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Camera-based time-correlated single photon counting (TCSPC) is a method where the position and the arrival time of the photons are recorded simultaneously. This has some advantages for fluorescence lifetime imaging (FLIM) with certain types of microscopy. It also allows FLIM with a macroscopic field of view.
We report on nanosecond fluorescence lifetime imaging (FLIM) combined with total internal reflection fluorescence (TIRF) microscopy based on a 40 mm diameter crossed delay line anode detector. It has a few 100 picoseconds time resolution, and is read out via three standard TCSPC boards. We apply this wide-field TCSPC detector to identify Förster Resonance Energy Transfer (FRET) in cell membrane proteins in TIR-FLIM microscopy.
In addition, we use a TCSPC single photon avalanche diode (SPAD) array as a macroscopic camera to visualise varnish removal on paintings.
Conference Presentation
(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.
Klaus Suhling,Jakub Nedbal,Louis Obeid Mogridge, andShirwa Awale
"Advances in wide-field camera-based time-correlated single photon counting fluorescence lifetime imaging", Proc. SPIE PC12848, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXXI, PC1284808 (13 March 2024); https://doi.org/10.1117/12.3001927
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Klaus Suhling, Jakub Nedbal, Louis Obeid Mogridge, Shirwa Awale, "Advances in wide-field camera-based time-correlated single photon counting fluorescence lifetime imaging," Proc. SPIE PC12848, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXXI, PC1284808 (13 March 2024); https://doi.org/10.1117/12.3001927