BrightEyes-FFS: an open-source platform for comprehensive analysis of fluorescence fluctuation spectroscopy experiments with small detector arrays

Abstract Fluorescence fluctuation spectroscopy (FFS) is an ensemble of techniques for the quantitative measurement of molecular dynamics and interactions. Recently, the introduction of small-format array detectors has opened up a new range of spatiotemporal information, enabling more detailed analysis of system kinetics. However, there is currently no open-source software available for analyzing the high-dimensional FFS data sets. We present BrightEyes-FFS, an open-source Python-based environment for FFS analysis with array detectors. The environment includes a Python package for reading raw FFS data, computing auto- and cross-correlations using various algorithms, and fitting the correlations to several models. A graphical user interface (GUI), available as a standalone executable, makes the analysis fast and user-friendly. An automated Jupyter Notebook writing tool enables users to transfer GUI-based analyses to Jupyter Notebook for custom analysis. We believe that BrightEyes-FFS will enable a wider community to study diffusion, flow, and interaction dynamics.