In response to Jan Nelsons question, We routinely run ISHAGE CD34PE/ CD45FITC with 7-AAD and Flowcount Fluorospheres, to obtain an absolute CD34 count on a single tube. On the Coulter XL the bead is counted in the third channel (625BP) and 7-AAD in the fourth channel (675BP). To reiterate Dave Coder's point compensation between FL4-FL2 is critical and should be done using a CD4 or CD8 PE to ensure you can see the spectral overlap from FL2 to FL4 (there are so few CD34+ cells in most samples it is not practical to compensate using this MoAb). Contact me directly if you would like more info Jan. Mike Keeney London Health Sciences Centre London, Ontario Canada 519 685 8300 ext 2187
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