Mitochondrial redox state in cardiac muscle was visualized by the ratio between fluorescence intensity of FAD and NADH. The ratio of FAD to NADH is an effective index for quantitative analysis of the redox state. Their concentration can be identified by their identical fluorescence properties with different excitation wavelength. Quantitative measurement is difficult to conduct imaging mitochondrial NADH as an index of the redox state, because of fluctuating condition during its measurement. Traditional approach with two cameras for observing fluorescence of each chemical substance had a problem of the parallax. Our new concept is taking two image data of FAD and NADH by single camera. Our system is composed of an EMCCD camera and two excitation light sources for FAD and NADH. The ratio between fluorescence intensity of these two successive video image data of two substances were processed every 1/15sec, and displayed on a monitor screen in real time together with heart image. Performance of this system was successfully demonstrated with rat hearts.

View full abstract