Microscopy - L9

detect the lifetime of fluorescence (not intensity)

very sensitive to environment

decay measured for every pixel

decay is fitted to instrument response function (IRF)

measure interaction of molecules by light emission

donor absorbs high-energy light -> partially transmitts energy to acceptor -> acceptor emits light

energy transmission only possible in small distance -> distance measuring between 3-8 nm

acceptor bleaching increases donor emission

for accurate result: Donor only, Acceptor only, FRET experiment sample