MINSTED fluorescence localization and nanoscopy

Nature Photonics, 2021

Abstract: We introduce MINSTED, a fluorophore localization and super-resolution microscopy concept based on stimulated emission
depletion (STED) that provides spatial precision and resolution down to the molecular scale. In MINSTED, the intensity mini-
mum of the STED doughnut, and hence the point of minimal STED, serves as a movable reference coordinate for fluorophore
localization. As the STED rate, the background and the required number of fluorescence detections are low compared with most
other STED microscopy and localization methods, MINSTED entails substantially less fluorophore bleaching. In our implemen-
tation, 200–1,000 detections per fluorophore provide a localization precision of 1–3 nm in standard deviation, which in conjunc-
tion with independent single fluorophore switching translates to a ~100-fold improvement in far-field microscopy resolution
over the diffraction limit. The performance of MINSTED nanoscopy is demonstrated by imaging the distribution of Mic60 pro-
teins in the mitochondrial inner membrane of human cells.

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