Simulation of live-cell imaging system reveals hidden uncertainties in cooperative binding measurements

We propose a computational method to quantitatively evaluate the systematic uncertainties that arise from undetectable sources in biological measurements using live-cell imaging techniques. We then demonstrate this method in measuring biological cooperativity of molecular binding networks: in particular, ligand molecules binding to cell surface receptor proteins. Our results show how the non-statistical uncertainties lead to invalid identification of the measured cooperativity. Through this computational scheme, the biological interpretation can be more objectively evaluated and understood under a specific experimental configuration of interest.