Real-Time Visualization and Quantification of Contractile Ring Proteins in Single Living Cells

Methods Mol Biol. 2016:1369:9-23. doi: 10.1007/978-1-4939-3145-3_2.

Abstract

Single-cell microscopy provides a powerful tool to visualize cellular and subcellular processes in wild-type and mutant cells by observing fluorescently tagged proteins. Here, we describe three simple methods to visualize fission yeast cells: gelatin slides, coverslip-bottom dishes, and tetrad fluorescence microscopy. These imaging methods and data analysis using free software make it possible to quantify protein localization, dynamics, and concentration with high spatial and temporal resolution. In fission yeast, the actomyosin contractile ring is essential for cytokinesis. We use the visualization and quantification of contractile ring proteins as an example to demonstrate how to use these methods.

Keywords: Contractile ring; Cytokinesis; Fimbrin; Fission yeast; GFP; Gelatin; ImageJ; Microscopy; Quantification; Septin; Tetrad fluorescence microscopy.

Publication types

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

MeSH terms

  • Contractile Proteins / metabolism*
  • Microscopy, Fluorescence*
  • Molecular Imaging / methods*
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins / metabolism*

Substances

  • Contractile Proteins
  • Schizosaccharomyces pombe Proteins