通过使用CellProfiler的RNAscope捕获的相邻组织部分跨越多个标记物的空间分析协议.
Katherine Copeland1, Ariestya Indah Permata Sari2, Justin Copeland1
1Science Division, Mahidol University International College, Mahidol University, Salaya, Phutthamonthon, Nakhon Pathom 73170, Thailand.
STAR protocols
|June 22, 2025
概括
这项研究引入了一种新的协议,用于使用先进的成像分析量化单细胞中的基因表达. 该方法有助于比较转移性和非转移性癌症样本之间的基因表达.
科学领域:
- 生物医学成像技术 生物医学成像技术
- 分子生物学分子生物学
- 癌症研究 癌症研究
背景情况:
- 在单细胞分辨率下精确量化基因表达对于理解复杂的生物过程至关重要.
- 在转移性与非转移性癌症中区分基因表达模式是针对性治疗的关键.
研究的目的:
- 以单细胞空间分辨率呈现使用光现场杂交 (FISH) 量化基因表达的详细协议.
- 为了使癌症样本与转移和没有转移之间的基因表达进行比较分析.
主要方法:
- 使用基于CellProfiler软件的成像分析管道进行基因表达量化.
- 采用RNAscope多重光测试来检测目标转录.
- 利用Vectra Polaris自动定量病理成像用于可视化.
- 提供了分析邻近组织部分的信号相互作用的选项.
主要成果:
- 该协议为高分辨率基因表达分析提供了一种标准化的方法.
- 促进与转移相关的差异性基因表达模式的识别.
- 能够在组织微环境中对基因表达和转录相互作用进行空间分析.
结论:
- 该协议为单细胞空间基因表达分析提供了一个强大的框架.
- 它是癌症研究的宝贵工具,特别是在了解转移方面.
- 该方法可以适应各种生物研究,需要精确的空间转录基因数据.
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