Real-time image resolution measurement for single molecule localization microscopy

Abstract

Recent advancements in single molecule localization microscopy (SMLM) have demonstrated outstanding potential applications in high-throughput and high-content screening imaging. One major limitation to such applications is to find a way to optimize imaging throughput without scarifying image quality, especially the homogeneity in image resolution, during the imaging of hundreds of field-of-views (FOVs) in heterogeneous samples. Here we introduce a real-time image resolution measurement method for SMLM to solve this problem. This method is under the heuristic framework of overall image resolution that counts on localization precision and localization density. Rather than estimating the mean localization density after completing the entire SMLM process, this method uses the spatial Poisson process to model the random activation of molecules and thus determines the localization density in real-time. We demonstrate that the method is valid in real-time resolution measurement and is effective in guaranteeing homogeneous image resolution across multiple representative FOVs with optimized imaging throughput.