Centrosome and Centriole

1st Edition, Volume 129 - July 10, 2015
Editors: Renata Basto, Karen Oegema
Language: English
Hardback ISBN:
9 7 8 - 0 - 1 2 - 8 0 2 4 4 9 - 2
eBook ISBN:
9 7 8 - 0 - 1 2 - 8 0 2 6 3 8 - 0

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 biogen… Read more

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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.

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

Life Sciences

Physical Sciences & Engineering

Social Sciences & Humanities

Health

Centrosome and Centriole

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Renata Basto

Karen Oegema