Fast stimulated emission nanoscopy based on single molecule localization.

For super-resolution microscopy methods based on single molecule stochastic switching and localization, to simultaneously improve the spatial–temporal resolution, it is necessary to maximize the number of photons that can be collected from single molecules per unit time. Here, we describe a novel approach to enhance the signal intensity (collected photons per second) from fluorescence probes by introducing a stimulated emission (SE) optical process. This process is based on the following two properties: first, with reasonable parameters, the photon emission rate can be significantly increased with SE; and second, the SE photons, which are spatially coherent with the stimulation beam, are more favorable for collection than fluorescence. Theoretical results have shown that signal intensity from a single fluorescent molecule can be greatly improved with SE. We therefore showed, using SE in combination with single molecule localization methodology, that fast imaging at a rate of 0.05 s per reconstructed image with lateral resolutions of ∼30  nm can be obtained.