Seminarium Instytutu Niskich Temperatur i Badań Strukturalnych PAN

MINFLUX nanoscopy and related matters

Prof. Dr. Stefan W. Hell

Instytut Maxa Plancka Nauk Multidyscyplinarnych w Getyndze oraz z Instytut Maxa Plancka Badań Medycznych w Heidelbergu w Niemczech

Seminarium odbędzie się w aplikacji Microsoft Teams (dla osób z domeny intibs.pl kod dostępu to: to8aqyj). Pozosotałe osoby zainteresowane seminarium proszone są o kontakt z dr. hab. Adamem Pikulem ().

The seminar will be held in the Microsoft Teams application (for people from the intibs.pl domain, the access code is: to8aqyj). Other people interested in the seminar are asked to contact dr. hab. Adam Pikul ().

Streszczenie:

I will show how an in-depth description of the basic principles of diffraction-unlimited fluorescence microscopy (nanoscopy) [1-3] has spawned a new powerful superresolution concept, namely MINFLUX nanoscopy [4]. MINFLUX utilizes a local excitation intensity minimum (of a doughnut or a standing wave) that is targeted like a probe in order to localize the fluorescent molecule to be registered. In combination with single-molecule switching for sequential registration, MINFLUX [4-7] has obtained the ultimate (super)resolution: the size of a molecule. MINFLUX nanoscopy, providing 1–3 nanometer resolution in fixed and living cells, is presently being established for routine fluorescence imaging at the highest, molecular-size resolution levels. Relying on fewer detected photons than popular camera-based localization, MINFLUX and related MINSTED [8,9] nanoscopies are poised to open a new chapter in the imaging of protein complexes and distributions in fixed and living cells. MINFLUX is also set to transform the single-molecule analysis of dynamic processes, as already demonstrated by tracking in detail the unhindered stepping of the motor protein kinesin-1 on microtubules at up to physiological ATP concentrations [10], and providing answers to longstanding questions with respect to the kinesin-1 mechanochemical cycle.

[1] Hell, S.W., Wichmann, J. Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt. Lett. 19, 780-782 (1994).
[2] Hell, S.W. Far-Field Optical Nanoscopy. Science 316, 1153-1158 (2007).
[3] Hell, S.W. Microscopy and its focal switch. Nat. Methods 6, 24-32 (2009).
[4] Balzarotti, F., Eilers, Y., Gwosch, K. C., Gynnå, A. H., Westphal, V., Stefani, F. D., Elf, J., Hell, S.W. Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes. Science 355, 606-612 (2017).
[5] Eilers, Y., Ta, H., Gwosch, K. C., Balzarotti, F., Hell, S. W. MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution. PNAS 115, 6117-6122 (2018).
[6] Gwosch, K. C., Pape, J. K., Balzarotti, F., Hoess, P., Ellenberg, J., Ries, J., Hell, S. W. MINFLUX nanoscopy delivers 3D multicolor nanometer resolution in cells. Nat. Methods 17, 217–224 (2020).
[7] Schmidt, R., Weihs, T., Wurm, C. A., Jansen, I., Rehman, J., Sahl, S. J., Hell, S. W. (2021) MINFLUX nanometer-scale 3D imaging and microsecond-range tracking on a common fluorescence microscope. Nat. Commun. 12:1478.
[8] Weber, M., Leutenegger, M., Stoldt, S., Jakobs, S., Mihaila, T. S., Butkevich, A. N., Hell, S. W. MINSTED fluorescence localization and nanoscopy. Nat. Photon. 15, 361-366 (2021).
[9] Weber, M., von der Emde, H., Leutenegger, M., Gunkel, P., Sambandan, S., Khan, T. A., Keller-Findeisen, J., Cordes, V. C., Hell, S.W. MINSTED nanoscopy enters the Ångström localization range. Nat. Biotechnol. (in press, 2022).
[10] Wolff, J. O., Scheiderer, L., Engelhardt, T., Engelhardt, J., Matthias, J., Hell, S. W. MINFLUX dissects the unimpeded walking of kinesin-1. bioRxiv doi.org/10.1101/2022.07.25.501426 (2022).