Ratiometric pH Sensing in Living Cells Using Carbon Dots

The ability to precisely sense physiological pH changes in the cellular environment is exceedingly difficult. Novel technologies are thus required to address this challenge. Fluorescent nanomaterials can be exploited to this effect because their optical properties can exhibit strong pH dependence. Herein, an intracellular pH-sensing probe is developed via a facile microwave-reaction synthesis method for the preparation of carbon dots (CDs) using glutathione and formamide. The CDs possess unique optical properties allowing for concomitant fluorescence in the blue and red regions of the spectrum. These dots are investigated as pH-sensors using the red fluorescence signatures at 650 and 680 nm. The two fluorescence bands respond differently following pH changes in their environment and could thus be used for ratiometric measurements. Cytotoxicity studies of the CDs in glioblastoma cells show no decrease in cell viability up to 100 mu g mL(-1) (24 h). Fluorescence imaging reveals that the dots localize in lysosomal compartments. Moreover, they can sense changes in lysosomal pH in response to serum and amino acid starvation, as well as administration of diclofenac and metformin, drugs currently in clinical trials for combination treatments of cancer. These CDs offer a new self-referencing approach for live intracellular pH sensing in 2D- and 3D-cell models.