RT Journal Article SR Electronic T1 MINSTED tracking of single biomolecules JF bioRxiv FD Cold Spring Harbor Laboratory SP 2023.09.15.557902 DO 10.1101/2023.09.15.557902 A1 Scheiderer, Lukas A1 von der Emde, Henrik A1 Hesselink, Mira A1 Weber, Michael A1 Hell, Stefan W. YR 2023 UL http://biorxiv.org/content/early/2023/09/15/2023.09.15.557902.abstract AB 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 spatio-temporal precision. By updating the position for each detected photon, MINSTED recognizes fluorophore steps of 16 nm within < 250 microseconds using about 13 photons. The power of MINSTED tracking is demonstrated by quantifying side-steps of the motor protein kinesin-1 walking on microtubules and switching protofilaments.Competing Interest StatementThe Max Planck Society holds patents on selected procedures and embodiments of MINSTED, benefitting S. W. H., H. v. d. E., and M. W..