Bleaching-corrected fluorescence microspectroscopy with nanometer peak position resolution

In brief

Fluorescence microspectroscopy (FMS) with environmentally sensitive dyes provides information about local molecular surroundings at microscopic spatial resolution. We showed that careful analysis of data acquired by stochastic wavelength sampling enables nanometer spectral peak position resolution even for highly photosensitive fluorophores with small spectral shifts, which are commonly used in bulk spectrofluorimetry but largely avoided in microspectroscopy.