Department of Cardiovascular Sciences Laboratory Safety Handbook The booklet is provided to help you to work safely, and produced to comply with the Health and Safety at Work Act (1974), the Control of Substances Hazardous to Health Regulations (1988), and the University's Statement on Health and Safety Policy and Statement of Organisation and Arrangements (2007). A breach of the rules, instructions and guidelines in this document could be an offence under the Health and Safety at Work Act and the Control of Substances Hazardous to Health Act. These Acts, among other things, require all employees to co-operate with their employer so far as it is necessary to enable the legal duties of the employer to be met. A breach of the University's safety policy could lead to disciplinary action. This handbook should be read in conjunction with the Departmental Safety Handbook. This document is supplemented by separate safety handbooks containing detailed rules, codes of practice and guidance on specific topics. All documents are available online from the Safety Services website. Safety publications from the University form part of the Department’s health and safety rules.

All staff, students and others working in the Department are responsible for ensuring they are aware of, and comply with, safety regulations. Health and safety within the University depends on the acts and omissions of all individuals. Everyone in the Department has a role to play.

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CVS Laboratory Safety Handbook | University of Leicester

Safety Information & Health and Safety Induction The Information in the handbook covers the areas occupied by the Department of Cardiovascular Sciences. The areas where the Department has laboratory space are: 

Leicester Royal Infirmary, Robert Kilpatrick Clinical Sciences Building, 2nd and 4th floor



Glenfield Hospital, Clinical Sciences Wing (CSW) and Cardiovascular Research Centre (CRC).

All laboratory workers must familiarise themselves with the hazards associated with their work. The relevant risk assessment forms, including COSHH assessments, must be completed before work begins. All workers must read the Departmental Safety Handbook, the Laboratory Safety Handbook, the site relevant emergency plan and any other appropriate safety literature and codes of practice e.g. the Departmental Tissue Culture Facility Code of Practice. ALL new staff, students and temporary workers MUST receive an induction before starting work in the laboratory. Fire procedures will be covered in detail during the general induction to the Department.

Departmental Management of Safety Day-to-day supervising and implementation of health and safety policy in laboratories rests with Local Safety Supervisors and technical members of staff within the Department. The Departmental Safety Officer has been appointed to assist the Head of Department with the responsibility for managing safety, and to advise members of the Department on the implementation of safety policy and safe working practices. The Departmental Safety Officer coordinates a rolling programme of health and safety selfinspections for all areas of the Department. There is also a Building Safety Officer for each building (RKCSB, CSW and CRC). Their role is to assist in the management of matters which are not specific to single Department but concern the building I which they are placed.

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CVS Laboratory Safety Handbook | University of Leicester

Local Safety Supervisors and designated technical support The Head of Department has overall responsibility for the provision of health and safety information in the Department. Health and safety information within the Department is disseminated by the Local Safety Supervisors, members of the Departmental technical team and the Departmental Safety Officer.

