Hi Flowers - I've attached a jpeg with some simple staining but strange looking plots. Setup: mouse SC from peptide-DC vaccinated mice were stimulated 4h with peptide+BFA, then stained for anti-CD62L FITC and anti-CD8 PE, then fix/permed and stained with anti-IFNg APC. Cells were immediately run on a FACSCalibur and analyzed using FlowJo (compensation done on FlowJo as well). On the jpeg I have my compensation controls (top row), lymphocyte gate on FSC v SSC (middle row) and CD8 PE v IFNg APC on the bottom row (IFNg APC FMO on the left, actual IFNg stain on the right). For simplicity, I did not inculde the numerous biological controls that were run, but all of the PE v APC plots look similar. I also didn't include the PE v FITC, which looks fine. My question: why does it look as though the APC fluorescence on the PE+ population is so much lower than that of the PEneg population? Did I do something wrong? To me, this data is interpretable, but I don't know how to explain the low APC fluorescence of my PE+population. Any comments would be appreciated. Thanks Diana Diana L. Martin Post-Doctoral Fellow Center for Tropical and Emerging Global Diseases University of Georgia Athens, GA 30602 (706)542-3396 This attachment - 'flowusergroup.jpg' - 489.76 KBytes - can be viewed at http://www.cyto.purdue.edu/MD-parts/de9b21f505512b281b0c85b2ae889e5842dad1c3.jpgReceived on Fri Sep 30 13:38:00 2005
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