In the living system, tissue water is considered to be composed of both free and bound water. Bound water encompasses the structural water of the cell wall and of various biological substances of high molecular weight, such as proteins and polypeptides. The present study was designed to measure thermoanalytically free and bound water on a quantitative basis in fresh brain of rats using differential scanning calorimetry (DSC). Our intention was to determine the fraction of freezable water in tissue. Freezable water in a tissue represents the fraction of free water. In the present study, freezing was conducted at a constant rate of -10 degrees C/min from room temperature to -75 degrees C by a SSC/560 S (Seiko Electronics). The system allows for calculation of the amount of free water from the differential scanning calorimetry curve employing a coloric constant of 79.4 cal/mg. Aluminium oxide was used as calorimetric reference. The fraction of bound water was calculated by subtraction of the amount of free water from that of total tissue water. Water binding to solid tissue component was estimated from tissue dry weight and the bound water fraction. Mean water content of normal gray matter in adult Wistar rats was 76.9 +/- 1.4% (SD). 88.9% of total tissue water was free whereas 11.1 +/- 2.8% (SD) was bound. Bound water of brain parenchyma amounted to 0.44 +/- 0.12 mg/mg dry weight. As compared to other tissues such as cardiac muscle and liver, brain parenchyma obviously exceeded in free water content. The total water content of serum was 94.4 +/- 1.2%; 90.7 +/- 2.6% was free and 9.3 +/- 2.6% (SD) was bound.(ABSTRACT TRUNCATED AT 250 WORDS)