SiFLIM
Single-image frequency-domain FLIM provides fast and photon-efficient lifetime data
Marcel Raspe (Nederlands Kanker Instituut - Antoni van Leeuwenhoek ziekenhuis)
Katarzyna M. Kedziora (Nederlands Kanker Instituut - Antoni van Leeuwenhoek ziekenhuis)
Bram Van Den Broek (Universiteit van Amsterdam, Nederlands Kanker Instituut - Antoni van Leeuwenhoek ziekenhuis)
Qiaole Zhao (TU Delft - ImPhys/Quantitative Imaging)
Sander De Jong (Lambert Instruments B.V.)
Johan Herz (Lambert Instruments B.V.)
Marieke Mastop (Universiteit van Amsterdam)
Joachim Goedhart (Van Leeuwenhoek Centre for Advanced Microscopy, Universiteit van Amsterdam)
Theodorus W.J. Gadella (Universiteit van Amsterdam, Van Leeuwenhoek Centre for Advanced Microscopy)
Ian T. Young (TU Delft - ImPhys/Quantitative Imaging)
Kees Jalink (Universiteit van Amsterdam, Van Leeuwenhoek Centre for Advanced Microscopy, Nederlands Kanker Instituut - Antoni van Leeuwenhoek ziekenhuis)
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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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