title: Real-Time Visualization and Quantification of Contractile Ring Proteins in Single Living Cells.
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.
authors.affiliation: Department of Molecular Genetics, The Ohio State University, 615 Biological Sciences Building, 484 W. 12th Ave, Columbus, OH, 43210, USA.
authors.forename: Reshma
authors.initials: R
authors.lastname: Davidson
authors.affiliation: Department of Molecular Genetics, The Ohio State University, 615 Biological Sciences Building, 484 W. 12th Ave, Columbus, OH, 43210, USA.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Molecular Genetics, The Ohio State University, 615 Biological Sciences Building, 484 W. 12th Ave, Columbus, OH, 43210, USA.
authors.forename: Kenneth S
authors.initials: KS
authors.lastname: Gerien
authors.affiliation: Department of Molecular Genetics, The Ohio State University, 615 Biological Sciences Building, 484 W. 12th Ave, Columbus, OH, 43210, USA. wu.620@osu.edu*|Department of Molecular and Cellular Biochemistry, The Ohio State University, 333 Hamilton Hall, 1645 Neil avenue, Columbus, OH, 43210, USA. wu.620@osu.edu*
authors.forename: Jian-Qiu
authors.initials: JQ
authors.lastname: Wu
mesh_terms: D003285:Contractile Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods*; D012568:Schizosaccharomyces / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*
chemical_list: D003285:Contractile Proteins; D029702:Schizosaccharomyces pombe Proteins
keywords: Contractile ring; Cytokinesis; Fimbrin; Fission yeast; GFP; Gelatin; ImageJ; Microscopy; Quantification; Septin; Tetrad fluorescence microscopy