Fluorometry of Intracellular Redox State
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Reduced pyridine nucleotides (NADH & NADPH) emit blue fluorescence (peaking at 480 mμ) upon ultraviolet irradiation (366 mμ). This phenomenon has been succesfully utilized for the following of intracellular oxidation-reduction (redox) state in vivo, because NAD is one of the key substances in the glycolytic and respiratory control mechanisms. The microfluorometric apparatuses ever developed, however, had one common defect which made it impossible to assay any metabolic event in the organs with circulatory blood. Schnitger et al.(1965) observed 40-60% hemodynamic artifact contained in rat-liver fluorescence.<BR>The purpose of this study was to devise a new instrument capable of a precise fluorometric recording of pyridine nucleotides in situ. Fluorescence emitted from the blood-circulating organs involves hemodynamic artifact as well as metabolic signals. Optical artifact corresponds with the blood content in the tissues which is measured in a similar manner as in photoelectric plethysmography.<BR>For realizing its cancellation, the authors adopted d. c.-lighting of the ultraviolet source (high-pressure mercury arc) in place of a. c. -lighting combined with an opticalchopper method, developed by Chance (1962). The blood volume changes were detected as light-scattering by another photomultiplier exclusively sensitive to the red light peaking at 720 mμ. Hemodynamic artifact in fluorescence could be cancelled electronically by that scattering signal.<BR>These modifications brought about satisfactory results concerning in situ monitoring of intracellular redox state in rat kidney cortex, rat liver, rabbit thyroid, and rabbit parathyroid.
- Japan Society of Clinical Chemistryの論文
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