We analyzed the isotope dilution residue function from a single cardiac chamber for an arbitrary inlet distribution of tracer and arbitrary mixing within the chamber, and established a general relationship between cardiac output and the chamber residue function. In our experiments, we made simultaneous temperature measurements in three left ventricular chamber subregions of the dog subjected to left and right atrial injections of chilled saline. Flow-proportional tracer labeling always occurred at the left ventricular inlet when injection was into the right atrium. This state almost never obtained, however, with direct left atrial injection, although it was approximated most closely when multiple side hole cathers were used. We also demonstrated that imperfect tracer mixing in the normal ventricle can lead to significant regional temperature inequalities during tracer passage. These inequalities are more pronounced in the ventricle with compromised function, but in both normal and compromised ventricles they are minimal several beats after tracer concentration peaks if injection is into the right atrium.