Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes

• DOI: 10.1126/science.aak9913
• Publication Date: 2016-12-23T22:10:42Z
• Authors: Francisco Balzarotti; Yvan Eilers; Klaus C. Gwosch; Arvid H. Gynnå; Volker Westphal; Fernando D. Stefani; Johan Elf; Stefan W. Hell
• Keywords: Photoactivated localization microscopy; Tracking (education); Temporal resolution; STED microscopy; Point spread function

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