The regulations cover basic chemistry, radioactive material, organic chemistry and work with microorganisms and GMO work, but only at a general level.
It is impossible for this website to cover everything, since many different techniques are being used. It is therefore the duty of every group leader (lecturer/professor) and supervisor to provide instructions for the safe use of special techniques. Furthermore, it is the duty of everybody working in the laboratory to seek the information required to work in a safe manner.
However, all safety rules are useless if not all employees make a continuous effort to maintain a safe work environment.
In connection to this, the following general principles must be made clear:
In some situations, it may be something simple so that you can do it yourself, e.g. wiping up a spill. In more complicated situations, help should be sought from one of the persons responsible for the room or the apparatus or one of the Work Environment Representatives.
All information in this website can also be found in this pdf with General Safety Regulations of the iNANO House.
This edition of the safety regulations is a thoroughly revised version of that from 2008.
The calling system is prepared for the house staff and cleaning staff in connection with accidents, incidents or other irregularities.
In the event of a FIRE or ACCIDENT (and after activating the fire alarm), call both numbers!
Alarm center (fire, ambulance, police)
AU's common alarm system
8715 1617 (24-hour staff)
Casualty ward (Skejby Hospital)- remember CPR number
(emergency doctor or treatment nurse on this number must ALWAYS be contacted before contacting the emergency room)
Guard staff; In case of irregularities in buildings or inventory that need urgent attention, call:
- Within normal working time
Building inspector Ove Fjordside
Alternative repair, maintanance:
- Outside normal working hours and only in emergency situations
Dansikring (access control)
Building inspector Ove Fjordside
Skadeservice Danmark (damage service)
Construction damage service, Safety Partner
iNANO building service, service mail
Building services, hotline
All accidents - both large and small - must be notified to the safety representative.
Near miss accidents must also be registered. A near miss is an event that could have developed into a work accident, but where nothing happened to the employee.
Near-miss accidents are characterized by the fact that the accident was avoided because of pure luck.
Land line phone: Dial 0; wait for the tone, then dial 112. Cell phone: Dial 112
When you are connected, state clearly:
Frostbite can be divided into three categories:
In all categories, first aid involves immersion in 38 degrees warm water. Be aware that this can be very painful. When the skin becomes red, a sterile bandage should be applied, carefully avoiding any pressure or cooling. For second and third degree burns, seek a doctor.
A First Aid box is to be found in the laboratories on each floor and in the student helper office. Make a note where they are to be found before you need them. Inform your Occupational Health and Safety representative/supervisor if something is missing from the box.
In severe etching, a reflective eye closure follows, so the affected person cannot adequately flush his/her own eye. The nearest one has to help.
The liquid in the eye wash bottles must always be sterile - see the instructions on the bottle.
Call the fire service by pressing 0 - 112 and give one of the following addresses to the alarm center:
Be prepared to provide information about the injured, what is burning and what phone number that is calling.
Quenching a fire is the task of the fire brigade, but it is important to act properly until the fire department reaches out to save any threatened people and limit the amount of fire.
NOTE! It may be fatal to enter smoking and burning buildings. It should be left to the fire service's specially trained crew.
Make sure that the fire service is met outside the building and is immediately informed about the
Remember to notify the switchboard/security personnel that the fire service is alerted.
Emergency shower is located above the door in most laboratories.
Fire blankets are available in all laboratories or just outside the laboratory.
CO2-extinguishers and hand sprayers/fire hoses are placed on all floors.
Familiarize yourself with the location and use of these before it's too late!
It is important that lab work is carefully prepared, with regard to both safety and the end result. A continuously updated lab logbook is an invaluable and indispensable tool in this respect.
An evaluation of the risk and safety precautions is an important part of the preparation for your work. For example, it can be necessary seek information on the properties of a substance: State, reaction with water, combustibility (kindling temperature, explosion limit), caustic and corrosive properties, odour, toxic properties and especially long-term effects, penetration of rubber and plastic (gloves) its possible hygienic threshold.
For many known chemical reactions and products, much of this information is unavailable. Only typical physical and chemical properties are registered. However, various types of literature have articles describing substances that are hazardous to health and the environment.
When preparing for lab work (whether experimental or routine) each person must consider the risk involved.
All work with substances and reactions that give rise to hazardous or malodorous gases or vapours must be carried out in a fume hood. As a general rule, the fume hood should be used for all forms of chemical work whenever possible.
The degree of safety provided by the fume hood depends partly on its technical and construction conditions and partly on personal and actual conditions, namely:
The temperature inside the fume hood.
The Danish Working Environment Authority (”Arbejdstilsynet”) demands that fume cupboards be fitted alarms that are activated when the airflow is inadequate.
Each fume cupboard has its own alarm, which sounds and shows a red light when the airflow falls below a certain level.
