EMERGENCIES
Police, Fire, Ambulance, Transportation
Dial 911
Put on CLEAN gloves. Determine if a foreign particle, such as glass, is present.
If no foreign particle or broken bone is suspected, cover with a clean cloth or towel and apply direct pressure to the wound for 5-15 minutes. If the wound is large, press the sides of the wound together. Lay the victim down and elevate the wound if possible. For transportation to medical aid, the cover over the wound should be held in place by a firm bandage that extends well beyond the edges of the wound.
If a foreign particle or broken bone is suspected, CALL 911. Cover the wound and apply pressure on each side of the wound, pressing the sides of the wound together firmly, but gently. If bleeding still continues or direct pressure cannot be applied, apply pressure to the artery supplying the affected limb, but no longer than 15 minutes.
Extinguish any flames with water or a blanket or coat (not a lab coat, which may melt and cause more damage!). A fire blanket is available in the metal cabinet in the hall on each floor. Some fire extinguishers contain chemicals that will cause harm if used on a person.
Application of an ice bag or cold water will cool tissues and reduce swelling. There are showers in each corridor. The burned area may also be placed under a gentle stream of cold running water in a sink. Treatment with cold water should be continued for 15-20 minutes if possible. The victim should then be placed in a reclining position, with the wound lightly covered with a clean cloth, and taken to the emergency room or urgent care center.
Speed is essential and WATER is best. Use showers or eyewash stations located in each corridor. Remove contaminated clothing if necessary. Thoroughly and continuously flood with water. Do NOT use neutralizing agents.
Eyes: If chemicals have been splashed into the eyes, thoroughly flush with water for 15 minutes. Make note of the type of chemical and then transport the victim to the emergency room so that a physician can promptly evaluate any possible damage.
AVOID EYE INJURIES! WEAR SAFETY GLASSES!
Any contact with phenol will ultimately produce a burn, even if the affected area is washed with soap and water. After thorough rinsing with cool water, treat all areas of the skin that have been exposed to phenol by gently applying glycerol (a good solvent for phenol that will not damage the tissue); cover the area with fresh glycerol for about 20 minutes. In the case of more extensive exposure to phenol, seek medical aid immediately.
Do NOT touch the victim directly until the current has been interrupted, or you may get the same degree of shock.
Break the electrical current by
Electrical shock may stop the heart and/or breathing and may require CPR. Check for this possibility and get help immediately.
Use caution if there are aqueous spills in the area.
For All Injuries
Report all injuries to the McArdle Departmental Office (Room 1010).
Refer to page 6E of the U.W. Chemical Safety and Disposal Guide for more information on emergency procedures.
All laboratories are required to have a Laboratory Emergency Information Notice posted outside each room as explained on page 3E of the U.W. Chemical Safety and Disposal Guide. See Tom Hart for blank forms.
Good laboratory technique includes the adequate protection of all personnel and of the environment during all laboratory work. This protection requires containment of possible hazards (radioisotopes, carcinogens, mutagens, other toxic chemicals, and viruses or bacteria that may be pathogenic for humans or laboratory animals). Another important requirement for laboratory safety is careful thought regarding accidents that could happen during performance of laboratory procedures. Seek advice and consider possible hazards, especially with new procedures. Remember that in a laboratory the safety of others is also your responsibility. Try to prevent accidents by others as well as by yourself. Safe science and good science are inseparable.
Use common sense, such as:
The few major accidents that have occurred at McArdle could have been avoided or minimized by forethought and use of protective devices. For example, one student lost the use of an eye in an explosion; this serious loss could have been prevented had he been wearing eye protection. Another worker received acid burns after a desiccator fell; her eyes were protected by glasses, and quick use of a nearby safety shower minimized the extent of the burns.
State regulations require the use of eye protection for workers in any laboratory. If you do not wear prescription glasses, you must wear glasses with nonrefracting lenses with side shields or goggles when you are in the laboratory. Contact lenses offer no protection and may cause serious problems if a chemical or biological agent gets into the fluid between the lens and the eye; for this reason, contact lenses are forbidden in the BL3 facility. Glasses or goggles must be worn by individuals wearing contact lenses. Full-time employees may obtain nonrefracting safety glasses without cost through the Safety Department. Those who wear prescription glasses need only supply their prescription to obtain glasses with hardened glass lenses free of charge; details are available in the departmental office (Room 1010).
Other safety aids are plastic goggles and glasses (available from the storeroom, Room 110), which provide a very high degree of protection from broken glass or flying objects (one kind also protects against ultraviolet rays); glass or plastic safety shields (order from scientific firms); and the safety glass windows in hoods. USE EYE PROTECTION AT ALL TIMES AND REMIND OTHERS.
Sonicators are high-frequency sound generators used to disrupt cells or shear nucleic acids. Wear earphone-type sound mufflers to protect your hearing while sonicating. If possible, have the sonicator located in a "sound-proof" cabinet while sonicating. Do not sonicate in a room containing people not wearing ear protection. Shut doors of room where sonication is taking place.
See pages 11D through 12D of the U.W. Chemical Safety and Disposal Guide for information on hand protection and glove selection.
Smoking is not permitted in the McArdle Laboratory. Eating and drinking should occur only in designated areas. A lunchroom is available on the first floor (Room 122); lab coats should not be worn in the lunchroom or in non-lab areas such as the reading room, 10th floor offices, copy room, etc. Food must not be stored in refrigerators or cold rooms used for experimental work.
Always wash your hands after working in a laboratory and before handling food or cosmetics. Laboratory glassware should not be used as containers for food. A microwave oven is available in Room 122 for heating of food. Do not use lab microwave ovens to heat food.
The following solvents are flammable (flash points below 27ºC): acetone, carbon disulfide, ethyl ether, ethyl acetate, ethanol, methanol, petroleum ether (hexanes, pentanes), benzene, and toluene. Carbon disulfide is spontaneously ignitable at steam bath temperatures. Many other organic solvents such as butyl and amyl alcohols, kerosene, xylene, etc., also burn readily, although their flash points are higher. Flammable solvents should be dispensed and used in a chemical fume hood.
