We report the imaging of the cell-substrate adhesion of a single cell with subcellular spatial resolution. Osmotic pressure was introduced to provide a controllable mechanical stimulation to the cell attached to a substrate, and high-resolution surface plasmon resonance microscopy was used to map the response of the cell, from which local cell-substrate adhesion was determined. In addition to high spatial resolution, the approach is noninvasive and fast and allows for the continuous mapping of cell substrate interactions and single-cell movements.