Optimized Stem Cell Detection Using the DyeCycle-Triggered Side Population Phenotype

Figure 2

Impact of dye concentration and staining cell density on the separation of DCV-SP cells. A2780V cells were stained either with different DCV concentrations at a fixed cell density (i.e., 10⁶ cells/mL; upper panels) or with a fixed DCV concentration (i.e., 10 μM) at different cell densities (lower panels). Both a high DCV concentration and a low staining cell density promote the separation of SP cells, as outlined by the increasing sizes of the rectangular gates. Note that panels “10 μM” and “1 Mio/mL” correspond to each other (10 μM DCV and 10⁶ cells/mL, resp.). Importantly, improved separation of SP cells at higher dye concentration or lower cell number is primarily due to increased fluorescence of NSP cells in the long-wave range of DCV emission (“DCV red”) (bar graphs).