LRI – Robert Kilpatrick Clinical Sciences Building Laboratory Room Number

LSS

222, 234, 241

John McDonald

219, 229

Sonja Khemiri

202,203, 216, 238, 242

Jonathan Barber

218, 220A, 232, 239

Barbara Horley

Glenfield – Clinical Sciences Wing Laboratory Room Number

LSS

CS002, CS004, CS006

Helen Rosendale

CS042

Dawn Smallwood

CS011, CS013, CS070

Malcolm Oswin

CS024, CS056

Julie Chamberlain, Martha Hardy

CS042a

Martha Hardy

CS036, CS065/6/7, CS076

Hash Patel

CS009, CS010, CS017

Paulene Quinn, Jodie Sandhu

Glenfield – Cardiovascular Research Centre

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Laboratory Room Number

LSS

0.01/C

Shireen Kharodia

1.03/A/B, 1.05

Laurence Hall

0.06, 1.04/A/B

Julie Chamberlain, Martha Hardy

CVS Laboratory Safety Handbook | University of Leicester

“Scientific” visitors and Work Placement Students Visitors and students from outside the University wishing to carry out research work or gain work experience in a scientific environment are permitted to do so. The Department Manager and Departmental Safety Officer must be informed prior to the visit. Relevant Insurance must be in place and a Risk Assessment be made of the anticipated work to be carried out before any work is started. In consultation with the supervisor, the Chief Technician will assess the visitor’s training requirement. The visitor must be made aware of potential risks and hazards associated with any work. The appropriate Local Safety Supervisor should be informed of any visitors intending to do laboratory work and a full induction must be given before any work is started. Prior notice of any visit to RKCSB must be made to John McDonald and Janet Harris and for the Glenfield site to Tim Barnes and Eleanor Jackson.

“Non-Scientific” visitors to the Department Visitors to the Department with no scientific training, including contractors, service engineers, company representatives, film crews, friends and relatives, should normally not enter laboratory areas. Where necessary, entry of "non-scientific" visitors will only be permitted when accompanied by a member of the Department. For servicing of equipment, engineers should not enter the laboratories without prior permission from appropriate Professional Services staff. It is important to ensure that the equipment is made safe before any work is started. Decontamination certificates (area-clean certificates) should be completed by the LSS. Contractors engaged in maintenance or work on the services or fabric of the building must comply with the University’s permit to work system and the University’s General Code of Safe Practice for Construction, Maintenance, Installation and Repair. The University Maintenance Department, the contractor, and the Department are all represented on these forms. Where work is to be carried out on Hospital property (CRC/CSW) the necessary permits and isolations must be obtained from Interserve (UHL estates contractor) representatives.

Unattended apparatus and experiments Many pieces of laboratory equipment, such as freezers and incubators, are designed to run continuously. Other pieces of apparatus operating between 6:00 p.m. and 8.00 a.m. (Monday to Friday) or during weekends should only be left on if: i) The person responsible is fully satisfied that it is safe to leave unattended. ii) It is left with a note saying "Please Leave On" giving details of how to shut down the equipment safely and an emergency contact telephone number and name.

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CVS Laboratory Safety Handbook | University of Leicester

Good Laboratory Practice Workers should ensure that they work under clean conditions and that where spills occur they are cleared up immediately. Work stations should be left in a condition such that other members of staff are not exposed to any risk. This applies particularly to balance areas where substances may be dispersed by static electricity.

Spillages Chemical Spillages of hazardous chemicals, or solutions containing hazardous chemicals, should be cleared immediately, whilst taking full safety precautions for the particular substance. Notwithstanding this, if the chemical is volatile, or you are in any doubt about how to deal with the spillage safely, your first priority should be to evacuate all personnel from the vicinity and secure the area.

Hazardous biological agent Specific advice for dealing with spillages of hazardous biological agents will be found in the individual risk assessment for that agent. As a general guide: 1. Disinfect the contaminated area. The Department recommends 1% Distel for routine disinfection of bench tops and other surfaces, 10% Distel for spillages of blood, or spillages where the presence of infectious agents is suspected. 2. Ensure contaminated material is rendered non-infective before disposal. 3. Take measures to prevent contamination of other workers.

Genetically modified organism Specific advice for dealing with spillages of genetically modified organisms will be found in the individual GM project document. The general advice given for hazardous biological agents (above) should be considered as a starting point.

Radioactive material 1. If the spillage is not contained behind screens i.e. in a work station, evacuate the area and physically isolate the spill if at all possible. 2. Deal with contamination of people first: remove any contaminated clothing and wash the affected area with water. 3. Report the incident to the Departmental Radiation Protection Officer (DRPO) or the Radiation Protection Supervisor (RPS) who will decide on an appropriate course of action. [If radiation levels are excessive do not attempt to clean up the area.] 4. Mop up liquid with tissues or other absorbent material, then wipe the area with 70% IMS and 5% Decon and continue until radiation levels are within background levels (see calibration label on minimonitor). If this process is unsuccessful, shield the area, label it as contaminated and consult the DRPO.

