Fig. 1From: The Effect of Bcr-Abl Protein Tyrosine Kinase on Maturation and Proliferation of Primitive Haematopoietic CellsStrategy for flow cytometric analysis of normal bone marrow (NBM) and chronic myeloid leukaemia (CML) samples. Mononuclear cells were stained with CD34+, lineage marker, and Thy antibodies as described in “Materials and Methods.” Gates were set to sort live (propidium iodide excluding), lineage-negative cells as shown (A and B). Further gating, as shown in (C) was applied to define the CD34+Lin-Thy+ and CD34+Lin-Thy-populations revealed by staining with CD34 allophycocyanin (APC) and Thy phycoerythrin (PE). Using this four-parameter approach, CD34+Lin-Thy-(D) and CD34+Lin-Thy+ (E) populations were sorted to >90% purity. The sorted cells were then stained with rhodamine-123 (Rh-123) and Hoechst 33342 to simultaneously analyze the cell cycle and Rh-123 retention profiles of the sorted populations (F) and (G). The quadrant marker separates high levels of rhodamine-123 (Rh-123hi) from low levels of rhodamine-123 (Rh-123lo) cells and cycling cells from non cycling cells. A sample of normal bone marrow is depicted throughout this figure; cells from CML patients were processed using an identical strategy.Back to article page