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  • Visualization of cortical microtubule networks in plant cells by live imaging and immunostaining.

Visualization of cortical microtubule networks in plant cells by live imaging and immunostaining.

STAR protocols (2021-02-09)
Fei Du, Feng Zhao, Jan Traas, Yuling Jiao
ABSTRACT

Cortical microtubules (CMTs) play pivotal roles during plant cell growth and division. The organization of CMTs undergoes important changes during different cellular and developmental processes. Here, we describe two methods for the visualization of CMT organization in plant cells using confocal laser scanning microscopy. CMT networks in the outer tissue layers can be directly visualized by live imaging of a fluorescent reporter line, and a protocol combining sectioning and immunostaining is applied for visualization of CMTs throughout tissues. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2020).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Paraformaldehyde, powder, 95%
Sigma-Aldrich
Glutaraldehyde solution, 50 wt. % in H2O
Sigma-Aldrich
Glycine, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Nicotinic acid, ≥98%
Sigma-Aldrich
Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Thiamine hydrochloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture
Sigma-Aldrich
Propidium iodide, ≥94.0% (HPLC)
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Phytagel, suitable for plant cell culture, BioReagent, powder
Sigma-Aldrich
Triton X-100, for molecular biology
Sigma-Aldrich
Potassium phosphate monobasic, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%