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CVS Laboratory Safety Handbook | University of Leicester

Chemical and biological spill kits These are located in a number of laboratories on each site, with further safety cupboards containing more extensive sets of cleaning and protective equipment to be used for larger spills. These latter are located in labelled cupboards in communal areas – as noted below.

Type of Spill Kit

Location

LRI – Robert Kilpatrick Clinical Sciences Building Biological spills

242

Basic Chemical and Biological spills kits

222

Safety Cupboard – respirator, safety specs, chemical spills, mercury collector, insulated gloves

222

Basic Chemical and Biological spills kits

219

Basic Chemical and Biological spills kits

238

Glenfield – Clinical Sciences Wing Larger Chemical and Biological Spills

CS011 – Autoclave Room

Basic Chemical and Biological spills

CS024 – Tissue Culture

Basic Chemical and Biological spills

CS004 – Laboratory

Glenfield – Cardiovascular Research Centre Basic Chemical and Biological spills

1.03 – Molecular Biology

Basic Chemical and Biological spills

1.04 – Tissue Culture

Spill kits are situated in the cupboards under the steel laboratory sinks. Ensure replacement of any materials used. Where a large spill of uncontaminated water occurs, there is a wet vacuum available from the Porters in CSW at Glenfield to rapidly clear this up with.

General Laboratory Cleanliness At the end of each period of work staff should remove their gloves and laboratory coat then wash their hands before leaving the laboratory. Laboratory coats and disposable gloves should not be worn in non-working areas. Disposable gloves should be removed before leaving the immediate work area and MUST NOT be worn outside the laboratory even if they are considered clean. If it is necessary to wear gloves for carrying an item between labs you must either seek help or have one ungloved hand for opening doors etc. Gloves are worn to protect the user from contamination, or to protect material from contamination by the user. In both cases it is inappropriate to wear gloves outside laboratories, since there is a danger of spreading contamination to others (via door handles etc) or of contaminating clean gloves.

Eating, Drinking and Smoking 6

CVS Laboratory Safety Handbook | University of Leicester

Under no circumstances should food and drink be taken into, stored or consumed in laboratories. Cosmetics should not be applied in laboratories. Smoking is prohibited in all areas of the building.

Use of Mobile Phones Use of mobile phones is not allowed in laboratories.

Personal Protective clothing (PPE) The protective clothing issued by the Department is for your protection against all manner of incident. Protective clothing available within the Department includes; Howie style laboratory coats, eye protection, face masks, rubber gloves, safety glasses and thermal gloves. It is essential that the correct PPE is used where necessary, for instance when indicated by the COSHH assessment form.

PPE

Equipment/Use

Laboratory coats

Any laboratory work

Gloves

Most bench work

Safety spectacles

Where indicated by COSHH assessment, for example acids.

Full Face Safety Visors

Liquid nitrogen / homogenising samples

UV certified visors

UV lightboxes

Dust masks

Allergy protection/weighing out of dusty solids

Thermal gloves

Liquid nitrogen and other cryogenic storage

Laboratory coats A white lab coat, buttoned at the front, should be worn at all times in the laboratories. Full length white coats with buttons or "popper" fastening should be of the "Howie" safety pattern (high neck, cuffed sleeves and wrap- over front) and should be fastened up, so that coat edges are not caught in doors, machinery or open flames. A properly fastened laboratory coat must be worn at all times when working in the laboratory - even over theatre-wear. Robert Kilpatrick Clinical Sciences Building: Laboratory coats are available from the scientific store cupboard Room 214, please see Lab Assistant for issue. Laboratory coats that require cleaning should be placed in the blue laundry bin (blue) located under sink area room 237, which is collected weekly (Thursday) for being laundered. Laboratory coats MUST NOT to be worn in room 236 and offices. Cardiovascular Research Centre / Clinical Sciences Wing: Laboratory coats are issued by Local Safety Supervisors and kept on hangers next to entry doors to laboratories. Laundry is carried out via Hash Patel.

