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斑马鱼端脑解剖和分离用于单细胞测序的方案。

Protocol for Dissection and Dissociation of Zebrafish Telencephalon for Single-Cell Sequencing.

机构信息

German Center for Neurodegenerative Diseases (DZNE) Dresden, Helmholtz Association, Tatzberg 41, 01307 Dresden, Germany.

Center for Molecular and Cellular Bioengineering (CMCB), TU Dresden, Fetscherstr. 105, 01307 Dresden, Germany.

出版信息

STAR Protoc. 2020 Jun 19;1(1):100042. doi: 10.1016/j.xpro.2020.100042. Epub 2020 May 29.

Abstract

Single-cell sequencing (sc-Seq) is a powerful tool to investigate the molecular signatures of cell types in a complex mixture of cells. A critical step in sc-Seq is preparing a single-cell suspension with a high number of viable cells. Here, we show how to dissect zebrafish telencephalon and how to dissociate it into a single-cell suspension. This is followed by flow cytometry-based sorting to enrich for neural progenitor stem cells. Our technique typically yields 70,000 live cells from one zebrafish telencephalon. For complete details on the use and execution of this protocol, please refer to Cosacak et al. (2019).

摘要

单细胞测序(sc-Seq)是一种强大的工具,可用于研究复杂细胞混合物中细胞类型的分子特征。sc-Seq 的一个关键步骤是用大量存活细胞制备单细胞悬浮液。在这里,我们展示如何解剖斑马鱼端脑,以及如何将其解离成单细胞悬浮液。接下来是基于流式细胞术的分选,以富集神经祖细胞干细胞。我们的技术通常可从一只斑马鱼端脑中获得 70,000 个活细胞。有关该方案的使用和执行的完整详细信息,请参阅 Cosacak 等人。(2019 年)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eafa/7580211/af17c11cdc62/fx1.jpg

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