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A key limitation of electron paramagnetic resonance (EPR), an established andpowerful tool for studying atomic-scale biomolecular structure and dynamics isits poor sensitivity, samples containing in excess of 10^12 labeledbiomolecules are required in typical experiments. In contrast, single moleculemeasurements provide improved insights into heterogeneous behaviors that can bemasked by ensemble measurements and are often essential for illuminating themolecular mechanisms behind the function of a biomolecule. We report EPRmeasurements of a single labeled biomolecule that merge these two powerfultechniques. We selectively label an individual double-stranded DNA moleculewith a single nanodiamond containing nitrogen-vacancy (NV) centers, andoptically detect the paramagnetic resonance of NV spins in the nanodiamondprobe. Analysis of the spectrum reveals that the nanodiamond probe has completerotational freedom and that the characteristic time scale for reorientation ofthe nanodiamond probe is slow compared to the transverse spin relaxation time.This demonstration of EPR spectroscopy of a single nanodiamond labeled DNAprovides the foundation for the development of single molecule magneticresonance studies of complex biomolecular systems.

NV Center Electron Paramagnetic Resonance of a Single Nanodiamond Attached to an Individual Biomolecule