This paper investigates the mechanism of bitumen surface ageing, which was validated utilizing the atomic force microscopy and the differential scanning calorimetry. To validate the surface ageing, three different types of bitumen with different natural wax content were conditioned in four different modes: both ultraviolet and air, only ultraviolet, only air and without any exposure, for 15 and 30 days. From the atomic force microscopy investigation after 15 and 30 days of conditioning period, it was found that regardless the bitumen type, the percentage of microstructure on the surface reduced with the degree of exposure and time. Comparing all the four different exposures, it was observed that ultraviolet radiation caused more surface ageing than the oxidation. It was also found that the combined effect was not simply a summation or multiplication of the individual effects. The differential scanning calorimetry investigation showed that the amount of crystalline fractions in bitumen remain constant even after the systematic conditioning. Interestingly, during the cooling cycle, crystallization of wax molecules started earlier for the exposed specimens than the without exposed one. The analysis of the obtained results indicated that the ageing created a thin film upon the exposed surface, which acts as a barrier and creates difficulty for the wax induced microstructures to float up at the surface. From the differential scanning calorimetry analysis, it can be concluded that the ageing product induced impurities in the bitumen matrix, which acts as a promoter in the crystallization process.