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Development of a TIRF-FLIM microscope for biomedical applications

Total internal reflection fluorescence microscopy (TIRFM) is a powerful optical technique to observe fluorescence close to surfaces. Associated with fluorescence lifetime imaging, TIRFM enables to measure contrasts independent of fluorophores concentration and reveal intracellular activity with subwavelength axial resolution. We developed an original setup which allows, thanks to a wide-field time resolved detection, to measure nanoseconds fluorescence lifetimes of membrane receptors in order to apprehend their signalisation pathway.