Summary: A microscopic reflectance spectrophotometer was constructed to obtain the spectra of single pial ves sels and of a region containing only capillaries (capillary region). The difference in the oxygen saturation (S02) of hemoglobin between the regional arteriole and venule [R(A -V)] and that between the regional arteriole or capillaries [R(A -C)] were calculated. The reduction of cytochrome aa3 was also estimated in the capillary re gion. This method was applied to the brain surface of spontaneously breathing rats subjected to hypoxic and anemic hypoxia. On decreasing the inhaled O2 from 100 to 15%, elevation of R(A -V) and R(A -C) with slight arteriolar dilatation (though statistically not significant) was observed. Below 10% O2 (especially at 4 and 3% O2), Optical techniques provide valuable information for the study of tissue oxygen metabolism (Chance, 1991). The advantages of such methods include a capability of continuous monitoring in living ani mals with minimal damage to the tissue. Therefore, many optical methods have been developed, e.g., determination of the redox state of NADH by ultra violet fluorometry (Chance, 1954) and monitoring of the O2 saturation of flowing blood cells and/or the redox state of cytochrome aa3 by visible (Millikan, 1942; Sato et aI., 1979; Seiyama et aI., 1991) and infrared (Jobsis, 1977; Hazeki and Tamura, 1988) spectroscopy.Macroscopic reflectance spectroscopy measures the degree of oxygenation in blood and tissue. Thus, it has been conventionally used for studying Received June 24, 1992; final revision received April 29, 1993; accepted May 4, 1993. Address correspondence and reprint requests to Dr. T. Shiga at Department of Physiology, Medical School, Osaka Univer sity, 2-2 Yamadaoka, Suita, Osaka 565, Japan.Abbreviations used: Hb, hemoglobin; Ht, hematocrit; OD, op tical density; R(A -V), regional arteriole-venule difference in O2 saturation; R(A -C), regional arteriole-capillary difference in O2 saturation.
75the R(A -V) and R(A -C) decreased in spite of signif icant arteriolar dilatation with progressive reduction of cytochrome aa3, indicating suppression of oxygen trans port to mitochondria. In the case of hemodilution down to 37% hematocrit (Ht), elevation of R(A -V) and R(A -C) occurred with a slight tendency toward arteriolar di latation. Below 32% Ht, the R(A -V) decreased but the R(A -C) remained steady, while reduction of cy tochrome aa3 progressed. Altogether, the S02 in the cap illary region decreased and the reduction of cytochrome aa3 progressed with the decline of arteriolar O2 supply in both hypoxic and anemic hypoxia.