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Title: Immunocytochemical distinction between primary and secondary granule formation in developing human neutrophils: correlations with Romanowsky stains

Abstract

Electron-microscopic studies with peroxidase cytochemistry have shown that primary (azurophilic) granules in human neutrophils are synthesized in promyelocytes, while secondary (specific) granules are formed in myelocytes. However, these studies were limited by the lack of specific markers for secondary granules and by the inability to make direct comparisons of electron-microscopic morphology with the light-microscopic appearance of the same cell. Thus secondary granules cannot be identified reliably and the relevance of these findings to the light-microscopic interpretation of clinical bone marrow specimens cannot be evaluated. To circumvent these problems, we developed a method to permit immunofluorescent demonstration of primary and secondary granule markers in cells stained with Romanowsky agents. Normal human marrow cells were stained with May-Gruenwald-Giemsa and photographed. The slides were decolorized in buffered glycerine and saline for 24 hr and then stained with fluorescein- and rhodamine-conjugated monospecific antisera to human granulocyte myeloperoxidase, cathepsin G, elastas, lysozyme, and lactoferrin. The same cells were then located and examined for conjugate binding. Double stains with fluorescein- and rhodamine-labeled antisera offered the additional advantage of simultaneous identification of primary and secondary granules. This approach confirmed the partition of primary and secondary granule proteins and related their appearance to the maturation of developing neutrophilsmore » in normal human bone marrow.« less

Authors:
; ; ;
Publication Date:
Research Org.:
Univ. of North Carolina, Chapel Hill
OSTI Identifier:
6165852
Alternate Identifier(s):
OSTI ID: 6165852
Resource Type:
Journal Article
Journal Name:
Blood; (United States)
Additional Journal Information:
Journal Volume: 53:2
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; LEUKOCYTES; CYTOLOGICAL TECHNIQUES; BONE MARROW CELLS; CYTOLOGY; EXPERIMENTAL DATA; IMMUNE REACTIONS; ISOLATED VALUES; MAN; MICROSCOPY; STAINS; ANIMAL CELLS; ANIMALS; BIOLOGICAL MATERIALS; BIOLOGY; BLOOD; BLOOD CELLS; BODY FLUIDS; CONNECTIVE TISSUE CELLS; DATA; DATA FORMS; INFORMATION; MAMMALS; NUMERICAL DATA; PRIMATES; SOMATIC CELLS; VERTEBRATES 550300* -- Cytology

Citation Formats

Pryzwansky, K.B., Rausch, P.G., Spitznagel, J.K., and Herion, J.C. Immunocytochemical distinction between primary and secondary granule formation in developing human neutrophils: correlations with Romanowsky stains. United States: N. p., 1979. Web.
Pryzwansky, K.B., Rausch, P.G., Spitznagel, J.K., & Herion, J.C. Immunocytochemical distinction between primary and secondary granule formation in developing human neutrophils: correlations with Romanowsky stains. United States.
Pryzwansky, K.B., Rausch, P.G., Spitznagel, J.K., and Herion, J.C. Thu . "Immunocytochemical distinction between primary and secondary granule formation in developing human neutrophils: correlations with Romanowsky stains". United States.
@article{osti_6165852,
title = {Immunocytochemical distinction between primary and secondary granule formation in developing human neutrophils: correlations with Romanowsky stains},
author = {Pryzwansky, K.B. and Rausch, P.G. and Spitznagel, J.K. and Herion, J.C.},
abstractNote = {Electron-microscopic studies with peroxidase cytochemistry have shown that primary (azurophilic) granules in human neutrophils are synthesized in promyelocytes, while secondary (specific) granules are formed in myelocytes. However, these studies were limited by the lack of specific markers for secondary granules and by the inability to make direct comparisons of electron-microscopic morphology with the light-microscopic appearance of the same cell. Thus secondary granules cannot be identified reliably and the relevance of these findings to the light-microscopic interpretation of clinical bone marrow specimens cannot be evaluated. To circumvent these problems, we developed a method to permit immunofluorescent demonstration of primary and secondary granule markers in cells stained with Romanowsky agents. Normal human marrow cells were stained with May-Gruenwald-Giemsa and photographed. The slides were decolorized in buffered glycerine and saline for 24 hr and then stained with fluorescein- and rhodamine-conjugated monospecific antisera to human granulocyte myeloperoxidase, cathepsin G, elastas, lysozyme, and lactoferrin. The same cells were then located and examined for conjugate binding. Double stains with fluorescein- and rhodamine-labeled antisera offered the additional advantage of simultaneous identification of primary and secondary granules. This approach confirmed the partition of primary and secondary granule proteins and related their appearance to the maturation of developing neutrophils in normal human bone marrow.},
doi = {},
journal = {Blood; (United States)},
number = ,
volume = 53:2,
place = {United States},
year = {1979},
month = {2}
}