Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking DyesJoseph D. Tario Jr. 1, Kristen Humphrey 1, Andrew D. Bantly 2, Katharine A. Muirhead 3, Jonni S. Moore 4, Paul K. Wallace 1
1Department of Flow and Image Cytometry, Roswell Park Cancer Institute, 2Flow Cytometry & Cell Sorting Resource Laboratory, University of Pennsylvania , 3SciGro, Inc., 4Department of Pathology and Laboratory Medicine, University of Pennsylvania
Successful use of cell tracking dyes to monitor immune cell function and proliferation involves several critical steps. We describe methods for: 1) obtaining bright, uniform, reproducible label-ing with membrane dyes; 2) selecting fluorochromes and data acquisition conditions; and 3) choosing a model to quantify cell proliferation based on dye dilution.