Application of novel low-intensity nonscanning fluorescence lifetime imaging microscopy for monitoring excited state dynamics in individual chloroplasts and living cells of photosynthetic organisms

Hann-Jörg Eckert; Zdeněk Petrášek; Klaus Kemnitz

doi:10.1117/12.685958

25 October 2006 Application of novel low-intensity nonscanning fluorescence lifetime imaging microscopy for monitoring excited state dynamics in individual chloroplasts and living cells of photosynthetic organisms

Hann-Jörg Eckert, Zdeněk Petrášek, Klaus Kemnitz

Author Affiliations +

Proceedings Volume 6372, Advanced Photon Counting Techniques; 637207 (2006) https://doi.org/10.1117/12.685958
Event: Optics East 2006, 2006, Boston, Massachusetts, United States

Abstract

Picosecond fluorescence lifetime imaging microscopy (FLIM) provides a most valuable tool to analyze the primary processes of photosynthesis in individual cells and chloroplasts of living cells. In order to obtain correct lifetimes of the excited states, the peak intensity of the exciting laser pulses as well as the average intensity has to be sufficiently low to avoid distortions of the kinetics by processes such as singlet-singlet annihilation, closing of the reaction centers or photoinhibition. In the present study this requirement is achieved by non-scanning wide-field FLIM based on time- and space-correlated single-photon counting (TSCSPC) using a novel microchannel plate photomultiplier with quadrant anode (QA-MCP) that allows parallel acquisition of time-resolved images under minimally invasive low-excitation conditions. The potential of the wide-field TCSPC method is demonstrated by presenting results obtained from measurements of the fluorescence dynamics in individual chloroplasts of moss leaves and living cells of the chlorophyll d-containing cyanobacterium Acaryochloris marina.

Citation Download Citation

Hann-Jörg Eckert, Zdeněk Petrášek, and Klaus Kemnitz "Application of novel low-intensity nonscanning fluorescence lifetime imaging microscopy for monitoring excited state dynamics in individual chloroplasts and living cells of photosynthetic organisms", Proc. SPIE 6372, Advanced Photon Counting Techniques, 637207 (25 October 2006); https://doi.org/10.1117/12.685958

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