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Lifetime estimation on moving sub-cellular objects in frequency domain FLIM imaging

Abstract : Fluorescence lifetime is usually defined as the average nanosecond-scale delay between excitation and emission of fluorescence. It has been established that lifetime measurement yields numerous indications on cellular processes such as inter-protein and intra-protein mechanisms through fluorescent tagging and Förster resonance energy transfer (FRET). In this area, frequency domain fluorescence lifetime imaging microscopy (FD FLIM) is particularly well appropriate to probe a sample non-invasively and quantify these interactions in living cells. The aim is then to measure fluorescence lifetime in the sample at each location in space from fluorescence variations observed in a temporal sequence of images obtained by phase modulation of the detection signal. This leads to a sensitivity of lifetime determination to other sources of fluorescence variations such as intracellular motion. In this paper, we propose a robust statistical method for lifetime estimation on both background and small moving structures with a focus on intracellular vesicle trafficking.
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Submitted on : Friday, December 18, 2015 - 4:31:27 PM
Last modification on : Wednesday, February 2, 2022 - 3:53:03 PM
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Philippe Roudot, Charles Kervrann, Cedric Blouin, Francois Waharte. Lifetime estimation on moving sub-cellular objects in frequency domain FLIM imaging. Journal of the Optical Society of America. A Optics, Image Science, and Vision, 2015, 32 (10), pp.15. ⟨10.1364/JOSAA.32.001821⟩. ⟨hal-01246392⟩



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