Advanced Bioimaging Unit, Institut Pasteur of Montevideo & Universidad de la República, Montevideo, Uruguay.
Departamento de Fisiopatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
Methods Mol Biol. 2023;2564:53-74. doi: 10.1007/978-1-0716-2667-2_3.
Fluorescent proteins are standard tools for addressing biological questions in a cell biology laboratory. The genetic tagging of protein of interest with fluorescent proteins opens the opportunity to follow them in vivo and to understand their interactions and dynamics. In addition, the latest advances in optical microscopy image acquisition and processing allow us to study many cellular processes in vivo. Techniques such as fluorescence lifetime microscopy and hyperspectral imaging provide valuable tools for understanding fluorescent protein interactions and their photophysics. Finally, fluorescence fluctuation analysis opens the possibility to address questions of molecular diffusion, protein-protein interactions, and oligomerization, among others, yielding quantitative information on the subject of study. This chapter will cover some of the more important advances in cutting-edge technologies and methods that, combined with fluorescent proteins, open new frontiers for biological studies.
荧光蛋白是细胞生物学实验室解决生物学问题的标准工具。将感兴趣的蛋白质与荧光蛋白进行基因标记，为我们提供了在体内追踪它们的机会，从而有助于我们了解它们的相互作用和动态。此外，光学显微镜图像采集和处理的最新进展使我们能够在体内研究许多细胞过程。荧光寿命显微镜和高光谱成像等技术为理解荧光蛋白相互作用及其光物理特性提供了有价值的工具。最后，荧光波动分析为解决分子扩散、蛋白质-蛋白质相互作用和寡聚化等问题提供了可能性，为研究对象提供了定量信息。本章将介绍一些前沿技术和方法的重要进展，这些技术和方法与荧光蛋白结合，为生物学研究开辟了新的领域。
