Supplementary Figure 5: Flow-cytometric analysis of control samples and gating out autofluorescence.
From: A high-throughput in vivo micronucleus assay for genome instability screening in mice
Unstained samples are used to centralise the main cell population within the side scatter (SSC) versus forward scatter (FSC) plot (data not shown). The main population is gated (erythrocytes), and within the propidium iodide (PI) versus FITC-conjugated transferrin receptor (CD71-FITC) plot a quadrant gate is drawn. Single-stain controls are then analysed to determine the correct voltage and gain settings, which ensures the majority of events fall within the scale of the X and Y axes, and within the correct quadrant gate. a) Unstained control sample and b) CD71-FITC alone. Auto-fluorescence can be detected with samples stained with PI alone, either within the transferrin receptor FITC-conjugated (CD71-FITC) channel (c; indicated by arrowhead), or within the higher logarithmic scale (Log) of PI channel (d; indicated by arrowhead). To remove the effect of auto-fluorescence, Region gate R3 is created within the PI versus side scatter (SSC) plot (d).
