Development of an Optical Triplicator for intravital video microscopy of oxygen saturation

The Optical Triplicator produces three copies of a portion of a microscopic image and places them side-by-side on the face of a video image tube, so that all three images can be viewed simultaneously in each video frame. The Optical Triplicator was used in an intravital microscopy assembly to obtain simultaneous images of a microvessel at three visible wavelengths selected to enable the accurate determination of oxygen saturation in microvessels of the hamster retractor muscle. An image processing system was used to obtain light intensity and optical density from video recordings made using the triplicator. Lumenal oxygen saturation profiles were determined using the measured intensity values and a published three wavelength photometric method.