When a fume cupboard is closed there is still a slight suction. When the window is opened, the airflow is increased, but when the height of ca. 40 cm is reached, the alarm is activated. Even before this height is reached, air movement around the opening can reduce the effectiveness of the air flow, especially if more than one fume cupboard in a laboratory is open at the same time, as there is a limit to the total capacity for suction.
This section mostly deals with the protection of eyes, skin and respiratory organs.
It is mandatory to wear safety goggles or a face mask when working with liquid nitrogen. It is also required to wear safety goggles or a face mask when working with anything that can splash when boiling, or can splinter, when working with strong acids, bases or radioactive materials.
When wearing contact lenses: be extremely careful when working with strong acids, bases or poisonous solutions. If any of these substances come into contact with the eye they may come underneath the contact lens and damage the eye. Therefore, always work in the fume cupboard and wear safety goggles. In the event of an accident, it is extremely important to remove the contact lens so the eye can be thoroughly washed.
Safety goggles are found in various qualities and sizes. They must have side protection so that there is less likelihood of particles coming into the eye from the side. Some have adjustable side lengths, some are adjustable up or down so that they fit the individual. Usually they are made from strong plastic material (e.g. polycarbonate) - their weakness being that some organic solvents can dissolve the surface so they become opaque.
Safety gloves are used when needed to protect the hands from substances that can damage the skin, either directly or by penetrating the skin and cause damage elsewhere.
N.B. There may be a risk of allergy even from powder-free gloves.
Before putting on gloves:
Only under very special circumstances, e.g. in the event of accident, will it be necessary to use respiratory aids (gas or dust masks). Respiratory organs are primarily protected by avoiding situations likely to give rise to hazardous gases, vapours or dust, such as working in a fume hood.
Gas cylinders with pressurised gas are frequently used in all types of laboratory work and carry many risk factors. Damage to a gas cylinder can cause it to explode because the gas is under high pressure. A broken valve can result in such a violent rush of gas that the cylinder becomes a projectile. The escaping gas from a damaged cylinder or a badly conducted experiment can cause an explosion and fire, poisoning, corrosion or choking, depending on the type of gas. A list over the most commonly used pressurised gases is found below:
The pressure at 20°C in the cylinders we use:
150 - 200 atm
10 - 60 atm
Carbon dioxide (56 atm)
Working with and storing liquid nitrogen and dry ice involves the risk of frostbite (N2, -196 ° C ; CO2, -78 ° C) and for asphyxiation (choking) (1 L liquid nitrogen at 20 °C, 1 atm will have a volume of ca. ¾ m3). Carbon dioxide also has a physiological effect and can cause immediate death in concentrations of 10 - 20%. Liquid nitrogen is widely used in cold traps and can cause atmospheric oxygen condensation both within the trap and in the liquid nitrogen tank. This oxygen can cause violent explosions in the presence of oxidizable substance, e.g. organic compounds.
All 3 classes are classified in subclass 1 for not water miscible and subclass 2 for liquids that are water miscible under all conditions
Max. storage in
Approved plastic or metal container
no restrictions up to 25 L
no restrictions up to 125 L
no restrictions up to 1250 L
Plastic packaging over 125 ml must always approved by the Danish Emergency Management Agency (”Beredskabsstyrelsen”).
The given amounts refer to the total amount of stock, usage and waste.
Altogether, there should be no more than 50 storage units per laboratory
Containers with inflammable liquids of Class I-1, I-2, II-1 and III-1 must not be placed in any of the escape routes (corridors, stairways, etc.).
Avoid working near open flames or where there is a risk of sparks.
NB: Explosive substances eg. diethyl ether and petroleum ether should not be stored in a normal refrigerator.
See also the KIROS database (https://www.kiros.dk/Web/)
Avoid contact with chemicals in all situations where chemicals are handled: Weighing out, pouring, routine lab work, transporting, cleaning up and disposal of waste.
For substances or containers with possible explosive qualities, age, storage temperature, light and air can be crucial for stability, so it is of utmost importance that these substances are not bought and stored in large quantities.
The Danish Emergency Management Agency (”Beredskabsstyrelsen”) has complete information on peroxides and formic acid.
If in doubt about high peroxide contents then be very careful about carrying and opening the container. A simple method of checking whether there are peroxides in e.g. ether is to mix a couple of mls with a potassium-iodide solution, add a couple of drops of diluted HCl and shake. The brown colour of iodine is a sure sign of peroxide.
The peroxide content can be checked with Peroxide Strips (Merck 1.10081.0001, level 1- 100 mg/L H2O2). Most peroxide-forming chemicals carry a stabiliser when delivered and chemical companies usually guarantee the shelf-life in unopened containers for three to five years from the production date. For chemicals not containing added stabilisers, there is a shorter shelf life.
