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

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 li...

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Detalles Bibliográficos
Autores principales: Sanchini, Caterina, Rosito, Maria, Comincini, Alessandro, De Panfilis, Simone, Bartolini, Francesca, Di Angelantonio, Silvia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10448205/
https://www.ncbi.nlm.nih.gov/pubmed/37573502
http://dx.doi.org/10.1016/j.xpro.2023.102499
Descripción
Sumario: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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