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The ability to manipulate environmental conditions while obtaining nanoscale information about a sample is invaluable. The current article describes the construction and use of a chamber to control the relative humidity while using the scanning probe microscope (SPM) to monitor changes in sample stiffness. Although the experimental setup described is simple, this work shows that the complex relationship between temperature and relative humidity needs to be fully understood for successful operation of the apparatus. In TappingMode™, increases in relative humidity caused the free amplitude of the oscillating cantilever to decrease. Force–distance measurements taken on glass, however, showed no changes in detector sensitivity, suggesting that force–distance mode could be used to measure changes sample stiffness as a function of relative humidity. Using the SPM in force–distance mode, three moisture-sensitive materials (gelatine, wool fibers, and cotton fibers), were examined as the relative humidity was gradually increased from ambient conditions to just under 100%. The results showed that each of the materials softened as the humidity increased and that the experiments were reproducible. From the data, it was estimated that gelatine passed through a glass transition at about 86% relative humidity at room temperature. Wool passed through a glass transition at about 85% relative humidity, which is consistent with previous findings. © 2002 American Institute of Physics.