Veronese S, Gambacorta M, Falini B
Istituto di Anatomia Patologica, Ospedale di Niguarda, Milan, Italy.
J Clin Pathol. 1989 Aug;42(8):820-6. doi: 10.1136/jcp.42.8.820.
Immunohistochemical staining with anti-bromo-deoxyuridine (BrdU) monoclonal antibody was performed on a variety of human tissues following in vitro incubation with BrdU. The effect of different fixatives and DNA denaturation techniques on the reactivity with anti-BrdU was investigated. Optimal preservation of the antigenicity of BrdU incorporated into the DNA of proliferating cells was seen in tissues fixed in Bouin's fluid, while samples which had been fixed with cross-linking reagents, such as formalin, were usually unreactive. Positivity for BrdU was restored in formalin fixed tissues after digestion with pepsin, but this was usually associated with loss of morphological details. Acid and thermal DNA denaturation techniques gave similar results. It is concluded that Bouin fixation followed by acid or thermal denaturation of DNA is the method of choice for the in situ detection of cells in S-phase using anti-BrdU monoclonal antibody.
在用溴脱氧尿苷（BrdU）体外孵育各种人体组织后，使用抗BrdU单克隆抗体进行免疫组织化学染色。研究了不同固定剂和DNA变性技术对抗BrdU反应性的影响。在Bouin氏液固定的组织中，掺入增殖细胞DNA的BrdU抗原性得到了最佳保存，而用福尔马林等交联试剂固定的样本通常无反应性。用胃蛋白酶消化后，福尔马林固定的组织中BrdU的阳性反应得以恢复，但这通常伴随着形态细节的丢失。酸和热DNA变性技术给出了相似的结果。结论是，对于使用抗BrdU单克隆抗体原位检测S期细胞，Bouin固定后进行DNA的酸或热变性是首选方法。
