Single molecule orientation and localization microscopy

Single Molecule Localization Microscopy (SMLM) offers enhanced spatial resolution in optical microscopy, providing detailed insights into the spatial organization of proteins in cells at the nanoscale. Over the past decade, SMLM has progressively incorporated the capability to retrieve the orientations of single molecules using their polarized dipolar emission pattern. This Review explores recent advancements in Single Molecule Orientation and Localization Microscopy (SMOLM), which yields super-resolved images of molecular 3D orientations, wobble, and 3D positions. This advancement opens possibilities to explore the nanoscale organization and conformation of biological molecules as well as to monitor and design local 3D optical fields in nanophotonics. The Review covers the principles of SMOLM, discusses recent advances and applications in biology and photonics, and finally highlights exciting future directions and challenges in the field.