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目的：探索一种简单有效的行BrdU 免疫组织化学染色抗原热修复的方法。方法：本实验探索了三种抗原热修复方法：1）漂片煮沸法，2）贴片煮沸法，3）贴片改良法。将贴有冰冻组织切片的玻片置于恒温封闭体系中进行热修复。之后行BrdU 免疫组化染色，采集图像对这三种热修复方法进行比较。针对贴片改良法行BrdU 与NeuN、p-CERB 和DCX 的荧光双标，采集图像以观察该方法在免疫荧光双标实验中应用的可行性。结果：1）漂片煮沸法脑组织切片在经过热修复处理后，切片卷起皱缩，不能进行后续实验步骤；2）贴片煮沸法可见少量BrdU 阳性细胞，但背景深，且少数脑片起泡，假阳性现象严重；3）贴片改良法BrdU 免疫组化及与NeuN、p-CERB 和DCX 的免疫荧光双标染色背景浅，阳性细胞明显。结论：采用改良的热修复法能充分暴露BrdU 抗原， DAB 显色及免疫荧光染色结果理想，此方法为一种较好的行BrdU 免疫组织化学染色抗原热修复方法。

英文摘要:

Objective: To study a convenient and effective technique for heat induced antigen retrieval in BrdU immunohistochemical staining. Methods: We explored three heat induced antigen retrieval methods in this experiments: 1) Boiled floating sections method; 2) Boiled patches method; 3) Improved patches method. Put the glass slides posted with frozen sections into a kind of thermostatic and closed device we designed for antigen retrieval. Then we did the BrdU immunohistochemical staining. Pictures were collected to compare the three heat induced antigen retrieval methods. And we did the BrdU immunofluorescent co-labelling with NeuN, p-CERB, DCX and collect pictures to find out whether the improved patches method could be used in BrdU immunofluorescental staining. Results: 1) Boiled floating sections method. The frozen sections were too shrunken to be used in the following steps after retrieved the antigen in this way. 2) Boiled patches method. Several BrdU labeled neural stem cells could be confirmed, but the background was too dark. And some of the patches blistered, then a great deal of false positive cells were found in the blistered areas. 3) Improved patches method. The positive products of BrdU immunohistochemistry and immunofluorescent co-labeling with NeuN, p-CERB, DCX were distinct and the background was controlled well. Conclusion: Our antigen retrieval technique could fully expose the BrdU antigen , the DAB staining and immunohistochemical staining was controlled well. Therefore, it is an improved technique of antigen retrieval in BrdU immunohistochemical staining.

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