title: siFLIM: single-image frequency-domain FLIM provides fast and photon-efficient lifetime data.
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.
authors.affiliation: Department of Cell Biology, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Marcel
authors.initials: M
authors.lastname: Raspe
authors.affiliation: Department of Cell Biology, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Katarzyna M
authors.initials: KM
authors.lastname: Kedziora
authors.affiliation: Department of Cell Biology, The Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Bram
authors.identifier: 0000-0003-1037-907X
authors.initials: B
authors.lastname: van den Broek
authors.affiliation: Department of Imaging Physics, Delft University of Technology, Delft, the Netherlands.
authors.forename: Qiaole
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: Lambert Instruments B.V., Groningen, the Netherlands.
authors.forename: Sander
authors.initials: S
authors.lastname: de Jong
authors.affiliation: Lambert Instruments B.V., Groningen, the Netherlands.
authors.forename: Johan
authors.initials: J
authors.lastname: Herz
authors.affiliation: Swammerdam Institute for Life Sciences, Section of Molecular Cytology, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Marieke
authors.initials: M
authors.lastname: Mastop
authors.affiliation: Swammerdam Institute for Life Sciences, Section of Molecular Cytology, University of Amsterdam, Amsterdam, the Netherlands.|Van Leeuwenhoek Centre for Advanced Microscopy, Amsterdam, the Netherlands.
authors.forename: Joachim
authors.initials: J
authors.lastname: Goedhart
authors.affiliation: Swammerdam Institute for Life Sciences, Section of Molecular Cytology, University of Amsterdam, Amsterdam, the Netherlands.|Van Leeuwenhoek Centre for Advanced Microscopy, Amsterdam, the Netherlands.
authors.forename: Theodorus W J
authors.initials: TW
authors.lastname: Gadella
authors.affiliation: Department of Imaging Physics, Delft University of Technology, Delft, the Netherlands.
authors.forename: Ian T
authors.initials: IT
authors.lastname: Young
authors.affiliation: Department of Cell Biology, The Netherlands Cancer Institute, Amsterdam, the Netherlands.|Swammerdam Institute for Life Sciences, Section of Molecular Cytology, University of Amsterdam, Amsterdam, the Netherlands.|Van Leeuwenhoek Centre for Advanced Microscopy, Amsterdam, the Netherlands.
authors.forename: Kees
authors.initials: K
authors.lastname: Jalink
mesh_terms: D002118:Calcium / Q000378:metabolism; D002138:Calibration; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D007089:Image Enhancement / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D017785:Photons; D059008:Time-Lapse Imaging / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D006632:Histamine; D002118:Calcium