Cytoskeleton Methods and Protocols

In this cutting-edge book, internationally renowned experts present techniques which reflect many of the recent technological advances in experimental tools for cytoskeleton research. There is emphasis on animal, plant, protist, and fungal model systems.

In the ten years since the publication of the first edition, great advances in fluorescent labeling, optics, and sample preparation have significantly improved the imaging capability of microscopy, allowing for a continual refinement of our understanding of the cytoskeleton as a dynamic synergy of components. In Cytoskeleton Methods and Protocols, Second Edition , internationally renowned experts present techniques which reflect many of the recent technological advances in experimental tools for cytoskeleton research with emphasis on animal, plant, protist, and fungal model systems. This cutting-edge volume contains methods for live-cell imaging, fluorescence microscopy, electron microscopy, analysis of cell and organelle motility, isolation of cytoskeleton components, and proteomics, amongst other topics. As a volume in the highly successful Methods in Molecular Biology ™ series, chapters incorporate introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes that provide unpublished technical information on troubleshooting and avoiding known pitfalls.

Up-to-date and comprehensive, Cytoskeleton Methods and Protocols, Second Edition serves as an ideal guide to scientists who wish to continue this fruitful and important biological research.

Covers a wide range of live-cell imaging techniques Detailed methods for SEM and TEM analysis of cytoskeleton components Techniques for imaging the plant cytoskeleton Cutting-edge protocols for quantitative fluorescence microscopy Presents proteomic tools for cytoskeleton research Comprehensive protocols for qualitative and quantitative analyses of cell and organelle motility Includes supplementary material: sn.pub/extras

Inhalt
Live-Cell Imaging of the Cytoskeleton.- High Resolution Multimode Light Microscopy of Cell Migration: Long-Term Imaging and Analysis.- Imaging the Cytoskeleton in Live Xenopus laevis Embryos.- Live-Cell Imaging of the Cytoskeleton and MitochondrialCytoskeletal Interactions in Budding Yeast.- Dynamics of Actin Filaments and Microtubules.- Jasplakinolide: An Actin-Specific Reagent that Promotes Actin Polymerization.- Analysis of Centrosome Function and Microtubule Dynamics by Time-Lapse Microscopy in Xenopus Egg Extracts.- Fluorescence Microscopy Protocols for Investigating Cytoskeleton Components.- Quantitative Fluorescence Microscopy Techniques.- A FRET-Based Approach for Studying Conformational Changes of a Cytoskeleton-Related Tumor Suppressor Molecule.- Sample Preparation for Fluorescence Imaging of the Cytoskeleton in Fixed and Living Plant Roots.- Imaging of the Cytoskeleton and Mitochondria in Fixed Budding Yeast Cells.- Electron Microscopy Protocols for Imaging the Cytoskeleton.- Imaging Cytoskeleton Components by Electron Microscopy.- Purification and Localization of Intraflagellar Transport Particles and Polypeptides.- High Pressure Freezing, Electron Microscopy, and Immuno-Electron Microscopy of Tetrahymena thermophila Basal Bodies.- Strategies for Imaging Microtubules in Plant Cells.- Analysis of Cell and Organelle Motility.- A Rapid Tracking Method for the Quantitative Analysis of Organelle Streaming Velocity.- Melanosome Motility in Fish Retinal Pigment Epithelial Cells.- Analysis of Properties of Cilia Using Tetrahymena thermophila.- Application of Cell Traction Force Microscopy for Cell Biology Research.- 2D and 3D Quantitative Analysis of Cell Motility and Cytoskeletal Dynamics.- Detergent-Extracted Models for the Study of Cilia or Flagella.- Geneticand Proteomics Approaches for Cytoskeleton Protein Function.- The Analysis of Intermediate Filament Dynamics Using Transfections and Cell Fusions.- Functional Analysis of Actin-Binding Proteins in the Central Nervous System of Drosophila.- Proteomic Tools for the Analysis of Cytoskeleton Proteins.