A method of calibrating a glucose sensor, which method comprises: (a) preparing a first glucose-containing calibration solution by combining water or an aqueous solution with alpha and beta glucose, the alpha and beta glucose being provided in solid form; (b) exposing a glucose sensor to said first glucose-containing calibration solution and determining the sensor output; (c) exposing the glucose sensor to one or more further calibration solutions having different glucose concentrations from each other and from said first glucose-containing calibration solution and determining the sensor output for the or each calibration solution; and (d) determining a calibration curve from the sensor output data collected in steps (b) and (c).