Stomatal regulation is a key determinant of plant photosynthesis and water relations, influencing plant survival, adaptation, and growth. Stomata sense the surrounding environment and respond rapidly to abiotic and biotic stresses. Stomatal conductance to water vapour (g(s)) and/or transpiration (E) are therefore valuable physiological parameters to be monitored in plant and agricultural sciences. However, leaf gas exchange measurements involve contact with leaves and often interfere with leaf functioning. Besides, they are time consuming and are limited by the sampling characteristics (e.g. sample size and/or the high number of samples required). Remote and rapid means to assess g(s) or E are thus particularly valuable for physiologists, agronomists, and ecologists. Transpiration influences the leaf energy balance and, consequently, leaf temperature (T-leaf). As a result, thermal imaging makes it possible to estimate or quantify g(s) and E. Thermal imaging has been successfully used in a wide range of conditions and with diverse plant species. The technique can be applied at different scales (e.g. from single seedlings/leaves through whole trees or field crops to regions), providing great potential to study plantenvironment interactions and specific phenomena such as abnormal stomatal closure, genotypic variation in stress tolerance, and the impact of different management strategies on crop water status. Nevertheless, environmental variability (e.g. in light intensity, temperature, relative humidity, wind speed) affects the accuracy of thermal imaging measurements. This review presents and discusses the advantages of thermal imaging applications to plant science, agriculture, and ecology, as well as its limitations and possible approaches to minimize them, by highlighting examples from previous and ongoing research.