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FAQ's
How do I get a Sort?
- Get in touch with Bob to set up a time to sort:
Bob: 404-712-4429
rkaraff@emory.edu
pager 404-686-5500, 14161
We will discuss your experimental designs as they relate to the use of the sorter. A mutually agreeable date and time will be set.
- The price of sorting is listed here
- What will I need to get my cells sorted?
- Your sample should be concentrated to approximately 1 X 10e8 cells/mL.
- 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.
- 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.
- A small amount of your sorted sample (~100uL) will be taken for post sort analysis.
- 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.
- If you have further questions, please direct them to Bob Karaffa at the above contact info.
Start Up Procedures for the Analyzers
- Turn on the cytometer.
- Check the cytometer
- Is the Sheath tank full?
- Is the Waste tank empty?
- Flip the pressure toggle*
*black toggle switch between the Sheath and Waste tanks.
- 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
- Turn on the computer and LOG IN (use your login/password issued at training).
- 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?
- Go through the start up procedure, again.
- Try this at least 3 times
- Call for help.
What if I get a clog?
- Remove your sample
- 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.
- Try your sample again.
Still Clogged?
- Prime again
- Run the cleaning sequence with 10% bleach and water.
- Try your sample again.
Still Clogged?
- Get help.
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