Some peroxide-forming substances can reach explosive peroxide levels without a concentration of the solvent, and the general rule for substitution means that there should be a special reason for using diisopropylether.
According to the Danish “ADR” rules for transporting dangerous material, many of the ethers used routinely are classified as class 3 inflammable liquids. These rules (article 188.8.131.52.1) state that Class 3 liquids that easily form peroxides can only be transported by road when the peroxide content is no more than 0.3%, i.e. 3000mg/L. Such a high peroxide content will seldom be found in a laboratory, and unused peroxide-forming substances will normally be disposed of via the waste disposal system.
If in doubt – or it is known – that a substance has a high peroxide content (limit 100ppm), contact an Occupational Health and Safety representative/supervisor for further information, e.g. it could mean destroying the peroxides with an acid solution of ferrous- sulfate.
Containers with unstable chemicals should be labelled with date of purchase, date of opening, stability control, location, etc.
Unless the mixing process is under control do not mix:
Please note: Concentrated or fuming nitric acid + ethanol must not be used for cleaning glassware because of the danger of explosion.
Water-reacting substances react radically with water, often producing a lot of heat and in many cases producing gases. When mixing, pour the substance carefully into the water – never the other way round.
Examples of such substances are:
Hazardous substances are defined as those substances that are dangerous for health and the environment.
Information can be found in Kiros and in the "Order of Dangerous Substances List", volumes 1-3, which are found in most laboratories. You can contact your environmental representative for an updated version.
According to the new Globalized Harmonized System (GHS) hazard scheme, hazards are classified as seen in the figure below. A full description of hazard and precautionary statments can be found in the General safety Regulations at iNANO.
It is your duty to obtain all the necessary information about the chemicals you need before you start an experiment. They are found in Kiros and in the supplier instructions.
Substances labelled with hazard symbols, as well as powders for media that can cause allergy, must always be handled in a fume cupboard.
The damage caused by chemicals - acids, bases or special cell poisons, depends on the nature of the substance, also on the concentration of the substance in the tissue and the time in which the substance is allowed to affect it. Therefore
Remember that gloves only gives limited protection. Some substances penetrate certain gloves very quickly. There are many different types of gloves, see the section Personal valuables or ask the safety representative or the laboratory technician.
Waste on tables and floors must be immediately dried up. Dirty glass is rinsed with plenty of water before being sent to dishwashing.
The scientific employee ordering or otherwise bringing a chemical into the laboratory must check and take responsibility for the availability of the chemical instruction manual and the necessary protective measures, including labelling of the substance, and the instruction of anyone who is to handle the substance as well as waste disposal.
Injury to the skin: If phenol comes into contact with the skin, wash immediately with plenty of water, after which the skin must be wiped for at least 15 min with gauze or a cloth soaked in a mixture of polyethylene glycol (PEG 400) and ethanol in the ratio 7:3. This should continue until every trace of solidified phenol is removed. Wash again with plenty of water. Clothes spotted with phenol must be removed immediately. The person giving aid must wear gloves.
Wherever phenol is used there must always be found a clearly labelled bottle of polyethylene glycol 400/ethanol in the ratio 7:3.
If this mixture is not to be found, wash with plenty of water for at least 15 min. If the injury is of a serious nature, take the victim to a hospital and give a detailed description of the type of accident and the procedures taken.
Injury to the eyes: If phenol splashes into the eyes, the phenol must be thoroughly washed away with a mixture of polyethylene glycol 400 and water in the ratio 1:1. Thereafter, wash with water for 5-10 min.
Wherever phenol is used there must always be found a clearly labelled bottle of polyethylene glycol 400/water in the ratio 1:1.
If this mixture is not to be found, wash with plenty of water for at least 15 min. Take the victim to a hospital, possibly to the eye department. Washing must be continued during transportation with the aid of an eye bottle, until a doctor has taken over.
Injury to the mouth, throat, etc.: Phenol in the mouth should be washed out with water, and followed by a couple of spoonfuls of edible oil. Do not try to induce vomiting.
A clearly labelled bottle of edible oil should always be found wherever phenol is used.
If the victim is unconscious and is not breathing, artificial respiration should be administered. If the victim is unconscious but breathing normally, then treat according to general first aid for shock: Turn the person on one side with head lower than the rest of the body and keep warm with a blanket or coat. Unconscious persons must never be given anything to drink.
After necessary first aid, the victim should be taken to a hospital and the staff be informed about the type of accident and the procedures taken.
See also workplace guidelines for Phenol.
Acrylamide is a white crystalline powder that is easily taken up through the skin, the lungs, and the wall of the intestine. Acrylamide is carcinogenic and can affect the nervous system even when only small amounts are ingested.
Acrylamide must therefore be handled with extreme caution, both in crystalline form and when in solution so that there is minimal risk for skin contact or ingestion by nose or mouth.
