A method for measuring the rate of oxygen release from single microvessels

Abstract

A system determining the rate of oxygen release from erythrocytes flowing in single microvessels was constructed with an inverted microscope by connecting 1) a scanning/grating spectrophotometer equipped with a photon-counting detector through a thin light guide, to obtain the visible absorption spectrum of a spot (5 μm in diameter) focused on a microvessel, 2) two photomultipliers (connected to a microcomputer via an analog-to-digital converter) through two light guides, to determine the flow velocity of erythrocytes by calculating the cross correlation between the light-intensity changes of two spots (3 μm in diameter, 5 μm apart from each other) focused on the microvessel, and 3) an image processor through a video camera, to estimate the diameter of microvessel from the digitized video images. The rate of oxygen release from single microvessels 7-25 μm in diameter in rat mesentery was measured under the superfusion of deoxygenated solution: 1) The maximal rate was obtained in capillaries, and the rate in arterial microvessels was larger than that in venous microvessels, when similar diameters were compared. 2) The rate was maximum at pH 7.0-7.2, and it decreased in more acidic and alkaline pH values. 3) The rate decreased with a decrease in temperature. The reliability of the measurement using the present apparatus was tested in detail.