title: Low-temperature glycol methacrylate resin embedding method: A protocol suitable for bone marrow immunohistochemistry, PCR, and fish analysis.
abstract: 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.
authors.affiliation: Department of Hematology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, China.
authors.forename: Qiguo
authors.initials: Q
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Hongyan
authors.initials: H
authors.lastname: Wu
authors.forename: Le
authors.initials: L
authors.lastname: Zhang
authors.forename: Jinyong
authors.initials: J
authors.lastname: Zhou
authors.forename: Qing
authors.initials: Q
authors.lastname: Ye
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Shao
authors.forename: Chaoyang
authors.initials: C
authors.lastname: Guan
authors.forename: Jingyan
authors.initials: J
authors.lastname: Xu
authors.forename: Yonggong
authors.initials: Y
authors.lastname: Yang
authors.forename: Rongfu
authors.initials: R
authors.lastname: Zhou
authors.forename: Jian
authors.initials: J
authors.lastname: Ouyang
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism; D018952:Antigens, CD34 / Q000737:chemistry / Q000378:metabolism; D001853:Bone Marrow / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000737:chemistry / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D053614:Janus Kinase 2 / Q000737:chemistry / Q000378:metabolism; D007938:Leukemia; D008689:Methacrylates / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D016610:Tissue Embedding / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D018952:Antigens, CD34; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008689:Methacrylates; C005044:hydroxyethyl methacrylate; D004247:DNA; C507924:JAK2 protein, human; D053614:Janus Kinase 2