Bayesian Classification and Modeling of Single Molecule Fluorescence Colocalization Images

Abstract

Multi-wavelength single-molecule fluorescence colocalization methods allow elucidation of complex biochemical reaction mechanisms. However, analysis of colocalization data is an intrinsically challenging and time-consuming problem for multiple reasons. First, to minimize dye photobleaching, images frequently are collected at low signal-to-noise ratios, making it challenging to discriminate real fluorescent spots from noise. Second, transient non-specific interactions of the binder molecule with the surface of the microscope slide can give rise to both false-positive and false-negative detection.