title: Spatially resolved recording of transient fluorescence-lifetime effects by line-scanning TCSPC.
abstract: We present a technique that records transient changes in the fluorescence lifetime of a sample with spatial resolution along a one-dimensional scan. The technique is based on scanning the sample with a high-frequency pulsed laser beam, detecting single photons of the fluorescence light, and building up a photon distribution over the distance along the scan, the arrival times of the photons after the excitation pulses and the time after a stimulation of the sample. The maximum resolution at which lifetime changes can be recorded is given by the line scan period. Transient lifetime effects can thus be resolved at a resolution of about one millisecond. We demonstrate the technique for recording photochemical and nonphotochemical chlorophyll transients in plants and transient changes in free Ca(2+) in cultured neurons.
authors.affiliation: Becker & Hickl GmbH, Nahmitzer Damm 30, 12277, Berlin, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Becker
authors.forename: Vladislav
authors.initials: V
authors.lastname: Shcheslavkiy
authors.forename: Samuel
authors.initials: S
authors.lastname: Frere
authors.forename: Inna
authors.initials: I
authors.lastname: Slutsky
mesh_terms: D000818:Animals; D002118:Calcium; D002478:Cells, Cultured; D002734:Chlorophyll / Q000737:chemistry; D005453:Fluorescence; D009474:Neurons / Q000737:chemistry*; D010777:Photochemistry; D017785:Photons; D010944:Plants / Q000737:chemistry*
chemical_list: D002734:Chlorophyll; D002118:Calcium
keywords: Ca2+ imaging; FLIM; FLITS; TCSPC; chlorophyll transients; fluorescence lifetime