OBJECTIVE The purpose of this study was to identify through infrared thermal imaging technology the facial surface temperature (FST) of laying hens in response to the variations in their thermal environment, and to identify the regional differences in FST to determine the most stable and reliable facial regions for monitoring of thermoregulatory status in chickens. METHODS Thirty Hy-Line Brown hens (25-week-old) were sequentially exposed to three different thermal conditions; optimal (OT, 22°C±2°C), low (LT, 10°C±4°C), and high temperature (HT, 30°C±2°C). The mean values of FST in five facial regions including around the eyes, earlobes, wattles, beak and nose, and comb were recorded through infrared thermography. The maximum FST (MFST) was also identified among the five face-selective regions, and its relationship with temperature-humidity index (THI) was established to identify the range of MFST in response to the variations in their thermal environment. RESULTS Hens exposed to OT condition at 15:00 displayed a higher temperature at wattles and around the eyes compared to other regions (p<0.001). However, under LT condition at 05:00 to 08:00, around the eyes surface temperature showed the highest value (p<0.01). In HT, wattles temperature tended to show the highest temperature over almost time intervals. Main distribution regions of MFST were wattles (63.3%) and around the eyes (16.7%) in OT, around the eyes (50%) in LT, and wattles (62.2%) and comb (18.3%) in HT. The regression equation between MFST and THI was estimated as MFST = 35.37+ 0.2383×THI (R2 = 0.44; p<0.001). CONCLUSIONS The FST and the frequency of MFST in each facial region of laying hens responded sensitively to the variations in the thermal environment. The findings of this experiment provide useful information about the effect of the thermal conditions on the specific facial regions, thus offering an opportunity to stress and welfare assessment in poultry research and industry.