Safety spectacles or goggles Our eyes are highly vulnerable to damage by a variety of hazards e.g. irritant chemicals, flying material, UV radiation, hot or cold materials and constitute a possible route of infection. Despite the sometimes devastating consequences of this damage, eye protection is routinely under-utilised in laboratories. Safety spectacles are provided in each laboratory and they should be worn to protect 7

CVS Laboratory Safety Handbook | University of Leicester

against splashes and light flying material. Some hazards require a higher level of protection to be worn. For example spray or "bottle bursting" as in autoclaving, use of liquid nitrogen and exposure to UV light require the use of a full face mask constructed from an appropriate material. Further information can be found on the University Safety Services Webpage, Eye Protection (2002) General safety spectacles are available in all laboratories.

Gloves Three types of protective glove are available in the Department: i) Rubber (Household type) gloves, used for washing up and for non-hazardous procedures where reuse is acceptable. ii) Disposable nitrile gloves, for use with material that is infective, toxic by absorption, irritant or radioactive. These gloves should be disposed of by autoclaving and incineration, see Glove Selection Guide. Latex gloves have been associated with the development of allergy, see Policy and Code of Practice for the Prevention of Latex Allergy (2001). As a consequence, their use is derogated and they are not provided by the Department for laboratory work. Note that all types of gloves should be inspected during use for the development of pinholes, particularly when using toxic chemicals such as phenol and acrylamide. iii) Thermal gloves, used to protect from heat e.g. items removed from microwave ovens, autoclaves or hotplates and from extreme cold e.g. liquid nitrogen, dry ice and the contents of -80°C freezers. It is essential that these gloves are coated with a non-absorbent material. Blue gloves are suitable down to -80°C, leather gloves for lower temperatures (liquid nitrogen).

Shoes These should be of an enclosed design to afford some protection to feet against spills. Open toed shoes or sandals are not suitable. Footwear must also give a secure grip on the floor; hence high heels are not suitable.

Laboratory Hazards Laboratories contain many potential hazards. Certain procedures and techniques are widely used in the Department; this section gives health and safety information on some of the more commonly encountered of these. In some cases this is in the form of a standard operating procedure (SOP). SOPs are for guidance only, if in doubt seek advice from those experienced in the technique, and consult relevant manuals and literature.

Standard Operating Procedures Autoclaves Centrifuges Class II Safety Cabinets Cold Rooms, Freezers and Fridges Dry Ice Electrophoresis Fume Cupboards Gas Cylinders Heating devices, distillation equipment and water baths 8

CVS Laboratory Safety Handbook | University of Leicester

Liquid Nitrogen Liquid Nitrogen (GGH) Microwave Ovens UV light Waste disposal – RKCSB Waste disposal – Glenfield

Chemical Hazards The University regulations on Hazardous Chemicals (1991) (available from Safety Services) list four general principles when dealing with chemical hazards: i.

Hazardous chemicals should be handled only if their use is justified by the importance of the experiment and if no safer alternative exists. Hazardous chemicals should be handled only in suitable areas with adequate equipment for their containment and for the protection of staff. Work with hazardous chemicals should be carried out only by users who are properly informed and have been given adequate training. Work with hazardous chemicals, including the disposal of waste, should be carried out according to clearly understood procedures.

ii. iii. iv.

The regulations also state that all persons dealing with hazardous substances must be aware of:    

hazards and how to recognise them; possible consequences of exposure or misuse; precautions to be taken; action in case of an emergency.

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CVS Laboratory Safety Handbook | University of Leicester

Hazard Symbols found on chemical containers Hazards encountered in the laboratory when using chemicals, biological agents, and electrical items of equipment present particular problems.

