Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Flemingovo nám. 2, 166 10, Prague, Czech Republic.
Chemistry. 2020 Aug 6;26(44):9945-9953. doi: 10.1002/chem.202001290. Epub 2020 Jul 14.
Fluorescent probes that light-up upon reaction with complementary bioorthogonal reagents are superior tools for no-wash fluorogenic bioimaging applications. In this work, a thorough study is presented on a set of seventeen structurally diverse coumarin-tetrazine probes that produce fluorescent dyes with exceptional turn-on ratios when reacted with trans-cyclooctene (TCO) and bicyclononyne (BCN) dienophiles. In general, formation of the fully aromatic pyridazine-containing dyes resulting from the reaction with BCN was found superior in terms of fluorogenicity. However, evaluation of the probes in cellular imaging experiments revealed that other factors, such as reaction kinetics and good cell permeability, prevail over the fluorescence turn-on properties. The best compound identified in this study showed excellent performance in live cell-labeling experiments and enabled no-wash fluorogenic imaging on a timescale of seconds.
荧光探针在与互补的生物正交试剂反应时会发光，是免洗荧光生物成像应用的优秀工具。在这项工作中，对一组十七种结构多样的香豆素-四嗪探针进行了深入研究，这些探针与反式环辛烯 (TCO) 和二环壬烯 (BCN) 亲二烯试剂反应时会产生具有出色开启比的荧光染料。一般来说，与 BCN 反应生成完全芳香的哒嗪含染料在荧光性方面表现更优。然而，在细胞成像实验中对探针进行评估后发现，其他因素，如反应动力学和良好的细胞通透性，比荧光开启特性更为重要。在这项研究中确定的最佳化合物在活细胞标记实验中表现出色，能够在数秒内实现免洗荧光生物成像。
