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Measuring the diffusion of fluorescent dye or protein inside living cells

机译:测量荧光染料或蛋白质在活细胞内的扩散

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摘要

Information about the space and time inside a living cell is important to fully understand the different molecular events that occur in the cell. The cell cytoplasm, interior of a red blood cell or mitochondrial matrix are spaces crowded with macromolecules like proteins, for example. What are the consequences of such molecular crowding on the rate of transport of solutes or metabolites within the cell is not clear. Is diffusion alone sufficient to take small solutes to different corners of the cell? Fluorescence microscopy provides a non-invasive approach to understand the inner workings of a living cell. Techniques like fluorescence recovery after photobleaching and time-resolved fluorescence anisotropy have made it possible to observe diffusion of small dyes like BCECF and average size proteins like GFP inside living cells in real time. In this article, we introduce fluorescence microscopy, its advantages/limitations and briefly highlight a few observations of diffusion inside living cells.
机译:有关活细胞内部空间和时间的信息对于全面了解细胞中发生的不同分子事件非常重要。例如,细胞质,红细胞内部或线粒体基质是充满大分子(如蛋白质)的空间。这种分子拥挤对溶质或代谢物在细胞内运输速率的后果尚不清楚。单独的扩散是否足以将小溶质带到细胞的不同角落?荧光显微镜提供了一种非侵入性的方法来了解活细胞的内部运作。光漂白后的荧光恢复以及时间分辨的荧光各向异性等技术使活细胞内实时观察小染料(如BCECF)和中等大小的蛋白(如GFP)的扩散成为可能。在本文中,我们介绍了荧光显微镜及其优势/局限性,并简要介绍了活细胞内扩散的一些观察结果。

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