Dual-Color and 3D Super-Resolution Microscopy of Multi-protein Assemblies

Methods Mol Biol. 2018:1764:237-251. doi: 10.1007/978-1-4939-7759-8_14.

Abstract

Breaking the resolution limit of conventional microscopy by super-resolution microscopy (SRM) led to many new biological insights into protein assemblies at the nanoscale. Here we provide detailed protocols for single-molecule localization microscopy (SMLM) to image the structure of a protein complex. As examples, we show how to acquire single- and dual-color super-resolution images of the nuclear pore complex (NPC) and dual-color 3D data on actin and paxillin in focal adhesions.

Keywords: Focal adhesions; Nuclear pore complex; PALM; Photoswitchable fluorescent protein; STORM; Single-molecule localization microscopy; Super-resolution microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism*
  • Actins / ultrastructure
  • Color
  • Fluorescent Dyes / metabolism
  • Focal Adhesions / ultrastructure*
  • HEK293 Cells
  • Humans
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Molecular Imaging / methods*
  • Multiprotein Complexes / metabolism*
  • Multiprotein Complexes / ultrastructure
  • Nuclear Pore / metabolism*
  • Nuclear Pore / ultrastructure
  • Paxillin / metabolism*
  • Paxillin / ultrastructure

Substances

  • Actins
  • Fluorescent Dyes
  • Multiprotein Complexes
  • Paxillin