We suggest that all groups buy acrylamide-bisacrylamide as ready-made solutions.
Thorough cleaning is essential so that others are not subjected to contact with acrylamide.
All glassware must be carefully rinsed.
Always use nitrile-disposable gloves (see section on personal safety equipment), also after polymerisation.
See also workplace guidelines for Acrylamide.
From 1 January 2009, SYBR Safe is to be used instead of Ethidium Bromide – see website for further details.
In special cases, if it is not possible to replace ethidium bromide with SYRBR Safe. Ethidium bromide is a powerful mutagen, possibly also carcinogenic, and must be handled with extreme caution. The following guidelines are suggested in order to minimise the risk involved in working with this substance.
See also workplace guidelines for ethidium bromide.
1) Destaining Bags from CLP (VWR-Bie & Berntsen, CLP 5459.25)
2) Active carbon filter, Carbon Cap 75 (Frisenette APS, 67047500)
Disposal of both Destaining Bags and Carbon Cap 75 is treated as B-waste, i.e. in closed plastic bags.
In the event of large spills of ethidium bromide solutions, use absorbing material as described in this leaflet.
Below, some general and practical advice is given concerning the handling of isotopes which are used in our laboratories at present. Whenever a new isotope is introduced, it will be included in the collection.
It is taken for granted that when working with isotopes, the safety rules which apply for working with hazardous chemicals also apply here, i.e., wear lab coat, gloves, safety goggles (when required), etc. Furthermore a thermoluminescent dosimeter (TLD/TL dosimeter) must be worn, either a personally registered dosimeter with quarterly/monthly checking, or an extra dosimeter that can be used by other persons as required and then sent for checking after use. The Government Bill 823 of 31 October 1997 form the basis for these rules. In contrast to the earlier Bill, the maximum allowed dose per person has been lowered to 20 mSv, and there are special rules for working with radioactive substances while pregnant (see next page).
Units of Activity: 1 mCi = 37 MBq 1μCi = 2.2 x 106 dpm (decay per minute)
1 MBq = 27 μCi 1 Bq = 1 dps (decay per second)
List of isotopes emitting β-particles
After storage for 10 half-lives, the radioactivity is decreased to about 1/1000. This will often be an appropriate time for contaminated glassware and other highly radioactive waste.
In the INANO house, p.t. (July 2015) only isotope labeled with 32P isotope
The INANO house has a S1 1/10 permit. This permission entitles to storage of up to 500 MBq 32P
For β-particles, the maximum range is dependent on the particle's energy. The particle is slowed down, and the heavier the braking substance, the quicker the particles are slowed down. Please be aware that the absorption of particle radiation in a heavy absorber gives a more powerful braking radiation than absorption in a light absorber. Plexiglas therefore offers better protection against 32P than ordinary glass.
Range and necessary shielding for selected isotopes:
1 cm perspex
1 cm Perspex
1 cm Perspex
0-1 cm perspex
3 mm lead or lead glass
Radioactive material must be stored in the refrigerator or freezer in an isotope laboratory, and must be registered in the isotope inventory database (The “HOT” database). The cabinets are fitted with locks. In the INANO House, 32P material is stored in room 1592-319, which functions as S1/10 laboratory. Niels Sandal (MBG) has the overall responsibility.
When planning to purchase radioactive material, the purchaser is responsible for making sure that the total allowable inventory for each isotope is respected. This information can be found in the database, KIROS, which is the department’s chemical registration system (https://www.kiros.dk/Web/). The purchaser is also responsible for registering all new purchases in this database, and all users must make sure that they register in the database every time they use any of the stored isotope.
If there is a C-laboratory in the building, work with S1/100 quantities can be done in all other laboratories. If activity levels exceeding the S1/100 limits are to be applied, it is possible to apply for dispensation for S1/10 classification. An application is required for each isotope.
32P, 14C, 33P
MBq / μCi in use at any one time simple wet procedures
Procedures with dry material
50 / 1350
5 / 135
0,5 / 13,5
Below are examples of what can be defined as:
· "simple wet procedures": Extraction from stock solutions, dilutions.
· "wet procedures": Normal experiments.
· "procedures with dry material": Work involving a certain risk: chromatograms, evaporation, dry gels.
The term ”in use at any one time” means the maximum amount of activity that can go on in any one laboratory at any given time. If several experiments are going on at the same time in the same laboratory, then the maximum amount per experiment must be reduced to compensate for an increased risk of contamination and accident.
The maximum amount of radionucleotide that can be stored in a laboratory is the same as that stated for a wet procedure.
· Upon extraction from vials with rubber cap: Always insert a needle with a cotton wool in the ampoule before use to equalise pressure differences (many compounds have been on dry ice and therefore assume considerable pressure by warming to room temperature).
