Cd metal and fresh whole blood staining issue
We are losing the Cd metal stain when staining fresh whole blood.
protocol is straightforward comparing 3 staining protocols.
A fresh whole blood sample split into 3 aliqutes:
1. WB--> smartube fix--> freeze -80--> thaw and lyse (smarttube reagent)--> heparin/Fc block--> extra stain--> BD fix/perm--> intra stain--> fix&Ir--> Helios.
2. WB--> Fc block--> extra stain-->Biolegend RBC lysis--> BD fix/perm--> intra stain--> fix&Ir--> Helios.
3. WB-->Fc block--> extra stain--> smartube fix--> freeze -80--> thaw and lyse (smarttube reagent)--> BD fix/perm--> intra stain--> fix&Ir--> Helios.
We use CAS plus with cell acquisition.
In the attached pdf live intact cells are plotted.
The Cd stain is normal with ST (1) but lost when we are staining prior to fixation (2, 3).
Any one else encountered this?
I'd appreciate any thoughts on this?
Thanks