BrdU免疫组织化学染色方法改进(漂片法).docVIP

BrdU免疫组织化学染色方法改进(漂片法) 　　[摘要] 目的 改进用漂片法进行BrdU免疫组织化学染色时的封闭液，以减少组织切片破损。 方法 用5-溴-2-脱氧尿嘧啶核苷（5-bromo-2-deoxyuridine, BrdU）标记大鼠踏转轮运动中所引起增殖细胞，通过 ABC免疫组织化学方法染色。在染色过程中，比较不同的封闭液（羊血清、牛血清白蛋白、胰酶抑制剂）减少脑片破损的效果。 结果 踏转轮运动可使大鼠海马齿状回BrdU阳性细胞数明显多于对照组。加入不同的封闭液后，组织片破损率明显下降，尤其是加入牛血清白蛋白和胰酶抑制剂组，但胰酶抑制剂组有非特异性染色。 结论 牛血清白蛋白是较理想的封闭液。 　　[关键词] BrdU；免疫组织化学染色；胰酶抑制剂；牛血清白蛋白；漂片法 　　[中图分类号] R392-33 [文献标识码] A [文章编号] 1673-7210（2012）02（b）-0016-03 　　 　　Improvement in technique of BrdU immunohistochemical staining by bleach section 　　GUO Jingjing1 SHAN Lidong2 WU Gang3 　　1.Department of General Surgery, Huashan Hospital Baoshan Branch of Fudan University, Shanghai 200431, China; 　　2.Department of Neurobiology and Medical Psychology, School of Preclinical Medicine and Biological Sciences,Medical School of Suzhou University, Jiangsu Province, Suzhou 215123, China;3.Department of General Surgery, Huashan Hospital Affiliated to Fudan University, Shanghai 200040, China 　　[Abstract] Objective To reduce the tissue slice breakage, explore the technique of 5-bromo-2-deoxyuridine (BrdU) immunohistochemical staining by blocking buffer in bleach section. Methods BrdU, which was detected by ABC immunohistochemistry on bleach section of brain slides, was used to incorporate the new born cells which were promoted proliferation by motor-driven wheel running. The different blocking effects of Goat serum, trypsin inhibitor and bovine serum albumin were compared in the process of immunohistochemistry. Results In the group of motor-driven wheel running in the dentate gyrus of the hippocampus in rats, the number of BrdU-positive cells was more than the control group. The breaking rate of slides was reduced obviously, especially in the groups with bovine serum albumin and trypsin inhibitor, after being added different blocking reagents. But cells in trypsin inhibitor group were appeared without specific staining. Conclusion Bovine serum albumin is preferable blocking buffer. 　　[Key words] BrdU; Immunohistochemical stainin