Trade-offs between structural integrity and acquisition time in stochastic super-resolution microscopy techniques

Abstract

The applicability of widefield stochastic microscopy, such as PALM or STORM, is limited by their long acquisition times. Images are produced from the accumulation of a large number of frames that each contain a scarce number of super-resolved localizations. We show that the random and uneven distribution of localizations leads to a specific type of trade-off between the spatial and temporal resolutions. We derive analytical predictions for the minimal time required to obtain a reliable image at a given spatial resolution. We find that the image completion time scales logarithmically with the ratio of the image size to the spatial resolution volume, with second order corrections due to spurious localization within the background noise. We validate our predictions against experimental localization sequences of labeled microtubule filaments obtained by STORM. Our theoretical framework makes it possible to compare the efficiency of emitters, define optimal labeling strategies, and allow implementation of a stopping criterion for data acquisitions that can be performed using real-time monitoring algorithms.