Working in classified biological laboratories

At iNANO we have three levels of clasificated laboratories for biological work. Most laboratories are class 0 for all work without any risk while we have a few class 1 laboratories typically for bacterial expression and other types of work with very low risk. Finally, we have class 2 laboratories for special work that includes a little risk. 

Rules for genetic engineering work in class 1 laboratories

The following rules only apply to work in Class 1 laboratories. By biologically active material is meant living organisms, cells or viruses which contain DNA or RNA derived from genetic engineering. Isolated DNA, RNA or protein produced by genetic engineering is thus not covered by this definition.

When working with biologically active material, this must take place in areas that are classified by the Danish Working Environment Authority (class 1).

• Bags, coats and other extraneous material must not be present in classified premises.

• It is forbidden to use a private mobile phone in class 1 laboratories.

• Access by unauthorized persons to classified laboratories must be restricted.

• Laboratory gowns must be worn in classified laboratories. These should be yellow, green or white with the special yellow gene technology label on the chest pocket. This also applies to visitors and craftsmen.
• Regular laboratory hygiene must be comply: Food and drink must never be present in the laboratory. Smoking is strictly prohibited. Always remember to wash your hands before leaving the laboratory.

• Mouth pipetting is not allowed.

• All bottles, petri dishes, tubes etc. containing biologically active material must be marked with a yellow marker when leaving the laboratory.

• Transport of biologically active material outside the designated laboratories or areas must take place in containers marked with genetic engineering warning signs (yellow labels or yellow marker). Closed Eppendorf tubes can thus be transported in marked racks, agar plates in marked bags or plastic trays. Glass bottles can be transported in marked plastic trays on a trolley, but be aware that any likely spillage must be collected in the tray.

• All waste containing biologically active material, ie. living organisms cells or viruses containing DNA or RNA derived from genetic engineering must be collected in labelled containers and disinfected. Solid waste is collected in labelled autoclave buckets which are disinfected by autoclaving and then disposed of as ordinary waste. Liquid waste is disinfected with 1% Virkon, which must stand 10-30 min before it can be poured into the drain.

The liquid waste can also be disinfected by autoclaving after which it can be poured into the sink.

• Replacement of filters in the LAF benches and filters in ventilation systems is done by a technician. The technicians must wear respiratory protection and wear work clothes and gloves. The used filters are placed in an autoclave bag and thereafter closed. Autoclaving is then performed by the normal laboratory staff. After autoclaving, filters are disposed of as normally flammable material.

• Glassware and the like, contaminated with biologically active material, must be autoclaved or disinfected with 1% Virkon for about 15 minutes before being sent to normal washing up.

• Waste for autoclaving is collected and transported for autoclaving in closed, labelled steel buckets.

• Needles, scalpel blades and other sharp objects are placed in the special cannula boxes, autoclaved and disposed of as H-solid waste.

• When working with biologically active material, procedures must be limited as much as possible. Possibly. Aerosol-inducing procedures take place in fume cupboards or in LAF benches (vertical airflow cabinets).

• The workplace must be kept tidy and cleaned and disinfected daily. Waste of biologically active material must be immediately dried and washed with 70% ethanol. In the event of a major accident, the safety representative is contacted / notified. 

• Disposable gloves that have been used for work with biologically active material in classified premises are autoclaved.

• Hands must be washed after contamination with biologically active material as well as during breaks at work and at the end of working hours. 

• Paper should not be left free or suspended freely in the laboratory. Recipes etc. must be covered with plastic. Cardboard boxes must not be stored in classified premises - use plastic boxes instead.

• Note-taking can take place at the workplace either on areas that are delimited with tape or on pull-out plates where these are found. Optionally, the window frames can be used.

• Remember that paper / recipes / books must be kept separate from laboratory settings.

• In the event of accidents, measures are taken in consultation with the person responsible for the project or other competent personal. The safety organization must be informed as soon as possible.

• A logbook must be kept of all accident situations that have given rise to a risk to the safety of persons or to the external environment. The logbook is available at the end of the classified laboratory.

• Signs: There must be signs marked "Genetic engineering laboratory class 1" on doors to the classified areas.

• The responsible head of the laboratory is obliged to instruct the employees and students to carry out the projects properly and in accordance with the existing safety regulations for work in the classified laboratories. It must always be possible to summon a professionally competent person. The telephone lists are hung at the entrance to the classified laboratories.

