Fig. 1

Measuring the distance of individual organelles to the nucleus is robust to changes in cell and organelle morphology in silico. Principle of analyzing organelle position using intensity, i.e., perinuclear index; I_p…intensity within a perimeter equidistant to the nucleus, I_t…intensity outside of the perimeter (A) or via measuring the distance of individually detected lysosomes from the nucleus (B). C Simulated cell for testing the robustness of organelle positioning measurements (blue: nucleus, magenta: organelles, white: perimeter for intensity ratio, green: cell perimeter). D–F Measuring the distance of individual organelles is robust to changes in cell size when normalized by Feret’s diameter and intensity-based measurements (E, F, green line) are more error prone than distance measurements (E, F, red line). G–I Distance measurements of individual organelles are robust to fluctuations in background fluorescence. J–L Measuring the distance of individual organelles is robust to changes in organelle size