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Abstract Understanding tree transpiration variability is vital for assessing ecosystem water‐use efficiency and forest health amid climate change, yet most landscape‐level measurements do not differentiate individual trees. Using canopy temperature data from thermal cameras, we estimated the transpiration rates of individual trees at Harvard Forest and Niwot Ridge. PT‐JPL model was used to derive latent heat flux from thermal images at the canopy‐level, showing strong agreement with tower measurements (R² = 0.70–0.96 at Niwot, 0.59–0.78 at Harvard at half‐hourly to monthly scales) and daily RMSE of 33.5 W/m²(Niwot) and 52.8 W/m²(Harvard). Tree‐level analysis revealed species‐specific responses to drought, with lodgepole pine exhibiting greater tolerance than Engelmann spruce at Niwot and red oak showing heightened resistance than red maple at Harvard. These findings show how ecophysiological differences between species result in varying responses to drought and demonstrate that these responses can be characterized by deriving transpiration from crown temperature measurements.