1. Calibration is performed prior to each acquisition with CaliBRITE 3 beads and addition of CaliBRITE APC beads, according to the manufacturer's protocol.
2. FACSCalibur is prepared for acquisition by the launching of Cell Quest Pro software. After the flow sheath and waste levels are checked (recommended), the appropriate acquisition template and instrument settings are selected.
3. Before acquisition of samples, standardization is performed with QuantiBRITE PE beads, which are reconstituted with 1 mL of flow PBS. Five thousand events are acquired on the FACSCalibur. An example of a result is shown in Fig. 1.
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