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Enzyme histochemistry and immunohistochemistry on biopsy specimens of
pathologic human bone marrow
JH Beckstead, PS Halverson, CA Ries and DF Bainton
We have systematically investigated a variety of fixation and plastic
embedding procedures and arrived at a method that allows processing of
approximately 2-micron sections of bone marrow biopsies for examination by
light microscopy. More importantly, 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 degrees C. Decalcification was unnecessary. Sections
2-micron 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 hr.
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. For example, we have discovered that early
myelofibrosis is accompanied by a marked increase in the number of
alkaline-phosphatase-positive reticulum cells.
Volume 57,
Issue 6,
pp. 1088-1098,
06/01/1981
Copyright © 1981 by The American Society of Hematology
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