A Unified Platform for FCS and RICS Analysis with Advanced Statistical Inference

Fluorescence correlation spectroscopy (FCS) and raster image correlation spectroscopy (RICS) are powerful techniques for measuring molecular diffusion, concentration, and dynamics in biological systems, yet current analysis tools lack unified frameworks that combine advanced statistical methods with high-performance computing. We present an open-source Python platform, IOCBIO FCS, that integrates FCS and RICS analysis with GPU-accelerated autocorrelation function calculation, robust statistical inference, and realistic optical modeling. The platform uniquely provides capabilities absent from existing open-source tools: direct incorporation of experimentally measured 3D point spread functions into fitting procedures, comprehensive statistical frameworks encompassing Bayesian inference alongside generalized, weighted, and ordinary least-squares methods for rigorous uncertainty quantification, and combined multiple-angle RICS analysis for characterizing anisotropic diffusion in complex biological systems. Additional features include image partitioning for spatial parameter mapping, advanced filtering strategies for data quality control, and comprehensive visualization of fitted results, residuals, posterior distributions, and parameter maps. This platform establishes a reproducible workflow bridging modern fluorescence microscopy with quantitative analysis of molecular transport across biophysics, biochemistry, and cell biology research.