The aim of this study was to comprehensively describe the physiological responses to an acute bout of mild cold in young lean men (n = 11, age: 23 ± 2 years, body mass index: 23.1 ± 1.2 kg/m2) to better understand the underlying mechanisms of non-shivering thermogenesis and how it is regulated. Resting energy expenditure, substrate metabolism, skin temperature, thermal comfort perception, superficial muscle activity, hemodynamics of the forearm and abdominal regions, and heart rate variability were measured under warm conditions (22.7 ± 0.2°C) and during an individualized cooling protocol (air-conditioning and water cooling vest) in a cold room (19.4 ± 0.1°C). The temperature of the cooling vest started at 16.6°C and decreased ~ 1.4°C every 10 minutes until participants shivered (93.5 ± 26.3 min). All measurements were analysed across 4 periods: warm period, at 31% and at 64% of individual´s cold exposure time until shivering occurred, and at the shivering threshold. Energy expenditure increased from warm period to 31% of cold exposure by 16.7% (P = 0.078) and to the shivering threshold by 31.7% (P = 0.023). Fat oxidation increased by 72.6% from warm period to 31% of cold exposure (P = 0.004), whereas no changes occurred in carbohydrates oxidation. As shivering came closer, the skin temperature and thermal comfort perception decreased (all P<0.05), except in the supraclavicular skin temperature, which did not change (P>0.05). Furthermore, the superficial muscle activation increased at the shivering threshold. It is noteworthy that the largest physiological changes occurred during the first 30 minutes of cold exposure, when the participants felt less discomfort.