Fluorescent tools have revolutionized our ability to probe biological dynamics, particularly at the cellular level. Fluorescent sensors have been developed on several platforms, utilizing either small-molecule dyes or fluorescent proteins, to monitor proteins, RNA, DNA, small molecules, and even cellular properties, such as pH and membrane potential. We briefly summarize the impressive history of tool development for these various applications and then discuss the most recent noteworthy developments in more detail. Particular emphasis is placed on tools suitable for single-cell analysis and especially live-cell imaging applications. Finally, we discuss prominent areas of need in future fluorescent tool development-specifically, advancing our capability to analyze and integrate the plethora of high-content data generated by fluorescence imaging.

中文翻译：

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活细胞成像荧光工具的批判性比较研究。

荧光工具彻底改变了我们探测生物动力学的能力，尤其是在细胞水平上。已经在几种平台上开发了荧光传感器，利用小分子染料或荧光蛋白来监测蛋白质，RNA，DNA，小分子，甚至细胞特性，例如pH和膜电位。我们简要总结了针对这些各种应用程序的令人印象深刻的工具开发历史，然后更详细地讨论了最新的值得注意的开发。特别强调适用于单细胞分析，尤其是活细胞成像应用的工具。最后，我们具体讨论了未来荧光工具开发中的重要需求领域，从而提高了我们分析和整合由荧光成像生成的大量高含量数据的能力。