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FAQ's

How do I get a Sort?

  1. Get in touch with Bob to set up a time to sort:
    Robert Karaffa (rkaraff@emory.edu)
    Sommer Durham (sdurha4@emory.edu)
    Voice: 404-712-4429

    We will discuss your experimental designs as they relate to the use of the sorter. A mutually agreeable date and time will be set.
  2. The price of sorting is listed here
  3. What will I need to get my cells sorted?
    1. Your sample should be concentrated to approximately 1 X 10e8 cells/mL.
    2. We will need a single-stained sample of all the fluorophores you would like to use (positive controls), including an unstained sample (negative control). We will do a short test-sort using a small aliquot of your sort sample to verify the sort set-up.
    3. You should test your samples on an analyzer cytometer prior to the sort (FACSCalibur, FACScan, etc.). This will give an idea of the percentage of positive/negative events, quality of stain, etc.
    4. A small amount of your sorted sample (~100uL) will be taken for post sort analysis.
  4. The speed of the sort will be determined by the concentration of your cells. The general rule is:
    1000 events/sec = 1 X 10e6 cells/ml.
    2 ml of cells at a concentration of 2 X 10e7/mL will take 30 minutes to an hour to sort.
  5. If you have further questions, please direct them to Bob Karaffa at the above contact info.

Start Up Procedures for the Analyzers

  1. Turn on the cytometer.
  2. Check the cytometer
    1. Is the Sheath tank full?
    2. Is the Waste tank empty?
    3. Flip the pressure toggle* 
      *black toggle switch between the Sheath and Waste tanks.
    4. Put a tube of dH2O on the sample introduction probe (SIP), put the machine in "Run" mode and allow to warm up for 2-3 minutes.*
      *This will be about the amount of time that it will take the computer to start up
  3. Turn on the computer and LOG IN (use your login/password issued at training).
  4. When you are logged in and the computer and cytometer are connected,
    YOU ARE READY TO GO!

What if the machine won't connect to the computer?

  1. Go through the start up procedure, again.
  2. Try this at least 3 times
  3. Call for help.

What if I get a clog?

  1. Remove your sample
  2. Put a tube of dH2O water on the SIP and hit "Prime"(FACSCalibur) or "Backflush" (FACSort/FACScan) 
          Look for bubbles in the tube.
          Do this 2X.
  3. Try your sample again.

Still Clogged?

  1. Prime again
  2. Run the cleaning sequence with 10% bleach and water.
  3. Try your sample again.

Still Clogged?

  1. Get help.

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