Chromium Release CTL Assay with Ova 9-26-12

2015-01-13

azim58 - 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.

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