Bio-Rad S3™ Cell Sorter - Manual (Page 62)

52 | S3 and S3e Cell Sorters
Acquisition
Note: Cycle mode can be enabled or the refresh button can be used to refresh the data
displayed on the screen as adjustments are being made.
When running multicolored experiments, compensation is necessary. In order to use the Auto
Compensation wizard, single stained compensation control samples must be saved in addition
to the multicolored stained samples.
13. Run single stained compensation control for first parameter.
14. Set event limit high enough to visualize positive population; collect and save file.
15. Repeat and collect samples for each parameter.
16. Use the Auto Compensation wizard to set compensation for multicolored samples.
Refer to Section 5.2.1, Auto Compensation Wizard, for instructions on compensation methods.
Depending on the fluorophores used in the experiment, filters may need to be optimized in
order to capture signal properly. Please refer to Section 5.4, Optical Filters, for information on
how to create custom filter configurations.
Refer to Section 3.2, Control Panel, for an overview of the instrument control functions.
5.1.1 Target Event Rate
The event rate is related to the sample concentration. Generally, for every 1,000,000 cells per
ml sample concentration, the event rate may be increased by 1,000 events per second. As an
example, if the sample is concentrated to 10,000,000 cells per ml, the maximum event rate
should be 10,000 events per second.
The target event rate is dependent on the trigger and threshold settings. A target event rate
can be entered into the event rate box in the instrument control panel (Figure 54). The software
will maintain the entered rate throughout the acquisition or sort if the sample concentration is
appropriate for the target event rate. The actual target event rate is shown to the right of the
target event rate and on the status bar at the bottom of the software screen. The maximum
target event rate while acquiring is 100,000 events per second.
The button for preset sample event rate can also be used. The sample pressure is preset for
low/med/high rates. The sample rate will be dependent upon the concentration of the sample.
Fig. 54. Target event rate.