Imaging of the Actin Cytoskeleton and Mitochondria in Fixed Budding Yeast Cells

Methods Mol Biol. 2016:1365:63-81. doi: 10.1007/978-1-4939-3124-8_3.

Abstract

The budding yeast Saccharomyces cerevisiae is widely used as a model system to study the organization and function of the cytoskeleton. In the past, its small size, rounded shape, and rigid cell wall created obstacles to explore the cell biology of this model eukaryote. It is now possible to acquire and analyze high-resolution and super-resolution multidimensional images of the yeast cell. As a result, imaging of yeast has emerged as an important tool in eukaryotic cell biology. This chapter describes labeling methods and optical approaches for visualizing the cytoskeleton and interactions of the actin cytoskeleton with mitochondria in fixed yeast cells using wide-field and super-resolution fluorescence microscopy.

Keywords: Actin; Cytoskeleton; Deconvolution; Fluorescent proteins; Live-cell imaging; Microscopy; Mitochondria; Ratio imaging; Vital staining; Yeast.

Publication types

  • Research Support, American Recovery and Reinvestment Act
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / metabolism*
  • Cell Adhesion / drug effects
  • Cell Nucleus / genetics
  • Cell Wall / metabolism
  • DNA, Mitochondrial / metabolism
  • Fluorescent Antibody Technique
  • Fluorescent Dyes / metabolism
  • Hydrolases / metabolism
  • Microscopy, Fluorescence / methods*
  • Mitochondria / metabolism*
  • Phalloidine / metabolism
  • Polylysine / pharmacology
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / drug effects
  • Spheroplasts / cytology
  • Staining and Labeling

Substances

  • DNA, Mitochondrial
  • Fluorescent Dyes
  • Phalloidine
  • Polylysine
  • zymolyase
  • Hydrolases