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用于胚胎活细胞显微镜观察的 SNAP 及 Halo 标记和染料引入方案。

SNAP- and Halo-tagging and dye introduction protocol for live microscopy in embryos.

机构信息

Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1085, USA.

Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1085, USA.

出版信息

STAR Protoc. 2022 Aug 18;3(3):101622. doi: 10.1016/j.xpro.2022.101622. eCollection 2022 Sep 16.

DOI:10.1016/j.xpro.2022.101622
PMID:36035797
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9405085/
Abstract

Traditional fluorescent proteins exhibit limitations in brightness and photostability that hinder optimal characterization of the dynamic cellular behavior of proteins of interest. SNAP- and Halo-tagging are alternatives to traditional fluorescent protein tagging utilizing bright, stable chemical dyes, which may improve signal-to-noise ratio. However, there has been limited use of this approach in developing organisms. Here, we present a protocol for implementing SNAP- and Halo-tagging in gastrula-stage embryos for live confocal microscopy. For complete details on the use and execution of this protocol, please refer to Varadarajan et al. (2022).

摘要

传统荧光蛋白在亮度和光稳定性方面存在局限性，这阻碍了对感兴趣的蛋白质的动态细胞行为进行最佳表征。SNAP 和 Halo 标记是利用明亮、稳定的化学染料替代传统荧光蛋白标记的方法，这可能会提高信噪比。然而，这种方法在发育生物中应用有限。在这里，我们提出了一种在原肠胚期胚胎中进行 SNAP 和 Halo 标记以进行活共聚焦显微镜的方案。有关此方案的使用和执行的完整详细信息，请参阅 Varadarajan 等人。（2022 年）。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/c4aee4d6c278/gr10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/f4bed18c60ac/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/4e210b396b1d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/7ec626c1a1a5/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/2f30bc60dbf8/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/4cee581163af/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/e341f58dc390/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/7e4de92a0b74/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/4cac9c229ab0/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/0a5ff6072e5c/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/2dda6d507395/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/c4aee4d6c278/gr10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/f4bed18c60ac/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/4e210b396b1d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/7ec626c1a1a5/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/2f30bc60dbf8/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/4cee581163af/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/e341f58dc390/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/7e4de92a0b74/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/4cac9c229ab0/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/0a5ff6072e5c/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/2dda6d507395/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c376/9405085/c4aee4d6c278/gr10.jpg

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