Label and quantify mRNA molecules in live cell experiments using SunRISER and dNEMO.
Label and quantify mRNA molecules in live cell experiments using SunRISER and dNEMO.
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DOI：
10.1016/j.xpro.2022.101630
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发表时间：
2022-09-16
期刊：
影响因子：
--
通讯作者：
Lee, Robin E. C.
中科院分区：
文献类型：
--
作者：
Guo, Yue;Kowalczyk, Gabriel J.;Lee, Robin E. C.
单细胞中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时间推移成像来标记、可视化和量化mRNA分子的方案，该方案能够在几小时到几天的持续时间内解析mRNA分子。我们提供了到信使核糖核酸标记质粒的链接，以及用于半自动图像分析管道的免费软件。有关本议定书的使用和执行的完整细节，请参阅和。Sunriser是一个针对单个RNA活细胞成像实验进行优化的工具箱快速检测荧光显微镜图像中标记的单个mRNA分子自动化单细胞分割和mRNA定量出版商备注：执行任何实验方案都需要遵守当地实验室安全和伦理方面的机构指南。单细胞中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时间推移成像来标记、可视化和量化mRNA分子的方案，该方案能够在几小时到几天的持续时间内解析mRNA分子。我们提供了到信使核糖核酸标记质粒的链接，以及用于半自动图像分析管道的免费软件。
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 and. SunRISER is a toolbox optimized for single mRNA live-cell imaging experiments Rapid detection of labeled single mRNA molecules in fluorescence microscopy images Automated single-cell segmentation and mRNA quantification Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.
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DOI：
10.1083/jcb.201004104
发表时间：
2010-05-31
期刊：
The Journal of cell biology
影响因子：
--
作者：
Linkert M;Rueden CT;Allan C;Burel JM;Moore W;Patterson A;Loranger B;Moore J;Neves C;Macdonald D;Tarkowska A;Sticco C;Hill E;Rossner M;Eliceiri KW;Swedlow JR
通讯作者：
Swedlow JR
影响因子：
56.9
作者：
Femino, A;Fay, FS;Singer, RH
通讯作者：
Singer, RH
影响因子：
16
作者：
Bertrand, E;Chartrand, P;Long, RM
通讯作者：
Long, RM
影响因子：
48
作者：
Schneider CA;Rasband WS;Eliceiri KW
通讯作者：
Eliceiri KW
影响因子：
64.5
作者：
Raj A;van Oudenaarden A
通讯作者：
van Oudenaarden A