For muscle heat measurements the methods available are sensitive and rapid, and the heat is related to the chemical changes in a manner that provides a firm outline for understanding the mechanism of contraction. For example linear dependence of the shortening heat on the sarcomere length has shown that the rate of turnover of cross-bridges increases during shortening. However, heat is bound to lack specificity. In order to cope with this problem, various methods such as rigorous chemical analyses, phosphorus NMR and microcalorimetry have been introduced. As a result of ultra-rapid freezing and chemical analysis by D. R. Wilkie (Gilbert, Kretzchmar, Wilkie and Woledge, 1971), the energy balance discrepancy between (heat + work) and the amount of phosphocreatine (PCr) split emerged, i.e. the unexplained enthalpy. Calcium ions move from the sarcoplasmic reticulum to the calcium-receptive proteins in the sarcoplasm during contraction. In an attempt to find the cause of the unexplained enthalpy, microcalorimetry of calcium binding to calcium-receptive proteins has been performed. The results have shown that calcium ions dislocated between sites within the sarcoplasm on activation may produce about 1/3 of the unexplained heat. In addition calcium pump should operate by consuming PCr to relocate the calcium after the contraction. Time-resolved phosphorus NMR has also shown that a certain amount of PCr splitting continues during early minute of recovery period after the contraction without Pi released. This delayed splitting of PCr is most likely caused by the kinetic properties of the contractile proteins and can explain another 1/3 of the unexplained enthalpy. The mechanism of how muscle is regulated is another important question. Studies of calcium binding to calcium-receptive proteins in the sarcoplasm by using titration microcalorimetry has shown that troponin C has a characteristic single calcium-binding site that is most likely to be involved in the regulation of contraction.