Multiplexed In Vivo Imaging with Fluorescence Lifetime‐Modulating Tags

Abstract: Fluorescence lifetime imaging microscopy (FLIM) opens new dimensions for highly multiplexed imaging in live cells and organisms using differences in fluorescence lifetime to distinguish spectrally identical fluorescent probes. Here, a set of fluorescence‐activating and absorption‐shifting tags (FASTs) capable of modulating the fluorescence lifetime of embedded fluorogenic 4‐hydroxybenzylidene rhodanine (HBR) derivatives is described. It is shown that changes in the FAST protein sequence can vary the local environment of the chromophore and lead to significant changes in fluorescence lifetime. These fluorescence lifetime‐modulating tags enable multiplexed imaging of up to three targets in one spectral channel using a single HBR derivative in live cells and live zebrafish larvae. The combination of fluorescence lifetime multiplexing with spectral multiplexing allows to successfully image six targets in live cells, opening great prospects for multicolor fluorescence lifetime multiplexing.

Object Details

• Location: Deutsche Nationalbibliothek Frankfurt am Main
• Extent: Online-Ressource
• Language: Englisch

References and Relationships

• Bibliographic citation: Multiplexed In Vivo Imaging with Fluorescence Lifetime‐Modulating Tags ; day:20 ; month:06 ; year:2024 ; extent:12 ; Advanced science ; (20.06.2024) (gesamt 12)

Contributors, Places and Time

• Creator: El Hajji, Lina ; Lam, France ; Avtodeeva, Maria ; Benaissa, Hela ; Rampon, Christine ; Volovitch, Michel ; Vriz, Sophie ; Gautier, Arnaud

Further information

• DOI: 10.1002/advs.202404354
• URN: urn:nbn:de:101:1-2406211403171.101766646125
• Rights: Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Last update: 14.08.2025, 11:02 AM CEST