ChemoMetec NC-3000 - Manual (Page 8)

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Enter an estimated compensation factor into the matrix box associated
with the two channels of interest and click ‘Preview’. Inspect the cell pop-
ulation to ascertain that cell population is spread horizontally across the
graph area. If the compensation 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 steps 1-7 for each individual fluorophore.
If multiple samples have been analyzed, enter the row number of the sam-
ple with the corrected compensation factor into the ‘Master Row’ box situ-
ated 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 fluo-
rescence into the orange channel. An increase in orange fluorescence is seen
as green fluorescence increases.