Epicardial Oxygen/Metabolism by Pd Porphyrin and NADH Imaging

Clyde H. Barlow, Dawn A. Rorvik, and Jeffrey J. Kelly

Barlow Scientific, Inc., Olympia, WA 98502
The Evergreen State College, Olympia, WA 98505

C.H. Barlow, D.A. Rorvik and J.J. Kelly  (1988)  Epicardial oxygen/metabolism by Pd porphyrin and NADH imaging.  The Journal of Cell Biology  107(6), 213a. NADH fluorescence imaging provides a method of visualizing and evaluating mitochondrial bioenergetics. Recent advances in spectroscopic oxygen determination by palladium porphyrin (PP) phosphorescence quenching makes possible high resolution studies of oxygen and NADH distribution in tissues. We have developed a computer-controlled videofluorometer to collect and analyze images of NADH fluorescence and PP phosphorescence. NADH and PP emission images of buffer-perfused rat heart epicardium were recorded with a CCD camera using filtered xenon flash excitation. Images, 512x480 pixels, were recorded at 50x40 µm pixel resolution. Xenon flash excitation and image acquisition were synchronized with the cardiac cycle and digitized with a 10 MHz, 8-bit frame grabber. A fluorescent reference was included in all images as a basis to normalize for variation in flash intensity. Relative oxygen concentration and epicardial distribution were estimated from the Stern-Volmer relationship applied to oxygen quenching of PP phosphorescence. Microcomputer analysis permitted image processing to evaluate metabolic parameters over the visible epicardial surface.

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Last modified: 01/08/2000