Sugihara H, Hattori T, Fukuda M
Histochemistry. 1986;85(3):193-5. doi: 10.1007/BF00494803.
Bromodeoxyuridine (BrdUrd) immunohistochemistry has recently been introduced for the visualization of DNA-synthesizing nuclei. In order to detect the BrdUrd incorporated into nuclear DNA in formalin-fixed, paraffin-embedded tissues, we tested several different pretreatment procedures including digestion with proteinase and hydrolysis with HCl, prior to immunoperoxidase staining. In order to determine the optimal conditions for detecting nuclear BrdUrd, mice were given BrdUrd and 3H-thymidine simultaneously, and the autoradiographic and immunohistochemical results obtained in BrdUrd-stained sections were compared. It was found that digestion with 0.05% proteinase at 37 degrees C for 20 min and hydrolysis with 1N HCl at 37 degrees C for 20 min was sufficient to detect BrdUrd immunoreactivity in 3H-thymidine-labelled nuclei, the results being virtually unaffected by the orders in which the two pretreatments were performed. Our method extends the range of application for BrdUrd immunohistochemistry in cell-kinetic studies.
溴脱氧尿苷（BrdUrd）免疫组织化学最近已被用于可视化DNA合成细胞核。为了检测福尔马林固定、石蜡包埋组织中掺入核DNA的BrdUrd，我们在免疫过氧化物酶染色之前测试了几种不同的预处理程序，包括蛋白酶消化和盐酸水解。为了确定检测核BrdUrd的最佳条件，给小鼠同时注射BrdUrd和3H-胸腺嘧啶核苷，并比较在BrdUrd染色切片中获得的放射自显影和免疫组织化学结果。结果发现，在37℃用0.05%蛋白酶消化20分钟和在37℃用1N盐酸水解20分钟足以检测3H-胸腺嘧啶核苷标记细胞核中的BrdUrd免疫反应性，结果几乎不受两种预处理执行顺序的影响。我们的方法扩展了BrdUrd免疫组织化学在细胞动力学研究中的应用范围。
