The cytotoxicity of various surface-functionalized gold nanowires with different aspect ratios is investigated by (3-(4,5-dimethylthiazol-2-yl)2,5- diphenyltetrazolium bromide) (MTT) assays for two cell lines, fibroblast and HeLa. It is found that functionalized gold nanowires with a diameter of 200 nm and lengths up to a few micrometers can be readily internalized by both types of cells regardless of the type of surface functionalization. However, the cytotoxicity of the gold nanowires is observed to depend on their surface modification. Serum-coated gold nanowires are the least toxic, whereas more than 50 % of the cells are damaged in the presence of mercapto-acid-modified gold nanowires even at very low concentrations (103 nanowires mL -1). Nanowires with different aspect ratios exhibit the same cytotoxicity within limits of experimental error. However, the uptake efficiency is found to be higher for shorter nanowires as compared to their longer counterparts. Therefore, we conclude that internalized nanowires with high aspect ratios are more toxic to cells than nanowires with low aspect ratios. Positively charged aminothiol-rnodified gold nanowires are employed to deliver both plasmid DNA and probe molecules into cells without compromising the viability of the cells. The local environment of individual nanowires within the cells is studied by monitoring the fluorescence signal from probe molecules attached to the nanowires.