使用 SunRISER 和 dNEMO 在活细胞实验中标记和定量 mRNA 分子。
Label and quantify mRNA molecules in live cell experiments using SunRISER and dNEMO.
机构信息
Department of Computational and Systems Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15260, USA; Department of Physics and Astronomy, University of Pittsburgh, Pittsburgh, PA 15260, USA.
Department of Computational and Systems Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15260, USA.
出版信息
STAR Protoc. 2022 Aug 18;3(3):101630. doi: 10.1016/j.xpro.2022.101630. eCollection 2022 Sep 16.
Visualization of mRNA molecules in single cells has revealed their core mechanisms as well as sources of cell-to-cell and spatiotemporal heterogeneity. Here, we describe a protocol to label, visualize, and quantify mRNA molecules by time-lapse imaging with the capability of resolving mRNA molecules over durations of hours to days. We provide links to mRNA-labeling plasmids as well as free software for a semi-automated image analysis pipeline. For complete details on the use and execution of this protocol, please refer to Guo and Lee (2022) and Kowalczyk et al. (2021).
单细胞内 mRNA 分子的可视化揭示了它们的核心机制以及细胞间和时空异质性的来源。在这里，我们描述了一种通过延时成像来标记、可视化和定量 mRNA 分子的方案，该方案具有解析数小时至数天内 mRNA 分子的能力。我们提供了 mRNA 标记质粒的链接以及用于半自动图像分析管道的免费软件。有关该方案使用和执行的完整详细信息，请参考 Guo 和 Lee（2022）以及 Kowalczyk 等人（2021）。
Zotero 插件