Superresolution Microscopy of the Nuclear Envelope and Associated Proteins

Methods Mol Biol. 2016:1411:83-97. doi: 10.1007/978-1-4939-3530-7_4.

Abstract

Superresolution microscopy is undoubtedly one of the most exciting technologies since the invention of the optical microscope. Capable of nanometer-scale resolution to surpass the diffraction limit and coupled with the versatile labeling techniques available, it is revolutionizing the study of cell biology. Our understanding of the nucleus, the genetic and architectural center of the cell, has gained great advancements through the application of various superresolution microscopy techniques. This chapter describes detailed procedures of multichannel superresolution imaging of the mammalian nucleus, using structured illumination microscopy and single-molecule localization microscopy.

Keywords: Linker of the nucleoskeleton and cytoskeleton (LINC) complex; Nuclear envelope (NE); Nuclear lamins; Nuclear pore complex (NPC); Photoactivated localization microscopy (PALM); Single-molecule localization microscopy (SMLM); Stochastic optical reconstruction microscopy (STORM); Structured illumination microscopy (SIM); Superresolution microscopy (SRM); Synaptonemal complex.

MeSH terms

  • Animals
  • Cell Cycle Proteins / metabolism
  • Cytoskeletal Proteins
  • Fibroblasts
  • Image Processing, Computer-Assisted
  • Imaging, Three-Dimensional
  • Male
  • Membrane Proteins / metabolism*
  • Membrane Proteins / ultrastructure*
  • Mice
  • Microscopy / methods*
  • Microscopy / standards
  • Nuclear Envelope / metabolism*
  • Nuclear Proteins / metabolism
  • Spermatocytes / metabolism

Substances

  • Cell Cycle Proteins
  • Cytoskeletal Proteins
  • KASH5 protein, mouse
  • Membrane Proteins
  • Nuclear Proteins