I was just curious as to whether anyone has tried doing intracellular cytokine staining on cells that had been frozen and then thawed. Can this be done? What problems do we need to be on the lookout for? We are interested in doing some ICS as part of a pathogenesis study where we will be taking samples from primates every day for six days. What we would like to do is look at gamma interferon/TNFalpha production by lymphocytes during the course of disease using whole blood similarly to what has been published by Picker & others. My thought was to remove the red blood cells and freeze the PBLs in freezing medium at -70 C. If we could freeze the cells and do the assay at a later time, that would greatly ease the workload during the study since other assays (looking for apoptotic cells, for example) must be done "fresh" and constraints on bench space and time exist as well. Your input would be greatly appreciated! Sincerely, Doug Reed Douglas S. Reed, Ph.D. Microbiologist Department of Aerobiology and Product Evaluation Division of Toxinology and Aerobiology U.S. Army Medical Research Institute of Infectious Disease 1425 Porter St. Ft. Detrick Frederick, MD 21702-5011 301-619-6728 301-619-2541 fax Doug.Reed@det.amedd.army.mil
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