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Welcome to the Jenner Flow Facility

The Jenner has a busy flow cytometry facility catering for over 100 scientists. For charging purposes the facility is registered with the University as a small research facility (SRF). It is currently equipped with one cell-sorter (Beckman Coulter MoFlo Legacy) and two analysers (one Becton Dickinson LSRII and a Beckman Coulter Cyan).

Cell sorting is provided as a service to users but the analysers can be used after a short training session.

As part of your access to the facility you will need to register yourself (see below for more details) and read the information provided in this pack for your own safety and ease of use.

Flow Facility User Information Form

When you first come for any kind of training in the facility you must bring the ‘User Information Form’ with you (click here to open). This allows a record to be kept of your details, which will need to include a grant code, for charging purposes, to enable you to use the flow facility.

Flow Facility Booking

Once training has been completed then you will be able to book time on the instruments using the web based Calpendo booking system, available at http://jenner.calpendo.com.

When you first use Calpendo you will need to register yourself using the above link. The grant code you provided in the ‘User Information Form’ will be associated with your Calpendo account. Flow usage charges are worked out on an end-of-month basis using Calpendo so please make sure your timings are accurate.

Flow Facility Charges

The current charges for the Jenner Flow Facility are given in the following table:

 

Machine and Location Use Rate per Hour  

Use of machine

 

Samples run by operator

(Drew Worth)

LSR II - ORCRB £50.00 £100.00
Cyan - ORCRB £50.00 £100.00
MoFlo - ORCRB N/A £100.00

 

Flow Facility Biosafety Risk Assessment

If you intend to do any Flow cytometry (sorting or analysis) you will need to make sure a Risk Assessment form for each separate project you undertake has been completed. The assessment takes into account the potential hazard of the cells, microorganisms, vectors, stains and chemicals that may be present in the samples. Once a form has already been submitted for a project you only need to know the risk assessment number which can then be entered into the Calpendo booking calendar or Sort request forms. It is not necessary for each individual in a group to risk assess the same type of sample.

Guidelines for using the Jenner Flow Facility

Training

Only trained staff may book and use equipment in the facility. Training can be arranged with the Flow Facility Manager - Drew Worth. An initial introductory session on the LSRII or CyAn ADP takes approximately 1.5 hours. Following training, users will be asked to supply a suitable grant code that will be entered into the Calpendo booking calendar for their FACS charges to be made against. Once an account has been set up in Calpendo then they will be able to use the facility equipment.

Booking instruments and sorts

To book an instrument please use Calpendo. There is high demand on the LSRII so it is essential to accurately estimate the time required to complete the work on this instrument. If in doubt please consult the flow facility manager for guidance. It is important to arrive in the facility on time and complete the work promptly. Users will be required to stop work immediately at the end of a booked slot if the next user is waiting. If it is necessary to cancel a booking this must be done at the earliest opportunity which may allow the slot to be re­booked by another user. Please inform other potential users of the cancellation by sending out an email to jennerflow@maillist.ox.ac.uk. For cell sorts it is necessary to liaise with the facility manager, Drew Worth, as his time will be required to perform the sorting of your prepared samples. There is also a ‘Sort Request Form’ (see appendix) to be completed.

Sort request forms

Sort request forms should be submitted to the facility as far in advance as possible, however due to the demand on the sorter the reservations must be as accurate as possible. Early notification of cancellation is essential and if a session is not used without prior cancellation it could result in charges being applied (see Jenner Flow Facility charges given in the outline pages). No sorts will be performed unless a risk assessment number is entered on the request form. The request form should include details of the panel of reagents used, the number of cells to be sorted and other details that will enable the facility manager to configure the cell-sorter accordingly. It is highly recommended that pilot experiments are carried out on the cell analysers to optimise conditions and ensure that the target cell populations can be resolved properly. Examples of staining profiles and gates should be attached to assist with instrument set up. It may be necessary to double the concentration of staining antibodies when running samples on the MoFlo as compared to the analysers as the MoFlo is slightly less sensitive, due to it being a stream-in-air system.

Flow Facility Safety Policy

The facility operates the same standard policy as the rest of the Jenner. In summary, when using the cytometers, lab coats and gloves must be worn. Operators loading samples must also wear eye protection. More detail can be found in “Flow Facility PPE Policy”, later in this document.

Biosafety

Risks associated with processing cells through the cell analysers (LSR II, Cyan ADP) are different to risks associated with cell sorter (MoFlo). The former are essentially closed systems and bio-containment is possible whereas cell sorters produce aerosols and containment is more difficult and complicated. Risks should always be minimized, so cells should be fixed prior to analysis. If fixation can be shown to have an adverse effect on data quality, it may be possible to analyse fresh cells subject to a risk assessment. The majority of sorts carried out require the cells to be unfixed. Mouse cells are normally pathogen free and associated risks are low. However in vitro manipulations or transformations may increase the associated risks of accidental contact and should be assessed carefully. Cells from non-human primates and from un-screened human samples are considered high risk and should only be handled on a sorter utilising appropriate bio-containment measures (normally CL3 or equivalent). Screened human samples may be handled at CL2 with additional aerosol containment precautions.

Flow Facility PPE Policy

The Flow Facility is essentially a clean area but the wearing of gloves, lab coats and appropriate eye protection is mandatory. All cells must be fixed1,2 prior to assay and consequently this minimises biological risks, however it is possible that hazards may remain in the form of chemicals and stains used (i.e. Propidium iodide, bromodeoxyuridine, formaldehyde etc.) or contamination found on the outside of tubes. Although this will not be the case for the majority of samples processed, there is a theoretical risk of contaminating equipment, keyboard, mouse etc. The wearing of gloves will not prevent this but the proper use of gloves and lab coats will minimize risks to individuals and prevent other areas being affected.

  • If possible, dispense cells into tubes prior to bringing them into the facility. (If it is necessary to dispense samples into tubes inside the facility do this away from the keyboards and wipe up any small spillages with a suitable disinfectant.)
  • Ensure that the outside of tubes are clean.
  • Carry the samples into the facility in suitable racks or boxes
  • On entering the facility wear a lab coat, suitable gloves and eye protection
  • Remove gloves before touching designated clean areas (e.g. telephone)
  • Contact the facility manager in the event of a spillage onto the equipment and decontaminate the affected area.
  • The flow facility manager will provide PPE to non-scientific staff (ie. IT staff) and engineers so that work can be carried out safely on the equipment

1. A suitable fixative known to be effective in eliminating infection risks from biological agents that may be present in the samples

2. Unfixed cells may only be assayed following a risk assessment. The Flow Facility Manager must receive a copy of the risk assessment and approve the work prior to it starting in the facility. A designated cytometer must be used and an appropriate disinfectant must be used to decontaminate waste.