Containers: The maximum allowable container sizes for flammables in labs are as follows:
Glass - 1 pint*
Metal or approved plastic - 1 gallon
Safety cans - 2 gallons
*Exception - Glass containers as large as 1 gallon may be used if required for purity.
Quantities: Not more than 10 gallons of flammables shall be stored in a single fire area outside of a separate approved cabinet. Each floor in McArdle is considered a fire area. Approved flammable liquid storage cabinets are strongly recommended, especially for users of a large variety of flammable liquids. Supply houses such as Fisher Scientific and Lab Safety Supply sell approved flammable liquid storage cabinets. At most, three cabinets are allowed on each floor. Not more than a total of 25 gallons of flammable liquids shall be stored in a single fire area. The storage room/solvent room by the freight elevator on the first floor is also available to store flammable liquids. If flammable liquids in the lab are no longer wanted, contact the Safety Department (262-9735). Their chemical safety staff will assist you in disposing of the chemicals.
Some refrigerators have been wired to permit storage of volatile flammable solvents in closed bottles. Most refrigerators and all McArdle cold rooms are not safe for this use, so that sparks from thermoregulators, motors, etc., could set off enormous explosions. Make sure the refrigerators in your laboratory are appropriately labeled and used.
Use steam, heating blocks, or, less preferably, electric hot plates, mantles, and oil baths. If possible, avoid use of open flames. When you must use open flames, always check first to be sure there are no flammable solvents or other flammable objects in the vicinity. Be sure to turn off the burner when you finish and before you leave the laboratory. Do not set the burner on a paper bench liner. Use proper Bunsen burner tubing.
When using microwave ovens to heat materials, be aware that water can easily superheat and boil over when its container is touched. Sealed bottles may explode - always heat an open container. Metal objects spark or diffract microwaves, ruining an oven. Never use an obviously damaged oven - it may leak microwaves. The Safety Department will check your microwave oven for leakage.
Don't forget to remove the stirbar from any flask you use in the microwave oven. Wear protective gloves when removing heated items from the microwave oven; a surge of active boiling may occur when you pick up the hot item. The boiling liquid may scald your unprotected hand and cause you to drop the container.
Turn off sterilizing flames not in active use. Two major fires in the McArdle Laboratory started from sterilizing flames left burning overnight.
Inspect dry glassware (especially large pieces and all glassware used for toxic, corrosive, or flammable chemicals) for cracks (hold up to the light) before use. Discard all glass desiccators or desiccator lids that have cracks or are chipped. Evacuated glass desiccators can implode and scatter glass and contents in all directions. Wire mesh guards for glass desiccators are recommended, especially if heat lamps are used to speed drying operations in these vessels. Flasks with large flat surfaces should not be evacuated unless they are the special thick-walled type (e.g., 125-ml Erlenmeyer flasks are OK, but for larger sizes a "suction flask" is necessary).
For introducing glass tubing into cork or rubber stoppers, be sure to lubricate the glass and stopper with oil or glycerol before inserting the tubing. Use a towel or cotton gloves to protect your hands.
Do not send to central dishwashing facility any glassware
For carrying glass bottles with liquid chemicals, especially concentrated acids and bases such as sulfuric acid, nitric acid, ammonia, sodium hydroxide solutions, solvents, phenol, etc., use rubber or plastic bottle carriers. Each lab should own such a carrier. Avoid carrying reagent bottles up and down stairs; use the elevator. Use carts for the transport of large glass carboys.
Hallways are not to be used as storage areas. Carts, autoclave buckets, tables, desks, chairs, etc., are not to be kept in hallways or stairwells. In case of emergency, movement through cluttered hallways would be difficult. If lights were to go out at night, safe passage would be impossible.
Use magnetic stirrers or electric induction or air motors whenever possible. Avoid use of open brush motors.
Electrical equipment must be adequately grounded via the plug receptacle. This grounding is especially important with equipment in contact with water or moist skin. Use water baths that shut off automatically when they overheat, e.g., owing to evaporation of the water. Particular care should be exercised with electrophoresis equipment. Such equipment should be equipped with a safety interlock designed to turn off the current when the cover is opened.
Use timers when running electrophoresis experiments. Choose proper current loads when running overnight electrophoresis separations - too much heat can dry up the gel and cause arcing of the current. Try to avoid overnight electrophoresis runs.
Use large shallow metal pans under chemical reactions, distillations, etc., to catch chemicals in the event of flask breakage, etc. Pans should always be used for work involving more than very small amounts of carcinogens, mutagens, radionuclides, or other toxic or corrosive chemicals so that the area can be readily decontaminated. For very small amounts and volumes, a disposable labmat can be adequate protection.
Before working with any chemical, learn its hazards. Every research group is required to keep a file of Material Safety Data Sheets from shipments of hazardous chemicals used in the lab. Material Safety Data Sheets and other information are also available from the Safety Department. The Safety Department home page (www.fpm.wisc.edu/safety) has links to Material Safety Data Sheet databases. Wisconsin law requires that supervisors and faculty train all their employees for work with hazardous chemicals.
Reagents such as fuming acids, peracids (especially perchloric acid), peroxides (as such, or formed in ethers), halogens, alkali metals, phenols, etc., can be hazardous. Read about their properties and seek advice on the safe handling of these chemicals. Solid residues of peracids (especially perchloric acid) and their salts can be detonated by a sharp blow if they are in contact with organic matter. Bottles of perchloric acid must be kept in glass or metal pans that do not contain other organic chemicals and that are washed periodically. Perchloric acid should not be evaporated in hoods, because the residues provide a potential hazard to workmen at a later time.
If a procedure requires the evaporation of large amounts of ethyl ether, be sure to start with low-peroxide ether. Peroxides accumulate in ether with time and may be concentrated on evaporation, making detonation more likely. Store ether only in the original metal cans in a hood. Ether can be treated to remove peroxides. For the method, see "Prudent Practices in the Laboratory," located on the safety shelf in the McArdle Reading Room. Also, refer to pages 2B and 20D through 23D of the U.W. Chemical Safety and Disposal Guide.
