Shreffler:Notebook/R03 Baso/2010/08/11

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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)

Flow Analysis

  • 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.
  1. Media:BasoSens10_20100811.pdf
  2. Media:BasoSens10_2010081!-Table.txt
  3. Media:BasoSens10_20100811-Table.pdf