Department of Biological Sciences, College of Science, University of Notre Dame, Notre Dame, IN, 46556, USA.
Mike and Josie Harper Cancer Research Institute, University of Notre Dame, South Bend, IN, 46617, USA.
STAR Protoc. 2021 May 13;2(2):100537. doi: 10.1016/j.xpro.2021.100537. eCollection 2021 Jun 18.
High dimensional compositional and transcriptional profiling of heterogeneous brain-infiltrating leukocytes can lead to novel biological and therapeutic discoveries. High-quality single-cell leukocyte preparations are a prerequisite for optimal single cell profiling. Here, we describe a protocol for epitope and RNA-preserving dissociation of adult mouse brains and subsequent leukocyte purification and staining, which is adaptable to homeostatic and pathogenic brains. Leukocyte preparation following this protocol permits exquisite single-cell surface protein and RNA profiling in applications including CyTOF and CITE-seq. For complete details on the use and execution of this protocol, please refer to Guldner et al. (2020) and Golomb et al. (2020).
对异质脑浸润白细胞进行高维组成型和转录组学分析可以带来新的生物学和治疗发现。高质量的单细胞白细胞制剂是进行最佳单细胞分析的前提。在这里，我们描述了一种用于保存抗原表位和 RNA 的成年小鼠大脑解离以及随后的白细胞纯化和染色的方案，该方案适用于稳态和病态大脑。遵循该方案的白细胞制备可实现包括 CyTOF 和 CITE-seq 在内的应用中的精细单细胞表面蛋白和 RNA 分析。有关该方案使用和执行的完整详细信息，请参阅 Guldner 等人（2020 年）和 Golomb 等人（2020 年）。
