Bio-Rad S3™ Cell Sorter - Manual (Page 61)

Instruction Manual | 51
Sample acquisition can be done independent of sorting. As a sample is run through the system,
data are gathered and can then be saved in FCS 3.1 format. These files can be loaded and
analyzed using third party software compatible with FCS 3.1 files. Often, setup samples — for
example, unstained samples or single-color compensation samples — are acquired prior to
sorting the actual sample of interest.
5.1 Acquisition Setup
Prior to running a sample, a protocol must be loaded or created. At least one histogram or
density plot is required to begin acquisition. Protocols can be created new or loaded using
a protocol file or from a previously acquired FCS file. Refer to Section 4.5, Protocols and
Workspace, for information on how to create or open protocols. Access all instrument controls
via the ProSort
™ instrument control panel.
To set up an acquisition or sort:
1. Create or open a protocol.
Note: At least one histogram or dot plot must be created to continue.
2. Load the sample onto the sample input position and move the loading stage into the
run position.
3. Detectors can be renamed for each parameter based on fluorophores or markers used.
4. Check cycle mode to allow refresh of the data displayed.
5. Define target event rate, trigger parameter, and threshold value.
6. From the instrument control panel, click the Start Sample Acquisition button.
7. Adjust PMT voltages to place populations.
8. Adjust target event rate, the trigger parameter, and threshold value.
9. Stop sample acquisition by pressing the same button.
10. Check and define event limit. Gate Limit can be chosen. Cycle mode can be unchecked at
this time.
Note: Data can be displayed in hyperlog if the compensation box is checked.
Optional: AutoSave can be enabled to automatically prompt to save file after event limit is reached.
11. Re-collect sample and save FCS file if AutoSave is not enabled.
12. Unlock and remove sample tube. Move the loading stage into the wash position before the
next sample is run.
Acquisition
5