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The assumptions that water vapor exchange occurs exclusively through stomata, that the intercellular airspace is fully saturated with water vapor, and that CO2gradients are negligible between stomata and the intercellular airspace have enabled significant advancements in photosynthetic gas exchange research for nearly 60 years via calculation of intercellular CO2(Ci). However, available evidence suggests that these assumptions may be overused. Here we review the literature surrounding evidence for and against the assumptions made by Moss & Rawlins (1963). We reinterpret data from the literature by propagating different rates of cuticular water loss, CO2gradients, and unsaturation through the data. We find that in general, when cuticle conductance is less than 1% of stomatal conductance, the assumption that water vapor exchange occurs exclusively through stomata has a marginal effect on gas exchange calculations, but this is not true when cuticle conductance exceeds 5% of stomatal conductance. Our analyses further suggest that CO2and water vapor gradients have stronger impacts at higher stomatal conductance, while cuticle conductance has a greater impact at lower stomatal conductance. Therefore, we recommend directly measuring Ciwhenever possible, measuring apoplastic water potentials to estimate humidity inside the leaf, and exercising caution when interpreting data under conditions of high temperature and/or low stomatal conductance, and when a species is known to have high cuticular conductance.HighlightLeaf water vapor and CO2exchange have been successfully used to model photosynthetic biochemistry. We review critical assumptions in these models and make recommendations about which need to be re-assessed.