Biology Laboratory Safety Course

Welcome to the Biology Department's Laboratory Safety Course
Jodie Deshler and Alicia Muelling

This course is created for students and employees of the biology department to assist in the general knowledge of basic safety procedures in the laboratory.

This course will introduce several safety concerns including hazard codes, protective equipment, chemical useage and compatibilities, microbial and biohazard materials, and emergency procedures. This knowledge will enable you to become familiar with and comfortable in your laboratory surroundings.

Upon completion of the course you may be required to pass the final quiz posted on WebCT. For further instructions regarding the quiz please see your instructor or supervisor.

For optimal viewing, maximize your screen to 800 X 600 True Color (24 bit). This will allow you to see the entire page without difficulty. The bold, blue underlined words are linked to their definition in the glossary. If you need to clarify a word, click on it, and resume the course by clicking the back button.

HAZARD CODES

Hazard codes are very important in a laboratory setting; they reinforce safety standards and protocols. The specific colors and codes on the labels indicate what the particular hazard is. This gives high visibility to the chemical or solution being used and ensures safety. You must understand the significance of codes on container labels.

There are several labels with these colored codes in the laboratory. These labels include the hazardous waste label, the National Fire Protection Association (NFPA) labels, and original labels on chemical containers. These labels detail the level of protection needed for a specific chemical or solution. These labels provide safety information on health (H), fire (F), reactivity (R), and personal protective equipment (PE) required to work with chemicals or solutions.

All hazard categories listed on each type of label are indexed according to health, flammability, and reactivity levels. These are rated on a 0 to 4 scale, with 0 the least hazardous and 4 the most hazardous. Protective equipment (PE) has its own graphics or letter codes to explain the personal protective equipment needed. If you do not know these systems of safety, you are taking an unnecessary risk. Memorize the rating scale so that you have, at a glance, a very good indication of the hazards of that substance. All of these ratings are available, for your reference, on the hazard rating index chart posted in each laboratory. Know where to find this chart in every laboratory you work in.

MSDS sheets include information on every chemical used in the department. These sheets can be found in the main biology office, room 414. They can also be found at Fisher Scientific MSDS on-line. If you need additional information on any chemical you can reference these sheets.

PROTECTIVE EQUIPMENT

Graphics, letters, or symbols are used to represent the type of safety equipment that is required for working with a specific chemical or solution. These symbols are found on the original container labels and NFPA labels (previous section). Understanding when and how to use protective equipment is also very important in the defense against exposure to hazardous materials.

The personal protective equipment used in the laboratory is critical for your safety. Protective equipment usage differs and depends on what chemical or solution you are using. Protective equipment is one of the best defenses against injury or infection. You must understand what equipment to wear or use, how to use that equipment, and when you should use that equipment. There are five major ways to protect yourself in the laboratory: aprons, gloves, goggles, masks, and fume hoods.

This protects against splatters and spills from any material used in the laboratory. You must wear an apron when working with the variety of different chemicals and solutions so knowledge of when to wear it is fundamental. This will protect you against direct contact with or residual contamination from the substance. Aprons and lab coats can be found in room 420 of the biology department. Before leaving the lab, always remove your lab coat or apron to prevent transfer of a substance outside of the laboratory.

Gloves are made out of latex, nitrile, or other forms of non-latex rubber. They fit skin- tight to allow free movement and dexterity of fingers. It is recommended that you wear gloves whenever working in the laboratory, but it is required that you wear gloves when working with hazardous materials. Most of the gloves in the department are powder-free, vinyl gloves that are hypoallergenic. If you know you have a laytex allergy, pay close attention to the type of glove you use. Gloves are most directly involved with safety because hands are most directly involved with chemicals and solutions. Gloves are located in every laboratory and are easily accessible.

