22 February 2017 Probing the intracellular fate of supramolecular nanocarriers and their cargo with FRET schemes
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Abstract
We designed a strategy to monitor self-assembling supramolecular nanocarriers and their cargo simultaneously in the intracellular space with fluorescence measurements. It is based on Fӧrster resonance energy transfer (FRET) between complementary chromophores covalently integrated in the macromolecular backbone of amphiphilic polymers and/or noncovalently encapsulated in supramolecular assemblies of the amphiphilic components. Indeed, these polymers assemble into a micelles in aqueous phase to bring energy donors and acceptors in close proximity and allow energy transfer. The resulting supramolecular assemblies maintain their integrity after travelling into the intracellular space and do not lose their molecular guests in the process. Furthermore, this mechanism can also be exploited to probe the fate of complementary nanoparticles introduced within cells in consecutive incubation steps. Efficient energy transfer occurs in the intracellular space after the sequential incubation of nanocarriers incorporating donors first and then nanoparticles containing acceptors or vice versa. The two sets of nanostructured assemblies ultimately co-localize in the cell interior to bring donors and acceptors together and enable energy transfer. Thus, this protocol is particularly valuable to monitor the transport properties of supramolecular nanocarriers inside living cells and can eventually contribute to the fundamental understating of the ability of these promising vehicles to deliver contrast agents and/or drugs intracellularly in view of possible diagnostics and/or therapeutic applications.
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Ek Raj Thapaliya, Ek Raj Thapaliya, Colin Fowley, Colin Fowley, Bridgeen Callan, Bridgeen Callan, Sicheng Tang, Sicheng Tang, Yang Zhang, Yang Zhang, John F. Callan, John F. Callan, Françisco M. Raymo, Françisco M. Raymo, } "Probing the intracellular fate of supramolecular nanocarriers and their cargo with FRET schemes", Proc. SPIE 10078, Colloidal Nanoparticles for Biomedical Applications XII, 100781B (22 February 2017); doi: 10.1117/12.2269514; https://doi.org/10.1117/12.2269514
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