Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes.
Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes.
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DOI：
10.1016/j.xpro.2021.100537
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发表时间：
2021-06-18
期刊：
影响因子：
--
通讯作者：
Zhang S
中科院分区：
文献类型：
--
作者：
Guldner IH;Golomb SM;Wang Q;Wang E;Zhang S
异质性脑浸润白细胞的高维组成和转录谱分析可以导致新的生物学和治疗发现。高质量的单细胞白细胞制剂是最佳单细胞分析的先决条件。在这里，我们描述了一个协议的表位和RNA保留解离成年小鼠大脑和随后的白细胞纯化和染色，这是适应于稳态和致病性的大脑。遵循该方案的白细胞制备允许在包括CyTOF和CITE-seq在内的应用中进行精细的单细胞表面蛋白和RNA分析。有关本方案使用和执行的完整详细信息，请参见和。小鼠脑解离成单细胞悬液用于下游单细胞分析富集脑驻留和低丰度脑浸润性白细胞经过测试可保留>40个免疫细胞表面表位的方案与scRNA-seq、流式细胞术和CyTOF兼容异质性脑浸润性白细胞的高维组成和转录分析可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最佳单细胞分析的先决条件。在这里，我们描述了一个协议的表位和RNA保留解离成年小鼠大脑和随后的白细胞纯化和染色，这是适应于稳态和致病性的大脑。遵循该方案的白细胞制备允许在包括CyTOF和CITE-seq在内的应用中进行精细的单细胞表面蛋白和RNA分析。
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 and. Mouse brain dissociation into single-cell suspension for downstream single-cell profiling Enrichment for brain-resident and low-abundant brain-infiltrating leukocytes Protocol tested to preserve >40 immune cell surface epitopes Compatible with scRNA-seq, flow cytometry, and CyTOF 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.
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