Antigen retrieval by microwave oven heating for immunohistochemical analysis of bone marrow trephine biopsies. 1996

W N Erber, and T A Gibbs, and J G Ivey
Hematology Department, Royal Perth Hospital, Perth, WA.

Immunohistochemical analysis of bone marrow trephine (BMT) biopsies with monoclonal antibodies for the analysis of hemopoietic disorders has been hindered by the fixation and decalcification regimens which mask or destroy tissue antigens. This study evaluated the effect of microwave oven treatment on the quality of immunostaining of fixed decalcified trephine biopsies. The aim was to establish whether this method of pre-treatment would enable additional antigens to be detected. Fifty-eight monoclonal and 4 polyclonal antibodies to hemopoietic antigens were assessed to compare no tissue pre-treatment, proteolytic (trypsin) enzyme digestion and microwave oven heating. The microwave heating of the sections was performed by placing them in a boiling solution of 0.01M tri-sodium citrate for a total of 10 mins. Following microwave heating 14 antibodies that previously showed no reactivity in BMT biopsies gave positive staining and 9 antibodies previously known to detect antigens in the absence of pre-treatment gave enhanced staining. Other antibodies showed no staining improvement with microwave heating and some failed to give a positive reaction by any of the pre-treatment methods. Antigen retrieval utilizing microwave oven heating can expose antigenic sites for antibody binding in bone marrow trephine sections. However not all antigens are retrieved and there is variation between epitopes on the one molecule and their ability to be exposed by microwave heating. Utilizing antigen retrieval methods, the range of antibodies applicable to BMT sections is greatly expanded enabling the immunophenotypic analysis of the majority of hemopoietic disorders.

UI MeSH Term Description Entries
D007150 Immunohistochemistry Histochemical localization of immunoreactive substances using labeled antibodies as reagents. Immunocytochemistry,Immunogold Techniques,Immunogold-Silver Techniques,Immunohistocytochemistry,Immunolabeling Techniques,Immunogold Technics,Immunogold-Silver Technics,Immunolabeling Technics,Immunogold Silver Technics,Immunogold Silver Techniques,Immunogold Technic,Immunogold Technique,Immunogold-Silver Technic,Immunogold-Silver Technique,Immunolabeling Technic,Immunolabeling Technique,Technic, Immunogold,Technic, Immunogold-Silver,Technic, Immunolabeling,Technics, Immunogold,Technics, Immunogold-Silver,Technics, Immunolabeling,Technique, Immunogold,Technique, Immunogold-Silver,Technique, Immunolabeling,Techniques, Immunogold,Techniques, Immunogold-Silver,Techniques, Immunolabeling
D008872 Microwaves That portion of the electromagnetic spectrum from the UHF (ultrahigh frequency) radio waves and extending into the INFRARED RAYS frequencies. EHF Waves,Extremely High Frequency Radio Waves,Micro Wave,Micro Waves,Ultrahigh Frequency Waves,Microwave Radiation,EHF Wave,Micro Waves,Microwave,Microwave Radiations,Radiation, Microwave,Ultrahigh Frequency Wave,Wave, EHF,Wave, Micro,Wave, Ultrahigh Frequency,Waves, Micro
D001853 Bone Marrow The soft tissue filling the cavities of bones. Bone marrow exists in two types, yellow and red. Yellow marrow is found in the large cavities of large bones and consists mostly of fat cells and a few primitive blood cells. Red marrow is a hematopoietic tissue and is the site of production of erythrocytes and granular leukocytes. Bone marrow is made up of a framework of connective tissue containing branching fibers with the frame being filled with marrow cells. Marrow,Red Marrow,Yellow Marrow,Marrow, Bone,Marrow, Red,Marrow, Yellow
D001855 Bone Marrow Diseases Diseases involving the BONE MARROW. Bone Marrow Disease,Disease, Bone Marrow,Diseases, Bone Marrow,Marrow Disease, Bone,Marrow Diseases, Bone
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D000941 Antigens Substances that are recognized by the immune system and induce an immune reaction. Antigen
D001706 Biopsy Removal and pathologic examination of specimens from the living body. Biopsies

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