This is one of the few discussion groups that is worth subscribing to, and it owes it success to the continued participation of real experts. It is always more interesting and illuminating when the experts don't agree. I thought Frank Tragonas had some interesting comments, and I eagerly await "Zen and the Art of (Old) Cytometer Maintenance." I am frequently called upon to review manuscripts containing flow data. In the past, my most frequent reasons for rejection of conclusions based on flow data have been bad compensation, randomly set integration gates, and no control for background fluorescence or non-specific staining. In the last year, I have been seeing a new problem that arises from multicolor analysis. Two parameter data are presented as a dot plot labeled by antibody or reagent, but not fluorochrome (e.g., tetramer-positive cells vs. interferon-gamma intracellular staining). However, that data is gated on CD3, CD8, and CD62L staining, none of which are shown. Moreover, the intracellular cytokine staining protocol uses monensin or some other inhibitor of protein export which affects traffic of all surface molecules including CD3, CD8, and CD62L. Often no information is presented on the fluorochromes used for these stains, but that hardly matters because the data are not shown. Minimal labeling of axes and display of all relevant data would make the interpretation of flow results possible. That now means FL1 .... FLn. Happy Holidays. Don Mosier _______________________________________________________________________________ Donald E. Mosier, PhD, MD Professor Department of Immunology, IMM-7 The Scripps Research Institute 10550 North Torrey Pines Road La Jolla, CA 92037, USA 858 784-9121 phone 858 784-9190 fax This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify Dr. Mosier by telephone or fax.
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