XRM2024 - Thu04K - "Molecule-scale resolution and dynamics in fluorescence microscopy"

I will show how an in-depth description of the basic principles of diffraction-unlimited fluorescence microscopy has spawned MINFLUX[1–4], a recent superresolution method that has reached the resolution of the size of a fluorophore molecule. Providing 1–3 nanometer resolution in fixed and living cells, as well as localization precisions in the Ångström range, MINFLUX and the related MINSTED concept[5,6] are being established for routine applications in the biomedical sciences[4]. Relying on fewer fluorescence photons than other methods, these techniques are also poised to characterize dynamic processes at the single protein level, as already demonstrated by tracking sub(nanometer) details of the unhindered stepping of the motor protein kinesin-1 on microtubules at up to physiological ATP concentrations[7]. References 1. Balzarotti, F., et al. (2017). Science 355, 606-6122. Eilers, Y., et al. (2018). PNAS 115, 6117-61223. Gwosch, K. C., et al. (2020). Nat. Methods 17, 217–2244. Schmidt, R., et al. (2021). Nat. Commun. 12:1478.5. Weber, M., et al. (2021). Nat. Photon. 15, 361-3666. Weber, M., et al. (2023). Nat. Biotechnol., 41, 569-576 (2023)7. Wolff, J.O., et al. (2023). Science, 379, 1004-1010 (2023)