Structured illumination superresolution imaging of the cytoskeleton

Methods Cell Biol. 2014:123:315-33. doi: 10.1016/B978-0-12-420138-5.00017-3.

Abstract

Structured illumination microscopy (SIM) with a 3-dimensional illumination pattern allows to double image resolution laterally and axially. For cell biologists, SIM may become an attractive tool for refined colocalization studies and to investigate the assembly of components at higher resolution. In this chapter, we focus on the use of a commercial available SIM setup and provide guidance on sample preparation and image acquisition. We present superresolution images of the cytoskeleton in fixed cells and discuss the potential and limitations for SIM in live imaging.

Keywords: Actin; Fluorescent protein; Immunocytochemistry; Microtubule; Refractive index; Spherical aberration; Structured illumination; Superresolution; Time-lapse imaging.

MeSH terms

  • Animals
  • Artifacts
  • Cell Line, Tumor
  • Fourier Analysis
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Microscopy, Fluorescence / methods
  • Microtubules / ultrastructure*
  • Neurons / ultrastructure
  • Refractometry
  • Time-Lapse Imaging