siFLIM: single-image frequency-domain FLIM provides fast and photon-efficient lifetime data.

Raspe, Marcel; Kedziora, Katarzyna M; van den Broek, Bram; Zhao, Qiaole; de Jong, Sander; Herz, Johan; Mastop, Marieke; Goedhart, Joachim et al. · Nat Methods · 2016

basic_science · Level V

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Abstract

We developed single-image fluorescence lifetime imaging microscopy (siFLIM), a method for acquiring quantitative lifetime images from a single exposure. siFLIM takes advantage of a new generation of dedicated cameras that simultaneously record two 180°-phase-shifted images, and it allows for video-rate lifetime imaging with minimal phototoxicity and bleaching. siFLIM is also inherently immune to artifacts stemming from rapid cellular movements and signal transients.

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