Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037, USA.
STAR Protoc. 2021 Dec 22;3(1):101064. doi: 10.1016/j.xpro.2021.101064. eCollection 2022 Mar 18.
Adaptive T and B lymphocytes expand, respond, and persist across a multitude of separable cell differentiation states. Small compartments of these cells present defined cell surface phenotype, but express potentially divergent immune functions. Here, we use high resolution flow cytometry to provide direct access to rare lymphocyte subpopulations for evaluation of steady-state or reactive transcriptional programs. We sort and index single cells by phenotype in 384-well format for quantification of targeted gene amplification through RNA sequencing (single cell qtSEQ). For complete details on the use and execution of this profile, please refer to Dufaud et al. (2021).
适应性 T 和 B 淋巴细胞在多种可分离的细胞分化状态下扩增、响应和持续存在。这些细胞的小隔间呈现出定义明确的细胞表面表型，但表达潜在不同的免疫功能。在这里，我们使用高分辨率流式细胞术直接访问稀有淋巴细胞亚群，以评估稳态或反应性转录程序。我们通过 RNA 测序（单细胞 qtSEQ）按表型对 384 孔格式中的单个细胞进行排序和标记，以定量靶向基因扩增。有关此分析方案的使用和执行的完整详细信息，请参阅 Dufaud 等人（2021 年）。