COSHH Nobody should use any substance unless they are satisfied that they can do so (from acquisition to disposal) without putting themselves or others at risk. The use of any chemical substance must be preceded by a risk assessment and in most cases this will take the form of a formal COSHH assessment. See the University Safety services information and advice on carrying out a COSSH assessment COSHH Assessment. RKCSB: Laboratory 237 has a filing cabinet containing completed COSHH forms and associated MSDS, along with a number of procedural COSHH assessment and lists of non-hazardous reagents. CRC/CSW: In each of the laboratories a filing cabinet containing completed COSHH forms and associated MSDS is kept for reference purposes.

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CVS Laboratory Safety Handbook | University of Leicester

Other Laboratory Hazards Flammable solvents

Liquid Nitrogen

Gas Cylinder

Hazards  Can easily be ignited by naked flames or sparks  In the case of a fire in the lab they can contribute to fire damage Precautions  Keep well away from naked flames, e.g. Bunsen burners  Solutions over 500mL kept in flame-tamer cabinet when not in use  The cabinet door should always be kept closed  Acids and other oxidising substances cannot be put in these cabinets as they are chemically incompatible  Keep the minimum amount that you need. Avoid keeping multiple bottles of the same chemical

Hazards o  At -196 C is capable of serious cold burns, especially to eyes  If not treated with proper care the individual 1.5mL cell storage vials can explode  Spillage of liquid nitrogen in an enclosed space can cause asphyxiation Precautions  You MUST have training before using liquid nitrogen  Use of a face visor and thermal gloves is essential  Doors to liquid nitrogen storage areas MUST be kept open as a priority to prevent gas build up unless there is adequate ventilation  Any quantity of liquid nitrogen should NEVER travel in a lift (elevator) or other confined space with people – two people should coordinate the transport between floors

Hazards  Impact from the blast of a gas cylinder explosion or rapid release of compressed gas  Contact with the released gas or fluid  Fire resulting from the escape of flammable gases or fluids  Impact from falling cylinders  Manual handling injuries  Leakage of gas can lead to asphyxiation Precautions  You must receive training before handling and using gas cylinders  Users should be able to carry out an external visual inspection of a cylinder, and any attachments  Correct, safe storage and transporting

Sharps

Lab Microwaves

Autoclaves

Hazards  Can cause cuts or puncture wounds which could lead to infection  Injury may result in contamination of undesirable lab organisms or chemicals Precautions  Never re-sheath needles  Dispose of all sharps immediately after use  Always dispose of sharps into a yellow Sharpak bin  Never dispose of sharps using ANY other method

Hazards  Heated substances can explode if not used carefully  Excessive heating, particularly of small volumes can create dangerous superheated liquids Precautions  Read the safety instructions on front of the microwave carefully  In particular ensure all tops on bottles are loosened

Hazards  Burn Hazards-pressurized heat an steam  Biohazards – content of the waste Never Put in Autoclave:  Items containing corrosives  Oxidizers  Solvents  Volatile substances  Radioactive materials Precautions Appropriate PPE:  Lab coat, safety goggles or face shield, heat resistant gloves are required

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CVS Laboratory Safety Handbook | University of Leicester

Hazardous Biological Agents Experimental use of any biological agent is potentially hazardous. The main risk of infection is through inoculation, exposure to aerosols, and direct contact with and transfer through the skin (via cuts, for instance). The transmission of tumour cells by inoculation is extremely rare. Fixed tissue samples also carry some risk, though this is more likely to be due to the toxic effects of the fixative. It is a University (and legal) requirement that a Risk Assessment must be made for all biohazards before any work can start (Hazardous Biological Agents (2011)). Risk Assessments must be made for all biological agents, which are defined as ‘living microscopic unicellular, multicellular microorganisms or subcellular biological entities capable of replication (including those that have been genetically manipulated); animal cell cultures; animal tissues and body fluids; microbial toxins.’

