Segnani Cristina, Ippolito Chiara, Antonioli Luca, Pellegrini Carolina, Blandizzi Corrado, Dolfi Amelio, Bernardini Nunzia
Unit of Histology and Medical Embryology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
Division of Pharmacology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
PLoS One. 2015 Dec 16;10(12):e0144630. doi: 10.1371/journal.pone.0144630. eCollection 2015.
Collagen detection in histological sections and its quantitative estimation by computer-aided image analysis represent important procedures to assess tissue localization and distribution of connective fibers. Different histochemical approaches have been proposed to detect and quantify collagen deposition in paraffin slices with different degrees of satisfaction. The present study was performed to compare the qualitative and quantitative efficiency of three histochemical methods available for collagen staining in paraffin sections of colon. van Gieson, Sirius Red and Sirius Red/Fast Green stainings were carried out for collagen detection and quantitative estimation by morphometric image analysis in colonic specimens from normal rats or animals with 2,4-dinitrobenzenesulfonic acid (DNBS) induced colitis. Haematoxylin/eosin staining was carried out to assess tissue morphology and histopathological lesions. Among the three investigated methods, Sirius Red/Fast Green staining allowed to best highlight well-defined red-stained collagen fibers and to obtain the highest quantitative results by morphometric image analysis in both normal and inflamed colon. Collagen fibers, which stood out against the green-stained non-collagen components, could be clearly appreciated, even in their thinner networks, within all layers of normal or inflamed colonic wall. The present study provides evidence that, as compared with Sirius Red alone or van Gieson staining, the Sirius Red/Fast Green method is the most sensitive, in terms of both qualitative and quantitative evaluation of collagen fibers, in paraffin sections of both normal and inflamed colon.
在组织学切片中检测胶原蛋白并通过计算机辅助图像分析进行定量评估，是评估结缔组织纤维的组织定位和分布的重要方法。人们已经提出了不同的组织化学方法来检测和定量石蜡切片中的胶原蛋白沉积，但其满意度各不相同。本研究旨在比较三种可用于结肠石蜡切片胶原蛋白染色的组织化学方法的定性和定量效率。对正常大鼠或经2,4-二硝基苯磺酸（DNBS）诱导结肠炎的动物的结肠标本进行了范吉森染色、天狼星红染色和天狼星红/固绿染色，以通过形态计量图像分析检测胶原蛋白并进行定量评估。进行苏木精/伊红染色以评估组织形态和组织病理学损伤。在这三种研究方法中，天狼星红/固绿染色能够最好地突出清晰的红色染色胶原蛋白纤维，并通过形态计量图像分析在正常和发炎的结肠中获得最高的定量结果。即使在较细的网络中，在正常或发炎结肠壁的所有层中，与绿色染色的非胶原蛋白成分形成对比的胶原蛋白纤维也能清晰可见。本研究提供的证据表明，与单独使用天狼星红或范吉森染色相比，在正常和发炎结肠的石蜡切片中，从天狼星红/固绿方法对胶原蛋白纤维的定性和定量评估来看，它是最敏感的。
