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Spatially resolved recording of transient fluorescence‐lifetime effects by line‐scanning TCSPC
Author(s) -
Becker Wolfgang,
Shcheslavkiy Vladislav,
Frere Samuel,
Slutsky Inna
Publication year - 2014
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.22331
Subject(s) - transient (computer programming) , fluorescence , line (geometry) , materials science , optics , analytical chemistry (journal) , chemistry , chromatography , physics , computer science , geometry , mathematics , operating system
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. Microsc. Res. Tech. 77:216–224, 2014 . © 2014 Wiley Periodicals, Inc.