Live imaging of Drosophila melanogaster neural stem cells with photo-ablated centrosomes

Drosophila neural stem cells (NSCs) divide asymmetrically to generate siblings of different sizes. This model system has proved helpful in deciphering the contribution of polarity cues and the mitotic spindle in asymmetric cell division. Here, we describe a technique we developed to flatten cultured Drosophila brain explants to accurately image the cytoskeleton in live NCSs. We also describe our approach to efficiently remove centrosomes by laser photo-ablation and to measure daughter cell size after NSC division. For complete details on the use and execution of this protocol, please refer to Thomas et al. (2021).