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bioRxiv · 10.64898/2026.04.08.717207

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

Slenders, E.·Perego, E.·Zappone, S.·Vicidomini, G.
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Abstract

Fluorescence fluctuation spectroscopy (FFS) is an ensemble of techniques for quantitative measurement of molecular dynamics and interactions. Recently, the introduction of small-format array detectors has opened up a new range of spatiotemporal information, allowing for 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 transition from the GUI to Jupyter Notebook for custom analysis. We believe that BrightEyes-FFS will enable a wider community to study diffusion, flow, and interaction dynamics.

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Slenders, E., Perego, E., Zappone, S., Vicidomini, G.. 2026-04-10. BrightEyes-FFS: an open-source platform for comprehensive analysis of fluorescence fluctuation spectroscopy experiments with small detector arrays. https://doi.org/10.64898/2026.04.08.717207

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