The monocytes contamination in a lymphoid gate is not resolved with a
CD45vSS, due to the fact that the altered monocytes have both the SS and
CD45 very similar to the lymphocytes; as opposed to granulocytic
contamination that can be solved by SSvCD45 gating.
If the lymphoid gate in FSvSS is contaminated by monocytes, you should
add a monocytoid line marker(CD14) to remove these cells.In any case a
2-3% of contamination is not a high value. I recommend you to do a
lymphoid gate in order to separate better the different populations from
a fresh sample.
The decrease problem expression can be solved titering the antibodies
and/or scanning the voltages of your cytometer.
The ideal panel to determine kappa/lambda is a matter of each particular
lab, but I strongly recommend you kappa/lambda/CD19 PerCP in the same
tube. We use kappa/lambda from Dako and CD19 from Becton Dickinson, and
we have great results.
Marco A. Fernández
e-Mail: reference@cinet.fcr.es