Method for Analysis of Excitation-Emission Spectral Scans to Characterize Fluorescent Probes in Confocal Microscopy

The main aim of the study was to use the spectral excitation-emission 2D scanning followed by 2D spectral
analysis in order to obtain an appropriate representation of the properties of the used dye in the original
image of a stained cell. We used Di-4-ANNEPS voltage sensitive dye (Invitrogen) binding to an adult rat
cardiomyocyte membrane to determine the stability of the dye and to characterize spatial differences of
its spectral properties.