The quantitative relation between stomatal aperture and gas exchange through the stomatal pore can be described by physical models derived from Fick's first law of diffusion. Such models, usually based on a simplified pore geometry, are used to calculate leaf conductance from stomatal pore dimensions or vice versa. In this study a combination of gas-exchange measurements and simultaneous microscopical observations of stomatal apertures was used to empirically determine this relationship. The results show a substantial deviation between measured stomatal conductance and that calculated from the simplified models. The main difference is a much steeper increase of conductance with aperture at small apertures. When the calculation was based on a realistic pore geometry derived from confocal laser scanning microscopy, a good fit to the experimentally found relationship could be obtained if additionally a significant contribution of a mesophyll diffusional resistance was taken into account.