High-content 3D multicolor super-resolution localization microscopy
Technologies: QuickPALM
Book chapter published in Methods in cell biology, January 2015
Publisher: Academic Press
The manuscript describes a protocol for high-content 3D multicolor super-resolution (SR) imaging using single-molecule localization microscopy (SMLM). It outlines the importance of automating the SR particle detection and localization process using ImageJ's macro scripting and choosing the appropriate detector for HRCSR. The authors also discuss challenges in single-molecule detection and localization, such as particle overlap, and the development of algorithms like SOFI, 3B, DAOSTORM, and ThunderSTORM for addressing these issues. The manuscript emphasizes the significance of SR drift correction and chromatic realignment using fiducial markers and elastic transformations during the reconstruction step. The manuscript also touches upon the advancements and challenges in the field of SR microscopy, particularly in HCS applications, and the role of computer control systems in microscopy.