These are protective eyewear. Eyewear can also include eyeglasses, but it is recommended that you wear goggles over eyeglasses to prevent splash accidents. You are required to wear protective goggles when working in the laboratory for certain experiments. Eyewear protects against contamination from substances that injure or infect the eyeball and cause vision loss. Goggles also protect against absorption of materials into the body via the eyeball. Contact lens wearers BEWARE. Contact lenses make it difficult to wash away substances accidentally introduced to the eye. They can also be ruined by chemicals and vapors. Contact lenses should not be worn in the laboratory. Protective glasses are available in most labs. If you have eyewear concerns, feel free to bring your own protective glasses.

Masks are fitted coverings that are placed over the mouth and nose. They are used to filter out small chemical particles and other reagents in the laboratory. Masks filter out hazardous materials that enter the body via the respiratory tract and lungs.

A fume hood consists of a vacuum and a hood. This sequesters hazardous vapors inside the fume hood and draws them into a filter. The vapors are filtered and then disposed of properly. This is very important when working with solutions and chemicals that need to be handled carefully. When using the fume hood the vertically sliding glass door should be pulled down as far as possible allowing only enough space for you to be able to work. The fume hood is found in the back of room 421 C in the biology department and should be ON AT ALL TIMES.

When working in the laboratory it is important to keep yourself and your work area clean to avoid accidents and residual contamination. Clear your workspace, leaving only what you need to complete your experiment. Keeping the clutter down will reduce the chance of spills and breakage of equipment. You may unknowingly come in contact with a chemical or evaporated solution on counter-tops or other surfaces, or put someone else in danger of contamination. If there is a residual chemical or solution on the your hands, you will be in danger of ingesting or introducing the material into your body by other means (eyes or mucous membranes). You must ALWAYS thoroughly wash and scrub your hands before you leave the laboratory. There are sinks with liquid hand soap and paper towels in every laboratory. Wash your hands, dry them with a paper towel and then turn the faucet off using the paper towel to avoid re-contaminating your hands.

The safety regulations for working in a lab include no eating, no drinking, and no smoking at any time. Any food or beverages must be kept outside of the door of the laboratory. There are tables and waste baskets outside of the lab for your use. Please DO NOT discard any food or beverage items in the trash cans found in the laboratory. These standards protect you from ingesting a hazardous substance.

Chemicals can enter your body through absorption by the skin and mucous membranes, inhalation, ingestion, and contact with your eyes. You will run across such chemicals in your laboratory experiences and need to know what your risks of exposure are. You must pay attention to your instructor's directions, warnings from your lab protocol, and hazard codes on the chemical's labels. Be sure to use all of the protective equipment required.

Some of the hazardous effects of chemicals found in thelaboratory include strong irritants, toxins, carcinogens and allergens. The following table highlights some of these chemicals, but there are also many more that you should be aware of in the laboratory.

CHEMICAL	TARGET ORGANS	HAZARDOUS EFFECTS
Acid and Alkaline solutions e.g. Sodium Hydroxide	Skin, membranes of eye, nose, throat, trachea, bronchi, and alveoli	Etches Tissue - sometimes leading to irreversible damage to tissue
Aliphatic Halogenated Hydrocarbons e.g. Chloroform	Liver and Kidneys	Necrotic cell death-- Redness, swelling, inflammation, CNS effects, PAIN!
Metals e.g. Lead and Silver	Bones	Sequestered for extended periods of time-- Long term toxic effects - enzyme inhibition
Amines e.g. Diphenylamine	Toxic to all tissues and adversely affect a number of organs	Strong irritant, corrosive, and carcinogenic
Amides e.g. Formamide	Central Nervous System, liver, kidneys	Very toxic-- Liver and Kidney toxicity CNS depression
Aldehydes e.g. Formaldehyde	Skin, eyes, mucous membranes	Severe irritant at just a few ppm as well as a strong sensitizer leading to-- Skin rashes, Puffy eyes, Headaches, Respiratory problems. Allergenic properties are worse than its potential carcinogenic properties.