Safety glasses or other eye protection should be worn at all times in the laboratory; this requirement is especially important when using corrosive chemicals. Alkali and phenol are particularly destructive to the lens of the eye.
These usually result from poorly connected water hoses or from plugged drains (often the result of loose paper towels). When condensers, etc., are used for long periods, especially overnight, be sure that the water hoses, connections, and drains are sound. Do not try to operate cold water faucets at low velocities, because some are equipped with anti-suckback devices, which will sometimes leak and cause floods. Use larger velocities and attach a by-pass (T-tube with clamp) to supply low velocity water to your equipment. If there is a flood in your lab, be aware of the dangers of wet electrical cords and equipment.
Water may be removed from a flooded floor with a vacuum device kept in the animal washroom (room 906). See departmental office (Room 1010) for a key to access the 9th floor. A mop and mop pail are available in Room B7.
People are reluctant to clean up spills in common-use areas if they do not know whether the spills are hazardous. If you spill liquids in elevators or hallways, clean them up.
Ultraviolet light, especially germicidal short wavelengths (~250 nm), gives rise to burns if used without adequate protection to the eyes and skin. An intensive eye itch and "foreign body" irritation in the eye are often the first hints of trouble a few hours after exposure. Seek medical aid. Some ordinary glasses (e.g., glasses with plastic lenses) do not protect the eyes from UV exposures; use special goggles. Skin protection can be achieved by use of a laboratory coat, gloves, glass shields, etc. (Note that polished metal surfaces can reflect sufficient UV light to be harmful.) Everyone using a UV microscope must wear protective goggles or glasses, and anyone else in the room during such use should also wear such goggles or glasses.
A special small room for dispensing liquid nitrogen has been constructed near the stockroom on the first floor. This room is open only from 8 AM to 4 PM Monday through Friday. It is equipped with a sign-out sheet, hearing protectors, a large plastic funnel, goggles, and gloves. All must be used whenever containers are being filled with liquid nitrogen! The door to the room must be open while nitrogen is being dispensed.
Liquid nitrogen (‑196ºC) can cause severe burns when splashed onto the skin or into the eyes. Glass or plastic vials sometimes explode when they are removed from liquid nitrogen, owing to rapid expansion of nitrogen that has leaked in during storage. Eye and hand protection should always be worn when vials are withdrawn. Plastic vials with O-rings explode less frequently and should be used whenever possible. If you see boiling within the vial as you withdraw it, quickly cover it with a heavy plastic beaker to contain a possible explosion. Be sure that student helpers know how to handle liquid N2 safely.
The cabinet for dry ice is near the stockroom on the first floor. Goggles or glasses must be worn when dry ice is handled, and the sign-up sheet must be filled out.
Cylinders must be chained or strapped to a firm support, such as a lab bench. Handle cylinders and valves gently; do not use excessive force with wrenches. Never attempt to remove main valves. Toxic gases (e.g., CO or HCl) should be ordered in small cylinders that could be housed in a hood with adequate ventilation; in the event of a leak, only the hood needs to be turned on. (This caution does not apply to CO2.) See Paul Weber in Room 110 about disposal of such a cylinder.
Reducing valves on cylinders that contain corrosive gases should be removed, flushed with water, and dried with a compressed air stream between uses; the valves last longer and are less likely to become stuck.
In turning off the flow of gas from a cylinder, close the main valve and then the reducing valve. When you start the gas flow, keep the reducing valve closed while you open the main valve; then adjust to the desired flow rate with the reducing valve. Needle valves in good condition do not require force to close them; they are damaged by use of force.
Gas cylinders should be capped during transport. Some types of rubber tubing (particularly those with thin walls) may burst if attached to a source of gas under high pressure.
Mouth pipetting is not permitted at any time. Various hand-operated pipetting devices are available from the storeroom and from commercial sources.
The laboratories are supplied with a house vacuum line connected to a large pump in the basement of the building. Whenever this vacuum line is used, the line should be protected by an appropriate trap or filter (a Gelman product is available from the Fisher catalog) to prevent any possibility of water, solvents, radioactivity, cells, or any other material getting into the line. This is important for the safety of the workmen who service the unit. In-line disposable HEPA filters are available to protect vacuum lines in the BL3 facility.
If you encounter an individual who does not belong in the building or who appears threatening to you or others, call Police and Security (911). For the protection of our personnel and equipment, the outside doors to McArdle are locked at 5:45 p.m. on weekdays and are locked at all times on weekends. These doors should never be blocked open. In addition, all of the doors to individual rooms (with a few exceptions) should be locked at night and on weekends if no one is in the immediate vicinity. At all times, keep your valuables either with you or in a locked place.
For the protection of the public and the experimental work, the laboratory areas of the building are not open to the general public. Visitors are welcome in specific areas at the discretion of the staff member in charge. Children under age 12 are not allowed in any of the laboratories or animal rooms. Keys to the McArdle Laboratory or to any part of it are a privilege accorded to facilitate work in the building. The keys should never be lent or copied and must be returned when employment is terminated. When you enter the building at night, don't let in strangers waiting by the doors.
These items provide protection to the skin in case of an accident. Buttoned laboratory coats or plastic aprons should be worn for most laboratory work. Bare arms and legs provide broad expanses for chemical burns and should be avoided. Feet should always be protected by closed-toe shoes (not sandals). Laboratory coats that have been used for laboratory work should not be worn in the lunch areas, restrooms, reading room, 10th floor offices, copy room, etc., or outside the building.
When plastic or rubber gloves are used to protect you from radioactive or toxic chemicals or infectious agents, they should be worn only when needed for protection and should be monitored while on your hands. Contaminated gloves must be removed and disposed of. Before answering the phone, pushing the elevator buttons, opening doors and refrigerators, etc., remove gloves to avoid possible dissemination of hazardous materials. Heavy rubber gloves or double plastic gloves should be used for work with high levels of radioactivity or toxic chemicals. Call the Safety Department (2‑8769) to determine the proper type of glove needed for specific chemicals.
are located near each end of each hall. After use, remove water from floor with vacuum device kept in the animal washroom, Room 906. You may borrow a key to the 9th floor from McArdle 10th floor office. A mop and mop pail are kept in Room B7.
are in each hall near the showers. (After use, notify Don Fechner or Tom Hart so that the unit may be refilled.)
