Training and guidelines
- Training is mandatory for all first-time users of the facility regardless of previous flow cytometry experience. Training is free (we only charge the hourly cost rate of the analyser).
- We help you with panel design using our experience in colour choices in order to get you on track faster and economically more efficient.
- A class II biosafety cabinet houses the BD FACS ARIA III U, for sterile sorting of Cat1/Cat 2 pathogens and only fixed Cat 3 pathogens.
Read the biosafety pages before using the facility for important information, and to download the forms required to use the facility.
Cell sorting service guidelines
Cell preparation for cell sorting
- Prepare your cells as you normally would for flow cytometer analysis (please consult us before purchasing your antibodies).
- To set up the machine, you need to provide 1x 105 cells/ml non-stained (negative control) cells in 0.5 ml volume.
- Provide single stained cells or comp beads to set up compensation in a minimum volume of 200 ul.
- For complicated multicolour stained samples, please also bring FMOs for gatings.
- It is essential to filter the cells through a cell strainer immediately before sorting. We provide the filter mesh needed for this.
- For sorting, labelled cells should be at 10 x 106 per ml in FACS Tubes in PBS (with 2-5% FCS Maximum, any more than this will cause the cells to clump more readily and cause the instrument to block).
- Two-way or four-way sorting: Cells are sorted into 1.5ml Eppendorf tubes, 5 ml FACS tubes, 15mL Falcon tubes containing medium(with antibiotics), serum, lysis buffer or Trizol.
- We can also sort into 96-well plates, 6 well plates, 12 and 24 well plates, microscope slides or dishes. For this, please state you require single cell sorting so that the Aria is set-up before your arrival.
- If you do not have the proper controls for your sort, we will highlight there is no guarantee regarding the outcome of the sort, during your session.
- You will still be charged for the set-up, cleaning and shutdown, but you are at liberty to cancel the sort at this point, so as not to incur any more unnecessary charges.
- Sort start times and end times will be strictly enforced barring any instrument malfunction.
- If you cancel less than 24 hours in advance, you will be charged for 50% of the time you reserved.
- Failure to inform us of a cancellation, or “no-shows” will be charged the full amount for the time booked.
Prior to using any of the facility instruments or equipment, a risk assessment form needs to be completed.