FAQs

Flowcore FAQs

• Q. What type of buffer should I prepare my cells in?

  A. All samples should be in FACS buffer. FACS buffer is PBS + 1-5% of protein of your choice (i.e., FBS, BSA, or serum) + 2mM EDTA (This helps keep the cells from forming clumps.)
  

• Q. Do my cells need to be filtered?

  A. Yes, all cells should be filtered before running on the machines. Cells run on the Fortessa should be run through at least a 70um mesh. Cells run on the sorter should be run through a 35um or 40um mesh. 5 ml round bottom tubes with strainer caps ( Falcon Tube #2235) are a great option. The core does not provide these tubes, but we do have some available in case you need them.
  

• Q. At what concentration should my cells be prepared?

  A. For the Fortessa: Your cells should be at a concentration of 20 million cells per mL. The smallest volume that is recommended for the Fortessa is 200uL. 

  For the Aria: Your cells can be at a concentration of 40 million cells per mL. The smallest volume that is recommended is 200uL. We always recommend that you bring extra FACS buffer so we can dilute the sample if we need to. It is easier for us to dilute than concentrate the sample as we currently do not have a centrifuge.
  

• Q. Do I need to bring a negative control and single-color controls for every experiment?

  A. Yes! It is very important to have a negative control so that we can see the background of your system. Single color controls are also very important as they help us to determine the amount of individual dye present on each cell. Without these proper controls it will be almost impossible for us to determine what is positive and negative and your true populations of interest.
  

• Q. If there are no available appointments, can I "squeeze" into the schedule?

  A. No. Everyone's experiment is treated the same. The lab is run on a first come, first-served basis. You are always welcome to check to see if anyone thinks they will finish early or is no longer able to use their time.

• Q. What if someone is using the instrument during time that I reserved?

  A. Any user has the right to insist that their experiment begin according to the published schedule. They may choose to give up some of their time, but it is not required.

• Q. Can I book an experiment after 5 p.m. or on the weekends or holidays?

  A. There are no Flow Facility staff in the lab after 5 PM to run samples. Investigators who need staff assistance beyond 5pm are encouraged to speak with staff in advance to coordinate a schedule. Independent users of the Fortessa or BioPlex can use the lab after-hours. Please refer back to our general policies (Link) to see how to apply for after-hours use.

• Q. Can I run the instruments without the flow operator's assistance?

  A. Independent users of the Fortessa and the BioPlex can use the instrument independently. Those who would like to be independent users of the Fortessa or BioPlex should sign up for training to become independent users, please see our Training page.

• Q. Do I need to put buffer in my collection tubes?

  A. Yes, it is strongly recommended that you do put buffer in your collection tubes. Your buffer can be anything that you choose for example, media, FACS buffer or RNA lysis buffer. Please be aware that any buffer you choose will be diluted by PBS from the instrument. For specific questions about how much it will be diluted please speak with staff.