Effective control of the surface properties of devices becomes more and more important to ensure their prolonged performance, as the size of devices diminishes to the nanoscale. At the nanoscale interfacial properties can differ from the macroscopic properties including conductivity, electron affinity, surface potential, and reactivity.[1] Here, we present a single-molecule sensor to detect molecular adsorption free energies in liquid under variable conditions. Utilizing atomic force microscopy (AFM), this approach provides nanoscale precision and excellent sensitivity to surface properties. This is demonstrated by the investigation of the aging of a diamond that was chemically modified to control its conductivity, surface potential, and wettability.Diamond exhibits extraordinary mechanical properties and is chemically inert as well as biocompatible. It is, therefore, suitable for biomedical applications [2,3] such as cell growth support [4,5] and as a refinement of medical indwelling devices. [6,7] In addition to the exceptional material characteristics, hydrogenated diamond exhibits p-type surface conductivity, whereas oxidized diamond is insulating.[8] These electronic properties have advanced the further development of diamond-based sensor devices, such as electrochemical sensors, [9] biosensors, [10,11] and ion-sensitive field effect transistors (ISFETs) in bioelectronics. [12][13][14] The application of diamond-based devices that take advantage of the unique surface properties of diamond, be it in air, in aqueous solution, or even in blood, necessitates stable surface conditions and interfacial properties in order to ensure the long lasting functionality of the device. Only the limited chemical stability under physiological conditions has hampered the implementation of silicon and other semiconductive materials into similar devices. Implanted medical devices should stay intact for months or even years without the need for removal due to restricted biocompatibility and foreign body reactions.We utilize single-molecule force spectroscopy (SMFS) in order to determine the adsorption free energy of poly(allylamine) (PAAm) on diamond surfaces of different termination and different age (Fig. 1A). In contrast to macroscopic setups, which suffer from bubbles and cavitation effects hampering equilibration, [15] our single-molecule approach allows for equilibration to be achieved on the experimental time scale. The forced desorption under equilibrium conditions is defined by a velocity-independent plateau of constant force over a separation defined by the polymer contour length and, thus, allows for a direct determination of the adsorption free energy per unit length from the area under the plateau curve (Fig. 1B).[16] The distribution of plateau forces for H-and O-terminated diamond are given in Figure 1C. The general observed trend of measuring higher adsorption forces and therefore higher free energies on freshly prepared, hydrophobic diamond compared to hydrophilic diamond is reversed after two months and can be r...