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MIRAVA POLYSCOPE – All in one and on for all: the perfect image
Science beyond Barriers

abberior instruments

Technology

2023
Small

Accelerated MINFLUX Nanoscopy, through Spontaneously Fast-Blinking Fluorophores

Authors:

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

Keywords:

Minflux, 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.

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Superresolution & Confocal Systems

  • Overview
  • MINFLUX
  • MIRAVA POLYSCOPE
  • INFINITY
  • FACILITY
  • STEDYCON
  • Software Overview
  • LiGHTBOX
  • STEDYCON smart control
  • iMSPECTOR

Superresolution & Confocal Modules

  • Overview
  • MINFLUX Module
  • MATRIX Detector
  • TIMEBOW Imaging
  • FLEXPOSURE Illumination
  • RAYSHAPE Mirror
  • TRUESHARP Deconvolution
  • EASY3D
  • RAINBOW Detection
  • STED Lasers
  • Autoalignment
  • Autofocus
  • Excitation Lasers
  • Accessories
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  • Overview
  • abberior STAR
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  • abberior FLUX
  • abberior CAGE
  • abberior Supplies

Shop

  • Dyes by Product Name
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  • Dyes by Label
  • Mounting Medium
  • Cells and Nanoparticles

Applications

  • Overview
  • Biophysics
  • Cell Biology
  • Live Cell Imaging
  • Material Science
  • Membrane biology
  • Microbiology
  • Neurobiology
  • Physiology
  • Plant Science
  • Virology
  • Zoology

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