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					MRC Clinical Sciences Centre Imperial College Faculty of Medicine Hammersmith Campus Du Cane Road London W12 0NN

RISK ASSESSMENT FOR A LABORATORY PROCEDURE
Technique Title

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IF staining. Staff involved

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All staff in room……………………………. Hazards involved

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Chemicals: PBS, BSA. DAPI stain. EtOH, Virkon Possible exposure to GMO’s and untreated Genetically modified waste: Biohazards: Manual Handling. Fire’s from HFC’s: RSI from pipetting: Cuts from glass slides: Route of exposure

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Skin, eyes, inhalation, ingestion. Harm

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Chemicals: The respective COSHH forms for the above substances must be read prior to this technique commencing so that their hazards and control measures are understood. Cuts: Broken, chipped or damaged glass slides can cut. Exposure to GMO’s: GMO’s could pose a threat to health. Biohazards: Cell lines need to be assessed for safety. Manual Handling: Overfilled waste boxes could be heavy and need care to be taken lifting them. Fire: Care needs to be taken when working with HFC’s as fires can stare easily in labs. RSI: Long term injury can result from using pipettes and other repetitive techniques.

Identify any persons and any special risks associated with the technique and steps that must be taken to minimise them.  Person carrying out the work: All chemicals, reagents and associated techniques used in this protocol will have their associated risks and control measures so their relevant COSHH forms should be read in conjunction with this protocol. There is a small risk of exposure to viable GMO’s but staff have been trained to work safely with GMO’s and biohazardous material and how to properly clean up spills should they occur. Glass slides can cut and in some cases inject their contents into the resulting wounds. GMO waste boxes will not be overfilled and so will not be excessively heavy; staff will be shown how to safely use centrifuges when they commence employment. Waste will be allowed to cool after autoclaving to eliminate the risk of burns and scalds. HFC’s can easily ignite in a laboratory area as often there are naked flames etc. present. RSI can develop from repeated use of pipettes. This needs to be monitored and considered for all work Other Persons: Small risk of exposure to chemicals and GMO/Biohazards from equipment door handles etc. This is unlikely though as all work takes place inside a TC cabinet. All spills inside cabinets are cleaned up immediately. They are only at risk if there is a spill of GMO’s which is very unlikely as these will be cleaned up immediately
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Risk Assessment Form for Laboratory Procedure MRC Clinical Sciences Centre Version 3 July 09

MRC Clinical Sciences Centre Imperial College Faculty of Medicine Hammersmith Campus Du Cane Road London W12 0NN

Identify any persons and any special risks associated with the technique and steps that must be taken to minimise them.  Controls 1 All staff will be given instruction of the hazards and control measures before the work commences by an experienced senior member of staff. Any spillages that occur will be cleaned up immediately. They will also receive appropriate Biological safety and Manual Handling training. Controls 2: Appropriate PPE is always worn as well (Nitrile or Latex Gloves and Lab coat, Cryogenic gloves, face protection and low O2 monitor). All laboratories have fully functional fume cupboards that are used when working with toxic or volatile chemicals. All Tissue culture cabinets in the department have the airflow and personal protection tests carried out twice and once a year respectively. Certain hoods will have airflow monitoring carried out monthly, depending on the application. No eating and drinking is allowed in CSC labs either so there is no real risk of ingestion of chemicals. After the waste has been autoclaved it will be allowed to cool before being emptied. The bins will have cooled to allow cleaning of the interiors also. Emergency Spillage Procedure   All chemicals in the protocol will have the relevant spillage listed in its own COSHH form and all staff should familiarise themselves with these procedures. Spills in Hoods will be immediately cleaned up with Virkon and/or 70% EtOH

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For a spill of a GMO/Biohazardous substance:  Make up 500 ml of 3% Virkon in a beaker, and a sprayer full of 3% Virkon.  Obtain the Spill kit (should hold posidrive screwdriver, forceps, a clinical waste bag, sharps bin and a towel roll).  Wearing lab coat and gloves spray the 3% Virkon liberally onto the spill.  Leave it for 30 minutes.  After 30 minutes; if there is broken glass, use forceps to pick up pieces and place them into the empty Sharpsafe. Mop up the spill with the paper towels and put them into a clinical waste bag  Dispose of the clinical waste bag as usual; place any remaining sharps into the sharps bin.  Inform your supervisor who should either complete a dangerous occurrence form in conjunction with the Dept Safety Co-ordinator (or the Divisional Safety Advisor if the DSC is away).  All accidents must be reported to the Lab Manager / Safety Supervisor or the departmental first aid personnel, and the safety unit via an accident report form: http://safety.csc.mrc.ac.uk/CSCAccidentReportForm.pdf Waste Disposal    Details of the proper means of disposal of chemical waste can be found in individual COSHH forms. After autoclaving, check the autoclave has cycled properly. If it has dispose of the waste in yellow clinical waste bags. Rinse out the autoclave box with tap water and dispose of the fluids down the provided sinks Tips eppendorf tubes etc. should all be disposed of as clinical waste. Include a brief stepwise description of the technique  When dealing with any GMO’s or Biohazardous material:  All solid waste must be soaked in a final concentration of 1% Virkon overnight and disposed of in the grey autoclave box. Autoclave all waste by the end of the week. (see “RA for waste disposal” ) All liquid waste must be made up to at least a final concentration of 1% Virkon overnight. Discard down the sink with excess water the following day.
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Risk Assessment Form for Laboratory Procedure MRC Clinical Sciences Centre Version 3 July 09

MRC Clinical Sciences Centre Imperial College Faculty of Medicine Hammersmith Campus Du Cane Road London W12 0NN

Include a brief stepwise description of the technique  All glassware and plastics must be soaked in at least a final concentration of 1% Virkon for at least 1hour. Rinse thoroughly and place into the washing-up bowls or yellow bins as appropriate.

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Put on your lab coat and gloves. Prepare and fix cells on PLL coated slides (see RA for PFA slide fixing) Treat with 50-100l of blocking solution in a humid box for 20-30mins at room temp. before ‘flicking’ off the excess. Treat with 50-100l of 1 antibody at appropriate dilution in blocking solution. Place in a humid box for 1 hour or overnight at 4C. Wash twice in 1xPBS for 5mins shaking gently at room temp. Treat with 50-100l labelled 2 antibody at appropriate dilution in 1xPBS. Place in a humid box for 20mins at room temp. Wash twice in 1xPBS for 5mins then once in ddwater, shaking gently at room temp. Add DAPI staining mounting media and cover slip – do not push cover slip onto slide.

Blocking solution =

10% serum X and 1% BSA in 1xPBS. (Note: serum X should match source of 2 antibody)

Due consideration should be made in this assessment for women of child-bearing years, and a full re-assessment should be carried out upon notification of pregnancy. This assessment has been carried out by the person responsible for the work. A copy of this assessment has been sent to CSC Safety.

RESPONSIBLE PERSON Person carrying out the assessment Principal Investigator
Group Safety Supervisor

NAME

SIGNATURE

DATE

Annual Review

Risk Assessment Form for Laboratory Procedure MRC Clinical Sciences Centre Version 3 July 09

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