Cell Tracking Using PKH-26

Marcia Woda

This figure illustrates the utility of the membrane labeling dye PKH-26 to determine the number of times a parent population has gone through the cell cycle. In this experiment, a fibroblast line was labeled with the red tracker dye and harvested 10 days later . Just prior to harvesting, the cells were exposed to 10 micromolar Hoechst 33342 for 30 min at 37deg C. Viable cells were run on a dual laser flow cytometer, with both the blue HO and red emmision collected in linear mode. Quantitative data about the number and percentage of active generations and proliferation index are determined by analysing the list mode data files with a proliferation modeler in conjunction with Modfit Software, Sigma, Inc and Verity House Software respectively. Addition of the HO dye allows for the cell cycle status of each generation.

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