CO2 fire extinguishers are in every laboratory and near each end of the hall on all floors. An ABL dry chemical fire extinguisher is in each hall on the laboratory floors. On the 10th floor, a water fire extinguisher, more suitable for paper fires, is in the hall. Learn how to operate the fire extinguishers - be sure to direct the stream at the base of the fire. Never store carts or tables in hallways blocking fire extinguishers. Every McArdle employee should attend yearly safety meetings to learn proper use of fire extinguishers as demonstrated by the Safety Department. (After use, notify the Safety Department so that the extinguishers will be recharged.)
Fire alarms are at each end of each hall.
If a fire cannot be immediately controlled with the extinguishers, UNLOCK AND CLOSE DOOR(S) OF AFFECTED ROOM, SOUND THE FIRE ALARMS (LOCATED AT THE ENDS OF EACH HALL), AND LEAVE THE BUILDING BY THE STAIRWELL. DO NOT USE THE ELEVATORS. Wait at the outside door to direct the firemen.
When the first alarm rings, all personnel should:
are placed in the halls next to some of the doors to the rooms. If a centrifuge goes off balance or other accident occurs, it may be desirable to turn off the electrical current at the circuit breaker. Learn where the circuit breakers for your room are located.
Shutoff valves for the hot and cold water lines and for the gas lines in the individual rooms are located on each floor in the vertical utility shafts. The doors to these shafts open into the hall and, if locked, can be unlocked by a utility key (see Don Fechner, Tom Hart, or 10th floor administrative office; call Police and Security in after-hour emergencies).
Chemical hoods should be used for all operations with noxious gases or other toxic or volatile chemicals. The safety glass in hood windows is a valuable safety shield. The maximum face velocity for the hoods in McArdle is marked on each hood. For some purposes (regulated and very potent chemical carcinogens, potent toxins such as di-isopropyl fluorophosphate, large amounts of radioisotopes, volatile nuclides, etc.) it is better to use the hood in room 920. Hoods for working with carcinogens and mutagens are in Room 5079 of the B-wing animal facility.
Vertical laminar-flow cabinets are available in a number of laboratories for containment of hazardous biological agents. Some of these hoods are connected to the building exhaust system so they can be used for biological experiments with toxic agents. Do not use these cabinets as chemical fume hoods. Training on proper use of bio-safety cabinets is important. See http://www.cdc.gov/od/ohs/biosfty/bsc/bsc.htm.
In addition to the vertical laminar-flow bio-safety cabinets, there are some horizontal laminar-flow cabinets. The horizontal-flow cabinets are useful for maintaining sterility of materials and can be used for purposes such as the preparation of culture fluids. The horizontal-flow cabinets must not be used with infectious agents, toxic chemicals, or mammalian cell lines, since the air from the hood is directed at the operator.
Disposable respiratory filters (for dusts) are available from the storeroom. Face respirators equipped with cartridges to remove dusts or certain chemicals are available from safety supply and scientific companies. Disposable HEPA filter respirators are available for partial protection against certain infectious agents transmitted via the respiratory route, but only when the agent cannot be manipulated within a bio-safety cabinet. Furthermore, fit testing of ALL respiratory protection devices is required by OSHA/Department of Commerce. Please contact Carla Alvarado (3-2177) or Bill Deppen (2-9179) of the Safety Department for further information about respiratory protection and fit testing.
The use of radioactive chemicals requires care to avoid contamination of personnel and the environment. The precautions required depend on the radioisotope, the nature of the chemical in which it is incorporated (e.g., chemically reactive molecules, volatile chemicals, etc.), and the amount handled. The responsibility for the safe use and disposal of radioisotopes rests with the individual using these materials and with the responsible staff member. No one is permitted to use radioactive materials until he/she is familiar with their safe handling and detection. A hands-on, 4-hour course must be completed prior to use of radioisotopes; see your major professor for details.
Any spill should be cleaned up with due consideration for the radioactivity (nuclide activity, chemical form) and of the chemicals (volatile, corrosive, etc.) involved. The best general approach is to (1)contain the spill, (2)move the items individually, after each has been demonstrated to be free of contamination, to a clean area, and (3)take care to avoid spreading the radioactivity with cloth, paper, etc. used for cleaning up the spill. Disposal of such clean-up materials should be in accord with University Health Physics standards (see pamphlets on the safety shelf in the McArdle Reading Room). Spills should be reported to the major professor and, if appropriate, to the Health Physicist at the Radiation Safety Office (2-8769).
Room 920 is available for work involving large amounts of radioisotopes. Be sure your group is authorized by the Radiation Safety Office to work in this room before you begin. The hood in Room 920 is designed for work with volatile radioisotopes.
Whether the work is carried out on an assigned workbench, in Room 920, an equipment room, or elsewhere, each worker is responsible for cleaning up and monitoring the work place. The kind of monitoring will depend on the level of radioactivity, the isotope, and the properties of the labeled material.
Remember that decomposition by yeast, bacteria, animals, etc., of chemicals and biological materials that contain 3H or 14C may yield 3H2O and 14CO2, both of which are volatile and will contaminate the atmosphere. Used culture media, dead animals, etc., that contain these isotopes should be properly packaged and refrigerated or frozen until they are removed for disposal.
See page 21 for information on the disposal of radioactive waste.
Wear disposable gloves, lab coat, and eye protection when working with any radioisotope. The gloves must be removed before transporting trays of samples to the counting room. Also, be sure to remove gloves when answering the phone, opening a door, or using shared lab equipment. Gloves as well as other radioactive waste must be disposed of properly - not tossed into trash cans (see page 21).
