Low-temperature glycol methacrylate resin embedding method: A protocol suitable for bone marrow immunohistochemistry, PCR, and fish analysis.
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.
Zhang
Qiguo
Q
Department of Hematology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, China.
Wang
Jing
J
Wu
Hongyan
H
Zhang
Le
L
Zhou
Jinyong
J
Ye
Qing
Q
Shao
Xiaoyan
X
Guan
Chaoyang
C
Xu
Jingyan
J
Yang
Yonggong
Y
Zhou
Rongfu
R
Ouyang
Jian
J
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*
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