Watching Moving Molecules in Real Time: Fluorescence Lifetime Imaging in Frequency-Domain Widefield Microscopes

Along the domain of spectroscopy and biophysics labs, fluorescence lifetime is rapidly becoming an accessible tool for “mainstream” molecular biology research involving live cell imaging. In measuring a different physical property of a fluorophore, fluorescence lifetime imaging (FLI) has advantages in measuring protein proximity, polymerisation, relative concentration of different molecules, separation of different labels with spectral overlap, ion concentration, and removal of autofluorescence.