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Analysis of intracellular protein dynamics in living zebrafish embryos using light-sheet fluorescence single-molecule microscopy
Biomedical Optics Express ( IF 2.9 ) Pub Date : 2021-09-13 , DOI: 10.1364/boe.435103
Matteo Bernardello 1, 2 , Radoslaw J Gora 2, 3 , Patrick Van Hage 3 , Gustavo Castro-Olvera 1 , Emilio J Gualda 1 , Marcel J M Schaaf 2, 3 , Pablo Loza-Alvarez 1, 2
Affiliation  

Single-molecule microscopy techniques have emerged as useful tools to image individual molecules and analyze their dynamics inside cells, but their application has mostly been restricted to cell cultures. Here, a light-sheet fluorescence microscopy setup is presented for imaging individual proteins inside living zebrafish embryos. The optical configuration makes this design accessible to many laboratories and a dedicated sample-mounting system ensures sample viability and mounting flexibility. Using this setup, we have analyzed the dynamics of individual glucocorticoid receptors, which demonstrates that this approach creates multiple possibilities for the analysis of intracellular protein dynamics in intact living organisms.

中文翻译:

使用光片荧光单分子显微镜分析活斑马鱼胚胎的细胞内蛋白质动力学

单分子显微镜技术已经成为对单个分子进行成像并分析它们在细胞内的动态的有用工具,但它们的应用主要限于细胞培养。在这里,提出了一种光片荧光显微镜设置,用于对活斑马鱼胚胎内的单个蛋白质进行成像。光学配置使许多实验室都可以使用这种设计,并且专用的样品安装系统可确保样品的可行性和安装灵活性。使用这种设置,我们分析了个体糖皮质激素受体的动力学,这表明这种方法为分析完整活生物体的细胞内蛋白质动力学创造了多种可能性。
更新日期:2021-10-01
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