Figure 6.

Approaches to SiMREPS data analysis. A TIRF microscopy movie is used to generate single-molecule time traces. These traces are then subjected to HMM and kinetic analysis (A) and accepted or rejected as kinetic fingerprints of analyte molecules. Alternatively, higher dynamic range can be achieved by performing a frame-by-frame subtraction to yield a framewise intensity change movie, which is then analyzed by super-resolution localization methods (B) to identify clusters of binding events indicative of the presence of analyte molecules. As a third alternative, the single-molecule traces are passed to an LSTM deep-learning classifier that was previously trained using control experiments (C) to score and reject or accept each trace.