At the most fundamental level, collagen fibrils are rope-like structures assembled from triple-helical collagen molecules. One key structural characteristic of the fibril is the 67 nm D-band pattern arising from the quarter-stagger packing of the molecules. Our current understanding of the structural changes induced by tensile loading of collagen fibrils comes mostly from atomistic molecular dynamics simulations and tissue level experiments. Tensile testing of individual fibrils is an upcoming field of investigation, and thus far structural analysis has always taken place after the fibrils have been ruptured or strained and subsequently dried. There is therefore a gap in understanding how the structure of collagen fibrils transforms under tension, and how this reorganization affects the functionality of collagen fibrils within tissues. In this study, atomic force microscopy based nanomechanical mapping is introduced to image hydrated collagen fibrils absorbed to an elastic substrate. Upon stretching the substrate between 5 and 30%, we observe a radial stiffening consistent with the fibrils being under tension. This is associated with an increase in D-band length. In addition the indentation modulus contrast associated with the D-band pattern increases linearly with D-band strain. These results provide direct confirmation of, and new information on the axially inhomogeneous structural response of collagen fibrils to applied tension as previously proposed on the basis of X-ray scattering experiments on stretched tissues. Furthermore our approach opens the road for studying the structural impacts of tension on cell-matrix interactions at the molecular level.