Genetically Modified Organisms All projects involving the use or creation of genetically modified organisms must be registered. This involves the performance of, and approval of, a risk assessment and the registration of workers in accordance with the Genetically Modified Organisms (Contained Use) Regulations (2000). For the purposes of regulation any biological entity capable of replication is included. This means that cultured cells fall under this provision. Guidance is available from Safety Services (Genetic Modification (2011)).

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CVS Laboratory Safety Handbook | University of Leicester

Radiation Safety Ionising Radiation Safety The University has detailed procedures controlling the use of ionising radiation contained in their publication “Radiation Protection Rules”. Workers who need to use ionising radiation must be registered, have a medical examination, and receive basic training, in the first instance, from the University Radiation Protection Officer (URPO). A separate induction and further training will be provided within the Department by the Departmental Radiation Protection Officer (DRPO), and Departmental Radiation Protection Supervisor. Contact the DRPO or DRPS for a copy of the registration form.

Role

Name

Email

Departmental Radiation Protection Officers (DRPO)

Dave Lambert (RKCSB)

[email protected]

Veryan Codd (Glenfield)

[email protected]

John McDonald (RKCSB)

[email protected]

Hash Patel (Glenfield)

[email protected]

Radiation Protection Supervisors (RPS)

All laboratory workers must be aware of the need to avoid radiation hazards. Radiation workstations and equipment are designated by the use of appropriate warning signs (“Long Term Health Hazard” symbol on symbols guide above). Only those workers specifically trained in the safe usage of radio-isotopes should work in these areas, use designated equipment or handle radioactive material itself (this includes un-packing deliveries). All workers should avoid leaning against benches where radio-isotope work is undertaken. When any package containing radioactive material is received, a qualified worker should be alerted immediately so that it may be logged and stored securely. If anyone is in doubt about any aspect of radiation safety, they should consult one of the Radiation Protection Supervisors, Dr. John McDonald (RKCSB) or Mr. Hash Patel (CRC/CSW), or the Departmental Radiation Protection Officers Prof. Dave Lambert (RKCSB) and Dr. Veryan Codd (CRC/CSW).

Waste Disposal Advice on disposal of laboratory waste can be found in the Waste Disposal Guides. There are different disposal procedures for the RKCSB and Glenfield sites, in our SOPs.

The table gives a general overview of different types of laboratory waste and routes for disposal.

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CVS Laboratory Safety Handbook | University of Leicester

Waste Disposal Disposal Item Autoclavable laboratory waste

Disposal Route Autoclave bags

General tissue culture waste and laboratory waste, tips, test tubes, gloves etc

Other Information Biohazard waste (e.g. cell culture, contaminated paper) via autoclave waste. Autoclave bags should be loosely tied with autoclave tape, labelled with the laboratory number and left in the demarcated area for the site – see LSS for details

Clinical waste / Sharps (non-infectious) Human sample Sharps in yellow ‘cin bin’ with label Limited chemical waste not suitable for autoclave bins waste – tissue, limited and items contaminated with Other clinical waste in yellow bags e.g. tri-reagent, acrylamide gels blood, plasma, urine or orange disposable bags

Glass waste (clean)

Glass bin

Wash out bottles before disposal When full, seal bin with tape and request porters to collect

Radioactive waste

Separate training and registration Contacts: required RKCSB – Dave Lambert (DRPO) / John McDonald (RPS) Glenfield – Veryan Codd (DRPO) / Hash Patel (RPS)

Chemical waste

See separate chemical waste disposal guide

Contacts: RKCSB – Barbara Horley Glenfield – Paulene Quinn

Plastic and paper packaging waste

Recycling bins are available for most items

Paper towels and domestic waste

Use black bags for any other waste to go via normal disposal route

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CVS Laboratory Safety Handbook | University of Leicester