title: Multifocal multiphoton excitation and time correlated single photon counting detection for 3-D fluorescence lifetime imaging.
abstract: We report a multifocal multiphoton time-correlated single photon counting (TCSPC) fluorescence lifetime imaging (FLIM) microscope system that uses a 16 channel multi-anode PMT detector. Multiphoton excitation minimizes out-of-focus photobleaching, multifocal excitation reduces non-linear in-plane photobleaching effects and TCSPC electronics provide photon-efficient detection of the fluorescence decay profile. TCSPC detection is less prone to bleaching- and movement-induced artefacts compared to wide-field time-gated or frequency-domain FLIM. This microscope is therefore capable of acquiring 3-D FLIM images at significantly increased speeds compared to single beam multiphoton microscopy and we demonstrate this with live cells expressing a GFP tagged protein. We also apply this system to time-lapse FLIM of NAD(P)H autofluorescence in single live cells and report measurements on the change in the fluorescence decay profile following the application of a known metabolic inhibitor.
authors: S
authors: S
authors: Kumar
authors: C
authors: C
authors: Dunsby
authors: P A A
authors: PA
authors: De Beule
authors: D M
authors: DM
authors: Owen
authors: U
authors: U
authors: Anand
authors: P M P
authors: PM
authors: Lanigan
authors: R K P
authors: RK
authors: Benninger
authors: D M
authors: DM
authors: Davis
authors: M A A
authors: MA
authors: Neil
authors: P
authors: P
authors: Anand
authors: C
authors: C
authors: Benham
authors: A
authors: A
authors: Naylor
authors: P M W
authors: PM
authors: French