Imaging of the Actin Cytoskeleton and Mitochondria in Fixed Budding Yeast Cells.
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.
Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University, 630 W. 168th Street, New York, NY, 10032, USA.
Ryo
R
Higuchi-Sanabria
Confocal and Specialized Microscopy Shared Resource, Herbert Irving Comprehensive Cancer Center, Columbia University, 1130 Saint Nicholas Ave, New York, NY, 10032, USA.
Theresa C
TC
Swayne
Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University, 630 W. 168th Street, New York, NY, 10032, USA.
Istvan R
IR
Boldogh
Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University, 630 W. 168th Street, New York, NY, 10032, USA. lap5@cumc.columbia.edu.|Confocal and Specialized Microscopy Shared Resource, Herbert Irving Comprehensive Cancer Center, Columbia University, 1130 Saint Nicholas Ave, New York, NY, 10032, USA. lap5@cumc.columbia.edu.
Liza A
LA
Pon
D008841:Actin Cytoskeleton / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D002467:Cell Nucleus / Q000235:genetics; D002473:Cell Wall / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000378:metabolism; D006867:Hydrolases / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D008928:Mitochondria / Q000378:metabolism*; D010590:Phalloidine / Q000378:metabolism; D011107:Polylysine / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000187:drug effects; D013104:Spheroplasts / Q000166:cytology; D013194:Staining and Labeling
D004272:DNA, Mitochondrial; D005456:Fluorescent Dyes; D010590:Phalloidine; D011107:Polylysine; C037833:zymolyase; D006867:Hydrolases
Actin; Cytoskeleton; Deconvolution; Fluorescent proteins; Live-cell imaging; Microscopy; Mitochondria; Ratio imaging; Vital staining; Yeast