Ethylated fluoresceins: assay of cytochrome P-450 activity and application to measurements in single cells by flow cytometry.

Publication Type:

Journal Article


Analytical biochemistry, Volume 133, Issue 1, p.46-57 (1983)


363, Animals, Aryl Hydrocarbon Hydroxylases, Cells, Cultured, Chromatography, Thin Layer, Cytochrome P-450 Enzyme System, Flow Cytometry Core Facility, Fluoresceins, Gas Chromatography-Mass Spectrometry, liver, Magnetic Resonance Spectroscopy, Male, MICE, Mice, Inbred C57BL


Four ethylated derivatives of fluorescein were synthesized and their metabolism by mouse liver homogenates and cultured mouse hepatoma cells was investigated. Two of the ethylfluoresceins are specifically metabolized by the polycyclic aromatic hydrocarbon inducible cytochrome P-450 to yield fluorescein, which is about 15 times more fluorescent than the ethylated compounds. One of these fluorogenic derivatives, ethoxyfluorescein ethyl ester, is useful for flow cytometric analysis and sorting of intact, viable cells on the basis of cytochrome P-450 activity. The structure-activity relationship of the ethylfluoresceins is discussed.