Bio-Rad S3™ Cell Sorter - Manual (Page 27)

Contaminant Testing
Instruction Manual | 17
Contaminant Testing
This protocol is designed to test the aerosol contamination levels that occur under normal
cell sorter operation while operating the biosafety system with only the aerosol evacuation
enabled (that is, without directional air flow). This test must be performed on an S3 biosafety
system that has already been installed enclosing the S3 or S3e cell sorter.
The suggested material used for this testing procedure is listed below and based on the
published protocol by Schmid et al. (2007). Please refer to this published protocol for preparing
the Glo Germs.
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Glo Germ Powder (Glo Germ Company, Cat. #GGP10, www.glogerm.com)
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A6 Single Stage Microbial Sampler (www.emlab.com)
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Microscope slides
Follow these setup procedures prior to performing the test. This will expose the cell sorter
completely, eliminating directional airflow of the Class I biosafety system.
1. Roll up both the front and the rear panels and lay them on top of the biosafety system.
2. Roll up both the left and right panels and lay them on top of the previously rolled panels.
3. Leave the main power on to the biosafety system.
4. Be sure to leave the evacuation hose attached to the rear of the cell sorter.
According to the published 2014 ISAC guidelines for cell sorting, this test and setup will fulfill
the requirements for verifying an aerosol evacuation system. Maintaining a log of containment
testing records is recommended in case there are any questions of biosafety and containment.
Testing Protocol
To set up an acquisition of the Glo Germ particles:
1. Create or open a protocol within ProSort software.
Note: At least one histogram or dot plot must be created to continue.
2. Place 4 ml diluted Glo Germ particles into a 5 ml tube.
3. Load the sample into the sample station.
4. Check Cycle Mode to allow refresh of the data displayed.
5. Define target event rate at 30,000 events per second (eps).
6. From the instrument control panel, click the Start Sample Acquisition button.
7. Set trigger to FSC and threshold value to 1.00.
8. Adjust PMT voltages to place populations on the screen.
9. Adjust target event rate, trigger parameter, and threshold value.
10. Stop Sample Acquisition by pressing the button.
11. Cycle Mode can be unchecked at this time.
Optional: AutoSave can be enabled to automatically prompt to save file after event limit is reached.
Note: Cycle mode can be enabled or the refresh button can be used to refresh the data as
adjustments are being made.