Department of Neurosurgery, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
STAR Protoc. 2022 May 25;3(2):101357. doi: 10.1016/j.xpro.2022.101357. eCollection 2022 Jun 17.
Here, we present a mass cytometry protocol optimized to examine the phenotype of immune cells within the mouse glioma microenvironment, using a Sleeping Beauty transposon-mediated mouse glioma model. We describe antibody conjugation and titrations for analysis of immune cells. We then detail mouse brain tumor tissue collection and processing, staining, followed by data acquisition, analysis, and gating strategy. This protocol can be applied to any brain tumor-harboring mouse model. For complete details on the use and execution of this protocol, please refer to Alghamri et al. (2021).
在这里,我们展示了一种优化的质谱细胞术方案,用于检测小鼠脑胶质瘤微环境中免疫细胞的表型,使用的是 Sleeping Beauty 转座子介导的小鼠脑胶质瘤模型。我们描述了抗体缀合和滴定分析免疫细胞的方法。然后,我们详细介绍了小鼠脑肿瘤组织的收集和处理、染色,以及随后的数据采集、分析和门控策略。该方案可应用于任何携带脑肿瘤的小鼠模型。有关该方案使用和执行的完整详细信息,请参阅 Alghamri 等人(2021 年)。