Deuterium MR spectroscopy was used for the determination of tissue blood flow (TBF). The tracer D2O was injected into the tissue of interest, and tracer washout was followed using a 4.7 T spectroscopy/imaging unit. Normal subcutaneous tissue in rats was studied, as well as tissue influenced by vasoactive agents (papaverine and adrenaline). The vasoactive agents introduced changes of 40% in TBF, compared with normal tissue. Normal tissue measurements were repeated using various D2O injection volumes (5–400 μl). The injection volume 5 μl gave TBF 11.7 ± 2.0 ml/100 g·min (mean ± 1 SD). This value was 40% higher than corresponding values observed at larger injection volumes (200–400 μl). This injection volume effect is probably partly due to a capillary dilution caused by tracer administration, and partly related to the non-physiological deuterium signal decrease observed in dead rats. Blood flow measurements in human colon tumours implanted in nude mice showed a rather poor reproducibility, not improved by the use of a multiple site injection technique.