If you are carrying out routine monthly surveys for possible laboratory contamination, treat the situation as though the samples being collected really contain significant counts; i.e., wear gloves, lab coat, and glasses. Student assistants must complete the 4-hour radioisotope training session before performing monthly contamination surveys.
If you are responsible for teaching new employees the proper use of radioactive material, be sure not to cut corners. For example, even though the isotopes 3H and 14C have relatively low energy and, therefore, little penetrating ability, instruct your trainees to wear gloves when using these isotopes. Do not tell them, "Just wash your hands before you go home." Cuts, scratches on hands, or irritated skin can allow internal contamination to take place even with 3H or 14C.
Some of the properties of the radionuclides commonly used in McArdle are listed below. For detailed information, see the University Radiation Safety Regulations and Radiation Safety for Radiation Workers. Copies of each of these booklets are available in the Reading Room and from each major professor. Booklets on the use of many radionuclides are available from the Safety Department.
This isotope emits a very weak b-particle with an average energy of 0.006 MeV and has a half-life of 12.6 years. There is no hazard from external radiation, but care should be taken not to inhale or ingest this isotope or to get it into an open cut. This precaution is particularly important for work with [3H]-nucleotides, which may become localized in DNA. Urine monitoring is required for persons working with 10mCi or more of unsealed tritium and is performed by the Safety Department. For this purpose, a background count must be taken within 1 week prior to tritium use, and further counts must be taken at least weekly during and after the procedure until background levels are again obtained.
This isotope has a weak b-particle emission of about 0.05 MeV and has a half-life of 5700 years. There is no hazard from external radiation, but care should be taken not to inhale or ingest this isotope or to get it into open sores. If ingested or inhaled as CO2, this isotope may be incorporated into bone mineral and remain there indefinitely.
35S emits a weak b-particle with an average energy of 0.05 MeV and has a half-life of 88 days. There is no hazard from external radiation, but the same precautions described for 3H and 14C should be applied to 35S.
This isotope emits a strong b-particle with an average energy of 0.69 MeV and has a half-life of 14 days. Shielding must be employed; you must wear glasses. It is best to store 32P in Plexiglas containers (usually 5/8 in. thick is adequate) rather than behind thin metal (such as lead). The reason for this recommendation is that when high-energy b-particles strike some materials, they produce secondary x-rays, which can penetrate most shields. Because of the possibility of the formation of secondary x-rays, storage areas for 32P should be monitored with a gamma-ray counter as well as with a b-particle counter. Personal dosimeters (obtainable from the departmental office) must be worn by persons exposed to 32P. Monitor your area frequently with a Geiger counter if you are working with 32P. 33P might be an alternative to 32P. 33P has lower energy and a longer half-life (25 days) than 32P.
125I has a half-life of 60 days and emits a gamma ray with an average energy of 0.035 MeV. 131I has a half-life of 8 days and emits a gamma ray with an average energy of 0.36 MeV. Persons working with either of these isotopes should wear personal dosimeters (obtainable from the departmental office). Protocols that may produce volatile iodine must be performed in the hood in Room 920. These isotopes should be stored and used with shielding. Isotopes of iodine pose the unique hazard of being concentrated in the thyroid gland. Therefore, thyroid scans are required of all persons who use 100 mCi or more of a volatile or dispersible radioiodine (such as NaI) or 1mCi or more of radioiodine bound to a nonvolatile agent. The background count must be taken within 6 months prior to the time that the iodine is used. A second count must be taken within 7 days after the iodine is received in the laboratory; if the iodine will not be used within 7 days from receipt, the Health Physicist must be called (2-8769) for an extension of this count. Whenever an iodination is carried out, the breathing zone air must be monitored for radioactive iodine with an apparatus available at the Safety Department.
Be sure to check the properties and the regulations that govern their use before working with any of these. Call Radiation Safety (2‑8769) to ask whether pamphlets are available.
Use of the irradiator is restricted to those who have completed a 2‑hour course given for new users by the Safety Department. Contact the Safety Department for the schedule, if you wish to participate.
If there are questions, confer with your major advisor, the University Health Physicist (2-8769), and/or a member of the McArdle Safety Committee.
Chemical carcinogens, mutagens, and teratogens are necessary experimental chemicals for many studies in McArdle. Basic precautions are necessary for the protection of the worker and others using these chemicals in the laboratory. The Chemical Safety and Disposal Guide gives general instructions for use of these chemicals. Staff members are responsible for ascertaining that appropriate procedures have been developed for the use of all chemical carcinogens, mutagens, and teratogens in their laboratories and for being sure that their personnel are instructed in appropriate procedures. Each worker should make sure that he/she understands the safety procedures to be followed.
Microorganisms or viruses that are pathogenic for humans or laboratory animals must be used and disposed of in a manner that will prevent human or animal exposures. The precautions depend on both the properties and pathogenicities of the agents and on the type of work being done. The best resource for procedures, equipment, and facility requirements can be found in the CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories. Also, a comprehensive listing of human pathogens according to Risk Group (1-4) is found in Appendix B of the NIH Guidelines for Research Involving Recombinant DNA Molecules. Both of these documents are accessible via the Office of Biological Safety Web Page: http://www.fpm.wisc.edu/biosafety or http://www.fpm.wisc.edu/safety (Government Web Resources).
The practice of Universal Precautions is required for handling human blood, tissues, and certain body fluids. This practice assumes that these materials are infections and are thus handled accordingly. The OSHA Bloodborne Pathogens (BBP) Rule applies to any person with a potential occupational exposure to certain materials of human origin. Included in the BBP rule are training, hepatitis B vaccination, exposure control plans, and post-exposure follow-up requirements. For more information, please contact Carla Alvarado (3-2177), Manager of the Occupational Health Program. http://www.fpm.wisc.edu/bbp
At a minimum, the use of organisms or viruses pathogenic for humans requires that mouth pipetting be forbidden, that precautions be taken to avoid the release of the agents into the air (for instance, through aerosols), that approved biological safety cabinets be used, and that all preparations containing these biological agents be decontaminated before disposal. The manner of decontamination depends on the organisms or viruses, but autoclaving (efficacy verified with spores, as needed) or use of 5% sodium hypochlorite (chlorine bleach) is widely applicable. Do not autoclave solutions containing bleach. In some cases, protective clothing should be used; in these cases, the protective clothing must not be worn outside the laboratory room. Materials that are to be autoclaved for decontamination must be transported to the autoclave in closed plastic autoclavable bags, closed metal containers, etc. Appendix B of the Guideines for Handling Pathogenic Microorganisms contains a listing of decontamination procedures and potential applications. Infected animal carcasses should be prepared for incineration as indicated in the University of Wisconsin Chemical Safety and Disposal Guide.
