BIO RAD 145-1001, Instruction Manual

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S3 Cell Sorter Instruction Manual | 65
Sorting
6
Sorting cell populations is highly dependent on several factors such as cell type, population
frequency in the starting sample, cell concentration, sample preparation, and sort logic
definitions. It is very important to start with a single cell suspension with minimal amount of
dead cells or debris contamination.
6.1 Sort Setup
The majority of sort setup procedure is automatic and is completed during daily QC. When
ready to sort, the desired populations must be selected using regions and gates. At this time
sort logic definitions must be set.
To start a sort:
1. Click
File > New Protocol or Open Protocol .
2. Click Home > Histogram or Density to add plots to the workspace.
3. Adjust temperature control for sample input station and sort collection area if desired.
4. Place filtered sample into the sample input station.
5. Move the loading stage into the run position.
6. Click
Start on the instrument control panel to acquire a small amount of the sample.
7. Set sorting regions and gates on plots with populations of interest.
8. Right click on the desired region to be sorted and select sort direction.
9. Repeat with a second region if two-way sorting is desired.
10. Click the
Sort Logic dropdown menu to select the collection vessel of choice.
11. Define the sort logic parameters in the Sort Logic window (Figure 70) by setting the sort
mode (purity, enrich, or single) and event limit for each collection position.
12. Define sort collection tube volume if you want to track collected volume to prevent
collection overflow.
13. Place sort collection vessel in the sort collection device at positions 1, 2, etc.
Note: 5 ml tubes should have at least 0.5 ml media and 1.5 ml tubes should have at least
0.1 ml filtered media to collect and cushion the sorted cells.