· General care must of course be shown. Always use plastic trays, gloves, etc. when working with isotopes.
· 32P-ampoules must always be stored in a lead container.
· All work with isotope amounts larger than 1 mCi (40 MBq) must be carried out in an isotope laboratory. Handling of the isotope must take place behind a plexiglass or a similar screen. Work with small amounts of isotopes can be carried out in S1 or S1/10 laboratories.
· Always use a plexiglass tray and use nitrile gloves.
· Avoid using glass when working with 32P, because “Bremsstrahlung” can be formed.
· Always keep a monitor beside you.
· Check yourself and the work top with the monitor frequently, e.g. every time you leave the area.
· The whole laboratory must be checked with a monitor once a week and the results recorded in a special protocol to be found in the laboratory.
· Use a plexiglass box or lead container for transport between laboratories.
· Liquid waste must be diluted to < 2.7 μCi (0.1 MBq) per litre and poured down the sink. Afterwards let water run down the sink for about 5 min. The maximum monthly limit that can be disposed of in the sink is 1.35 mCi (50 MBq) per permit.
· Liquid waste, which in addition to 32P contains organic solvents (phenol), should be collected in a fume cupboard (e.g. in a special plastic container together with other phenol waste) until the isotope has decayed. It is then disposed of according to the Departmental rules.
· All solid waste contaminated with 32P must be collected. Use the plexiglas containers in the isotope laboratory. Very weakly contaminated things (gloves, etc.), can be thrown into the usual hazardous waste boxes. All fluid waste, marked with group name and date, must be cleared away after six months.
· Contaminated objects can be washed with Decon (use only the sink in the isotope laboratory). When there is no trace of contamination, they can be sent for washing. If there is still contamination, put them to soak in a decontamination bath overnight. If this is still not sufficient, store the objects for 10 half lives.
· Cleaning-up the work area: Check the work area with a monitor and decontaminate any contaminated places. Remember that an isotope laboratory does not necessarily have to be contaminated!
· 32P waste from the isotope laboratory is stored in plastic container for approx. 10 half lives (5 months).
· The box is sent for incineration when the total radiation level is less than 1.3 mCi (50 Mbq) inside and the radiation is less than 5 μSv on the outside.
The working schedule for pregnant women should be such that an unborn child is not subjected to more than 1 mSv. The State Department for Radiation Safety recommends the following maximum limits:
32P 5 MBq ~ 135 μCi
3H, 14C, 33P and 35S 50 MBq ~ 1.35 mCi
dositometry films must be changed once a month.
If a pregnant woman works in a laboratory where colleagues are using open radioactive sources, then the dosage and risk should be seen as the total exposure. The Department of Occupational Medicine (Arbejdsmedicinsk Klinik) can be consulted regarding an evaluation of the risk involved.
When a woman is breast feeding at the time she is working with radioactive substances, she should be aware that in the event of accidental spill, radiation may be transferred to the child through the breast milk. If the amount used is less than the limits for an S1-classification, then the risk is very small.
It is the responsibility of the person who spills to make sure that the spill is cleaned up immediately and thoroughly. If the spill is considerable and over a large area, then an Occupational Health and Safety representative/supervisor and the isotope-responsible staff member must be contacted. A small spill of liquid radioactivity should be wiped up with absorbent paper (paper towel). Spills of powder or other dry material should be wiped with wet absorbent paper. Afterwards, wash with a carrier-solution, i.e., a non-radioactive solution of the labelled substance that was spilled. For 32P-spills, a potassium phosphate solution should be used.
All paper as well as other materials used for cleaning-up should be treated as solid radioactive waste.
After cleaning-up, check the area for radioactive contamination. 32P and 33P can be checked directly with a monitor, but because of the monitor’s low sensitivity for 33P, - beta radiation, an extra precaution for these is to wipe the area with a damp filter paper, dry the paper, put in a scintillation vial with 5 ml scintillation fluid and count in a scintillation counter.
Persons, who frequently work with or nearby 32P, must wear a dosimetry film. The Danish National Institute of Radiation Protection sends the dosimetry results every month and a yearly statement. The limit is 20 mSv/year, although during pregnancy, the limit is 1 mSv, (see Guidelines for working with radioactive substances while pregnant or breast feeding).
Gloves must always be worn when working with radioactive isotopes, and hands must be washed thoroughly afterwards. However, in the event of contamination of the skin, the area should be washed a number of times with a carrier-solution and then several times with soap and water. If there is still sign of contamination (monitor), you should go immediately to the hospital’s casualty ward.
If the skin is damaged, as well as radioactively contaminated (corrosion or sores), rinse liberally with water, possibly opening the sore to induce bleeding and cleansing. Immediately afterwards go to the hospital’s casualty ward.
Clothing which has become severely contaminated should be treated as radioactive waste.
