Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes Article Swipe
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· 2016
· Open Access
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· DOI: https://doi.org/10.1126/science.aak9913
· OA: W2554260837
Superresolution imaging in sharper focus An optical microscope cannot distinguish objects separated by less than half the wavelength of light. Superresolution techniques have broken this “diffraction limit” and provided exciting new insights into cell biology. Still, such techniques hit a limit at a resolution of about 10 nm. Balzarotti et al. describe another way of localizing single molecules called MINFLUX (see the Perspective by Xiao and Ha). As in photoactivated localization microscopy and stochastic optical reconstruction microscopy, fluorophores are stochastically switched on and off, but the emitter is located using an excitation beam that is doughnut-shaped, as in stimulated emission depletion. Finding the point where emission is minimal reduces the number of photons needed to localize an emitter. MINFLUX attained ∼1-nanometer precision, and, in single-particle tracking, achieved a 100-fold enhancement in temporal resolution. Science , this issue p. 606 ; see also p. 582
