Fluorescence Lifetime Multiplexing with Fluorogen-Activating FAST Protein Variants and Red-Shifted Arylidene–Imidazolone Derivative as Fluorogen

Авторы: 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.
Журнал: Biosensors
Том: 15
Номер: 5
Год издания: 2025
Издательство: MDPI
Местоположение издательства: Basel, Switzerland
Первая страница: 274
Последняя страница: 274
DOI: 10.3390/bios15050274
Аннотация: 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 N871b from the arylidene–imidazolone family. We showed that a series of FAST protein mutants exhibit similar steady-state optical properties in complex with N871b fluorogen but have different fluorescence lifetimes. The similar brightness and binding strength of pairs of these FAST protein variants with N871b allows them to be successfully used for multiplexing up to three intracellular structures of living cells simultaneously
Добавил в систему: Сидоренко Светлана Вадимовна

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