Spatially resolved recording of transient fluorescence-lifetime effects by line-scanning TCSPC

Wolfgang Becker*, Vladislav Shcheslavkiy, Samuel Frere, Inna Slutsky

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


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 Ca2+ in cultured neurons. Microsc. Res. Tech. 77:216-224, 2014.

Original languageEnglish
Pages (from-to)216-224
Number of pages9
JournalMicroscopy Research and Technique
Issue number3
StatePublished - Mar 2014


  • Chlorophyll transients
  • FLIM
  • Fluorescence lifetime


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