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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
- 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.
- Subjects :
- Indoles
Histology
Platelet Glycoprotein GPIIb-IIIa Complex
Biology
Immunofluorescence
chemistry.chemical_compound
medicine
Humans
DAPI
Fluorescein
Molecular Biology
Fluorescent Dyes
Ploidies
medicine.diagnostic_test
Bone Marrow Examination
DNA
Cell Biology
Flow Cytometry
Molecular biology
Staining
Medical Laboratory Technology
medicine.anatomical_structure
chemistry
Case-Control Studies
Myelodysplastic Syndromes
Monoclonal
biology.protein
Bone marrow
Ploidy
Antibody
Megakaryocytes
Fluorescein-5-isothiocyanate
Subjects
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