1. Always open chemical and solution bottles with the mouth of the bottle aimed away from your body and others.

3. There are two options for transporting chemicals or solutions.
A) Use a cart when there are a large number of bottles.
B) Carry one at a time and use both of your hands. Position one hand on the bottom and one hand on the mouth of the bottle or handle if there is one. Never pick up a bottle by its cover!

4. Always place chemical or solution bottles away from the edge of the lab bench to prevent accidents.

Some of the chemicals you will be working with will require proper disposal. Follow all directions and locate the bottle for your waste labeled with the UW-RF hazardous waste label as pictured above. Any bottle labeled as hazardous waste should always be in secondary containment and have a secure screw-cap cover. Always replace the cover when you are finished disposing of your waste, even if someone is waiting to use the bottle. If you are not absolutely sure of the proper disposal methods first look around the lab for a container labeled with the Hazardous Waste label and read what its contents are. If you are unable to find a waste container, ask your instructor. Never pour anything down the drain if you are at all unsure of a solution's hazardous waste status.

There are three major areas in the department where chemicals are stored, cabinets, refrigerators, and freezers. Room 421 houses molecular biology's chemicals, 422A has general use chemicals and microbiological media, and 426 (stockroom) houses the acids and flammables, each in their own cabinets. These places are organized by compatibility and by the storage codes found on their labels. The materials in each cabinet are are alphabetized. Chemicals used in the laboratory can react violently when mixed. To avoid any dangerous reactions chemicals are stored in specific places. What you need to remember is, simply, TO PUT THINGS BACK EXACTLY WHERE YOU FOUND THEM.

Microbial cultures and samples are a large part of the Biology Department at the University of Wisconsin-River Falls. Microorganisms are a hazardous material in the laboratory because if they are ingested or introduced into the body or the outside environment, they may cause a harmful effect.

Always handle a microbial culture as if it is an infectious material, or pathogen. Always wash your hands before leaving the laboratory.

All microbial cultures must be discarded on the Microbiology Discard Cart found in room 422 A.

Autoclaving is a sterilization technique that uses extreme vacuum pressure, 15-17 pounds per square inch, and extreme steam heat, 121 degrees Celsius, to disinfect bacterial or fungal samples. The autoclave is used to sterilize media. This kills bacterial and fungal samples, allowing them to be discarded in the garbage. To discard cultures and media:

2. Liquid and solid media must be separated on the Microbiology Discard Cart. Their respective sides of the cart are labeled.

3. Agar plates must be discarded in the orange biohazard bag. When the bag is 3/4 full, loosely tape the bag shut and place it on the bottom shelf of the cart. New bags are usually on the bottom shelf of the cart as well.

If you are unaware of the autoclaving procedure, you may endanger yourself and others by misusing the autoclave or not correctly sterilizing bacterial or fungal samples. Never use the autoclave without training from a biology staff member.

If a bacterial or fungal sample spill occurs, you should immediately pour a disinfectant, 5% Lysol ®, over the spill, let it disinfect for approximately ten minutes, and then wipe up spill with paper towels and throw in the Biohazard bag on the Microbiology Discard Cart. For large spills, follow with covering the spill area with 20% bleach, allow to disinfect for approximately ten minutes, and then wipe the area completely dry. Throw any contaminated articles into the Biohazard bag on Discard Cart. Pre-diluted Lysol is found on each bench in room 422 and 422 A and the bleach solution is found under the sink in 422 A.

