Bender MedSystems Technical Tipps for FlowCytomix

The Bender MedSystems Technical Service Team is always happy to answer any questions you may have about either the protocol of our FlowCytomix products or the information given here.

Contact: techserv@bendermedsystems.com

phone: +43 1 796 40 40-120

Please read these tipps carefully before you start:

1. Mixing:

When preparing the FlowCytomix bead mixture, pipette the bead solution all the way down to the bottom of the tube, so that you do not lose any material on the walls of the tube.

Inadequate mixing can lead to little or no beads being detected. Vortex the beads immediately before mixing with the standards or samples and again before analysis on the flow cytometer. Vortex each sample tube for 3-5 seconds before placing the tube on the flow cytometer as this will yield better discrimination of the bead populations in the FL3/FL4 channel.

2. To avoid double bead populations in the analysis plot:

When applying the FlowCytomix bead solution to the assay plate, put the beads into the standard- / sample solution in the well to make sure that the beads do not stick on the side of the tube/well.

(Beads which do not come in contact with the detection antibody during the incubation step will show a much lower PE signal. These beads will lead to a second, almost "PE-negative" population.)

3. To optimize FlowCytomix sensitivity:

It is critical that all steps after the addition of the detector reagents are protected from light to obtain maximum fluorescence of the PE reporter system.

When establishing the instrument settings with FlowCytomix Standard one (highest concentration), it is essential to place the bead populations at the very right margin of the acquisition plot. Thereby the distribution of all standard concentrations across all four decades in FL-2 will be optimized.

4. Blocked FlowCytomix filter plate:

Serum or plasma samples can sometimes block the filter plate during washing. If this occurs very carefully push a needle through the hole underneath the respective well of the plate. Do not stick the needle all the way through the filter as this will damage the membrane!