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

1. Turn off sample stream.
2. Remove probe from the 10L sheath tank and place into 10L EtOH tank.
3. Select "Prime after tank refill" (select sheath tank from the menu, repeat 2x).
4. Select "Fluidics startup" (this will flood plenums with 70%EtOH).
5. Start stream and load 70%EtOH sample, run sample for 5 min.
6. With stream still on, unload sample, shut down instrument by quitting the DiVa program.
7. Turn off the power to the instrument.

Aria Startup Procedure

1. Power up DiVa, turn on computer and start DiVa program.
2. Remove cart filter and replace with shunt.

Completely drain 70%EtOH from filter by loosening top stopcock

1. removing bottom stopcock
2. and unscrewing bottom outlet line
3. from filter, leave under Laminar flow hood and let dry until next day. See figure 9 below.
• Remove probe from 70%EtOH tank and let drain. Place probe into 10L sheath tank.
• Select "Prime after tank refill" (select sheath tank from menu, repeat 2x).
• 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.
• Select "Prime after tank refill" (sheath tank, repeat 2x).
• Loosen stopcock on cart filter and wait for sheath to run out (approx. 1 min).
• Run &quyot;Fluidics Startup", repeat 2x.
• Turn on stream.

Figure 9. Sheath Filter:
A= top stop-cock, B= lower stop-cock, C= Bottom outlet line

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