Bunsen burners are common in the laboratory setting. To prevent a fire, clear the space around your Bunsen burner of anything flammable (i.e. lab notebooks and paper towels). Avoid passing your arms or hands directly over the flame. Your hair will easily ignite. Don't lean over the flame and keep your hair tied back. Person's with long hair should always tie it back during lab. In the event of any fire, if possible, cover it to deplete the oxygen supply. If covering the flame isn't possible, evacuate the area, and if possible try to extinguish with a fire extinguisher found on the wall of every laboratory. Notify proper authorities as soon as possible. The fire alarms are located by each stair well of the fourth floor. If you catch on fire:

If any substance is introduced into your eyes IMMEDIATELY get assistance and go to the eyewash station which is located in the prep room of every laboratory. Holding your eyes open, flush for at least 15 minutes. Get medical attention as soon as possible. You will be instructed on how to use the emergency eyewash station in the introduction to your lab class. There are also instructions posted next to the station. If it is not covered in class, don't be afraid to ask.

If you get a substance on your hands or arms flush with cold water, over the sink, for 15 minutes. If there is a large amount of a substance spilled on you, IMMEDIATELY get assistance, remove any contaminated clothing, and use the emergency eyewash as a shower. Rinse yourself for at least 15 minutes. Get medical attention as soon as possible.

Handling glassware and pipets in the laboratory can be quite dangerous. Much of the laboratory glassware is delicate and easily broken, which contributes to cuts and punctures. Many glass cuts occur when attempting to force a pipet into a pipetor or force open a lid on a bottle or jar. Avoid the use of unnecessary force. Inspect glassware for chips and cracks which will weaken the glass. Discard the damaged glass in the broken glass box and ask for new glassware. Pay close attention to your instructor's demonstration on how to handle these items. If you are washing glassware always use the brushes to clean the inside. Avoid putting your hands inside beakers, graduated cylinders and other glass items. Always wear rubber gloves for additional protection. In the event of an accident there are first aid kits found in every prep room for your use. Control any bleeding and seek medical attention in case of an accident.

Sharps include razor blades, scalpels, and needles. Always remember to cut AWAY from your body. In the event of any injury, follow the instructions above.

If you break any type of glassware, take care of yourself first, then CLEAN UP the broken glass to prevent others from cutting themselves. A broom and dustpan are located in each laboratory. All broken glass is deposited in the blue and white broken glass box in the lab. Sharps are disposed of in a separate box. This box is red plastic and labeled for sharps disposal. It is usually located near the broken glass box in the lab.

1. Think safety first. Be prepared for lab. Determine what protective equipment you need; remember preventing and accident is easier than dealing with the consequences of one.

2. Know what you are working with. Read the container labels, reference the hazard rating index chart or the MSDS. Know where to dispose of that substance when you are done using it.

3. Follow all safety procedures. Carefully follow the protocol in your lab manual and pay close attention to your instructor's directions.

4. Practice good housekeeping and personal hygiene. Keep your area clean and uncluttered. Wipe up spills and use the trash cans! Wash your hands frequently.

5. Report dangerous activities and situations. Pay attention to the way other people in the lab are conducting themselves; someone else's mistakes or carelessness can hurt you. Be aware of your surroundings at all times.

6. Know emergency responses and exit routes. Look around the lab and locate fire extinguishers, eyewash stations, and first aid kits. These items are there for your use in the event of an accident. Report all injuries and accidents to your instructor or supervisor.

8. Notify your instructor of any allergies or health related concerns especially if you are pregnant, expecting to become pregnant, or nursing.

We thank the University of Wisconsin-River Falls Grants Office, Nan Jordahl and Bill Campbell, for their assistance with our grant. Funding for this WebCT project was provided by a UW-RF Instructional Technology Mini-Grant. We also thank Torey Kauth for the professional scientific modeling which he eagerly volunteered for and Karen Klyczek for her assistance and support with WebCT. We thank E. Katherine Miller for mini-grant authorship and editorial contributions. Finally, we thank Amanda S. Holder for her fabulous and continued help in the UW-RF IDe3A Center.

Fisher Scientific. Fisher Scientific Catalog. Illinois: Fisher Scientific, 2000.

Gove, Philip Babcock. Webster's Third New International Dictionary. Merriam-Webster Inc.,
Springfield Massachusetts: 1981.

