Single-molecule orientation and localization microscopy

Abstract

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. Here we explore recent advancements in single-molecule orientation and localization microscopy (SMOLM), which yields super-resolved images of molecular three-dimensional (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. We cover the principles of SMOLM, discuss recent advances and applications in biology and photonics, and finally highlight exciting future directions and challenges in the field. The Review discusses recent advances in single-molecule orientation and localization microscopy (SMOLM) along with remaining challenges and promises for future developments of the field.