In this paper, a new technique has been developed to enable fibre diameters to be measured in an aqueous environment using the OFDA 2000 instrument. The existing OFDA instrument has only been used to measure diameters of animal fibres under dry conditions. This new technique was utilised to assess the effects of pH and temperature on the diameters of merino wool fibres in aqueous environments. Significant changes in fibre diameter under aqueous conditions were found as a function of pH and temperature. Wool fibre diameters were at a minimum close to the wool isoelectric point (pH 4.8) and increased at both lower and higher pHs. Swelling of merino wool fibres was observed to increase linearly by around 15% as the temperature rose from ambient to 70°C.