Multicolor fluorescent probes based on small organic molecules have the advantages of low cost, good biocompatibility, easily modifiable molecular structures and adjustable fluorescence performance. In addition, small molecule multicolor fluorescent probes generally undergo multi-site or multi-step reactions, which means that they can be used for the specific detection of structurally similar substances in complex bio-systems. In this review, we focus on the design and application of multicolor fluorescent probes based on small organic molecules: single fluorophores with multiple reaction sites, multiple fluorophores with single reaction sites, or multiple fluorophores with multiple reaction sites. Moreover, a design strategy for multicolor fluorescent probes and its application in biological imaging are also summarized, providing a systematic plan for future research on fluorescent probes functionalized by small organic molecules. It will also play an important role in the development of additional functions for small organic molecule fluorescent probes.

基于有机小分子的多色荧光探针具有成本低，生物相容性好，分子结构易于修饰，荧光性能可调的优点。另外，小分子多色荧光探针通常会经历多位点或多步反应，这意味着它们可用于复杂生物系统中结构相似物质的特异性检测。在这篇综述中，我们专注于基于有机小分子的多色荧光探针的设计和应用：具有多个反应位点的单个荧光团，具有单个反应位点的多个荧光团或具有多个反应位点的多个荧光团。此外，还总结了多色荧光探针的设计策略及其在生物成像中的应用，为将来由小有机分子功能化的荧光探针的研究提供系统的计划。它还将在有机小分子荧光探针的附加功能开发中发挥重要作用。