Stochastic Optical Reconstruction Microscopy (STORM) Stochastic Optical Reconstruction Microscopy, STORM, is one of a family of super-resolution Single Molecule Localization Microscopies (SMLM) for the visualization of biological systems with an optical resolution measured in the tens of nanometers (nm) in the x, y, and z directions, pioneered in the laboratory of Xiaowei Zhuang at Harvard University.
STORM and other SMLMs are conceptually similar techniques: the photochemical properties of the fluorophore are exploited to induce a weakly emissive or non-emissive "dark" state. From the dark state, very small populations of (ideally) fluorophore are returned to an emissive state, excited, and detected. However, in order to be identified, emission profile must exhibit minimal overlap in each image. The centroid position of each identified molecule is statistically fitted, most often to a Gaussian function, and with a level of precision scaling with the number of detected photons. By imaging and fitting single emitters to a sub-diffraction limited area over many thousands of single images, eventually the user will have enough data to create a composite reconstruction of all identified emitters.