Densitometric studies show that the large flow measurement errors and inability to obtain reproducible densitometer calibrations reported with indocyanine green (ICG) in nearly isotonic saline may have arisen from two sources: (a) slowed optical stabilization, and (b) sedimentation of dye aggregates formed in the salt "solutions" of ICG, both of which are avoidable by preparing the dye in water. The assumptions of the widely used regional blood flow measurement technique using radionuclide-labeled microspheres are described. Simultaneous injection of 8mu and 15mu microspheres in turkeys and in dogs demonstrated the existence of at least 8 mu arteriovenous communications (AVCs) in the stomach and intestine, not previously described by this technique, which, in addition to their physiologic functions, may play a role in production of acute gastric mucosal ischemia and erosions. Similar AVCs, producing a lesser degree of "shunting," were also found in the heart. Loss of 8 mu relative to 15 mu microspheres continued with time in the gastrointestinal circulation.