Measurement of ceroid accumulation in macrophages by flow cytometry

Abstract

The accumulation in macrophages of ceroid, an sutofluorescent polymer composed of oxidised protein and lipid, can be monitored seiniquantitatively by staining techniques. However, such methods are crude and give little infor mation about the amount of ceroid within cells. Flow cytometry, however, can give a quantitative assessment of cellular ceroid accumulation in vitro. Recently, flow cytometry was explored as a method for measurement of the accumulation in macrophages of ceroid. The accumulation appeared to be diminished in the presence of the antioxidant, α‐tocopherol. This is consistent with the role of lipoprotein oxidation in ceroid accumulation. Here the optimum wavelengths of emission and excitation, using both conventional fluorescence spectroscopy of cellular ceroid and w cytometric measurements with a number of optical filters, are determined. The use of optimal wavelengths determined in these studies enhances overall sensitivity. The findings are discussed in the context of future investigation of cell‐mediated lipid oxidation and its potential antagonists. © 1994 Wiley‐Liss, Inc.

Discrimination of Human Coronary Artery Atherosclerotic Lipid-Rich Lesions by Time-Resolved Laser-Induced Fluorescence Spectroscopy

“…Also, 2 lipopigments, ceroid and carotenoids, were related to atherosclerotic lesion fluorescence emission. 15,31,32 Ceroid, an end product of lipoprotein oxidation, is found in macrophages and in lesions with lipid-rich necrotic cores. Carotenoids are present in lipid-rich lesion cores.…”

“…The main fluorescent components of normal and diseased arterial wall, together with their emission characteristics, are depicted in Table II (http://atvb.ahajournals.org). [25][26][27][28][29][30][31][32] The fluorescence of the structural proteins elastin and collagen was related primarily to artery fluorescence. Previous research 8,25 demonstrated that the fluorescence of elastic fibers within the internal lamina or media predominates over the emission of normal coronary artery or aorta, respectively, whereas the fluorescence of collagen (30% to 60% dry weight 3 ) prevails over the emission of advanced collagenous lesions (type V a or fibrous plaques).…”