Those who have questions with regard to the use or disposal of biological agents should first consult the staff member in charge of their work and then, if needed, other staff members such as Dr.Sugden or Dr.Lambert. The Biological Safety Officer of the Madison Campus is Dr.Joseph Kanabrocki (3-2037), who may also be consulted.
Additional procedures are required for tissue culture work with infectious forms of hepatitisB virus or human immunodeficiency virus. These procedures are outlined in the Biosafety Level3 instructions. All work with these agents is to be carried out in the BL3 laboratory. Anyone who plans to work with these agents must contact Don Kaehler for instructions on the use of the BL3 laboratory.
The McArdle Biosafety Level-3 (BL3) lab represents the level of containment recommended by the Center for Disease Control (CDC) and the National Institutes of Health (NIH) for the safe handling and containment of various human pathogens such as HIV. It incorporates features that restrict access to the facility, limit exposure to pathogenic agents, and prevent those agents from leaving the facility and presenting a potential risk to the community. Two design features define this level of biocontainment.
While a comprehensive description of BL3 facility recommendations can be found in BMBL http://www.cdc.gov/od/ohs/biosfty/bmb/bmb11-1.htm, the salient features include:
Federal guidelines: http://www4.od.nih.gov/oba/rac/guidelines/guidelines.html
Some experimentation involving recombinant DNA technology/genetic engineering requires prior approval by the University Institutional Biosafety Committee (IBC) and National Institutes of Health (NIH) Recombinant DNA Advisory Committee. Investigators planning to initiate molecular cloning experiments are obligated to familiarize themselves with the NIH guidelines (copies available from the Biological Safety Office, [3‑2037]) and determine their appropriate responsibilities. Research with transgenic animals is subject to IBC review. Any question regarding approval procedures or containment levels should be addressed to the above office.
For further information: http://www.fpm.wisc.edu/bbp/
Samples of blood, tissues, tissue culture cell lines and their supernatants, etc. from humans may be contaminated by viruses or bacteria. Such viruses and bacteria might be able to infect and clearly have the capacity to proliferate in human tissues. Accordingly, all work with such tissues should be carried out in a manner that will protect the user and others who might come in contact with the materials from inadvertent infections. For example, blood samples should not be allowed to come into contact with open cuts or skin abrasions, and they must be decontaminated before disposal. (Note: A fraction of persons who work with human blood samples develop hepatitis. This can be avoided by wearing gloves.) Researchers who use human blood, tissues, or certain human body fluids are subject to the OSHA Bloodborne Pathogens Rule. A requirement under this Rule is the creation of an Exposure Control Plan. Please consult with Carla Alvarado (3-2177), Occupational Health Manager.
The McArdle Laboratory has facilities for the care of rodents on the 9th floor of McArdle and on the 5th and 6th floors of the B wing of the Medical Science Center. The space in these facilities is assigned to specific staff members who have received an animal protocol number. Animals may be put in an assigned animal room only with the approval of the major professor. DO NOT put your animals in ANY room not assigned to you.
A manual describing the regulations for the use of experimental animals is being assembled (expected completion in Fall of 1999). All animal users must familiarize themselves with the contents of this manual. In the meantime, special instructions are available for the use of the animal rooms, or consult Norman Drinkwater or Alexandra Dove.
Animals must come from an approved source, and all orders must be placed through Patty Swan (phone 2-2820). In the event that someone requires animals from a different source, prior quarantine arrangements must be made. This regulation is required to avoid the introduction of a pathogen into the animal facilities. (Animals from many sources carry viruses such as mouse hepatitis virus, parasites such as mites and pinworms, or any of a number of other pathogens.) Once a pathogen is introduced into the facility, it is likely to spread to other rooms and may cause serious health and mortality effects. Even if overt disease does not occur, the loss in valuable research can be very high. Furthermore, the time, effort, and cost required to free the animal facility of a pathogen are enormous.
IN ADDITION, ANIMALS INOCULATED WITH ANY TISSUES OR MAMMALIAN CELLS, including hybridoma lines obtained from other labs, MUST BE QUARANTINED. Such material may contain viruses and must be treated with the same precautions as indicated for new animals. See a member of the Animal Care Committee for information on quarantine procedures.
Before radioactive materials are given to or injected into an animal, approval must be obtained from the University Radiation Safety Committee.
Blood from experimental animals, along with tubes, needles, etc., must be discarded in rigid, autoclavable containers which, when full, must be sealed and sterilized. (See section on sharps disposal, page 27.)
All animal byproducts should be assumed to be infectious.
For additional information please consult Section G of the U.W. Chemical Safety and Disposal Guide and the section on disposal in the Radiation Safety Regulations (both prepared by the U.W. Safety Department), or call the U.W. Safety Department.
Organic solvents, except for small amounts of water-miscible solvents, must be discarded in the 5-gallon waste solvent jugs supplied by the Safety Department. These solvents are sent to an incineration facility. The solvents must not contain unneutralized acids or bases or chemically reactive materials. They must not contain solids such as glassware, corks, major precipitates, etc. The solvent waste may contain water, but the amounts should be kept as low as possible. The solvent waste may also contain up to a total of 3mCi of 14C and/or 3H per 5-gallon jug. The solvent waste may contain carcinogens, mutagens, and other organic chemicals that can be totally combusted in an incinerator (2200°F).
If appreciable amounts of halogenated hydrocarbons are used, they should be disposed of separately from the other solvents. In this case, it may be necessary to have one 5-gallon carboy for regular solvent disposal and another for solvents containing halogenated hydrocarbons.
