BD FACSAria Skrócona instrukcja obsługi - Strona 5

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stream flowing into a collection tube. In this event hit the Emergency Stop (big

red button) to stop the stream. After a clog you will have to clean the sort

chamber, remove the nozzle, and sonicate it.

4. If the software freezes, first minimize and maximize the DIVA software window,

if this doesn't work push the Emergency Stop button.

Sort in plates

1. Place the ACDU adapter under the sort chamber.

2. Select a custom device with the correct number of wells from the bottom of the

Device dropdown menu in the upper left corner of the Sort Layout.

3. Align the far-left stream so it will fall vertically as much as possible. (Usually ~18)

4. Calibrate>Custom Device. This will default to the custom device selected in the

Sort Layout.

5. Go to Home. Align stream so it falls in the middle of A1 using the arrows and

squirt button in the middle of the Home device screen. Set Home.

6. Go to Farthest. Align stream so it falls in the middle of H12 using the arrows and

squirt button in the middle of the Home device screen. Set Farthest.

7. Apply>Close

8. It is usually useful to do a test sort to make sure the sort arm is well calibrated:

sort 100 beads or unstained cells in each corner of the plate or randomly assign

wells to be sorted all over the plate to test accuracy.

Cleaning and Shut Down (NEVER USE CONTRAD)

1. After the experiment, run bleach 10% for 5 minutes at flow rate of 11.

2. If you are the last user of the day, you must shutdown the instrument.

3. Select Fluidics Shutdown in the Instrument menu: as indicated, take out the

nozzle tip (there are no closed loop flow nozzles, ignore this step). Disconnect

the tubes from the Sheath tank and plug them on the EtOH tank and put a tube

of 1.5% Citronox in the sample loader. Follow the software instructions.

4. After the shutdown is complete, turn off the software and turn off the

instrument.