Magnetic resonance spectroscopy of single biomolecules under near-physiological conditions could substantially advance understanding of their biological function, but this approach remains very challenging. Here we used nitrogen-vacancy centers in diamonds to detect electron spin resonance spectra of individual, tethered DNA duplexes labeled with a nitroxide spin label in aqueous buffer solutions at ambient temperatures. This work paves the way for magnetic resonance studies on single biomolecules and their intermolecular interactions in native-like environments.

The in situ measurement of the distribution of biomolecules inside a cell is one of the important goals in life science. Among various imaging techniques, magnetic imaging (MI) based on the nitrogen-vacancy (NV) center in diamond provides a powerful tool for the biomolecular research, while the nanometer-scale MI of intracellular proteins remains a challenge. Here, we use ferritin as a demonstration to realize the MI of endogenous proteins in a single cell using the NV center as the sensor. With the scanning, intracellular ferritins are imaged with a spatial resolution of ca. 10 nm, and ferritin-containing organelles are colocalized by correlative MI and electron microscopy. The approach paves the way for nanoscale MI of intracellular proteins.

Despite the increasing number of regulatory proteins identified in clathrin-independent endocytic (CIE) pathways, our understanding of the exact functions of these proteins and the sequential manner in which they function remains limited. In this study, using the Caenorhabditis elegans intestine as a model, we observed a unique structure of interconnected endosomal tubules, which is required for the basolateral recycling of several CIE cargoes including hTAC, GLUT1, and DAF-4. SEC-10 is a subunit of the octameric protein complex exocyst. Depleting SEC-10 and several other exocyst components disrupted the endosomal tubules into various ring-like structures. An epistasis analysis further suggested that SEC-10 operates at the intermediate step between early endosomes and recycling endosomes. The endosomal tubules were also sensitive to inactivation of the Rab GTPase RAB-10 and disruption of microtubules. Taken together, our data suggest that SEC-10 coordinates with RAB-10 and microtubules to form the endosomal tubular network for efficient recycling of particular CIE cargoes.endosomal tubules | recycling | microtubule | clathrin-independent endocytosis | live worm imaging