Super-resolution Fluorescence Imaging with Single-molecule Localization

Fluorescence microscopy has made a breakthrough in its resolving power by an order of magnitude largely due to recent advances in the rapidly emerging field of super-resolution microscopy. Here, a form of super-resolution microscopy based on single-molecule localization of stochastically switching probes will be discussed. During the imaging process in the approach, referred to as STORM or (F)PALM, a small subset of photoswitchable probes is switched on to isolate single molecules and subsequently localize them at nanometer-scale precision. The facilitated access to single molecules allows for multicolor, three-dimensional visualization of molecules and ultrastructures in fixed or living cells. The single-molecule-based super-resolution method is beginning to provide new insights in cell biology, microbiology and neurobiology at levels of single molecules, structures and cells.