Advanced fluorescence imaging course on FLIM and FCS

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Authors

EŠNER Milan

Year of publication 2019
Type Workshop
MU Faculty or unit

Central European Institute of Technology

Citation
Description There are a lot of in vitro techniques to study small molecular interactions, but most of them fail to study the same under in vivo conditions. Advanced fluorescence imaging is an excellent for this purpose. Apart from the initially used property of photon intensity in fluorescence imaging studies, advanced imaging techniques use fluorescence properties such as fluorescence lifetime, correlation of movement of fluorescent molecules, total internal reflection or structured illumination approaches. Here we introduce the advanced fluorescent imaging techniques FLIM-FRET and FCS. FLIM-FRET is combination of fluorescence lifetime imaging microscopy (FLIM) and Förster resonance energy transfer (FRET). Fluorescent correlation and cross-correlation spectroscopy (FCS, FCCS) are techniques that allow us to study the fluorescent molecule concentration and diffusion at the single molecule level. FCCS is excellent for description of complex formation between fast moving molecules transferring across the confocal volume of a microscope.
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