0:00:22 Start of webinar
0:50:09 Q&A Session
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Stefan Hell, Max Planck Institute, Göttingen, Germany.

Prof. Stefan W. Hell was awarded the 2014 Nobel Prize in Chemistry, for a technique of using fluorescent molecules to bypass the inherent resolution limit in optical microscopy. The prize was awarded to Prof. Hell together with American chemist W.E. Moerner and American physicist Eric Betzig.

This talk is part of the Biophysical seminar/Journal Club series organised by Ilaria Testa and Erdinc Sezgin. The event is will be broadcast in Youtube via zoom.

Abstract

MINSTED fluorescence localization and nanoscopy
Michael Weber, Marcel Leutenegger, Stefan Stoldt, Stefan Jakobs, Tiberiu S. Mihaila, Alexey N. Butkevich, Stefan W. Hell

We introduce MINSTED, a stimulated-emission-depletion (STED) based fluorescence localization and super-resolution microscopy concept providing spatial precision and resolution down to the molecular scale. In MINSTED, the intensity minimum of the STED donut, 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 to most other STED microscopy and localization methods, MINSTED entails substantially less fluorophore bleaching. In our implementation, 200-1000 detections per fluorophore provide a localization precision of 1-3 nm in standard deviation, which in conjunction with independent single fluorophore switching translates to a ~100-fold improvement of far-field microscopy resolution over the diffraction limit. The performance of MINSTED nanoscopy is demonstrated by imaging the distribution of Mic60 proteins in the mitochondrial inner membrane of human cells.