Global climate change and intensifying heat islands have reduced human thermal comfort and health in urban outdoor environments. However, there has been little research that has focused on how microclimates affect human thermal comfort, both psychologically and physiologically. We investigated the effect of a range of landscape microclimates on human thermal comfort and health using questionnaires and physiological measurements, including skin temperature, skin conductance, and heart rate variability, and compared the results with the effect of prevailing climate conditions in open spaces. We observed that in landscape microclimates, thermal sensation votes significantly decreased from 1.18 ± 0.66 (warm–hot) to 0.23 ± 0.61 (neutral–slightly warm), and thermal comfort increased from 1.18 ± 0.66 (uncomfortable–neutral) to 0.23 ± 0.61 (neutral–comfortable). In the landscape microclimates, skin temperature and skin conductance decreased 0.3 ± 0.8 °C and 0.6 ± 1.0 μs, respectively, while in the control, these two parameters increased by 0.5 ± 0.9 °C and 0.2 ± 0.7 μs, respectively. Further, in landscape microclimates, subject heart rate variability increased significantly. These results suggest landscape microclimates improve human thermal comfort and health, both psychologically and physiologically. These findings can provide an evidence base that will assist urban planners in designing urban environments for the health and wellbeing of residents.