Centrosome and Centriole - 1st Edition - ISBN: 9780128024492, 9780128026380

Centrosome and Centriole, Volume 129

1st Edition

Serial Volume Editors: Renata Basto Karen Oegema
eBook ISBN: 9780128026380
Hardcover ISBN: 9780128024492
Imprint: Academic Press
Published Date: 10th July 2015
Page Count: 434
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Table of Contents

  • Series Editors
  • Preface
  • Chapter 1. Correlative light and electron microscopy analysis of the centrosome: A step-by-step protocol
    • Introduction
    • 1. Experimental Procedure
  • Chapter 2. Generation of a conditional analog-sensitive kinase in human cells using CRISPR/Cas9-mediated genome engineering
    • Introduction
    • 1. CRISPR/Cas System
    • 2. Analog-Sensitive Kinases
    • 3. Methods
    • 4. Functional Analysis
    • Conclusion
  • Chapter 3. Studying centrosome function using three-dimensional cell cultures
    • Introduction
    • 1. Methods
    • Conclusion
  • Chapter 4. Analysis of centrosomes in human cancer
    • Introduction
    • 1. Methods
    • Conclusions
  • Chapter 5. In vivo examination of the cortical cytoskeleton in multiciliated cells using electron tomography
    • Introduction
    • 1. Rationale
    • 2. Methods
    • 3. Materials
    • Conclusion
    • List of Abbreviations
  • Chapter 6. Tackling centrosome biology through gene targeting in chicken B cells
    • Introduction
    • 1. Methods
    • 2. Analysis of DT40 Cells
    • Conclusion
  • Chapter 7. Centriole biogenesis and function in multiciliated cells
    • Introduction
    • 1. Visualization of Centrioles and Related Structures in Xenopus MCCs
    • 2. Driving Specific Expression in Xenopus MCCs
    • 3. Generating Mosaic Xenopus Embryos to Circumvent Embryo Lethality
    • 4. Ciliated Epithelial Spheroid Culture
    • 5. Manipulating MCC Formation
    • 6. Fully-Automatic, Program-Assisted Counting of Centrioles in Confocal Images Using ImageJ
    • Conclusion
  • Chapter 8. Subdiffraction resolution microscopy methods for analyzing centrosomes organization
    • Introduction
    • 1. Methods
    • Summary and Outlook
  • Chapter 9. Probing mammalian centrosome structure using BioID proximity-dependent biotinylation
    • Introduction
    • 1. Materials
    • 2. Methods
    • 3. Large-Scale BioID Pulldown of BirA*-Fusion Proteins
    • 4. Data Analysis
    • Conclusion
  • Chapter 10. Purification of centrosomes from mammalian cell lines
    • Introduction
    • 1. Methods
    • Conclusions
  • Chapter 11. Isolation, cryotomography, and three-dimensional reconstruction of centrioles
    • Introduction
    • 1. Methods
    • Conclusions
  • Chapter 12. New insights into centrosome imaging in Drosophila and mouse neuroepithelial tissues
    • Introduction
    • 1. Imaging Centrosomes in Drosophila Neuroepithelial Cells
    • 2. High Spatial Resolution Imaging of Centrosomes in Neural Stem Cells of the Mouse Neocortex
    • Conclusion
  • Chapter 13. Microinjection techniques for studying centrosome function in Drosophila melanogaster syncytial embryos
    • Introduction
    • 1. Protocol
    • 2. Collection and Preparation of Drosophila Embryos
    • 3. Preparing Samples for Microinjection
    • 4. Mounting Embryos for Spinning Disc Confocal Microscopy
    • 5. Microinjection of Embryos
    • 6. Perturbation via Cold Treatment
    • 7. Imaging and Data Analysis
  • Chapter 14. A yeast two-hybrid approach for probing protein–protein interactions at the centrosome
    • Introduction
    • 1. Identifying Direct Protein Interactions Have Had a Profound Impact on Understanding Centrosome Function
    • 2. Challenges to Understanding Protein–protein Interactions in the Centrosome
    • 3. Dissection of Complex, Multicomponent Protein Machines Using Y2H
    • 4. Performing an Array Based Screen for Centrosome Protein Interactions
    • 5. Generating Specific, Separation of Function Mutations by Reverse Y2H
    • Summary
  • Chapter 15. Genome-wide RNAi screens in S2 cells to identify centrosome components
    • Introduction
    • 1. Methods
    • 2. Genome-Wide Screen
    • 3. Image Acquisition
    • 4. Scoring Phenotypes and Image Analysis
    • 5. Validation of Potential Hits
    • Discussion
  • Chapter 16. Time-lapse recording of centrosomes and other organelles in Drosophila neuroblasts
    • Introduction
    • 1. Primary Cultures
    • 2. Ex vivo Whole Mount
    • 3. Fluorescent Reporters
    • 4. Remarks and Troubleshooting
  • Chapter 17. Centrosomes and spindles in ascidian embryos and eggs
    • Introduction
    • 1. Methods
  • Chapter 18. Ultrastructural analysis of Caenorhabditis elegans cilia
    • Introduction
    • 1. Methods
    • Discussion
  • Chapter 19. Method: In vitro analysis of pericentriolar material assembly
    • Introduction
    • 1. Methods
    • Conclusion
  • Chapter 20. Monitoring SPB biogenesis in fission yeast with high resolution and quantitative fluorescent microscopy
    • Introduction
    • 1. Monitoring SPB Duplication in Fixed SPB-Labeled Strains
    • 2. Quantitative Analysis of SPB Biogenesis in Live Cells
    • Conclusion
  • Volumes in Series
  • Index

Description

This new volume of Methods in Cell Biology looks at methods for analyzing centrosomes and centrioles. Chapters cover such topics as methods to analyze centrosomes, centriole biogenesis and function in multi-ciliated cells, laser manipulation of centrosomes or CLEM, analysis of centrosomes in human cancers and tissues, proximity interaction techniques to study centrosomes, and genome engineering for creating conditional alleles in human cells.

Key Features

  • Covers sections on model systems and functional studies, imaging-based approaches and emerging studies
  • Chapters are written by experts in the field
  • Cutting-edge material

Readership

Researchers and students in cell, molecular and developmental biology


Details

No. of pages:
434
Language:
English
Copyright:
© Academic Press 2015
Published:
Imprint:
Academic Press
eBook ISBN:
9780128026380
Hardcover ISBN:
9780128024492

Reviews

Praise for the Series:
"The series is invaluable for workers at all levels of cell biology." --Nature


About the Serial Volume Editors

Renata Basto Serial Volume Editor

Affiliations and Expertise

Cell Biology Department, CNRS, Institut Curie, France

Karen Oegema Serial Volume Editor

Affiliations and Expertise

Ludwig Institute for Cancer Research, University of California, San Diego, USA