The florescence characteristics and the toxicities of carbon nanodots (CDs) are directly related to their elemental compositions. Fluorescent and non-toxic agent for imaging of biological systems was aimed. Sulfur and nitrogen co-doped CDs (S/N-CDs) was hydrothermally produced in an average size of 8 nm. S/N-CDs showed blue fluorescence under UV-light with an excitation wavelength of 365 nm. After 24 h, S/N-CDs was non-cytotoxic in HUVEC and L929 cells. S/N-CDs have a great potential to act as an alternative material for commercial fluorescent materials with its 85.5% of quantum yield. S/N-CDs was approved in vitro as an imaging agent for an ocular fundus angiography of rats.