Organic acids and bases may be put in the waste solvent jugs after neutralization, as indicated above. Inorganic acids and bases should be neutralized and washed down the drain with lots of water. For detailed information refer to pages 11G through 16G of the U.W. Chemical Safety and Disposal Guide.
(Additional details are contained in Section XIX of the University Radiation Safety Regulations.)
A. Disposal to the Radiation Safety Office (preferred). The Safety Department provides 5-gallon carboys for aqueous and organic 3H, 14C, and 35S wastes. In some cases, Safety also provides bottles for small volumes of liquid waste. All other waste containers and/or boxes must be provided by the user. Prepare and package the waste according to the waste type in steps 1-5. Follow steps 6-9 below for final preparation of all wastes for pickup. Wastes will not be picked up if improperly packaged. Authorized users must correct waste disposal inadequacies before additional pickups will be made.
1. Solids. Place solid wastes in a yellow plastic bag designated for radioactive waste, seal, and put in a labeled radioactive waste box.
a. Use strong packaging tape to secure the bottom and top of these boxes.
b. Package and seal sharps (syringes, blades, Pasteur pipette tips, etc.) inside an impenetrable container before adding them to the other solid waste.
c. Package small-volume, concentrated wastes (e.g., stocks, hot products or wastes, sealed sources, etc.) into small boxes separate from large volumes of dilute, contaminated wastes.
d. Then follow steps 6-9 of this section.
2. Lead Pigs. Do not put lead pigs in solid waste boxes. Lead is accepted with other waste only when needed to shield very hot, concentrated wastes in small boxes only. Meter lead pigs with an appropriate instrument to determine whether a. or b. applies.
a. Safety will pick up clean surplus lead for recycling.
(1) Pack in small box (i.e., <50 lb.).
(2) Write Lead Pigs on box.
(3) Write Lead Pigs on disposal form.
(4) Put out with other radioactive wastes.
b. Contaminated Lead Pigs
(1) Place in plastic bag, then separately pack contaminated pigs in small box.
(2) Identify the nuclide.
(3) Write Contaminated Lead on the box.
(4) Then follow steps 6-9 of this section.
3. Liquids. Keep aqueous and organic solvent wastes separate and collect in plastic or shatter-proof glass containers of at least 500 ml, but not more than 4 liters; use an appropriate size for the actual waste volume. Five-gallon carboys may be used for large volumes of dilute liquids; when requested, liquid waste containers are delivered during waste pickups.
a. Neutralize aqueous liquids; pH to be between 5.5 and 8.5.
b. Very concentrated wastes (e.g., stocks), not exceeding 50 ml, should not be diluted (and need not be neutralized). Tightly seal vials and package separately in a small box.
c. Document in proper section of disposal form (i.e., organic liquid or aqueous liquid).
d. Do not put solids in liquid waste containers.
e. Fill container 2/3 - 3/4 full; do not over-fill containers; allow room for thermal expansion.
f. Seal containers securely by wrapping parafilm around a tightly closed, twist-type cap (do not use foil, cracked caps, etc.).
g. Complete and attach a Radioactive Liquid Waste tag to each container >50 ml. List all constituents, including water, methanol, etc.
h. Cushion and place no more than four containers into each box.
i. Then follow steps 6-9 of this section.
4. Liquid scintillation cocktail (LSC) wastes. Sewer-disposable LSC solutions may be poured directly to the sewer (see Section B, Disposal to the Sanitary Sewer). For pickup by Safety, sewer-disposable LSC solutions may be either poured into aqueous liquid containers (see above) or kept in their vials and repackaged in cases (preferred). Organic hydrocarbon cocktails (e.g., toluene, xylene, pseudocumene, etc.) must be kept in their original counting vials for pickup by Safety. If you generate these cocktails in bulk, request an exception. When disposing of LSC wastes in vials, follow these procedures:
a. Keep organic hydrocarbon cocktails separated from sewer-disposable cocktails.
b. Keep vials separated by size and type (e.g., plastic, glass).
c. Place vials upright in trays and package in full cases only (20-ml vials, 500/case; mini-vials, about 1700/case).
d. On each case mark the LSC cocktail brand name and any biological or chemical hazard that might make sewer disposal inappropriate.
e. Then follow steps 6-9 of this section.
5. Animals. All animals that have been injected with or administered radioactive materials must be disposed through Safety when sacrificed or expired. Double-bag and box all animal tissue and contaminated bedding/waste (U.W. Stores stocks these supplies).
a. Activity limits: <15 mCi 3H and/or 14C (per package); <2 mCi any other nuclide.
b. Carcasses must be placed in a plastic bag, boxed, and frozen.
c. Weight limit is 50 lb/package (larger animals must be sectioned).
d. Blood, urine, and feces should be diluted and disposed via the sewer system (see Section B, Disposal to the Sanitary Sewer, page 24).
e. Then follow steps 6-9 of this section.
Call the Radiation Safety Department in advance to discuss any exceptions for animal disposals.
6-9.Final preparation of all wastes
6. Complete the Radioactive Waste label for each waste box. Complete a Radioactive Liquid Waste tag for each liquid container; tie or tape the tag securely to the container. Always report activity in millicurie (mCi) units. Do not use "trace" or "less than X". Note other hazardous chemicals, including biohazards and toxic materials, and state precautions needed for safe handling.
a. Measure or estimate the waste activity. The higher the activity, the more precise the estimate must be.
b. Waste activities should be accurately recorded if possible. Sub-sample, count, correct for efficiency (cpm to dpm), and convert from specific activity (mCi/ml or mCi/g) to total activity [specific activity times (x) volume].
c. Correct waste activities for decay when significant (i.e., 20% or more of the original activity); report decay in the lower right corner of the disposal form (Appendix J-1). Decay graphs for various isotopes are available from Radiation Safety.
d. CORD will subtract the activities disposed by a lab from that lab's received inventory balance.
