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Article Dans Une Revue RSC Advances Année : 2016

Dye-doped silica nanoparticle probes for fluorescence lifetime imaging of reductive environments in living cells

Luca Petrizza
  • Fonction : Auteur
Mayeul Collot
Ludovic Richert
  • Fonction : Auteur
Yves Mely
  • Fonction : Auteur
Luca Prodi
  • Fonction : Auteur

Résumé

Fluorescence detection sensitivity can be drastically improved by the application of nanoparticles (NPs)because of their superior brightness compared to organic dyes. Here, using dye-doped silica NPs(SiNPs), we developed FRET-based nanoparticle probes for the detection of reductive environments inliving cells. To this end, we designed three FRET acceptors based on black hole quenchers (BHQs). Theirpolarity was tuned by introducing hydroxyl, PEG and sulfate groups. To conjugate them to NPs, we usedan original pre-functionalization approach, where the quencher was coupled by a“click”reaction toPluronic F127 and further used for the preparation of silica NPs. This approach enabled easy preparationof silica NPs functionalized with varying amounts of quenchers by simple mixing of functionalized andparent Pluronic F127 in different mol%. The increase in the quencher concentration at the SiNPs surfaceproduced a rapid drop in thefluorescence intensity with 80% quenching and a 2-fold drop in theemission lifetime for 16 mol% of the quenchers. Then, to obtain turn-ON sensing of reductiveenvironments, the quenchers were coupled to the NPs through a disulfide linker using the same pre-functionalization strategy. The obtained nano-probes showed a >10-fold increase in theirfluorescencein the presence of reductive agents, such as tris(2-carboxyethyl)phosphine (TCEP) and glutathione.Remarkably, BHQ quencher bearing sulfate group showed the highest turn-ON response, probably dueto its superior capacity to escape from the NP surface after disulfide bond cleavage. The obtained bestnanoprobe was successfully applied for detection of reductive environments inside living cells usingfluorescence lifetime imaging (FLIM). This work provides insights for FRET acceptor design and itscontrolled grafting, which enables preparation of thefirst redox-sensitive silica nanoparticle probe forlifetime imaging.
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Dates et versions

hal-03283566 , version 1 (11-07-2021)

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Luca Petrizza, Mayeul Collot, Ludovic Richert, Yves Mely, Luca Prodi, et al.. Dye-doped silica nanoparticle probes for fluorescence lifetime imaging of reductive environments in living cells. RSC Advances, 2016, 6 (106), pp.104164-104172. ⟨10.1039/C6RA21427D⟩. ⟨hal-03283566⟩

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