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Comparison of the DNA content of megakaryocytes identified immunologically with that identified morphologically

Comparison of the DNA content of megakaryocytes identified immunologically with that identified morphologically

Authors :
Motomi Ikeda
Nobuhiko Uoshima
Koji Tanaka
Motoharu Kondo
Shinya Kimura
Tatuo Sugano
Masaru Ozawa
Satoshi Chikayama
Katuya Wada
Yufuko Takahashi
Naoyuki Maruo
Yutaka Kobayashi
Source :
Histochemistry and Cell Biology. 108:115-120
Publication Year :
1997
Publisher :
Springer Science and Business Media LLC, 1997.

Abstract

We devised a new microfluorometric method for determining the ploidy of megakaryocytes identified immunologically in bone marrow smears. The smears were immunostained by incubation with mouse monoclonal anti-glycoproteins (GP) IIb antibodies, followed by fluorescein isothiocyanate-conjugated goat antimouse IgG antibodies. They were then stained with 4',6-diamidino-2-phenylindole (DAPI). Megakaryocytes were identified by their GPIIb immunofluorescence using a microfluorometer and, after the filters were changed, their DNA content was assayed by measuring the intensity of DAPI fluorescence. This intensity was shown to be proportional to the DNA content when the aperture of the objective lens was reduced. We compared these results with those obtained when megakaryocytes were identified morphologically, using DAPI staining after Wright-Giemsa destaining. In all 12 normal controls, the ploidy peaks were shown to be 16N by both methods, and the mean ploidy detected by the immunological method was only reduced 0.961 times relative to the estimate from the morphological method. In contrast, in eight myelodysplastic syndrome (MDS) patients, the ploidy peaks were either 8N or 4N and the mean was reduced by 0.906 times (P = 0.018). Thus we could immunologically identify small megakaryocytes which we could not identify morphologically. Therefore, this method is useful for measuring megakaryocytic ploidy, especially in the pathological megakaryocytes of MDS patients.

Details

ISSN :
1432119X and 09486143
Volume :
108
Database :
OpenAIRE
Journal :
Histochemistry and Cell Biology
Accession number :
edsair.doi.dedup.....9dc176620017a03c29dd568167499e13
Full Text :
https://doi.org/10.1007/s004180050152