ENZYME-HISTOCHEMISTRY AND IMMUNOHISTOCHEMISTRY ON BIOPSY SPECIMENS OF PATHOLOGIC HUMAN-BONE MARROW

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  • 1 January 1981
    • journal article
    • research article
    • Vol. 57  (6) , 1088-1098
Abstract
Various fixation and plastic embedding procedures were investigated, and a method was arrived at that allows processing of .apprx. 2-.mu.m sections of bone marrow biopsies for examination by light microscopy. This method permits the use of enzyme histochemical and immunohistochemical procedures that are rapidly becoming mandatory in the diagnosis of hematologic malignancies. Over 200 full-length bone marrow biopsy specimens were fixed in a mixture of paraformaldehyde, glutaraldehyde and acrolein, dehydrated in acetone and embedded in a mixture of methyl and glycolmethacrylate. All procedures were carried out at 4.degree. C. Decalcification was unnecessary. Sections 2 .mu.m thick were cut and incubated for peroxidase, naphthol AS-D chloroacetate esterase, .alpha.-naphthyl butyrate esterase, acid phosphatase (with and without tartrate), or alkaline phosphatase and then examined by light microscopy. Specimens could be prepared for examination within 48 h. This approach, which provides definitive markers for various hematopoietic cell lines in intact tissues, is invaluable when aspirated material is unavailable. It is also useful in the analysis of focal lesions of bone marrow due to inflammation or neoplasia and shows potential as an investigative tool, e.g., in discovering that early myelofibrosis is accompanied by a marked increase in the number of alkaline-phosphatase-positive reticulum cells.