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J Appl Physiol 91: 321-327, 2001;
8750-7587/01 $5.00
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Vol. 91, Issue 1, 321-327, July 2001

Microvascular and interstitial PO2 measurements in rat skeletal muscle by phosphorescence quenching

Masahiro Shibata, Shigeru Ichioka, Joji Ando, and Akira Kamiya

Department of Biomedical Engineering, Graduate School of Medicine, University of Tokyo, Tokyo 113-0033, Japan

To clarify the transport of O2 across the microvessels in skeletal muscle, we designed an intravital laser microscope that utilizes a phosphorescence quenching technique to determine both the microvascular and tissue PO2. After we injected the phosphorescent probe into systemic blood, phosphorescence excited by a N2-dye pulse laser was detected with a photomultiplier over a 10 µm in diameter area. In vitro and in vivo calibrations confirmed that the present method is accurate for PO2 measurements in the range of 7-90 Torr (r = 0.958) and has a rapid response time. This method was then used to measure the PO2 of microvessels with different diameters (40-130 µm) and of interstitial spaces in rat cremaster muscle. These measurements showed a significant drop in PO2 in the arterioles after branching (from 74.6 to 46.6 Torr) and the presence of a large PO2 gradient at the blood-tissue interface of arterioles (15-20 Torr). These findings suggest that capillaries are not the sole source of oxygen supply to surrounding tissue.

oxygen transport; intravital microscope; microcirculation; palladium-porphyrin; cremaster muscle


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