To establish our framework for quantifying the heterogeneity of localisations at droplet interfaces, we first placed 100 neighboring but non-overlapping droplets of different diameters in a box and simulated a homogeneous distribution of proteins on each droplet (Figure ). We then separated each droplet based on the known ground truth (Figure ) and calculated the relative position distribution for each droplet. The histograms of two selected droplets show a local maximum for small distances representing a convolution of the localization precision σlp and the localization accuracy σac and a local maximum for large distances defined by the diameter of the droplet, further discussed below (Figure ). Normalizing the distance distributions over all droplets showed a smooth distribution with the local maxima identified earlier in the individual droplets (Figure ). The normalized histogram presents a fingerprint of expected distances.