Research on the setting of optimal conditions and adjustment of sorting parameters for BD Influx flow cytometer

Objective: To explore the optimal conditions and adjustment of sorting parameters for BD Influx flow cytometer. Methods: Using Rainbow Beads and Accudrop Beads as experimental materials, optimization of pinhole, stream, fluorescence channel, forward scatter signal, drop frequency, piezo amplitude and drop delay was conducted. Results: The optimal condition was as follows, using the 100 μm nozzle, the stream was centered on the pinholes and sharply focused, the stream was aimed at the drain, the brightest signal was in the center of the pinhole after optimizing the fluorescence channel, the brightest signal was in the center of the FSC pinhole after optimizing the forward scatter signal, the breakoff point was highest after adjusting the drop frequency, and the droplet shape remained constant. When the piezo amplitude was around 5.20±1.00 and the drop delay value was around 35.20±2.00, the adjustment efficiency achieved its optimal condition, with its purity at 99% and sorting efficiency at 95%. Conclusion: Rapid setting of optimal conditions and adjustment of sorting parameters for BD Influx flow cytometer guarantees the success of gathering cells of high purity and activity after cell sorting.