Cytoskeleton Methods and Protocols

Part I Imaging the Cytoskeleton in Live and Fixed Cells

1. Long-Term Live Cell Imaging of Cell Migration: Effects of Pathogenic Fungi on Human Epithelial Cell Migration
Torsten Wöllert and George M. Langford

2. Live-Cell Imaging of Mitochondria and the Actin Cytoskeleton in Budding Yeast
Ryo Higuchi-Sanabria, Theresa C. Swayne, Istvan R. Boldogh, and Liza A. Pon

3. Imaging of the Actin Cytoskeleton and Mitochondria in Fixed Budding Yeast Cells
Ryo Higuchi-Sanabria, Theresa C. Swayne, Istvan R. Boldogh and Liza A. Pon

4. Imaging of the Cytoskeleton Using Live and Fixed Drosophila Tissue Culture Cells
Derek A. Applewhite, Christine A. Davis, Eric R. Griffis, and Omar A. Quintero

5. Imaging Cytoskeleton Components by Electron Microscopy
Tatyana Svitkina

6. Purification and Localization of Intraflagellar Transport Particles and Polypeptides
Roger D. Sloboda

7. Fluorescence Imaging of the Cytoskeleton in Plant Roots
Julia Dyachok, Ana Paez-Garcia, Cheol-Min Yoo, Karuppaiah Palanichelvam, and Elison B. Blancaflor

8. Microtubules in Plant Cells: Strategies and Methods for Immunofluorescence, Transmission Electron Microscopy, and Live Cell Imaging
Katherine Celler, Miki Fujita, Eiko Kawamura, Chris Ambrose, Klaus Herburger, Andreas Holzinger, and Geoffrey O. Wasteneys

Part II Cytoskeleton Dynamics

9. Basic Methods to Visualize Actin Filaments In Vitro Using Fluorescence Microscopy for Observation of Filament Severing and Bundling
Shoichiro Ono

10. An In Vitro Model System to Test Mechano-Microbiological Interactions Between Bacteria and Host Cells
Luís Carlos Santos, Emilia Laura Munteanu, and Nicolas Biais

11. Reconstitution of a Minimal Actin Cortex by Coupling Actin Filaments to Reconstituted Membranes
Sven K. Vogel

12. Use of Nanobodies to Localize Endogenous Cytoskeletal Proteins and to Determine Their Contribution to Cancer Cell Invasion by Using an ECM Degradation Assay
Isabel Van Audenhove and Jan Gettemans

13. Actin-Dynamics in Plant Cells: The Function of Actin Perturbing Substances: Jasplakinolide, Chondramides, Phalloidin, Cytochalasins, and Latrunculins
Andreas Holzinger and Kathrin Blaas

Part III Cell and Organelle Motility

14. Quantitative Motion Analysis in Two and Three Dimensions
Deborah J. Wessels, Daniel F. Lusche, Spencer Kuhl, Amanda Scherer, Edward Voss, and David R. Soll

15. Measurement of Cell Motility Using Microgrooved Substrates James H-C. Wang, Guangyi Zhao, and Bin Li

16. The Study of Cell Motility by Cell Traction Force Microscopy (CTFM)
James H-C. Wang, Guangyi Zhao, Bin Li

17. Melanosome Motility in Fish Retinal Pigment Epithelial (RPE) Cells
Christina King-Smith

18. Analysis of Stem Cell Motility In Vivo Based on Immunodetection of Planarian Neoblasts and Tracing of BrdU-Labeled Cells After Partial Irradiation
Junichi Tasaki, Chihiro Uchiyama-Tasaki, and Labib Rouhana

19. Chemotaxis: Under Agarose Assay
Derrick Brazill

Part IV Genetic and Proteomic Protocols

20. Functional Analysis of Actin-Binding Proteins in the Central Nervous System of Drosophila
Qi He and Christopher Roblodowski

21. Proteomic Analysis of Cytoskeleton Proteins in Fish
Michael Gotesman, Simon Menanteau-Ledouble, Mansour El-Matbouli

22. Using a Hand-Held Gene Gun for Genetic Transformation of Tetrahymena Thermophila
Michael Gotesman and Selwyn A. Williams

23. Proteomic Tools for the Analysis of Cytoskeleton Proteins
Michael Scarpati, Mary Ellen Heavner, Eliza Wiech and Shaneen Singh

24. Homology Modeling Procedures for Cytoskeletal Proteins of Tetrahymena and Other Ciliated Protists
Giovanni Pagano, Linda Hufnagel, and Roberta King