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

观察固定和活体原代小胶质细胞中微管及其末端的实验方案

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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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