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MINFLUX, or minimal fluorescence photon fluxes microscopy, is a super-resolution light microscopy method that images and tracks objects in two and three dimensions with single-digit nanometer resolution.[1][2][3]
MINFLUX uses a structured excitation beam with at least one intensity minimum – typically a doughnut-shaped beam with a central intensity zero – to elicit photon emission from a fluorophore. The position of the excitation beam is controlled with sub-nanometer precision, and when the intensity zero is positioned exactly on the fluorophore, the system records no emission. Thus, the system requires few emitted photons to determine the fluorophore's location with high precision. In practice, overlapping the intensity zero and the fluorophore would require a priori location knowledge to position the beam. As this is not the case, the excitation beam is moved around in a defined pattern to probe the emission from the fluorophore near the intensity minimum.[1]
Each localization takes less than 5 microseconds,[1] so MINFLUX can construct images of nanometric structures or track single molecules in fixed and live specimens by pooling the locations of fluorescent labels. Because the goal is to locate the point where a fluorophore stops emitting, MINFLUX significantly reduces the number of fluorescence photons needed for localization compared to other methods.[2][4]
A commercial MINFLUX system is available from abberior instruments GmbH.[5]