7. Seal all containers. All wastes, except carboys, must be packaged in sealed boxes. Liquids must be cushioned.
8. Complete the appropriate Radioactive Waste Disposal form. Use the blue form for animal tissue and the orange form (Appendix J-1) for all other wastes (Appendix I, Radioactive Waste Disposal Guidelines).
a. One form may be used for all wastes being disposed. List each box on a separate line.
b. Keep the original for your records and attach Safety's copy to a waste container.
c. Go to Room 19 Biochemistry or call the Safety Department for additional disposal forms.
d. Maintain all receipt and disposal records for at least 3 years after final disposal.
9. Call the Safety Department (262-8769) or use the waste disposal request on the Safety Department web page, http://www.fpm.wisc.edu/safety, to schedule the waste pickup. Radioactive waste pickup schedule: animals on Wednesday and Friday mornings beginning at 8:30 AM, and all other waste on Monday and Wednesday afternoons beginning at 12:30 PM.
a. Lock animal waste in the freezer located beside the McArdle ice machine; lock non-animal waste in Room #B1260, located in the loading dock area of the Biotech Center. See Paul Weber (Room 110 McArdle) for keys.
b. After placing the lab's waste inside, insure the count rate is <650 cpm on contact with outside wall of cabinet or freezer.
c. Wastes that exceed the 650 cpm exposure limit or do not fit into the cabinet should be kept in your lab until Radiation Safety arrives. Inform Safety when you schedule the pickup that you need to be called because you have waste in your lab and give the reason. When called, promptly take waste to pickup area.
B. Disposal to the Sanitary Sewer. As part of the authorization process, authorized users may be allowed to dispose of up to 2 mCi/year to the sewer for all radionuclides combined. This limit may be increased if necessary or deemed practical; authorized users must apply for any exceptions. In addition, radioactive liquids must satisfy the following criteria for disposal to the sanitary sewer:
1. Concentrations must be at or below the limits listed in Table 9, Section XIX of the University Radiation Safety Regulations prior to discharge to the sewer. Keep records of concentrations and activities disposed.
2. Neutral pH: between 5.5 and 8.5.
3. Materials must be aqueous and readily soluble in water.
4. Other chemical and biological waste constituents must be safe for sewer disposal (check the U.W. Chemical Safety and Disposal Guide for chemical restrictions).
Always use the lab's designated (i.e., labeled) "hot" sink and run additional water during and after release to flush the drain and pipes. Survey the sink after use. Maintain a log of disposals at the sink (e.g., Appendix P, University Radiation Safety Regulations; Radioactive Material Record). Report releases to the sewer, at least monthly, in the lower right corner of the Waste Disposal form.
C. Exhausting Radioactive Material to the Atmosphere. Call Radiation Safety in advance if any procedure may result in a radioactivity release to the atmosphere. Releases to the atmosphere require evaluation of radionuclide air concentrations and special approval of the designated exhaust system (i.e., hood). Air monitors are installed in hoods approved for volatile iodine releases (see Section XIII, Part B, University Radiation Safety Regulations). The Safety Department measures and documents releases from procedures involving volatile iodine. All other releases must be measured and/or evaluated and documented by the authorized user. Report releases to the atmosphere, excluding iodine, in the lower right corner of the Waste Disposal form. Atmospheric releases must be reported within the same year as the release.
D. Natural Decay (not recommended for disposal option):
1. Decay. Users of radionuclides with short half-lives (e.g., 32P) who do not dispose of these wastes via routine pickups must periodically (at least four times per year) record disposal by decay. This is done by completing the lower right block (Other Disposal Methods) of the Radioactive Waste Disposal form.
2. Decay and disposal to normal trash. The amount of radioactive material on hand continually decreases because of radioactive decay. If the waste has a nuclide with a half-life less than 65 days (e.g., 32P, 125I), it may be convenient to wait until the natural decay process removes most of the radioactivity. As a rule, after ten (10) half-lives there will be approximately 0.1% of the original remaining. This may be low enough so that there is essentially no activity remaining; however, a 1-mCi solution of radioactive material will still have approximately 1 mCi of material remaining after 10 half-lives. It is crucial for disposal by natural decay to do the following:
a. Hold the waste for at least 10 half-lives.
b. Survey with a sensitive survey meter (see Section IX.B.1 or Section XI.A.1, University Radiation Safety Regulations) for detectable radiation and/or radioactivity before disposal via normal trash. Survey should show less than 100 cpm net when surveyed with a thin window GM or LEG survey meter in contact with the trash or less than 0.05 mR/hr (essentially background).
c. Document the decay (i.e., date the waste was generated and date of the survey; there must be at least 10 half-lives between the two dates) on the Radioactive Material Record form, the results of the survey (must be less than 100 cpm above background for b/g emitters), and the type and serial number of the meter used for the survey.
d. Deface/remove all radiation symbols before disposal to normal trash.
e. Report the decays to the Safety Office by completing the Radioactive Waste Disposal form at least quarterly.
E. Cleaning of glassware before sending it to the washroom. For the protection of the washroom personnel and for the protection of everyone's experiments, it is essential that all radioactive material be removed from all glassware before it is sent to the washroom. As a minimum, this requires adequate rinsing. Quite frequently hot detergent (such as Isoclean) is necessary. Monitor the glassware if there is any question of radionuclide contamination. Some phosphorus-containing compounds have affinity for glass surfaces, and 32P may disperse over several loads of glassware during washing. Be certain 32P has decayed or has been physically removed before sending glassware to the washroom.
Removal of other residues is also critical for protection of the experimental work in the laboratory (e.g., if silicones, Dowex resins, Sephadex, etc. are on glassware placed in the dishwashers, these materials are distributed over all the glassware as well as the machine and are very difficult to remove).
Glassware contaminated by pathogenic microorganisms must be decontaminated before it is sent to the washroom.
Wisconsin's Employe Right-to-Know Law is no longer applicable. It has been superseded by the Laboratory Standard or the Hazard Communication Standard (OSHA, Department of Commerce). The standard is explained in Appendix B of the U.W. Chemical Safety and Disposal Guide.
