Dear Andy, You are correct..CFSE is difficult to deal with in a multicolor strategy until about 24 hrs post labeling. This is partially because it takes some time for the staining of the cytosolic proteins to become labeled in a stable manner. A better choice would be to use PKH26 as a red tracker and PKH67 as a green tracker. Since the PKH dyes bind to lipid moieties, the staining is instantaneous and stable. You can control the intensity by concentration of dye. We have used these (and CFSE) in a variety of multicolor protocols with success and feel that for shorter time periods the PKH family is better...for longer time periods where you need to assess cell division, then more caveats enter into the equation. good luck Jonni >Dear Flow-ers, >I have a friend who wishes to identify adoptively transferred >murine cells approx. 12 hours after injection. He would like to track 2 >separately labelled, or "tagged" cell populations, staining with >multiple cell surface markers after isolating the cells from the >recipient, but isn't interested in cell division. Because of the >experimental setup congenic markers wouldn't be suitable. Am I right in >thinking that CFSE would be too bright at this timepoint? >Any info would be greatly appreciated. >Thanks, >Andy > >---------------------- >Andrew Herman >Department of Pathology & Microbiology >University of Bristol >University Walk >Bristol BS8 1TD >Tel. +44 117 928 7511 >Fax. +44 117 928 7896 >A.Herman@bristol.ac.uk -- Jonni S. Moore, Ph.D. Associate Prof. of Path. and Lab. Med. Director of Clinical and Research Flow Cytometry University of Pennsylvania Cancer Center University of Pennsylvania School of Medicine 203 John Morgan Bldg. Philadelphia, PA 19104-6082 Phone: 215-898-6853 FAX: 215-898-4227
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