: 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.
Protocol for observing microtubules and microtubule ends in both fixed and live primary microglia cells / Sanchini, Caterina; Rosito, Maria; Comincini, Alessandro; De Panfilis, Simone; Bartolini, Francesca; DI ANGELANTONIO, Silvia. - In: STAR PROTOCOLS. - ISSN 2666-1667. - 4:3(2023), p. 102499. [10.1016/j.xpro.2023.102499]
Protocol for observing microtubules and microtubule ends in both fixed and live primary microglia cells
Caterina SanchiniPrimo
;Maria Rosito;Silvia Di Angelantonio
Ultimo
2023
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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
