MINSTED nanoscopy enters the Ångström localization range

DOI

Super-resolution techniques have achieved localization precisions in the nanometer regime. Here we report all-optical, room temperature localization of fluorophores with precision in the Ångström range. We built on the concept of MINSTED nanoscopy where precision is increased by encircling the fluorophore with the low-intensity central region of a stimulated emission depletion (STED) donut beam while constantly increasing the absolute donut power. By blue-shifting the STED beam and separating fluorophores by on/off switching, individual fluorophores bound to a DNA strand are localized with σ = 4.7 Å, corresponding to a fraction of the fluorophore size, with only 2,000 detected photons. MINSTED fluorescence nanoscopy with single-digit nanometer resolution is exemplified by imaging nuclear pore complexes and the distribution of nuclear lamin in mammalian cells labeled by transient DNA hybridization. Because our experiments yield a localization precision σ = 2.3 Å, estimated for 10,000 detected photons, we anticipate that MINSTED will open up new areas of application in the study of macromolecular complexes in cells.

Identifier
DOI https://doi.org/10.17617/3.0YVCIK
Metadata Access https://edmond.mpg.de/api/datasets/export?exporter=dataverse_json&persistentId=doi:10.17617/3.0YVCIK
Provenance
Creator von der Emde, Henrik
Publisher Edmond
Publication Year 2022
OpenAccess true
Contact hvonder(at)mpinat.mpg.de
Representation
Language English
Resource Type Dataset
Version 1
Discipline Other