With interest we saw the results of your study on counting of CD4, CD8 and CD3 using a bead based single platform device.I donīt think that your underestimation has anything to do with the anticoagulants that you used in your study. We found the same 10 - 20% underestimation for such subclasses by comparing the results of a volumetrical flow cytometer measurement with those of a dual platform system. In case of CD34 cells, the loss was much more dramatic. Depending on the lysing reagent used, the underestimation was in the range of 34 - 45%. In a detailed study we recognized the lysing reagents as the reason for this cell loss, by membrane destruction. Using the dual platform mode, these effects are compensated by the hematology counter results in connection with the CD45 numbers. We found that the destruction of some leucocytes by different lysing reagents is well known in the literature, but obviously not enough being taken into consideration in the companies supported protocols. The best solution could be a non lyse protocol, we found one which obviously works well.
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:27 EST