Two-color nanoscopy of organelles for extended times with HIDE probes

Nat Commun. 2020 Aug 26;11(1):4271. doi: 10.1038/s41467-020-17859-1.

Abstract

Performing multi-color nanoscopy for extended times is challenging due to the rapid photobleaching rate of most fluorophores. Here we describe a new fluorophore (Yale-595) and a bio-orthogonal labeling strategy that enables two-color super-resolution (STED) and 3D confocal imaging of two organelles simultaneously for extended times using high-density environmentally sensitive (HIDE) probes. Because HIDE probes are small, cell-permeant molecules, they can visualize dual organelle dynamics in hard-to-transfect cell lines by super-resolution for over an order of magnitude longer than with tagged proteins. The extended time domain possible using these tools reveals dynamic nanoscale targeting between different organelles.

Publication types

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

MeSH terms

  • Cell Line
  • Fluorescent Dyes* / chemistry
  • HeLa Cells
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Imaging, Three-Dimensional
  • Microscopy, Confocal
  • Microscopy, Fluorescence / methods*
  • Nanotechnology / methods*
  • Organelles / metabolism*
  • Photobleaching
  • Time-Lapse Imaging

Substances

  • Fluorescent Dyes