArticle

Abstract

We report onwide-field time-correlated single photon counting (TCSPC)-based fluorescence lifetime imagingmicroscopy (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 microwattswith 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. This article is protected by copyright. All rights reserved.

Original languageEnglish
Article numbere201960099
JournalJournal of Biophotonics
DOIs
Publication statusE-pub ahead of print - 29 Oct 2019
Externally publishedYes

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