Deformation of lipid droplets in fixed samples
Deformation of lipid droplets in fixed samples
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DOI：
10.1007/s00418-002-0462-7
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发表时间：
2002-11-01
影响因子：
2.3
通讯作者：
Fujimoto, T
中科院分区：
文献类型：
--
作者：
Fukumoto, S;Fujimoto, T
尼罗红、苏丹III和油红O已被用于染色脂滴（LD）以用于荧光显微镜检查。我们注意到尼罗红标记的LD与后两种染料染色的LD在外观上不同。为了了解差异的原因，我们使用了顺序标记程序（第一次LD染色-摄影-猝灭-第二次LD染色-摄影），并检查了几个因素的影响。免疫荧光标记脂肪分化相关蛋白（ADRP），LD标记，也观察到与脂质染色比较。因此，我们发现，乙醇和异丙醇用于苏丹III和油红O染色，分别和甘油用于安装，导致相邻的LD融合，即使在戊二醛固定的样品。通过相同的处理，ADRP的免疫荧光标记移位到由于人工融合而形成的大LD的边缘。结果表明，用尼罗红比用苏丹Ⅲ或油红O更能观察到LD结构。
Nile red, Sudan III, and oil red O have been used to stain lipid droplets (LDs) for fluorescence microscopy. We noticed that LDs labeled by Nile red are different in appearance from those stained by the latter two dyes. To understand the cause of the difference, we used sequential labeling procedures (first LD stain-photography-quenching-second LD stain-photography), and examined the effect of several factors. Immunofluorescence labeling for adipose differentiation-related protein (ADRP), an LD marker, was also observed comparatively with the lipid stains. As a result, we found that ethanol and isopropanol used for Sudan III and oil red O staining, respectively, and glycerol used for mounting, cause fusion of adjacent LDs even in glutaraldehyde-fixed samples. By the same treatment, immunofluorescence labeling for ADRP was dislocated to the rim of large LDs that were formed as a result of the artifactual fusion. The result indicates that the LD structure can be better observed with Nile red than with Sudan III or oil red O.