Immunology Flow Cytometry Flow Cytometry IM-SOP#015

Rm 2D205

SAFE OPERATING PROCEDURE FOR
FLOW CYTOMETERS
BD FACSCanto II, BD FACSAria, BD Accuri C6 and AMNIS ImageStream X
Located in Room 2D 205
DOC # IM-SOP#015
S.O.P NUMBER / RISK ASSESSMENT NUMBER / RISK ASSESSMENT
Flow Cytometry
IM- SOP#015 / Flow Cytometry
IM-RA#015 / Low
DATE CREATED / REVIEWED / Employees Involved in SOP Creation / CONTACT PERSON
31.07.13 / Sheree Bailey/Nick vanHolst / Nick van Holst
Personal Protective Equipment Required
Closed footwear is to be worn in the Flow Cytometry Facility at all times
Before Work Commences
Users must have received authorised and documented training.Training is organised throughFlow Cytometry Unit.Contact Nick van Holst, ext. 64958
You must have read and understood this SOP and the associated Risk Assessment (Flow Cytometry IM-RA#015).
Instrument User’s Guides are available for each instrument and are located in Rm 2D205.
Hazards
Postural damage - Risk Assessment LOW
Eye damage from Laser exposure - Risk Assessment MEDIUM
Electrical shock - Risk Assessment MEDIUM
Exposure to Biohazardous reagents - Risk Assessment MEDIUM
Injury by Moving parts – Risk Assessment LOW
Risk of damage to equipment
1-Risk of damage to FACSAria flow cell.
Prevention: Ensure the laser beams are disabled when the stream is turned off.

TO USE

.You must comply with the “before work commences” criteria on the previous page

.Follow instructions below for booking the equipment, use of the flow cytometers and additional SOP requirements as described below.

.Do not attempt to modify any of the equipment. Maintenance to the equipment is only to be performed by authorised personnel (Flow Cytometry Facility staff and BD Biosciences service personnel).

1. BOOKINGS AND CANCELATIONS:

You must follow the rules for booking of the flow cytometers to ensure the flow cytometers are available for use;

.You must book your session.

Bookings are made online at:-

.If you can’t make it to a session that you have booked, it is your responsibility to ensure you have notified flow cytometry facility staff that your session is cancelled to allow other users availability

.The maximum time you may book the equipment for is 2.5 hours.If the equipment is available for a longer period on the day of use you may continue to use it.

2. USE OF THE FLOW CYTOMETER

Note: This document does not constitute a manual or training guide. Users of this equipment must receive authorised training and use this Safe Operating Procedure in conjunction with the Instrument User’s Guide and the risk assessment for the appropriate instrument.

.Check the Sheath and Waste levels before turning on the Flow Cytometer.

.Turn on the Flow Cytometer.

.Launch the BD FACSDiva (FACSCanto II and Aria), CFlow Plus (Accuri C6) or Inspire software and perform a fluidics start-up.

.Turn on the stream if required, verify the instrument configuration and optimise the instrument settings for your experiment.

.Create an experiment folder and workspace for your experiment.

.Install the sample tube onto the flow cytometer and acquire the data.

.Record and analyse the data.

.Export your data to the “//fusafmcf01/labs/Flow Cytometry” server.

3. ADDITIONAL SOP REQUIREMENTS FOR USE

.Maintain correct posture whilst working at the flow cytometer affiliated computer (refer to )

.Ensure optics covers and laser shields are installed, and instrument doors are closed during instrument operation. Under these conditions the instrument poses no danger of exposure to laser radiation.

.The nozzle, stream-charging wire and deflection plates in the FACSAria conduct electrical charge when the voltage is engaged. Do not touch these components when the voltage warning light is illuminated.

.To prevent exposure to biohazardous reagentshandle all biological specimens as if capable of transmitting infection. Expose waste container contents to bleach by adding bleach to the container immediately before disposal.

.The BD FACSAria can produce aerosols around the sample stream. When acquiring biohazardous samples, keep both the sort block and sort chamber closed during sorting andactivate the Aerosol Containment Unit next to the BD FACSAria.

.Sample tubes are to be disposed of in the biohazard bins or removed from in an appropriate container.

.Keep your hands and clothing away from moving mechanical parts to prevent injury to hands or fingers.

.Precaution labels appear on the Flow Cytometers to indicate a potential hazard.

.Copies of clearances from the relevant committees will be requested for projects involving the use of biohazardous or genetically modified organisms.

Fusafmcf01/Labs/Immunology/Lab General/Equipment-SOP/SOP List/IM-SOP#015