If you accidentally swallow a radioactive solution, vomiting should be induced immediately (finger down the throat) and go immediately afterwards to the hospital’s casualty ward.
The National Institute of Radiation Hygiene: 24-hour security 0 - 4494 3773, must be contacted for major accidents with radioactive isotopes.
No urgent inquiries and questions: Security Partner 0 - 70 113 113
Reference: "Vejledning om strålebeskyttelse ved arbejde med åbne radioaktive kilder", Statens Institut for Strålehygiejne, 2005 (Guidelines for protection from radiation while working with open radioactive sources), and "Bekendtgørelse om anvendelse af åbne radioaktive kilder på sygehuse, laboratorier m.v.", Bekendtgørelse nr. 954 af 23. oktober 2000 fra Sundhedsstyrelsen. (Act on the use of open radioactive sources in hospitals, laboratories, etc.).
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.
· 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.).
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.
When you enter a laboratory, you should behave as though everything is hazardous. Therefore:
· do not move anything
· do not touch anything
· do not attempt to smell the contents of bottles
If you accidentally knock something over, leave it as it is but inform one of the laboratory personnel.
The incident may appear to be harmless, but it can be dangerous, or the results of several days’ work. A knowledgeable person may be able to salvage the pieces, whereas everything will be lost if you attempt to put things right.
If you break a glass bottle and the contents run out, you must leave the laboratory at once. Do not attempt to wipe it up because the substance may be corrosive or poisonous. If you spill some on yourself, leave the room, and quickly wash with lots of water. Inform someone who can evaluate the situation.
If you come across something which makes you feel uncertain, e.g. a strange smell, sound, or smoke, go out of the room immediately and inform someone who can decide whether it is hazardous or not. Remember that the Occupational Health and Safety representative/supervisors are here to be helpful.
It is better to ask too often than too seldom!!
Do not work alone. If you must work in a room where nobody can see you, inform someone about what you are doing and tell them when you are finished.
In case of an accident, remember
· do not touch anything, but leave the room
· wash with lots of water if you spill something on yourself
· fetch laboratory personnel
When you enter a laboratory, which has a sign “Genteknologisk laboratorieområde - Klasse 1” you must wear a buttoned up yellow lab coat or a white one with the special yellow gene technology symbol on the chest pocket. Cleaning utensils that you use in these laboratories must not be used in other areas.
Persons you can contact:
Safety committee at iNANO
· Certain chemicals, except solvents, are received in postroom 2nd floor, 1590
· Solvents and chemicals are received in room 1593-218 (student helpers office) or room 1591-016 (chemical storage)
· Transport in the house must be done safely. Usually chemicals are shipped securely packaged and transported to the laboratory in the mail package; large and heavy gaskets with e.g. solvents, acids and bases are transported in a trolley.
· Many corrosive and hazardous chemicals can often be purchased in plastic bottles with plastic safety coating. If a bottle is dropped, the contents will not run out.
· All received chemicals must be labelled with a bar code and registered in KIROS. When requiring special precautions for a chemical, it must be indicated on the chemical labelling.
· KIROS works as a workplace manual for the individual chemicals and in KIROS there is also Material Safety Data Sheet. (MSDS)
· In the labelling and handling of chemicals we are considered as end users; we are therefore not obliged to change obsolete safety instructions and R and S phrases to H and P sentences
· Gases are delivered in an approved gas cage, where the pressure cylinders are securely fastened with a chain.
I. A common stock primarily for solvents in approved rooms
II. Local stocks in the research groups (possibly shared between several groups).
III. In the research and teaching laboratories.
· Solvents: The solvents are ordered in units that have a size so to avoid any kind of storage in the storage type I and II. Ie. that the units received can be used in the laboratories. The size of the units is determined by the solvents' classification.
· Explosion-proof cabinets should be used for larger storage of solvent in latex type I and II. Explosion-proof cabinets must not be located in laboratories.
· Other common chemicals: Store in stock I-III in enclosed ventilated enclosures.
· Acids, bases and highly oxidizing chemicals must always be stored separately from solvents
· Poisonous and licensed chemicals: Store in specially locked cabinets and keep accounts with the quantities.
· Pressurized gas cylinders are securely transported in special bottle carts and must always be securely stored in laboratories and in no larger quantities than necessary. Reduction valves must be approved for the gas in question. Outside the laboratory, an approved sign with the text must be fitted: “Trykflasker – Fjernes ved Brand” ("Pressure bottles - Remove when fire").
· Anyone working in a chemistry laboratory must have completed the Chemical Institute's safety course and fire extinguishing course before they get independent access to the laboratories and receive key cards and keys.
· Anyone who works in a chemistry laboratory must sign that he/she has read, understood and will comply with the safety rules for working with chemicals and equipment.
