State Key Laboratory of Organ Failure Research, Key Laboratory of Mental Health of the Ministry of Education, Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Guangdong-Hong Kong Joint Laboratory for Psychiatric Disorders, Guangdong Province Key Laboratory of Psychiatric Disorders, Guangdong Basic Research Center of Excellence for Integrated Traditional and Western Medicine for Qingzhi Diseases, Department of Neurobiology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
Methods Mol Biol. 2024;2794:341-351. doi: 10.1007/978-1-0716-3810-1_29.
Single-cell RNA sequencing (scRNA-seq) has been widely applied in neuroscience research, enabling the investigation of cellular heterogeneity at the transcriptional level, the characterization of rare cell types, and the detailed analysis of the stochastic nature of gene expression. Isolation of single nerve cells in good health, especially from the adult rodent brain, is the most difficult and critical process for scRNA-seq. Here, we describe methods to optimize protease digestion of brain slices, which enable yield of millions of cells in good health from the adult brain.
单细胞 RNA 测序（scRNA-seq）已广泛应用于神经科学研究，能够在转录水平上研究细胞异质性、鉴定稀有细胞类型，并深入分析基因表达的随机性。分离健康的单个神经细胞，特别是成年啮齿动物大脑中的单个神经细胞，是 scRNA-seq 中最困难和最关键的步骤。在此，我们描述了优化脑片蛋白酶消化的方法，该方法可从成年大脑中获得数以百万计的健康细胞。
