Bio-Rad S3™ Cell Sorter - Manual (Page 68)

58 | S3 and S3e Cell Sorters
Acquisition
6. Locate and select the single control FCS file for each parameter.
7. Select whether to use height or area for all parameters.
8. Click the forward blue arrow to open the Automatic Compensation Calculation Wizard
window (Figure 63).
9. Adjust the scatter, negative, and positive regions as necessary.
Note: The positive and negative population gates must not overlap.
10. Click the blue forward arrow to go to the next control.
11. Repeat steps 9 and 10 for remaining controls.
12. After all controls are adjusted, click the checkmark button. This will complete the automatic
compensation calculation.
13. Check the compensation box on the axis of the desired histogram or density plot to display
the compensated data and acquire or sort.
14. The compensation matrix may be viewed at this point if desired.
The hyperlog feature allows data that have values <0 to be correctly displayed. This data display
format is used only for compensated data (Figure 64). It converts a small portion of the scale
around zero to linear, and adds a small amount of negative log scaling to each access, preventing
negative events from piling up on the axis. This allows better visualization of compensated data.
Note: The area parameter is grayed out and cannot be changed because compensate is
checked. Compensation can only be applied to area or height depending on which selection
was chosen during auto compensation.
Fig. 63. Using the auto compensation wizard to adjust positive and negative gates.