MINFLUX nanoscopy delivers 3D multicolor nanometer resolution in cells

Nat Methods. 2020 Feb;17(2):217-224. doi: 10.1038/s41592-019-0688-0. Epub 2020 Jan 13.

Abstract

The ultimate goal of biological super-resolution fluorescence microscopy is to provide three-dimensional resolution at the size scale of a fluorescent marker. Here we show that by localizing individual switchable fluorophores with a probing donut-shaped excitation beam, MINFLUX nanoscopy can provide resolutions in the range of 1 to 3 nm for structures in fixed and living cells. This progress has been facilitated by approaching each fluorophore iteratively with the probing-donut minimum, making the resolution essentially uniform and isotropic over scalable fields of view. MINFLUX imaging of nuclear pore complexes of a mammalian cell shows that this true nanometer-scale resolution is obtained in three dimensions and in two color channels. Relying on fewer detected photons than standard camera-based localization, MINFLUX nanoscopy is poised to open a new chapter in the imaging of protein complexes and distributions in fixed and living cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Color*
  • Fluorescent Dyes / chemistry
  • Humans
  • Image Processing, Computer-Assisted
  • Microscopy, Fluorescence / methods*

Substances

  • Fluorescent Dyes