Etherton T D, Thompson E H, Allen C E
J Lipid Res. 1977 Jul;18(4):552-7.
Two methods are described for the study of adipose tissue cellularity and metabolism. In the first, 8 M urea was used to liberate osmium tetroxide-fixed adipocytes from the connective tissue matrix. In the smaller-sized cell ranges there was a significant reduction in apparent adipocyte number of rat, pig, and beef adipose tissue with 8 M urea treatment. This was attributed to solubilization of connective tissue debris that was counted as adipocytes in samples isolated without urea. There was no effect on the larger cell-size fractions with 8 M urea treatment. Eight molar urea had no effect on fixed adipocyte retention of radioactivity. The second method entailed the use of hydrogen peroxide to volatilize the black, osmium tetroxide-fatty acid complex of osmium tetroxide-fixed adipocytes, containing radioactivity, resulting in colorless lipid suitable for liquid scintillation counting. This latter technique permits incubation of unfixed adipose tissue slices with a radioactive substrate, followed by fixation with osmium tetroxide and subsequent separation of the adipocytes, by screening, into the desired size ranges. Adipocytes in various size fractions can then be counted, sized, and then decolorized with hydrogen peroxide in order to quantitate the amount of radioactivity within the adipocytes. There was no loss of radioactivity from the fixed cells with hydrogen peroxide treatment.
本文描述了两种用于研究脂肪组织细胞构成和代谢的方法。第一种方法是，使用8M尿素从结缔组织基质中释放四氧化锇固定的脂肪细胞。在较小尺寸的细胞范围内，经8M尿素处理后，大鼠、猪和牛肉脂肪组织中可见的脂肪细胞数量显著减少。这归因于结缔组织碎片的溶解，在未用尿素分离的样本中，这些碎片被计为脂肪细胞。8M尿素处理对较大尺寸的细胞组分没有影响。8M尿素对固定脂肪细胞的放射性保留没有影响。第二种方法是使用过氧化氢挥发四氧化锇固定的、含有放射性的脂肪细胞的黑色四氧化锇 - 脂肪酸复合物，从而得到适合液体闪烁计数的无色脂质。后一种技术允许用放射性底物孵育未固定的脂肪组织切片，然后用四氧化锇固定，随后通过筛选将脂肪细胞分离成所需的尺寸范围。然后可以对不同尺寸组分中的脂肪细胞进行计数、测量大小，然后用过氧化氢使其脱色，以定量脂肪细胞内的放射性量。用过氧化氢处理后，固定细胞的放射性没有损失。
