Department of Hematology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, China.
Microsc Res Tech. 2010 Oct;73(11):1067-71. doi: 10.1002/jemt.20836.
Molecular analyses such as fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) are demanded to improve diagnostic accuracy in addition to immunohistopathology of bone marrow (BM) trephine specimens. Conventional BM embedding method needs decalcification, and its procedure may impair tissue morphology and DNA quality. Here, we report an undecalcified method by which glycol methacrylate resin is polymerized at low temperature (4°C). Using this method, BM enzyme activity and antigenic determinants are well preserved, and moreover, DNA extracted from plastic embedding sections is suitable for PCR amplification and sequencing, FISH analysis can be well done because of the DNA integrity of BM sections. If working with BM trephine specimen, our protocol offers the possibility to combine superior morphology with modern molecular analysis.
除了骨髓（BM）活检标本的免疫组织病理学检查外，还需要进行荧光原位杂交（FISH）和聚合酶链反应（PCR）等分子分析，以提高诊断的准确性。传统的 BM 包埋方法需要脱钙，其过程可能会损害组织形态和 DNA 质量。在这里，我们报告了一种在低温（4°C）下聚合乙二醇甲基丙烯酸酯树脂的非脱钙方法。使用这种方法，BM 酶活性和抗原决定簇得到很好的保留，而且，从塑料包埋切片中提取的 DNA 适合 PCR 扩增和测序，由于 BM 切片的 DNA 完整性，FISH 分析也可以很好地进行。如果使用 BM 活检标本，我们的方案提供了将优异的形态与现代分子分析相结合的可能性。
