APPLICATION OF FLOW CYTOMETRY AND CELL SORTING TO MEGAKARYOCYTOPOIESIS

  • 1 January 1979
    • journal article
    • research article
    • Vol. 53  (4) , 732-745
Abstract
Flow cytometry (FCM) and cell sorting were employed 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 1st 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 the normal or perturbed population can now be studied rapidly and quantitatively.