Live-Cell Imaging of Mitochondria and the Actin Cytoskeleton in Budding Yeast

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1365; p. 25
• Main Authors: Higuchi-Sanabria, Ryo, Swayne, Theresa C, Boldogh, Istvan R, Pon, Liza A
• Format: Journal Article
• Language: English
• Published: United States 2016
• Subjects: Acetates - pharmacology; Actin Cytoskeleton - metabolism; Chromosomes, Fungal - genetics; Coloring Agents - metabolism; Genetic Loci - genetics; Genetic Vectors - genetics; Integrases - metabolism; Mitochondria - metabolism; Molecular Imaging - methods; Movement; Optical Imaging; Polymerase Chain Reaction; Saccharomycetales - cytology; Saccharomycetales - drug effects; Saccharomycetales - genetics; Sequence Analysis; Transformation, Genetic - drug effects; Deconvolution; Mitochondria; Yeast; Microscopy; Actin; Fluorescent proteins; Cytoskeleton; Live-cell imaging; Vital staining; Ratio imaging
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-3124-8_2

Maintenance and regulation of proper mitochondrial dynamics and functions are necessary for cellular homeostasis. Numerous diseases, including neurodegeneration and muscle myopathies, and overall cellular aging are marked by declining mitochondrial function and subsequent loss of multiple other cellular functions. For these reasons, optimized protocols are needed for visualization and quantification of mitochondria and their function and fitness. In budding yeast, mitochondria are intimately associated with the actin cytoskeleton and utilize actin for their movement and inheritance. This chapter describes optimal approaches for labeling mitochondria and the actin cytoskeleton in living budding yeast cells, for imaging the labeled cells, and for analyzing the resulting images.