Q1. How can I start using your facility?
In order to get access to the scheduler you will need to fill out the Work Authorization Form. Once complete, you can return to the address listed at the top of the form. I normally like to do instrument training with actual samples. That way we can tailor the training specific to your assay and set up a template and instrument settings with you. Once you know when your cells will be ready send me an email email@example.com and we will set up the appointment.
Q2.How can I get access to facility instruments after hours?
Once you are fully trained on a particular instrument you are eligible for after hours access. In order to get your ID card activated you need to go to the core facilities administration office located at SOM 5C124. Terra Curley (3-2926) or Audrey Grisham (3-2928) will need your ID number and also the 6 numbers following the 2* on the back of the card
Q3. How can I get involved with the Flowjo Site License?
***Please note that billing is now being done for all REGISTERED computers. It doesn’t make a difference if you are using your copy or not. If you have it registered, you will get billed.***
Please follow the steps below to register your computer.
1. Install Flowjo on a computer. Follow this link FLOWJO. Go to Download–>For Windows or Download–>For Mac. If you are using windows I recommend the newest Version X. If you are using a Mac I still really like the Version 9 or you can use the new Version X. Both downloads are available on that page. And you can have both versions downloaded if you want.
Once it is installed it will ask you to install a serial number after you agree to the license agreement. At the bottom of that screen will be the hardware address that needs to be registered.
2. To Register, follow this link ( http://www.flowjo.com/FLS/registration/utah.html ) DO NOT add spaces or colons when you enter your Hardware address.
Once registered, Install the Serial Number R596cZ4aYgpE9554. Once the serial number is installed, the program should work immediately. There may be issues with your firewall or proxy server (if you have one) at first. If you experience any problems using the software after registration please call or email me and I will get it worked out for you.
PRICING!!! We will hopefully have enough copies at the end of the year so that it comes out to about $250/yr. Basically how this works is that they send me a bill once a year and then I pay them the full amount. Then go out and collect from the registered users.
Please feel free to contact me with any questions that you may have.
Q4. Which cell sorter should I use.
Q5. What do I need to bring to sort cells?
1. Samples- Your samples can be in 5ml tubes or 15ml conical. You should aim for a concentration around 10 million/ml for cell lines or 15 million/ml for lymphocytes. If you don’t have nearly that many cells, don’t resuspend in less than 500ul. Buffer can be anything that the cells are happy in. More detailed info here
2. Controls- For fluorescent protein work we need an untransfected sample to set up the instrument. For multicolor experiments we need unstained in addition to single color controls.
3. Collection Tubes- We can collect into eppendorf, 5ml, 15ml 50ml or 96well plates. Each collection tube should have collection media already in the tube.
4. Extra media- In case there is a clog and a sample needs to be resorted.
5. Jump Drive- If you need your data files
6. Ice- If appropriate.
Q6. How should I acknowledge the facility in my publication?
The most important acknowledgement for the facility and university is referencing the shared instrumentation grant # IF YOU USED THE CELL SORTER for any work related to your publication. Something like this would work.
“Research reported in this publication was supported by the National Center
for Research Resources of the National Institutes of Health under Award
Also referencing the NCI Award Number 5P30CA042014-24 is helpful.
Q7. Whats the deal with this 96well single cell sorting business?
That is correct, we can sort single cells into a variety of 96 well plates, and also 384 well plates. This can be accomplished on the 5 laser Aria or the 4 laser Aria. The setup where we target the wells takes about 5-10 min and then it takes about 90 seconds to run a plate. A single cell will come out in about 2.3nl (very tiny droplets!!) so you definitely want to have media already in your plates. And there is generally no need to spin and wash when you are done.