MaCADEMIA: a software toolbox for single-molecule fluorescence image quantification

MaCADEMIA, a MATLAB toolbox for single-molecule localisation microscopy, achieves localisation accuracies down to 0.5 pixels RMSE across wide SNR ranges on standardised particle tracking benchmarks.

• Why it matters: SMLM can quantify copy number, stoichiometry, interactions, and dynamics, but analysis is fragmented across specialised software, limiting straightforward deep quantification.
• What they did: The toolbox uses ensemble particle detection and is benchmarked against 14 leading methods, with localisation accuracy reported as RMSE down to 0.5 pixels across wide SNR ranges.
• The result: Its integrated workflow supports molecular counting and stoichiometry via stepwise photobleaching, mobility via mean squared displacement, colocalisation analysis, and absolute protein copy number quantification in 3D cellular models.

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