Protocol for observing microtubules and microtubule ends in both fixed and live primary microglia cells

观察固定和活的原代小胶质细胞中的微管和微管末端的方案

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作者:Caterina Sanchini, Maria Rosito, Alessandro Comincini, Simone De Panfilis, Francesca Bartolini, Silvia Di Angelantonio

Abstract

Microtubule dynamics and orientation have crucial roles in many vital cellular processes. However, functional live imaging of microtubules and/or microtubule ends in primary microglia can be challenging. Here, we present a protocol for observing microtubules and microtubule ends in both fixed and live primary microglia cells. We describe steps for microglia culture and in vitro stimulation, SiR-tubulin labeling, lentivirus preparation, live imaging, immunostaining, and image acquisition. We also provide procedures for SiR-tubulin, EB3-EGFP, and EB1 analyses. For complete details on the use and execution of this protocol, please refer to Rosito et al. (2023).1.

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