Self-ignited fires at municipal solid waste (MSW) storage sites are relatively common. The minimization of the phenomenon of self-heating in the waste can reduce the risks for smouldering combustion. The purpose of this work was to develop a method that can be used to measure and characterize the heat production in MSW. The method is based on isothermal heat conduction microcalorimetry (IMC). The heat production in MSW was determined based on sampling from two sites in two different geographical locations in Sweden. Both the original waste and milled/homogenised waste were tested. The heat production was measured at different temperatures together with gas analysis using micro-gas chromatography. The activity in the waste, in terms of its heat flow, increased when the temperature increased up to 60 °C and decreased at higher temperatures, e.g., 70 and 80 °C. The consumption of oxygen and the production of carbon dioxide, together with the heat production, indicated that aerobic metabolism was responsible for the heat production. This is further strengthened by the marginal heat production observed for ultraviolet treated waste. The results showed that IMC is a valuable tool for characterising the self-heating in municipal and industrial waste.