Accelerated MINFLUX Nanoscopy, through Spontaneously Fast‐Blinking Fluorophores

Abstract: The introduction of MINFLUX nanoscopy allows single molecules to be localized with one nanometer precision in as little as one millisecond. However, current applications have so far focused on increasing this precision by optimizing photon collection, rather than minimizing the localization time. Concurrently, commonly used fluorescent switches are specifically designed for stochastic methods (e.g., STORM), optimized for a high photon yield and rather long on‐times (tens of milliseconds). Here, accelerated MINFLUX nanoscopy with up to a 30‐fold gain in localization speed is presented. The improvement is attained by designing spontaneously blinking fluorescent markers with remarkably fast on‐times, down to 1–3 ms, matching the iterative localization process used in a MINFLUX microscope. This design utilizes a silicon rhodamine amide core, shifting the spirocyclization equilibrium toward an uncharged closed form at physiological conditions and imparting intact live cell permeability, modified with a fused (benzo) thiophene spirolactam fragment. The best candidate for MINFLUX microscopy (also suitable for STORM imaging) is selected through detailed characterization of the blinking behavior of single fluorophores, bound to different protein tags. Finally, optimization of the localization routines, customized to the fast blinking times, renders a significant speed improvement on a commercial MINFLUX microscope.

Location
Deutsche Nationalbibliothek Frankfurt am Main
Extent
Online-Ressource
Language
Englisch

Bibliographic citation
Accelerated MINFLUX Nanoscopy, through Spontaneously Fast‐Blinking Fluorophores ; day:15 ; month:01 ; year:2023 ; extent:11
Small ; (15.01.2023) (gesamt 11)

Creator
Remmel, Michael
Scheiderer, Lukas
Butkevich, Alexey N.
Bossi, Mariano L.
Hell, Stefan W.

DOI
10.1002/smll.202206026
URN
urn:nbn:de:101:1-2023011614143622460574
Rights
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
Last update
15.08.2025, 7:36 AM CEST

Data provider

This object is provided by:
Deutsche Nationalbibliothek. If you have any questions about the object, please contact the data provider.

Associated

  • Remmel, Michael
  • Scheiderer, Lukas
  • Butkevich, Alexey N.
  • Bossi, Mariano L.
  • Hell, Stefan W.

Other Objects (12)