Real time monitoring and quantification of uptake carbon nanodots in eukaryotic cells

The real time monitoring and quantification of the concentration of highly fluorescent nitrogen-doped carbon nanodots (C-dots) in eukaryotic Tobacco bright yellow-2 (BY-2) plant cells was investigated by fluorescence and confocal microscopy. The quantitative measurement of their fluorescent emission intensity was possible because of the high photo-resistance, good water solubility and the absence of fading effect of the nanoparticles, which is frequent occurred problem of the conventional organic dyes. The microscopic analysis revealed that C-dots entered generally into the cells through endocytosis and caused negligible cytotoxicity. The multicolor cellular imaging of labeled Tobacco BY-2 demonstrates that the cells were in good health conditions and any blinking artifacts were not observed. The quantification of fluorescence emission intensity was carried out in the intracellular regions where the relationship between the C-dots concentration and relative emission was linear. Based on a control experiment with fluorescence liposomes with known dependence between C-dots concentration and emission, we were able to determine the amount of accumulated nanoparticles in the inner compartments of the eukaryotic cell through subsequent digital image analysis. The reported microscopic approach may be used for accurate testing and direct examination of the drug internalization mechanisms by C-dots as sensitive probes in single cells or tissues.