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基于核内免疫染色的胚胎皮质组织 FACS 方案。

Intranuclear immunostaining-based FACS protocol from embryonic cortical tissue.

机构信息

German Center for Neurodegenerative Diseases, 37077 Goettingen, Germany.

Ruhr University of Bochum, 44791 Bochum, Germany.

出版信息

STAR Protoc. 2021 Feb 2;2(1):100318. doi: 10.1016/j.xpro.2021.100318. eCollection 2021 Mar 19.

Abstract

Cell sorting can be used to purify cell populations for cell type-specific molecular probing. Fluorescence-activated cell sorting (FACS) coupled with high-throughput sequencing affords molecular signature identification for specific cell types. FACS has many challenges that limit comprehensive cell purification from the brain, leading to incomplete molecular characterization. Here, we present the intranuclear immunostaining-based FACS protocol with several modified steps, which allows optimized nuclei/cell sorting from mouse or human embryonic cortical tissue for distinct downstream molecular investigation of basal intermediate progenitors.

摘要

细胞分选可用于纯化具有特定细胞类型分子特征的细胞群体。荧光激活细胞分选(FACS)与高通量测序相结合,可鉴定特定细胞类型的分子特征。FACS 存在许多限制其从大脑中全面纯化细胞的挑战,导致分子特征分析不完整。本研究提出了一种基于核内免疫染色的 FACS 方案,对该方案进行了多项改进,可从小鼠或人胚胎皮质组织中优化核/细胞分选,以对基底中间祖细胞进行下游特定的分子研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c03e/7859298/93ff88d4e4ac/fx1.jpg

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