Cy5, Atto 655 and Atto 647N are nearly unresolvable spectrally however they are characterized by significantly different lifetimes. LINCam is capable of imaging Surface-immobilized single molecules in the most challenging for its (QY ~2%) far-red spectral region. In the top figure the

three fluorophores are imaged separately. The precision in lifetime determination is less than 0.2 ns.

Same three types (bottom figure) of fluorophores on the surface measured in a mix. Lifetime- based identification of the fluorophore type is straight- forward.

Read full story: https://doi.org/10.1021/acs.jpca.0c01513