(A): All nucleated cells (R1) and cells with relatively low SSC (R2) are gated. (B): Cells in R2 in panel (A) are displayed on a CD45/CD34 plot, and CD34+ cells with intermediate CD45 expression are gated (R3). (C): Cells in the R3 gate are plotted on a CD45/SSC display. In the CD45/SSC dot plot, we can identify a cell population consisting predominantly of CD34+ B-cell progenitor cells which formed an easily recognizable cluster (R5) that had the lowest SSC and relatively low CD45 expression. Other CD34+ cells (R4) showing more SSC and a wider distribution of CD45 expression consisted predominantly of myeloblasts. (D) All nucleated cells are plotted on a CD45/SSC display. Histogram (D1) SSC of lymphocytes (in red) and granulocytes (in gray). SSC peak channel values (SSC channel number where the maximum number of cells occurs) of both fractions were computed. Histogram (D2) shows CD45 expression of lymphocytes (in red) and CD34+ myeloblasts (in cyan). Mean fluorescence intensity (MFI) of CD45 of both populations are computed.

Figure 1: Ogata’s gating strategy cells stained with CD34 and CD45 antibodies [8-11].