Hello I have a microscopy/staining problem. I have CFSE labeled T cells and transferred them into hosts which were subsequently infected with Listeria monocytogenes. I then made spleen sections which were stained for: 1) anti B220 followed by anti rat alexa fluor 350 anti CD3 followed by biotin anti hamster Ig followed by SA alexa fluor 594 With this combination I can visualize green transferred T cells (CFSE), blue B cells and red endogenous T cells using a multi-filter Leica digital light microscope. The second stain combo is 2) anti CD11b (Mac-1) followed by anti rat alexa fluor 350 anti Listeria monocytogenes followed by biotin anti rabbit Ig followed by SA alexa fluor 594 With this combination I can visualize blue macrophages and red Listeria monocytogenes, but I CAN NOT visualize the green transferred T cells. Any thoughts as to why the second stain combination negates/dilutes/quenches the CFSE labeled T cells that are visible in the first stain combination? Thanks, Angie MobleyReceived on Thu Dec 2 13:58:00 2004
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