7
Enter an estimated compensation factor into the matrix box associated
with the two channels of interest and click ‘Preview’ or use the scale bar
for live updated adjustments. Inspect the cell population to ascertain that
cell population is spread horizontally across the graph area. If the compen-
sation factor is incorrect, click ‘Roll back’, enter a new value and inspect the
cell population again. Repeat this procedure until the correct compensation
factor is ascertained and click ‘Apply’.
Repeat step 1-7 for each individual fluorophore.
If multiple samples have been analyzed, enter the row number of the sample
with the corrected compensation factor into the ‘Master Row’ box situated
in the top left-hand corner of Plot Manager and click ‘Apply’. This will replicate
the compensation in all other samples selected in Plot Manager.
Tip:
after the final compensation has been set, a row with analysis including
all compensation can be used as master row in the setup of the protocol
(see Template Setup).
Figure 5
.
Scatter plot of cells expressing GFP showing spillover of green
fluorescence into the orange channel. An increase in orange fluorescence is
seen as green fluorescence increases.