· Anyone working in a molecular biology laboratory must have undergone safety regulations, waste management and general chemical handling before they gain independent access to the laboratories and receive key cards and keys.
The constituents of the waste: Buthanol, > 25% ethanol, > 25% acetic acid (with larger quantities also collected below 25%, applies to both ethanol and acetic acid), approx. 1% Coomassie blue, methanol, acetonitrile, propanol, tetrahydrofuran, acetone, dioxane, guanidine hydrochloride. Small amounts (about 1%) of paraformaldehyde, silver nitrate, phosphoric acid, trichloroacetic acid and other halogens below 1%.
Liquid waste must only be poured into approved 5 L containers, which are collected in room 1590-049.
Containers must only be filled to the top mark (See photo). The lid is screwed securely and tight with the special tool (found in room 1590-049). The waste containers are put into the correct transport bins in the waste room 1590-051. Filling and suggestions for marking as shown in photo.
Waste constituents: Gels, napkins, pipette tips, gloves and the like with very small amounts of chemicals. Cannula boxes for needles and scalpels.
Dry GMO waste must be autoclaved and disposed of as ordinary waste.
Waste constituents: Counting fluid OptiPhase Hisafe 3[JNP1] (including di-isopropyl naphthalene and polyethylene glycol mono-4-nonylphenyl ether)
Elisa plates with very small amounts of organic / inorganic chemicals (eg Bradford, Lowry).
Waste components: Chloroform, phenol, <10% isoamyl alcohol in tubes, trichloroacetic acid, sulfolane, acetone. Halogen over 1%. Notify your security representative if anything else is left in the list!
Waste constituents: Carbon filters and carbon bags with ethidium bromide residues REMEMBER: The carbon filters / carbon bags are wet and packed wet in plastic bag.
Waste constituents: Equal to or more than 50% solvent. Collected in smaller containers in the laboratory, disposed of in barrel in waste container. Do not contain halogens.
Waste constituents: Mixed laboratory waste from clean-up.
A barrel: for solid Z-waste - write consecutive numbers, the chemical name, the quantity (in%), your initials and group names in the logbook.
A barrel: for liquid, corrosive and acidic - as stated above, write in the logbook on the barrel.
A barrel: for liquid, corrosive and basic - as stated above, write in the logbook on the barrel.
Remember: O waste must always be packed in a barrel by itself.
You may pack 2 mercury thermometers or mercury lamps at the top of the barrel in a thick plastic bag, if you write it under “affaldets bestanddele” ("The components of the waste")
Waste components: Empty spray cans as well as empty small butane gas cans
Waste components: See the list of "O substances" (reactive substances)
It must be written in the book which substances come in the barrel, as the different types of reactive waste must be kept separate from each other.
An O-waste barrel, especially for hydrogen peroxide 20 - 60%
Waste constituents: Elisaplader with small amounts of heavy metals. Liquid nickel chloride and copper sulfate from columns. Notify your safety representative if any metals other than nickel and copper are being poured.
Waste components: Engine oil (not chlorinated oil, Fp over 61 ° C), oil from vacuum pumps.
There is a box in the student helpers office for used batteries
All solid waste must be autoclaved and disposed of as ordinary waste. Liquid waste can be autoclaved or treated with Virkon S (10/00) and then poured into the sink.
After storage for 10 half-lives, radioactivity has decreased to approx. 1/1000. This will often be an appropriate time for contaminated glassware and other highly radioactive waste.
32P waste must be stored for 5 months
35S and 33P waste must be stored for 30 months
125I waste must be stored for 20 months
After storage it can be considered as ordinary, non-radioactive waste. Waste after iodination (solid form e.g. in gel matrix or aspirated into absorption material or absorbent of the like) is stored in closed 1" steel tubes which are stored for at least 20 months.
These shortened guidelines are based on information from the website of Department of Molecular Biology and Genetics:
where there are illustrations of the correct plugs and sockets as well as wrong and dangerous plugs.
Safety when working with electricity must be taken very seriously.
Strict regulations were imposed in 1993 regarding:
· Equipment safety
· User familiarity with safety conditions
Discard any apparatus that fails to meet safety standards.
When in doubt, do not use it and ask a technician to take care of it.
· Electric shocks cause muscles to contract violently, resembling a cramp or seizure.
· The most serious electrical accidents lead to cardiac arrest, while a milder shock can cause significant discomfort.
· Never touch electrophoresis apparatus or the electric cables when the electricity is switched on. (Under proper conditions* this should not be possible).
· Most liquids in the laboratories conduct electricity. Many of them are even very good conductors and correspond to a direct connection with the connected core.
· Electrical apparatus and cables must always be clean and dry, without salt deposits. Never handle electrical equipment with wet hands or gloves (the thin rubber gloves do not protect against high voltage).
· The cold room increases the risk of electrical failure and accident because of condensing water.