Malachowski, M.J., and Arleen Goldberg. Health Effects of Toxic Substances,
Government Institutes, Rockville, Maryland: 1999.

Mercier, Paul. Laboratory Safety Pocket Handbook. Genium Publishing Corp., Schenectady, New York: 1996.

absorption -- the movement of a chemical from the site of initial contact with the biologic system, across a biologic barrier, and into either the bloodstream or the lymphatic system

acid -- a substance which dissolves in water and releases hydrogen ions (H+); a proton donor or an electron acceptor; acids cause irritation, burns, or more serious damage to tissue, depending on the strength of the acid, which is measured by the pH, 1 strongest to 6 weakest

autoclave -- an airtight chamber that can be filled with steam under pressure that is used for sterilizing requiring moist or dry temperatures above 100 degrees Celsius (212 degrees Fahrenheit) without boiling

Bunsen burner -- a burner used in the laboratory that consists of a straight tube four to five inches long with a gas orifice and holes near the bottom for admission of air, the mixture of gas and air forms burning at the top, with a feebly luminous but intensely hot flame

carcinogen -- a chemical or physical agent capable of causing cancer. Such an agent is often described as carcinogenic. The ability to cause cancer is termed carcinogenicity. Words with similar meaning include onogenic and tumorigenic

contamination -- the result of materials, chemicals, substances, or life forms located where they are not wanted

corrosive -- a liquid or a solid that causes visible destruction or irreversible alterations in human skin tissue at the place where it touches the skin

fire -- or flammability. The range of gas or vapor concentrations (percent by volume in air) that will burn or explode if an ignition source is present.

flammable -- catches on fire easily and burns rapidly. The National Fire Protection Agency and the U.S. Department of Transportation define a flammable liquid as having a flashpoint below 37.78 degrees Celsius ( 100 degrees Fahrenheit)

hazard -- that which can be hazardous to life upon exposure; typically toxic, flammable, explosive, and corrosive materials or unsafe living or working conditions

hazardous waste -- waste materials that are classified as toxic, corrosive, flammable, explosive, radioactive, or biological/infectious wastes

health -- or toxicity. The quality or degree of being poisonous or harmful to plant, animal, or human life; the degree of danger posed by a substance to life

Material Safety Data Sheets (MSDS) -- a document produced by chemical manufacturers that describes physical and chemical properties of chemicals, first aid information, personal protective equipment to use, and suggested means of disposal

microbiological media -- any nutrient system for the artificial cultivation of a bacteria or other organism or cells that is sometimes a simple substance but more commonly a complex of inorganic and organic materials in a fluid base or one rendered more or less solid by coagulation or by the addition of gelatin or agar

microorganisms -- any organism of microscopic size e.g. bacteria, fungi, protozoa, or virus

mucous membranes -- the moist, soft lining of the nose, mouth, throat, bronchus, and eyes

National Fire Protection Association (NFPA) -- organization that has developed a scale for rating the severity of fire, reactivity, and health hazards of substances. References to the ratings appear on chemical labeling and MSDS's.

necrosis -- the death of tissue, usually as individual cells, groups of cells, or in small localized areas

personal protective equipment -- eye, hand, and body protection as well as fume hood use requirement

ppm -- parts per million, a measure of concentrations, such as parts of a substance per million parts of air or water.

reactivity -- the ability of a substance to undergo a chemical reaction (such as combining with another substance). Substances with high reactivity are often quite hazardous

residual -- a residue of a substance left on surfaces even after the substance has been cleaned away

secondary containment -- a plastic container such as a tray or washtub that the waste bottle is placed into to prevent leakage or spills

sensitizer -- sensitization, a condition of being made sensitive to a specific substance such as protein or pollen, antigen or hapten, usually as a result of repeated exposure; activation of an immune response

solution -- a mixture in which the components are uniformly dispersed. All solutions consist of some kind of solvent (such as water or other liquid) which dissolves another substance, usually a solid