Intracellular imaging and concurrent pH sensing of cancer-derived exosomes using surface-enhanced Raman scattering

Exosomes have attracted significant attention as cancer diagnostic targets and therapeutic agents due to their unique biogenesis and structure. To clarify the biological activities of exosomes, it is important to obtain a picture of their intracellular distribution and how they evolve over time. In this work, a new kind of intracellular exosome imaging and concurrent pH sensing method is demonstrated by using the surface-enhanced Raman scattering (SERS) technique. Specifically, 4-mercaptobenzoic acid (4MBA)-tagged silver nanoparticles are attached onto the outer surfaces of exosomes, in which silver nanoparticles are employed as SERS generators. Raman agents 4MBA are susceptible to a specific intracellular stimulus, that is, undergo a protonation or deprotonation in response to intracellular pH variation, which correspondingly exhibit different vibrational spectra features. By using the SERS spectroscopy, tracking of the intracellular distribution of exosomes and the concurrent quantitative sensing of environmental pH were achieved, which demonstrated that, as time prolonged, exosomes first attached with the tumor cell surfaces, and then entered into the cells and accumulated in lysosomes. Such SERS-active hybridized exosomes, that are sensitive to discrete variations in intracellular pH, have proved their capability for the investigation of interactions between exosomes and cells. The spectral diversity and flexible surface modification of these hybridized exosomes are also highly expected in developing multifunctional exosome-based nanoplatforms, which offers great potential to promote the exosome-based therapeutics forward into an advanced stage.