Building Electromagnetic Hot Spots in Living Cells via Target-Triggered Nanoparticle Dimerization

Electromagnetic hot spots of surface-enhanced Raman scattering have been extensively employed for bioanalysis in solution or on a substrate, but building hot spots in living systems for probing targets of interest has not been achieved yet because of the complex and dynamic physiological environment. Herein, we show that a target-programmed nanoparticle dimerization can be combined with the background-free Raman reporters (alkyne, C≡C; nitrile, C≡N) for multiplexed imaging of microRNAs (miRNAs) in living cells. The in situ formation of plasmonic dimers results in an intense hot spot, thus dramatically enhancing the Raman signals of the reporters residing in the hot spot. More significantly, the reporters exhibit single nonoverlapping peaks in the cellular Raman-silent region (1800-2800 cm^-1), thus eliminating spectral unmixing and background interference. A 3D Raman mapping technique was harnessed to monitor the spatial distribution of the dimers and thus the multiple miRNAs in cells. This approach could be extended to probe other biomarkers of interest for monitoring specific pathophysiological events at the live-cell level.