sorting stream

Training

  • 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.

Please complete the form FACS user agreement (Word)  and send an electronic copy to Yanping Guo (yanping.guo@imperial.ac.ukto arrange the training and cell sorting service. 

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).

Cell collecting

  • 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.

 Policies

  •  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.

Risk assessment


Prior to using any of the facility instruments or equipment, a risk assessment form needs to be completed.

Please see Project registration and risk assessment for biological material (Word)