Fast and multiplexed superresolution imaging with DNA-PAINT-ERS

Nat Commun. 2020 Aug 28;11(1):4339. doi: 10.1038/s41467-020-18181-6.

Abstract

DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) facilitates multiplexing in superresolution microscopy but is practically limited by slow imaging speed. To address this issue, we propose the additions of ethylene carbonate (EC) to the imaging buffer, sequence repeats to the docking strand, and a spacer between the docking strand and the affinity agent. Collectively termed DNA-PAINT-ERS (E = EC, R = Repeating sequence, and S = Spacer), these strategies can be easily integrated into current DNA-PAINT workflows for both accelerated imaging speed and improved image quality through optimized DNA hybridization kinetics and efficiency. We demonstrate the general applicability of DNA-PAINT-ERS for fast, multiplexed superresolution imaging using previously validated oligonucleotide constructs with slight modifications.

Publication types

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

MeSH terms

  • Cell Line
  • Cytological Techniques / methods*
  • DNA / chemistry*
  • Humans
  • Image Processing, Computer-Assisted / methods
  • Microscopy, Fluorescence / methods*
  • Molecular Docking Simulation / methods*
  • Oligonucleotides
  • Staining and Labeling / methods

Substances

  • Oligonucleotides
  • DNA