COMPUTER-IMPLEMENTED PLATFORM FOR AUTOMATED FLUORESCENCE IMAGING AND KINETIC ANALYSIS

摘要

Automatically selecting time traces from a fluorescence experiment, in one aspect, may include capturing results of the fluorescence experiment in a moving image; localizing sources of fluorescence in the moving image; producing time traces of each fluorescent source by monitoring fluorescence intensity of said localized sources in the moving image over time; removing unuseful time traces from said produced time traces; and selecting useful time traces from said produced time traces based on one or more defined criteria. FRET traces from selected time traces may be further calculated and analyzed. A unified computer-implemented platform in one aspect may include tools to locate single molecules, extract traces, classify smFRET traces according to adjustable parameters, and quantify the kinetic parameters of FRET transitions using analytical procedures such as Hidden Markov Modeling (HMM) procedures.