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Low‐temperature glycol methacrylate resin embedding method: A protocol suitable for bone marrow immunohistochemistry, PCR, and fish analysis
Author(s) -
Zhang Qiguo,
Wang Jing,
Wu Hongyan,
Zhang Le,
Zhou Jinyong,
Ye Qing,
Shao Xiaoyan,
Guan Chaoyang,
Xu Jingyan,
Yang Yonggong,
Zhou Rongfu,
Ouyang Jian
Publication year - 2010
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20836
Subject(s) - bone decalcification , fluorescence in situ hybridization , polymerase chain reaction , microbiology and biotechnology , bone marrow , fish <actinopterygii> , trephine , in situ hybridization , chemistry , immunohistochemistry , materials science , biomedical engineering , biology , pathology , biochemistry , messenger rna , medicine , gene , chromosome , fishery
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. Microsc. Res. Tech. 73:1067–1071, 2010. © 2010 Wiley‐Liss, Inc.