title: Application of flow cytometry and cell sorting to megakaryocytopoiesis.
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: D000818:Animals; D002452:Cell Count; D002469:Cell Separation; D002498:Centrifugation; D005453:Fluorescence; D006410:Hematopoiesis; D008533:Megakaryocytes / Q000166:cytology; D051379:Mice; D012212:Rheology