Super-resolution imaging of microtubules in Medicago sativa
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
32896319
DOI
10.1016/bs.mcb.2020.03.004
PII: S0091-679X(20)30032-7
Knihovny.cz E-resources
- Keywords
- Airyscan microscopy, CLSM, Immunofluorescence labeling, Medicago sativa, Microtubules, Structured-illumination microscopy,
- MeSH
- Plant Epidermis cytology MeSH
- Microscopy, Confocal MeSH
- Plant Roots cytology MeSH
- Medicago sativa cytology metabolism MeSH
- Microtubules metabolism MeSH
- Seedlings metabolism MeSH
- Imaging, Three-Dimensional methods MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Study of microtubules on cellular and subcellular levels is compromised by limited resolution of conventional fluorescence microscopy. However, it is possible to improve Abbe's diffraction-limited resolution by employment of super-resolution microscopy methods. Two of them, described herein, are structured-illumination microscopy (SIM) and Airyscan laser scanning microscopy (AM). Both methods allow high-resolution imaging of cortical microtubules in plant cells, thus contributing to the current knowledge on plant morphogenesis, growth and development. Both SIM and AM provide certain advantages and characteristic features, which are described here. We present immunofluorescence localization methods for microtubules in fixed plant cells achieving high signal efficiency, superb sample stability and sub-diffraction resolution. These protocols were developed for whole-mount immunolabeling of root samples of legume crop species Medicago sativa. They also contain tips for optimal sample preparation of plants germinated from seeds as well as plantlets regenerated from somatic embryos in vitro. We describe in detail all steps of optimized protocols for sample preparation, microtubule immunolabeling and super-resolution imaging.
References provided by Crossref.org
Imaging plant cells and organs with light-sheet and super-resolution microscopy
Comparing Super-Resolution Microscopy Techniques to Analyze Chromosomes