The power-time curves of Tetrahymena thermophila exposed to tributyltin (TBT) were detected by microcalorimetry. Metabolic rate (r) decreased significantly while peak time (PT) increased with the enhancement of TBT level. Compared with the measured multibiomarker including catalase, lactate dehydrogenase, glutathione S-transferase, ATPase and membrane fluidity, PT and r could be sensitive biomarkers for assessing TBT toxicity at cellular level. The effective concentrations obtained by them were consistent to those obtained by the protozoan community toxicity test. As a result, the microcalorimetric assay of T. thermophila had a great potential in assessing TBT acute toxicity and monitoring TBT pollution in the freshwater ecosystem.