Protocol for Dissection and Dissociation of Zebrafish Telencephalon for Single-Cell Sequencing
Protocol for Dissection and Dissociation of Zebrafish Telencephalon for Single-Cell Sequencing
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DOI：
10.1016/j.xpro.2020.100042
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发表时间：
2020-06-19
期刊：
影响因子：
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通讯作者：
Kizil, Caghan
中科院分区：
文献类型：
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作者：
Cosacak, Mehmet Ilyas;Bhattarai, Prabesh;Kizil, Caghan
单细胞测序（sc-Seq）是研究复杂细胞混合物中细胞类型分子特征的有力工具。sc-Seq中的关键步骤是制备具有大量活细胞的单细胞悬浮液。在这里，我们展示了如何解剖斑马鱼端脑，以及如何将其分离成单细胞悬浮液。随后进行基于流式细胞术的分选以富集神经祖干细胞。我们的技术通常从一个斑马鱼端脑中产生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 et al.（2019）。
Single-cell sequencing (sc-Seq) is a powerful tool to investigate the molecular sig-natures of cell types in a complex mixture of cells. A critical step in sc-Seq is pre-paring 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).