Scanning microfluorometric measurement of cell constituents. Principles of the method and its application to the determination of NAD content and redox state of XTH-2 cells in culture.

Abstract

The scanning of fluorescence of a cell culture along a path several milimeters long, gives a series of signals, which allow calculation of the fluorescence emitted per cell. The emission at 450 +/- 10 nm, excited at 360 nm, provides a measure of NADH (and NADPH) per cell. Combination of this method with modifications in energy metabolism, allows determination in the same sample of total NAD content, of NAD redox state, content of NAD in the mitochondria and content of NAD plus NADP in the cytoplasm. The method can be extended to measure other cellular constituents by labelling with fluorescent markers, e.g. antibodies.