An enhanced immunocytochemical method for staining bone marrow trephine sections.
OBJECTIVE
The detection of cellular antigens in fixed decalcified bone marrow trephine (BMT) sections depends on the method of processing, the nature of the antigen and antibody, antigen retrieval techniques, and the sensitivity of the immunocytochemical method. This study evaluated a tyramide enhanced avidin-biotin immunostaining method on formalin fixed decalcified BMT sections to determine whether the method could detect previously undetectable antigens.


METHODS
Nineteen BMT biopsies from a range of haematological disorders were evaluated with 43 antibodies to haemopoietic antigens using horseradish peroxidase and alkaline phosphatase detection methods, using the tyramide enhanced avidin-biotin immunostaining method.


RESULTS
Compared with standard avidin-biotin immunostaining methods the tyramide enhanced immunostaining method showed enhanced signal intensity, gave positive labelling for antigens that require pretreatment by other methods, and previously unreactive antigens were detected. Primary antibodies could be used at up to 200 times higher dilutions.


CONCLUSIONS
The tyramide enhanced immunostaining method, while retaining specificity, is highly sensitive and enables an increased number and range of antigens to be detected than previously possible. The method could be applied to BMT sections for the routine diagnosis and classification of haematological disorders.
Erber
W N
WN
Haematology Department, Royal Perth Hospital, Australia. wenderbe@dunamis.rph.uwa.edu.au
Willis
J I
JI
Hoffman
G J
GJ
D000911:Antibodies, Monoclonal; D001706:Biopsy; D001853:Bone Marrow / Q000276:immunology*; D006402:Hematologic Diseases / Q000175:diagnosis* / Q000276:immunology; D016591:Histocytological Preparation Techniques; D006801:Humans; D007124:Immunoenzyme Techniques; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods
D000911:Antibodies, Monoclonal