A comparing study of quantitative staining techniques for retinal neovascularization in a mouse model of oxygen-induced retinopathy
A comparing study of quantitative staining techniques for retinal neovascularization in a mouse model of oxygen-induced retinopathy
复制标题
氧诱导性视网膜病变小鼠模型视网膜新生血管定量染色技术的比较研究
DOI：
10.3980/j.issn.2222-3959.2012.01.01
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发表时间：
2012-02-18
影响因子：
1.4
通讯作者：
Long, Liao-Xu
中科院分区：
文献类型：
--
作者：
Liang, Xiao-Ling;Li, Jie;Long, Liao-Xu
目的：探索一种高效、实用、客观的定量方法评价氧诱导性视网膜病变（OIR）小鼠模型视网膜新生血管的形成情况。方法：采用30只C57BL/6J小鼠进行OIR模型建立。摘除眼睛进行不同的染色方法，包括：（1）HE染色； (2)用加纳单叶凝集素(GSL)进行免疫组织化学分析； (3) FITC标记的CD31抗体进行免疫荧光检查； (4) 用纯化的CD31抗体进行两步免疫荧光； (5)FITC-葡聚糖灌注结合两步纯化-CD31免疫荧光。通过成像软件分析视网膜脉管系统的图像。结果：GSL 免疫组化可清晰显示深层和浅层毛细血管床。FITC 标记的 CD31 免疫荧光模糊，荧光背景高，难以区分某些区域的视网膜新生血管。两步纯化 CD31 免疫荧光组提供了新生血管和预先存在的视网膜血管的出色细节。结论：GSL 免疫组织化学可以清楚地显示深层和浅层毛细血管床中的新生血管簇。血管内皮特异性抗原CD31的免疫荧光可以选择性地标记小鼠视网膜的新生血管。结合计算机分析软件，是评价OIR小鼠模型视网膜新生血管形成的有效、客观的定量方法。
AIM: To explore an efficient, practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR).METHODS: Thirty C57BL/6J mice were explored in OIR model procedure. Eyes were removed for different staining methods including: (1) HE staining; (2) immunohistochemistry with Griffonia Simplicifolia Lectin (GSL); (3) Immunofluorescence with FITC labeled CD31 antibody; (4) Two-step immunofluorescence with purified-CD31 antibody; (5) FITC-Dextran perfusion combined with two-step purified-CD31immunofluorescence. Images of the retinal vasculature were analyzed by imaging software. "RESULTS: GSL immunohistochemistry could clearly demonstrate the deep and superficial capillary beds. FITC labeled CD31 Immunofluorescence was blurring with high fluorescence background which was hard to distinguish retinal neovascularization in some area. Excellent detail of neovascularization and preexistent retinal vessels was provided in two-step Purified-CD31 immunofluorescence group.CONCLUSION: GSL immunohistochemistry can clearly demonstrate neovascularization tufts in deep and superficial capillary beds. Immunofluorescence of specific antigen CD31 on vascular endothelium can selectively label the neovascularization of mouse retina. When combined with computer analysis software, it is an effective and objective quantitative method to evaluate the retinal neovascularization in OIR mouse model.