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

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Basophil Project
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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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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

Comments

 * 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]]


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