General Procedures
Decontamination Procedure (FACSAria)
Pre-requisite instructions
- Remove the bubble trap filter (see FACSAria instruction manual). This filter is used to remove micro-bubbles, however it will also act as a bacterial reservoir and trap and must be removed before using this procedure. A shunt can be clipped in place of this filter.
- 70% EtOH is stored and used in a 10L tank.
- Sheath tanks are autoclaved prior to use (weekly)
- Pre-packaged sterile sheath (PBS with 50mg/l of Gentamycin) is obtained in 20 liters IV bags (HyClone, http://hyclone.com). This is dispensed directly into sterilized 10L sheath tanks.
- Only four 10L tanks are used, one for the Sheath, one for the 70%EtOH, one for the waste and one in reserve. No other tanks are used in this procedure.
Aria Shutdown Procedure
- Turn off sample stream.
- Remove probe from the 10L sheath tank and place into 10L EtOH tank.
- Select "Prime after tank refill" (select sheath tank from the menu, repeat 2x).
- Select "Fluidics startup" (this will flood plenums with 70%EtOH).
- Start stream and load 70%EtOH sample, run sample for 5 min.
- With stream still on, unload sample, shut down instrument by quitting the DiVa program.
- Turn off the power to the instrument.
Aria Startup Procedure
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Power up DiVa, turn on computer and start DiVa program.
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Remove cart filter and replace with shunt.
Completely drain 70%EtOH from filter by loosening top stopcock
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removing bottom stopcock
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and unscrewing bottom outlet line
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from filter, leave under Laminar flow hood and let dry until next day. See figure 9 below.
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Remove probe from 70%EtOH tank and let drain. Place probe into 10L sheath tank.
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Select "Prime after tank refill" (select sheath tank from menu, repeat 2x).
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Replace shunt with 70%ETOH filter, which was drained from previous day. Note: Two ETOH filters are in rotation; one in use for the day and one drying from the previous day.
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Select "Prime after tank refill" (sheath tank, repeat 2x).
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Loosen stopcock on cart filter and wait for sheath to run out (approx. 1 min).
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Run &quyot;Fluidics Startup", repeat 2x.
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Turn on stream.
Figure 9. Sheath Filter:
A= top stop-cock, B= lower stop-cock, C= Bottom outlet line
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