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使用空间注释单细胞测序对肿瘤内异质性进行分析的方案。

Protocol for profiling in vitro intratumor heterogeneity using spatially annotated single-cell sequencing.

机构信息

Department of Molecular Genetics, Erasmus University Medical Center, 3000 CA Rotterdam, the Netherlands; Erasmus MC Cancer Institute, 3000 CA Rotterdam, the Netherlands.

Department of Molecular Genetics, Erasmus University Medical Center, 3000 CA Rotterdam, the Netherlands; Erasmus MC Cancer Institute, 3000 CA Rotterdam, the Netherlands; Oncode Institute, 3521 AL Utrecht, the Netherlands.

出版信息

STAR Protoc. 2023 Sep 15;4(3):102447. doi: 10.1016/j.xpro.2023.102447. Epub 2023 Jul 14.

DOI:10.1016/j.xpro.2023.102447
PMID:37453069
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10384656/
Abstract

Here, we present a protocol for spatially annotated single-cell sequencing, a technique for spatially profiling intratumor heterogeneity with deep single-cell RNA sequencing and single-cell resolution. By combining live-cell imaging and photopatterned illumination, we describe steps to identify regions of interest in an in vitro tumor model, label the selected cells with photoactivatable dyes, and isolate and subject them to scRNAseq. This protocol can be applied to a range of cell lines and could be expanded to tissue sections. For complete details on the use and execution of this protocol, please refer to Smit et al. (2022)..

摘要

在这里，我们提出了一种空间注释单细胞测序的方案，该技术通过深度单细胞 RNA 测序和单细胞分辨率对肿瘤内异质性进行空间分析。通过结合活细胞成像和光图案化照明，我们描述了在体外肿瘤模型中识别感兴趣区域、用光活化染料标记选定细胞以及分离和进行 scRNAseq 的步骤。该方案可应用于一系列细胞系，并可扩展到组织切片。有关此方案的使用和执行的完整详细信息，请参阅 Smit 等人（2022 年）。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e25c/10384656/487a0c90d724/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e25c/10384656/514263ac4fb9/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e25c/10384656/7b2f10c38fe1/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e25c/10384656/32e28235057d/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e25c/10384656/487a0c90d724/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e25c/10384656/514263ac4fb9/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e25c/10384656/7b2f10c38fe1/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e25c/10384656/32e28235057d/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e25c/10384656/487a0c90d724/gr3.jpg

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