Chromium+Release+CTL+Assay+with+Ova+9-26-12

Chromium Release CTL Assay with Ova 9-26-12

Planning started 9-26-12

Diluted SIINFEKL peptide without any DMSO (I diluted in 10% DMSO in the past). The peptide dissolved quite easily without any DMSO. Item: SIINFEKL 1000 ug/mL 9-26-12 in 1XPBS (no DMSO) at -20 freezer in tissue culture room

10-7-12 Danielle and I plan to start this assay tomorrow.

I plan to use the plate map found here: C:\kurt\storage\CIM Research Folder\DR\2012\8-16-12\ctl plate map\ctl plate map.xlsx

Danielle and I discussed having all of the controls on my plate, and then having fewer samples for her plate. Here is a potential example Danielle plate "C:\kurt\storage\CIM Research Folder\DR\2012\10-7-12\CTL\potential danielle ctl plate map.xlsx"

Here is a spreadsheet that contains info for both of us "C:\kurt\storage\CIM Research Folder\DR\2012\10-7-12\CTL\CTL plate maps 10-7-12.xlsx" here's a macro-enabled version of the spreadsheet "C:\kurt\storage\CIM Research Folder\DR\2012\10-7-12\CTL\CTL plate maps 10-7-12.xlsm"

10-8-12 Danielle and I did the chromium release assay protocol. We let the Luma plates dry overnight. Item: Ova mouse sera 10-8-12 at CTL 6-13-11

Images of counts from topcount here C:\kurt\storage\CIM Research Folder\DR\2012\10-9-12\CTL\Images

Results here "C:\kurt\storage\CIM Research Folder\DR\2012\10-9-12\CTL\CTL Results 10-9-12.xlsx"

There doesn't seem to be as much radioactivity detected as there was the first time that I did this experiment. Perhaps the batch of chromium is bad, or the batch of chromium is too old, or maybe the cells were not labeled well (too large a volume, not enough time, etc.). Maybe I should try to label some target cells with different conditions to see how the results vary.

The data also shows that certain columns have higher counts overall than other columns regardless of the sample (columns 1, 5, and 9). I think these columns were read first maybe? Although these counts are higher, the counts are so low that this may not be too much of an issue.