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A ratio-fluorescence assay was developed for on-line localization and quantification of lipid oxidation in living cells. The assay explores the oxidative sensitivity of C11-BOD-IPY 581a591 . Upon oxidation, the fluorescence of this fluorophore shifts from red to green. The probe incorporates readily into cellular membranes and is about twice as sensitive to oxidation as arachidonic acid. Using confocal microscopy, the cumene hydroperoxide-induced oxidation of C11-BODIPY 581a591 was visualized at the sub-cellular level in rat-1 fibroblasts. Preloading of the cells with tocopherol retarded this oxidation. The data demonstrate that C11-BODIPY 581a591 is a valuable tool to quantify lipid oxidation and anti-oxidant efficacy in single cells.z 1999 Federation of European Biochemical Societies.

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The decrease in fluorescence of conjugated polyenic acyl chains is used as a sensitive assay for lipid peroxidation. The fatty acid cis-trans-tru1ts-cis-9,11,13,15-octadecatetraenoic acid (cis-parinaric acid) is introduced into liposomal membranes as free fatty acid or, by using the PC specific transfer protein from bovine liver, as l-palmitoyl-2-c~-parinaroyl-sn-glycero-3-phosphocholine.The peroxidation process as monitored by the decrease in fluorescence intensity is compared with other peroxidation assay systems. Applications of the new assay system are discussed.