Volume-osmotic pressure relationships at equilibrium have been obtained in chick heart fibroblasts grown in slide-coverslip cultures in a fluid medium consisting of heparinized plasma and embryo extract. The refractive index of the fibroblast gives a direct measure of its solid concentration, and the volume is estimated as the reciprocal of concentration. The volume is found to be linearly related to the reciprocal of the osmotic pressure over a range from 130 to 587 m-osm, provided the measurements are carried out rapidly at 38°C. The isotonic water content of the cells derived from the gradient of the regression line on the basis of the simple Boyle-van’t Hoff Law was found to be less than actual water content obtained by direct refractometry, i. e. the value of Ponder’s ℛ was 0⋅94 (s. d. 0⋅04). In cultures grown in a simple saline medium and measured at 22°C the volume was related linearly to the reciprocal of the osmotic pressure only between the limits of 330 and 191 m-osm. Outside these limits the volume was greater than expected and this was attributed to alterations in the semi-permeable properties of the cell membrane. The value of Ponder’s ℛ in these cultures was 1⋅15. The importance of the quantity, ℛ, as applied to cells other than the erythrocyte, is indicated. The value, 0⋅94 (s. d. 0⋅04), obtained in fibroblasts under physiological conditions is not explicable on the basis of the probable osmotic properties in vitro of the cell proteins. The discrepancy is within the experimental error, but it may also be due to abnormal osmotic behaviour of the cell proteins resulting from some form of intermolecular structure in the cytoplasm.