Combined orcein and martius scarlet blue (OMSB) staining for qualitative and quantitative analyses of atherosclerotic plaques in brachiocephalic arteries in apoE/LDLR(-/-) mice.
Combined orcein and martius scarlet blue (OMSB) staining for qualitative and quantitative analyses of atherosclerotic plaques in brachiocephalic arteries in apoE/LDLR(-/-) mice.
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DOI：
10.1007/s00418-017-1538-8
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发表时间：
2017-06
影响因子：
2.3
通讯作者：
Chlopicki S
中科院分区：
文献类型：
--
作者：
Gajda M;Jasztal A;Banasik T;Jasek-Gajda E;Chlopicki S
在动脉粥样硬化斑块的切片中应分析大量的细胞和细胞外成分，以评估动脉粥样硬化的进展和易损性。在这里，我们结合了弹性纤维的地衣红（O）染色和马歇斯猩红蓝（MSB）多色染色，以显示apoE/LDLR−/−小鼠头臂动脉（BCA）中斑块的各种形态学内容。弹性纤维（包括断裂的弹性层和“埋藏”的纤维帽）被染成紫色，它们可以很容易地与胶原纤维（蓝色）区分开来。即使是最细的弹性纤维，地衣红也能清楚地识别出来。红细胞被染成黄色，并且它们可以容易地与成熟纤维蛋白（红色）区分开。旧纤维蛋白倾向于获得蓝色。OMSB染色方法简单，耗时不到1 h，可适用于自动染色机。最重要的是，颜色分离足够好，以允许组织切片中特定成分的数字自动分割和斑块成分的定量分析。使用OMSB比较apoE/LDLR−/−小鼠BCA近端和远端区域的动脉粥样硬化斑块。总之，OMSB染色代表了一种新型染色，可常规用于动脉粥样硬化病变的福尔马林固定和石蜡包埋切片的定性和定量显微镜评估。
Numerous cellular and extracellular components should be analyzed in sections of atherosclerotic plaques to assess atherosclerosis progression and vulnerability. Here, we combined orcein (O) staining for elastic fibers and martius scarlet blue (MSB) polychrome to visualize various morphological contents of plaque in brachiocephalic arteries (BCA) of apoE/LDLR−/− mice. Elastic fibers (including broken elastic laminae and ‘buried’ fibrous caps) were stained purple and they could be easily distinguished from collagen fibers (blue). Orcein allowed clear identification of even the finest elastic fibers. Erythrocytes were stained yellow and they could easily be discerned from mature fibrin (red). Old fibrin tends to acquire blue color. The method of OMSB staining is simple, takes less than 1 h to perform and can be adapted to automatic stainers. Most importantly, the color separation is good enough to allow digital automatic segmentation of specific components in tissue section and quantitative analysis of the plaque constituents. OMSB was used to compare atherosclerotic plaques in proximal and distal regions of BCA in apoE/LDLR−/− mice. In conclusion, OMSB staining represents a novel staining that could be routinely used for qualitative and quantitative microscopic assessments of formaldehyde-fixed and paraffin-embedded sections of arteries with atherosclerotic lesions.