1. Total renal blood flow and its distribution within the renal cortex of the conscious rabbit were studied with radioactive microspheres of 15 and 25 μm diameter.
2. The reliability of the microsphere technique was influenced by microsphere diameter and number (dose). The optimum microsphere diameter for determination of flow distribution in the rabbit kidney was 15 μm and dose 100–150 000 spheres.
3. Spheres of 15 μm nominal diameter were randomly distributed within the renal cortex of adult rabbits. The larger spheres in batches nominally 15 μm in diameter in young rabbits and 25 μm diameter in adult rabbits were preferentially distributed to the superficial cortex.
4. In adult rabbits 15 μm diameter spheres lodged in glomerular capillaries. Larger spheres occasionally lodged in interlobular arteries causing intrarenal haemorrhage.
5. Microspheres of 15 μm caused a decrease in renal clearance of creatinine and of p-aminohippurate when the total injection dose was about 200 000 spheres. These effects were greater when the injection dose was increased to 500 000 spheres.
6. The reduction in total renal blood flow observed with large doses of spheres largely reflected decreased outer cortical flow, as measured by a second injection of spheres, and confirmed by a decrease in p-aminohippurate extraction.
7. The reproducibility of multiple injection studies was limited by these intrarenal effects of microspheres.
8. Total renal blood flow measured in six rabbits in acute experiments by the microsphere technique was 107 ± 12 (mean±sd) ml/min and by p-aminohippurate clearance was 100 ± 10 ml/min.
9. Total renal blood flow in twelve conscious, chronically instrumented rabbits was 125 ± 11 ml/min, of which 92 ± 6 ml/min was distributed to the superficial cortex and 33 ± 4 ml/min to the deep cortex.