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使用 MADM-CloneSeq 原位绘制克隆相关细胞的谱系和细胞类型身份图谱的方案。

Protocol for mapping cell lineage and cell-type identity of clonally-related cells in situ using MADM-CloneSeq.

机构信息

Institute of Science and Technology Austria (ISTA), Am Campus 1, 3400 Klosterneuburg, Austria.

Institute of Science and Technology Austria (ISTA), Am Campus 1, 3400 Klosterneuburg, Austria.

出版信息

STAR Protoc. 2024 Sep 20;5(3):103168. doi: 10.1016/j.xpro.2024.103168. Epub 2024 Jul 4.

Abstract

The lineage relationship of clonally-related cells offers important insights into the ontogeny and cytoarchitecture of the brain in health and disease. Here, we provide a protocol to concurrently assess cell lineage relationship and cell-type identity among clonally-related cells in situ. We first describe the preparation and screening of acute brain slices containing clonally-related cells labeled using mosaic analysis with double markers (MADM). We then outline steps to collect RNA from individual cells for downstream applications and cell-type identification using RNA sequencing. For complete details on the use and execution of this protocol, please refer to Cheung et al..

摘要

克隆相关细胞的谱系关系为研究健康和疾病状态下大脑的发生和细胞结构提供了重要的线索。在这里,我们提供了一个同时评估原位克隆相关细胞的谱系关系和细胞类型身份的方案。我们首先描述了使用双标记马赛克分析(MADM)标记的包含克隆相关细胞的急性脑切片的制备和筛选。然后,我们概述了从单个细胞收集 RNA 的步骤,以便进行下游应用和使用 RNA 测序进行细胞类型鉴定。有关此方案的使用和执行的完整详细信息,请参阅 Cheung 等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c3f8/11452916/9470cfc7a9b5/fx1.jpg

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