UNLABELLED At least two quantitative rCBF measurements are needed to evaluate rCBF changes with pharmacological intervention. We have developed the split dose 123I-IMP SPECT method, which enables measurement of rCBF to be repeated in a short time. METHODS Thirty-one cerebrovascular disease patients were investigated to assess reproducibility and vasoreactivity to acetazolamide. During 44-min dynamic SPECT imaging, 123I-IMP injection and respective arterial sampling were performed twice at an interval of about 25 min. The rCBF values were calculated using a microsphere model in which the washout of 123I-IMP from the brain can be negligible in the first several minutes after injection. For the second rCBF measurement, the remaining activity due to the first 123I-IMP injection was estimated and subtracted from the total brain activity. RESULTS In ten patients, two consecutive resting mean rCBF values in the MCA territory (CBF1 and CBF2) had good correlation (CBF1 = 47.4 +/- 4.0 (ml/min/100 ml: mean +/- s.d.), CBF2 = 45.2 +/- 8.2, CBF2 = 0.900*CBF1 + 2.9, r = 0.915). In 11 patients with occlusive lesions in the unilateral ICA system, mean rCBF in the MCA territory was increased by only 27.7% +/- 14.0% in the affected side by a 1-g intravenous acetazolamide injection, while 44.5% +/- 12.3% increase was found in the nonaffected side. In 10 patients without a major arterial lesion, a 49.7% +/- 17.0% increase of rCBF was demonstrated. CONCLUSIONS This split dose method 123I-IMP SPECT can be useful to estimate vascular reserve.