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FIGURE 1. Representative flow cytometric analysis of V79 cells from multicellular cluster containing 50% unlabeled cells and 50% cells labeled with 4.25 mBq per cell of 131IdU. The cells containing 131IdU were dyed with CFDA-SE. (A) Side scatter (SSC) vs. forward scatter (FSC) dot plot that was obtained with FACS Vantage cell sorter. Debris and aggregates were gated out as indicated by rectangle denoted P1. (B) P1-gated SSC vs. FL-1 height. (C) Number and percentage of events in each gate relative to parent gate and mean FL-1 height. (D) Distribution of undyed (P2 gate; unlabeled) and dyed (P3 gate; labeled) cells within P1 gate. Analysis indicated that, within P1 gate, 46.2% of cells were undyed, 42.8% were dyed, and remaining 11% were outside P2 and P3 gates. Dyed (P3) and undyed (P2) cells were automatically and separately seeded by FACS Vantage cell sorter in known numbers into 6-well plates. Cells outside P2 and P3 gates were discarded to ensure high degree of certainty in purity of dyed and undyed populations.
