Light microscopy offers many advantages for the investigation of biological samples. However, traditional methods are limited by diffraction to a resolution of a few hundred nanometers. Due to this limitation, these methods cannot discern many intracellular structures and activities of critical importance to contemporary life science research.
Super-resolution techniques can surpass the diffraction limit to enable imaging of specifically labeled molecules well beyond the optical diffraction limit, down to 20 nm laterally. With this enhanced resolving power, super-resolution microscopy (SRM) methods allow the imaging and quantitative analysis of cellular dynamics and nanostructures that were previously inaccessible. As a result, SRM overcomes some key challenges associated with nanoscale biological imaging.
This guide focuses primarily on the single-molecule localization microscopy (SMLM) method of SRM.