Anurans use physical and biological microhabitat resources at a fine scale, giving each species unique adaptive, morphological, physiological, and behavioural traits. Here I evaluate the microhabitat use of the treefrog Dendropsophus luddeckei in pasture agro-ecosystems in Villa de Leyva Municipality, Boyaca Department, Colombia. Visual Encounter Surveys were conducted during both day and night to obtain data on a series of microhabitat variables, including environmental temperature, relative humidity, substrate type, and horizontal position. I obtained 64 records of D. luddeckei using a total sampling effort of 72 person-hours. The species’ environmental temperature range suggests that its thermal biology has considerable plasticity, while ranges for relative humidity, substrate type, and horizontal position may be related to reproductive activity patterns, use of shelters, and morphological traits. This research contributes to our knowledge of the ecophysiology and ecomorphology of D. luddeckei and supplies relevant information for its conservation in highland ecosystems in the Colombian Andes.