Fluorescence-Raman dual-mode quantitative detection and imaging of small-molecule thiols in cell apoptosis with DNA-modified gold nanoflowers.

The monitoring of small-molecule thiols (especially glutathione) has attracted widespread attention due to their involvement in numerous physiological processes in living organisms and cells. In this work, a dual-mode nanosensor was designed to detect small-molecule thiols, which is based on the "on-off" switch of fluorescence resonance energy transfer (FRET) and surface-enhanced Raman scattering (SERS). Briefly, DNA was modified by Cy5 (signal probe) and disulfide bonds (recognition element). Gold nanoflowers (AuNFs) were used as the fluorescence-quenching and SERS-enhancing substrate. However, small-molecule thiols can cleave disulfide bonds and release short Cy5-labeled chains, causing the recovery of the fluorescence signal and a decrease of the SERS signal. The nanosensor showed a sensitive response to small-molecule thiols represented by GSH, with a linear range of 0.01-3 mM and a detection limit of 913 nM. In addition, it competed with other related biological interferences and presented good stability and better selectivity towards small-molecule thiols. Most importantly, the developed nanosensor had been successfully applied to in situ imaging and quantitative monitoring of the concentration of small-molecule thiols which changed during T-2 toxin-induced apoptosis in HeLa cells. Meanwhile, nanosensors are also versatile with their potential applications and can be easily extended to the detection and imaging of other human cell lines. The proposed method combines the dual advantages of fluorescence and SERS, which has broad prospects for in situ studies of physiological processes involving small-molecule thiols in biological systems.