| Chapter title |
Microtubules in Plant Cells: Strategies and Methods for Immunofluorescence, Transmission Electron Microscopy, and Live Cell Imaging
|
|---|---|
| Chapter number | 8 |
| Book title |
Cytoskeleton Methods and Protocols
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3124-8_8 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3123-1, 978-1-4939-3124-8
|
| Authors |
Katherine Celler, Miki Fujita, Eiko Kawamura, Chris Ambrose, Klaus Herburger, Andreas Holzinger, Geoffrey O. Wasteneys |
| Editors |
Ray H. Gavin |
| Abstract |
Microtubules (MTs) are required throughout plant development for a wide variety of processes, and different strategies have evolved to visualize and analyze them. This chapter provides specific methods that can be used to analyze microtubule organization and dynamic properties in plant systems and summarizes the advantages and limitations for each technique. We outline basic methods for preparing samples for immunofluorescence labeling, including an enzyme-based permeabilization method, and a freeze-shattering method, which generates microfractures in the cell wall to provide antibodies access to cells in cuticle-laden aerial organs such as leaves. We discuss current options for live cell imaging of MTs with fluorescently tagged proteins (FPs), and provide chemical fixation, high-pressure freezing/freeze substitution, and post-fixation staining protocols for preserving MTs for transmission electron microscopy and tomography. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 59 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 11 | 19% |
| Student > Ph. D. Student | 11 | 19% |
| Student > Master | 6 | 10% |
| Student > Bachelor | 4 | 7% |
| Other | 4 | 7% |
| Other | 9 | 15% |
| Unknown | 14 | 24% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 21 | 36% |
| Biochemistry, Genetics and Molecular Biology | 15 | 25% |
| Physics and Astronomy | 1 | 2% |
| Chemistry | 1 | 2% |
| Medicine and Dentistry | 1 | 2% |
| Other | 1 | 2% |
| Unknown | 19 | 32% |