Procedure for upgrading from Class 0 to Class 1

· The Danish Working Environment Authority (”Arbejdstilsynet”) must be notified when a laboratory is upgraded. This contact is made via the department’s OHS organisation. Upgrading of a laboratory can only take place after permission has been given.

· A person is selected to have the overall responsibility in cooperation with the HSO representative and they apply to the Danish Working Environment Authority for a Class I approval. The person’s name is given to the Danish Working Environment and written in the logbook for the laboratory.

· At the entrance to each classified room is a notice with the telephone number of the person responsible for that room.

· For cleaning purposes, classified laboratories/areas should contain only the most items. All items standing on the floor must be removable (on wheels).

· All persons working in a classified laboratory /area must wear buttoned-up lab coats.

· Use of the logbook is a requirement from the Act relating to gene technology. The date of the upgrading is to be entered into the logbook.

· A warning sign stating: “Genteknologisk laboratorieområde - Klasse 1” (Class I laboratory for Gene Technology) must be found at the entrance to the laboratory. 

· Round plastic containers for solid hazardous waste, labelled autoclave buckets, and containers for sharp objects must be found in classified laboratories

· Bottles with 70% ethanol must be found in classified laboratories.

· Lab coats must be hung on hooks just inside the classified laboratories.

· Cleaning personnel must wear lab coats when working in Class I laboratories. The person responsible for a laboratory must inform the OHS Committee (Secretariat) whenever an upgrading occurs. Laboratory personnel must carry out the daily cleaning and disinfection of the working area and the items used.

· Students working with biologically active material must be supervised by competent persons. 

· Stainless steel buckets for autoclaving must be placed in upgraded laboratories. These buckets are to be used for the collection and decontamination of items used while working with biologically active material (disposable pipettes, centrifuge tubes, etc.).

Cleaning instructions for classified laboratories

The cleaning staff must wear a buttoned up yellow or special white lab coats with a yellow gene technology mark on the chest pocket when cleaning classified laboratories bearing a notice “Gene technology laboratory Class I”.

At INANO, a yellow lab coat labelled “Rengøring” (”Cleaning”) is to be found on each floor in all the buildings. The lab coat hangs on a hook labelled “Rengøring” and is to be used only in classified laboratories.

When the lab coat needs washing, place it in the laundry basket in the laundry room unless it is contaminated with active biological material. In this case, the kit must be placed in a gelatine bag which is dissolved during the washing process

The cleaning staff’s daily cleaning routine consists of mopping the floor and removing ordinary trash. Note that the trash is removed together with the waste bag, and that waste must not be poured from one bag to another. The waste bags must be closed before they are removed from the laboratory.

In addition to the daily cleaning, the floor must be washed 2-3 times a week. Trolleys and hazardous waste boxes must be moved out when the floor is being washed.

At iNANO, floor cloth, soapy water and bucket are only to be used in the classified laboratories and the water is to be poured in the sink. The bucket and floor cloth may thus not be used anywhere else. When the floor cloth is to be washed, it must be autoclaved first. This is done by putting the cloth in an autoclave bag, closing it with autoclave tape and labelling it with yellow tape. After autoclaving, the cloth is washed in the usual way.

In addition, the cleaning staff is responsible for taking care of the following tasks:

·  Dusting and vacuuming elevated ventilation channels and electrical fixtures every six months

·  Cleaning the cooling systems in the laboratories every six months

·  Cleaning under refrigerators and freezers twice a year (laboratory personnel must pull them out and put them back)

·  Cleaning all stationary furniture /equipment below table/bench height. 

The dates for the thorough cleaning must be entered into the logbook. 

The laboratory personnel are responsible for the daily cleaning of laboratory sinks, window sills, tables/benches, fume cupboards, sterile benches and other work places, as well as a monthly cleaning of shelves, cupboards, bottles, electric supplies, free-standing apparatus and furniture.

In connection with the up- or downgrading of a laboratory, all the above cleaning procedures must be carried out. 

In the event of an accident, a notice must be placed on the door to the laboratory and the person responsible for the laboratory must be informed. The name of the person responsible for the project is to be found at the entrance to the laboratory. The cleaning must be discontinued.