Basophil Sensitization (10) - 8/11/10
- Had starting cell count of approx. 6 million PBMCs per sample (250 million total)
- Running low on H12 and P4E (rDerp2-specific IgE), so used half the volume described in protocol (instead of 80uL H12 + 80uL P4E + 640 uL medium + 800 uL cells for 1600 uL total, used 40+40+320+400=800)
- Cells clumped slightly, but wasn't a huge problem as with some past experiments (didn't have to resuspend with pipettor before every step)
- Basos were nonresponding...again
- Event rate was good
- Due to repeated nonresponders, Wayne suggested we stim/stain blood straight from buffy, then run in flow before starting brunt of experiment.