The possibility to detect and quantify protein-protein interactions with good spatial and temporal resolutions in live cells is crucial in biology. Number and brightness is a powerful approach to detect both protein aggregation/desegregation dynamics and stoichiometry in live cells. Importantly, this technique can be applied in commercial set ups: both camera based and laser scanning microscopes. It provides pixel-by-pixel information on protein oligomeric states. If performed with two colours, the technique can retrieve the stoichiometry of the reaction under study. In this review, we discuss the strengths and weaknesses of the technique, stressing which are the correct acquisition parameters for a given microscope, the main challenges in analysis, and the limitations of the technique.

中文翻译：

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检测活细胞中的蛋白质聚集和相互作用：数量和亮度指南

在活细胞中以良好的空间和时间分辨率检测和量化蛋白质-蛋白质相互作用的可能性在生物学中至关重要。数量和亮度是检测活细胞中蛋白质聚集/分离动力学和化学计量的有效方法。重要的是，这种技术可以应用于商业设置：基于相机和激光扫描显微镜。它提供有关蛋白质寡聚状态的逐像素信息。如果使用两种颜色进行，该技术可以检索所研究反应的化学计量。在这篇综述中，我们讨论了该技术的优点和缺点，强调了给定显微镜的正确采集参数、分析中的主要挑战以及该技术的局限性。