A wide-field multi-parameter FLIM and FRAP setup to investigate the fluorescence emission of individual living cyanobacteria

Marco Vitali; Matthias Reis; Thomas Friedrich; Hann-Jörg Eckert

doi:10.1117/12.871520

24 November 2010 A wide-field multi-parameter FLIM and FRAP setup to investigate the fluorescence emission of individual living cyanobacteria

Marco Vitali, Matthias Reis, Thomas Friedrich, Hann-Jörg Eckert

Author Affiliations +

Proceedings Volume 7376, Laser Applications in Life Sciences; 737610 (2010) https://doi.org/10.1117/12.871520
Event: Laser Applications in Life Sciences 2010, 2010, Oulu, Finland

Abstract

Multi-parameter fluorescence lifetime imaging microscopy is a powerful tool to investigate the spatial dependence of the fluorescence decay signal of chromophores in living cells. A multi-channel detection system based on a position sensitive Quadrant Anode photomultiplier and time-correlated single photon counting (TCSPC) was applied to monitor the fluorescence decay in individual living cells of the cyanobacterium Thermosynechococcus elongatus. The fluorescence lifetime imaging system was used to detect at a frame-rate of 1 Hertz the recovery after photobleaching of Phycobilisomes (PBS), a photosynthetic pigment-protein complex with light harvesting functions. Simultaneous monitoring of the fluorescence decays of the PBS- and the Chlorophyll a (Chl a)-containing antenna systems unveils a heterogeneity of the cyanobacterial population with respect to the fluorescence lifetime. Furthermore the FLIM images clearly show that the decay of the fluorescence signal in the targeted area becomes longer after the bleaching at 633 nm for the whole measuring time. The results suggest that the diffusing PBSs, which replace the bleached ones in the targeted area, can only partially reestablish excitation energy transfer to Chl a.

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Marco Vitali, Matthias Reis, Thomas Friedrich, and Hann-Jörg Eckert "A wide-field multi-parameter FLIM and FRAP setup to investigate the fluorescence emission of individual living cyanobacteria", Proc. SPIE 7376, Laser Applications in Life Sciences, 737610 (24 November 2010); https://doi.org/10.1117/12.871520

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