Hunt J V, Bottoms M A, Skamarauskas J, Carter N P, Mitchinson M J
Department of Pathology, Cambridge, United Kingdom.
Cytometry. 1994 Apr 1;15(4):377-82. doi: 10.1002/cyto.990150414.
The accumulation in macrophages of ceroid, an autofluorescent polymer composed of oxidised protein and lipid, can be monitored semiquantitatively by staining techniques. However, such methods are crude and give little information 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, alpha-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 flow 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.
脂褐素是一种由氧化蛋白质和脂质组成的自发荧光聚合物，巨噬细胞中脂褐素的积累可以通过染色技术进行半定量监测。然而，这些方法比较粗糙，几乎无法提供细胞内脂褐素含量的相关信息。而流式细胞术能够对体外细胞脂褐素积累进行定量评估。最近，人们探索将流式细胞术作为一种测量巨噬细胞中脂褐素积累的方法。在抗氧化剂α-生育酚存在的情况下，脂褐素的积累似乎有所减少。这与脂蛋白氧化在脂褐素积累中的作用是一致的。在此，通过使用细胞脂褐素的传统荧光光谱以及配备多个光学滤光片的流式细胞术测量，确定了发射和激发的最佳波长。这些研究中确定的最佳波长的使用提高了整体灵敏度。将在细胞介导的脂质氧化及其潜在拮抗剂的未来研究背景下讨论这些发现。
