Fluorescence bleach rate imaging

Abstract

Summary Bleach rate imaging on a (cooled) CCD can be easily achieved using a confocal microscope with bilateral scanning and detection coupled to a workstation; it is as easy as acquiring regular fluorescence images. Several analysis and display methods for bleach rate imaging are presented such as the bleach map (and its inverse) and a matrix‐based decomposition method for multi‐labelled specimens based on the bleach rate differences between the dyes used. With these tools, bleach‐rate‐based imaging can become a viable alternative to multiple labelling techniques for component identification in fluorescent specimens.