title: Functional in vivo imaging using fluorescence lifetime light-sheet microscopy.
abstract: Light-sheet microscopy has become an indispensable tool for fast, low phototoxicity volumetric imaging of biological samples, predominantly providing structural or analyte concentration data in its standard format. Fluorescence lifetime imaging microscopy (FLIM) provides functional contrast, but often at limited acquisition speeds and with complex implementation. Therefore, we incorporate a dedicated frequency domain CMOS FLIM camera and intensity-modulated laser into a light-sheet setup to add fluorescence lifetime imaging functionality, allowing the rapid acquisition of volumetric data with concentration independent contrast. We then apply the system to image live transgenic zebrafish, demonstrating the capacity to rapidly collect volumetric FLIM data from an in vivo sample.
authors.forename: Claire A
authors.initials: CA
authors.lastname: Mitchell
authors.forename: Simon P
authors.initials: SP
authors.lastname: Poland
authors.forename: James
authors.initials: J
authors.lastname: Seyforth
authors.forename: Jakub
authors.initials: J
authors.lastname: Nedbal
authors.forename: Thomas
authors.initials: T
authors.lastname: Gelot
authors.forename: Tahiyat
authors.initials: T
authors.lastname: Huq
authors.forename: Gerhard
authors.initials: G
authors.lastname: Holst
authors.forename: Robert D
authors.initials: RD
authors.lastname: Knight
authors.forename: Simon M
authors.initials: SM
authors.lastname: Ameer-Beg
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D008856:Microscopy, Fluorescence / Q000379:methods*; D013997:Time Factors; D015027:Zebrafish / Q000235:genetics