MINSTED tracking of single biomolecules.

Scheiderer, Lukas; von der Emde, Henrik; Hesselink, Mira; Weber, Michael; Hell, Stefan W · Nat Methods · 2024

basic_science · Level V

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Abstract

Here we show that MINSTED localization, a method whereby the position of a fluorophore is identified with precisely controlled beams of a STED microscope, tracks fluorophores and hence labeled biomolecules with nanometer/millisecond spatiotemporal precision. By updating the position for each detected photon, MINSTED recognizes fluorophore steps of 16 nm within <250 μs using about 13 photons. The power of MINSTED tracking is demonstrated by resolving the stepping of the motor protein kinesin-1 walking on microtubules and switching protofilaments.

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