Single-molecule based super-resolution imaging

Fluorescence microscopy is the method of choice to study biological samples in a comparatively non-invasive way. The field has received a powerful boost with the development of super-resolution imaging methods, which overcome the limitations imposed by optical diffraction.
A very strong class of these novel techniques can be merged under the generic term single-molecule localization microscopy (SMLM), which relies on the detection of single-molecules, their precise localization and the reconstruction of an artificial super-resolution image. It can improve on the ~200 nm resolution limit by a factor of ten and more, and thus have opened the door for the study of finer cellular ultrastructure.
While in the early days the focus was on methodological advancement, such as characterization of fluorophores and engineering of multicolour and 3D imaging capabilities, in recent years SMLM is increasingly applied in biology and medicine. Besides its high spatial resolution SMLM also provides access to quantitative information and thus enables the study of structure-function relationships.
I will provide a concise overview of the development and underlying principles of SMLM with an emphasis on dSTORM, describe novel technical developments and conclude with applications in the field of neurobiology.