Super-resolution fight club - assessment of 2D and 3D single-molecule localization microscopy software

Authors: Daniel Sage, Thanh-An Pham, Hazen Babcock, Tomas Lukes, Thomas Pengo, Jerry Chao, Ramraj Velmurugan, Alex Herbert, Anurag Agrawal, Silvia Colabrese, Ann Wheeler, Anna Archetti, Bernd Rieger, Raimund Ober, Guy M. Hagen, Jean-Baptiste Sibarita, Jonas Ries, Ricardo Henriques, Michael Unser, Seamus Holden
Technologies: NanoJ-SRRF and QuickPALM
Paper published in Nature Methods, April 2019
Publisher: Nature Publishing Group US New York

The manuscript describes the results of a large community effort to assess and rank the performance of software packages for single-molecule localization microscopy (SMLM) in both two-dimensional (2D) and three-dimensional (3D) imaging modalities. The study generated simulated datasets for popular SMLM modalities and evaluated 36 participant packages against these data. The results provide insights into the current limits of the field and highlight the importance of efficient and automated image processing for maximizing spatial resolution and reducing imaging time. The study identified several classes of algorithms used in the participant software and found that the choice of localization software greatly affects the quality of final super-resolution data. The manuscript also acknowledges funding sources and thanks the localization microscopy challenge participants and sponsors. Additionally, the manuscript discusses the importance of wobble correction in analyzing localization microscopy simulations using experimentally derived PSFs. The study involved simulated and experimental datasets, with the latter consisting of fluorescent structures with varying noise levels and spot densities. The results were made available on the competition website, along with the software and datasets.