Adaptation of aberration-tolerant 3D single-molecule localisation for flexible analysis

A 3D single molecule localisation setup and workflow exists (Palm3d, [1]) that produces localisations with a relative insensitivity to the aberrations of the optical system and without requiring a model of the noise in the images recorded. However, this workflow results only in histogram reconstructions that are inefficient for repeated reconstructions and for sub-regions of the FOV, with limited criteria available for selection of the localisations, and with no other output of final localisation data to the user. Therefore, we have developed a new user interface for filtering, efficient reconstruction and extraction for downstream quantitative analysis of the localisation data. Some examples of its use are given, including processing of 3D PALM data that reveals the 20-nm repeating structure of  α-actinin2 in the cardiomyocyte Z-disc.