Malone Mary K, Ujas Thomas A, Cotter Katherine M, Britsch Daimen R S, Lutshumba Jenny, Turchan-Cholewo Jadwiga, Stowe Ann M
Department of Neurology, Department of Neuroscience, The University of Kentucky, Lexington, KY, USA.
Methods Mol Biol. 2023;2616:213-229. doi: 10.1007/978-1-0716-2926-0_17.
Flow cytometry enables the multi-parametric quantification of cell types, especially in immunophenotyping of unique immune cell subsets that can either contribute to or ameliorate pathology. For tissues to be used in such analyses, single-cell suspensions must be created. Here we describe protocols for preparing single-cell suspensions of mouse spleen and brain tissue, as well as the steps for fluorescently activated cell staining/sorting (FACS). Specifically, this protocol enables the isolation of lymphocytes for the study of immune responses during various diseases, such as long-term neuroinflammation following ischemic stroke.
流式细胞术能够对细胞类型进行多参数定量分析，尤其是在对独特免疫细胞亚群进行免疫表型分析时，这些亚群可能会导致病理变化或改善病理状况。对于用于此类分析的组织，必须制备单细胞悬液。在此，我们描述了制备小鼠脾脏和脑组织单细胞悬液的方案，以及荧光激活细胞染色/分选（FACS）的步骤。具体而言，该方案能够分离淋巴细胞，用于研究各种疾病期间的免疫反应，如缺血性中风后的长期神经炎症。
