High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events | Zendy

David M. Grant | Zendy; James McGinty | Zendy; Ewan J. McGhee | Zendy; Tom D. Bunney | Zendy; Dylan M. Owen | Zendy; Clifford Talbot | Zendy; W. Zhang | Zendy; Sunil Kumar | Zendy; Ian Munro | Zendy; Peter M. P. Lanigan | Zendy; Gordon T. Kennedy | Zendy; Christopher Dunsby | Zendy; Anthony I. Magee | Zendy; Patrick Courtney | Zendy; Matilda Katan | Zendy; Mark A. A. Neil | Zendy; P. M. W. French | Zendy

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High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events

Author(s) -

David M. Grant,

James McGinty,

Ewan J. McGhee,

Tom D. Bunney,

Dylan M. Owen,

Clifford Talbot,

W. Zhang,

Sunil Kumar,

Ian Munro,

Peter M. P. Lanigan,

Gordon T. Kennedy,

Christopher Dunsby,

Anthony I. Magee,

Patrick Courtney,

Matilda Katan,

Mark A. A. Neil,

P. M. W. French

Publication year - 2007

Publication title -

optics express

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.394

H-Index - 271

ISSN - 1094-4087

DOI - 10.1364/oe.15.015656

Subject(s) - live cell imaging , fluorescence lifetime imaging microscopy , microscopy , förster resonance energy transfer , microscope , materials science , confocal microscopy , confocal , fluorescence , fluorescence microscope , optics , light sheet fluorescence microscopy , biological imaging , cell , chemistry , physics , biochemistry

We present a time domain optically sectioned fluorescence lifetime imaging (FLIM) microscope developed for high-speed live cell imaging. This single photon excited system combines wide field parallel pixel detection with confocal sectioning utilizing spinning Nipkow disc microscopy. It can acquire fluorescence lifetime images of live cells at up to 10 frames per second (fps), permitting high-speed FLIM of cell dynamics and protein interactions with potential for high throughput cell imaging and screening applications. We demonstrate the application of this FLIM microscope to real-time monitoring of changes in lipid order in cell membranes following cholesterol depletion using cyclodextrin and to the activation of the small GTP-ase Ras in live cells using FRET.

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