Application of flow cytometry and cell sorting to megakaryocytopoiesis.
Blood, 1979/4;53(4):732-45.
Nakeff A, Valeriote F, Gray JW, Grabske RJ
PMID: 154895
Impact factor: 25.476
Abstract
We have employed flow cytometry (FCM) and cell sorting to quantitate and study megakaryocytes in mouse and rat femoral marrow following their 20- to 30-fold concentration by centrifugal elutriation (CE). This enrichment of megakaryocytes permitted the first determination of their DNA-related fluorescence by FCM analysis following DNA staining. Fluorescence distributions of CE-enriched cell fractions following supravital staining with Hoechst 33342 were similar to those following chromomycin A3 staining of ethanol-fixed cells. Microscopic examination of cells sorted onto glass slides on the basis of their DNA-related fluorescence following supravital staining together with specific acetylcholinesterase staining for megakaryocytes indicated that megakaryocytes generally increased in cell size with increasing DNA content. This technologic application represents a significant advance in the study of megakaryocytopoiesis, since the kinetics of either the normal or perturbed population can now be studied rapidly and quantitatively.
MeSH terms
Animals; Cell Count; Cell Separation; Centrifugation; Fluorescence; Hematopoiesis; Megakaryocytes; Mice; Rheology
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