6. Fluorescence Lifetime Imaging (FLIM)
The setup produces an image based on the differences in the excited state decay time of fluorophores accumulated in different regions in a sample. The image contrast is based on the lifetime of individual fluorophores rather than their emission spectra.
Integrated in SP8 STED
Fluorescence resonance energy transfer measurements using FLIM can provide a method to discriminate between different fluorophores with strongly overlapping spectra or fluorophores in a different environment.