Gilvanov Aidar R, Myasnyanko Ivan N, Goncharuk Sergey A, Goncharuk Marina V, Kublitski Vadim S, Bodunova Daria V, Sidorenko Svetlana V, Maksimov Eugene G, Baranov Mikhail S, Bogdanova Yulia A
Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, Moscow 117997, Russia.
Laboratory of Medicinal Substances Chemistry, Institute of Translational Medicine, Pirogov Russian National Research Medical University, Ostrovitianov 1, Moscow 117997, Russia.
Biosensors (Basel). 2025 Apr 29;15(5):274. doi: 10.3390/bios15050274.
Fluorescence-lifetime imaging microscopy (FLIM) is a powerful technique for highly multiplexed imaging in live cells. In this work, we present a genetically encoded FLIM multiplexing platform based on a combination of fluorogen-activating protein FAST and red-shifted fluorogen from the arylidene-imidazolone family. We showed that a series of FAST protein mutants exhibit similar steady-state optical properties in complex with fluorogen but have different fluorescence lifetimes. The similar brightness and binding strength of pairs of these FAST protein variants with allows them to be successfully used for multiplexing up to three intracellular structures of living cells simultaneously.
荧光寿命成像显微镜（FLIM）是一种用于活细胞中高度多重成像的强大技术。在这项工作中，我们基于荧光团激活蛋白FAST和来自亚芳基咪唑酮家族的红移荧光团的组合，提出了一种基因编码的FLIM多重检测平台。我们表明，一系列FAST蛋白突变体与荧光团复合时表现出相似的稳态光学性质，但具有不同的荧光寿命。这些FAST蛋白变体对具有相似的亮度和结合强度，这使得它们能够成功地同时用于对活细胞的多达三种细胞内结构进行多重检测。
