To directly determine the pressure-diameter relationship of individual pulmonary microvessels, it is necessary to measure the width of the column of blood in the vessel because microvascular walls are invisible when using intravital microscopy. To identify the margins of the blood column accurately, we developed a method for computer enhancement and measurement of vessel images. After recording microvessels on videotape, consecutive frames from the videotape were digitized by a computer. Pixels that changed from frame to frame (moving erythrocytes) were turned white, and unchanging pixels were turned black. In this way an image of the erythrocyte column with distinct edges was produced. The width of this column was measured with a heuristic technique involving interactions between the computer and the user. The measurements were reproducible and accurate. This technique has been used to measure microvascular diameters over a range of well-defined microvascular pressures and construct precise pressure-diameter curves.