A carbon-based fluorescent nanoparticle is considered to be a new generation nontoxic nanoprobe suitable for various bioimaging and sensing applications. However, the synthesis of such a high-quality nanoparticle is challenging, and its application potential is mostly unexplored. Here we report a vitamin B1 carbonization-based approach for blue and green fluorescent carbon nanoparticles of <10 nm size with a fluorescence quantum of up to 76%. We found that carbonization of vitamin B1 in the presence of phosphate salt at ∼90-130 °C for about 2 h produces highly fluorescent carbon nanoparticles of 1-6 nm size. The particle size and fluorescence property can be controlled by varying the reaction temperature and nature of phosphate salt. Elemental analysis shows the incorporation of a large percentage (up to 48 wt %) of other elements (such as nitrogen, oxygen, phophorus, and sulfur) in the carbon matrix. The chemical structure of vitamin B1 (thiamine) is unique in a sense that it consists of a large number of heteroatoms along with unsaturated bonds and offers low-temperature carbonization with the formation of a nanoparticle having an optimum ratio of sp2 and sp3 carbon atoms. These carbon nanoparticles have high colloidal stability and stable fluorescence and have been used as fluorescent imaging probes. © 2014 American Chemical Society.