· The two most dangerous currents are 230 volt mains electricity and that from high voltage electrophoresis apparatus.
If in doubt whether the apparatus is safely set up, think about this basis rule:
* The apparatus must be protected against accidental touching of live current in any situation!
All electrical equipment must be earthed via the mains cable. This provides better protection from electric shock and accidents even when there may be some leakage in the apparatus.
Therefore: All laboratory apparatus must have a Danish 3-pin mains plug that is earthed, 2 round pins and one flat “earth “pin below!
· Mains cables with 2-pin plugs are not earthed and must NOT be used (see picture). Throw them away. The correct plugs can be obtained through the electronic workshop (Incuba)
· Plugs and cables must be in perfect condition. Users must make sure that the mains connections, i.e., mains socket and apparatus socket are not damaged (the plug pins are intact and the cable insulation is not torn, etc.).
Note also that:
· cables must be of robust quality, without holes or burn marks from hot plates, etc.
· cables must be attached firmly to the mains and to the apparatus.
When in doubt about the quality of the mains cable, discard it and get a new one.
Please note: Leakage circuit breakers cannot handle everything. They can only ensure against leakage from 230 volt mains connection to earth, and may thus prevent an electric shock. Contact with both “active” mains cables will give many kilowatts for hours. Leakage circuit breakers do not protect against errors in the high voltage output.
· Cables must be of high quality and always have approved “safety plugs”
· Cables and gel apparatus must have insulation against more than 1000 Volt (even 1500 Volt.)
In other words:
· Silicon rubber cables that give a continuous heat should be used
· Avoid rubber tubing (they crack) and thin plastic tubing that melts in contact with the edge of a hot plate.
· Never use transformers, adjoining cables or adaptors that can transform the approved safety sockets to those that are less safe. They are dangerous and must be discarded.
· Cables and gel apparatus must be capable of taking the same current as the attached safety plug.
Only two types of safety plugs are allowed. Other types are not allowed in the laboratories.
Note that both types have solid insulation sleeves so that the plug pins cannot be touched directly.
· Ordinary 4 mm safety plugs that are standard for most equipment. They are internationally approved and are suitable for about 1000 volt when they are dry.
· 2 mm safety plugs with a longer, stronger covering, suitable for up to 1500 volt.
Please note that plugs with resilient coverings are unsuitable as they are only for low voltage (to minimise short circuit). Under no circumstance must they be used or be available! The same applies for old-fashioned banana plugs (as well as sockets and extension cables). They must be discarded as they do not meet the regulations and are far too dangerous.
· Power supplies must of course have the correct sockets, corresponding to the correct safety plug.
· 4 mm safety sockets have mouldings suitable for the plugs covering.
· 2 mm safety sockets are deeper, corresponding to the long plug’s bigger covering.
· Equipment with old-fashioned sockets for banana plugs must be discarded.
· Power supplies with high voltage current must be insulated with respect to “earth” or have a safety circuit that breaks when earthed (this can be checked by technicians).
All new equipment must be CE-marked (this has been obligatory since 1997). However, a CE mark is not an approval (as many think), but merely the factory’s statement that the equipment is safe to use. You must therefore be critical and evaluate the equipment before buying!
Note, that Bio-Rad, for example, uses a “longer, modified, banana plug” without a solid covering, with accompanying lowered sockets in their power supplies.
Maybe this looks reliable when their equipment is assembled, but it is especially dangerous when used together with other equipment having the correct safety sockets.
· banana plugs with exposed electrical parts must be discarded immediately
· plugs with resilient coverings must not be used
· high voltage plugs and sockets must be the standard safely type with solid coverings
· all electric laboratory apparatus must be earthed, i.e., mains plugs must be Danish with 3pins
· you should contact the electronic technicians before buying new equipment to make sure that it is in agreement with the above rules.
Names of contact persons are to be found by each apparatus. Ask them for advice about the use of apparatus.
Students, trainee technicians, and all other users who do not have a thorough knowledge about the use of the apparatus, must - before attempting to use it - have clear instructions. It is the responsibility of each supervisor or project leader to ensure that each student or new person is well-instructed in the use of the apparatus. Instructions regarding use can be found in the "Instruction Manual" beside the machine. If in doubt, ask one of the contact persons.
Please notify the responsible contact person for the apparatus.
It is not a matter of doing a spring clean, but the obvious and necessary cleaning that must follow every time it has been used. Each and every user must make sure that the apparatus is cleaned!
In the event of spillage of genetically modified material, the spill must be wiped up and the area disinfected with 70% alcohol.
Rotors must NOT be washed with ordinary soap with a basic pH. A bottle of this should be found by each centrifuge.
Remember that centrifuges must be closed in such a way that the lid cannot be lifted while a rotor is in action. Rotors must never be stopped by hand.