Module II

NOTE: This Module is currently being updated.

II. LAB SAFETY

INTRODUCTION

This module will introduce you to safety concerns and safety procedures to be followed during your work in the Techniques Facility. Most of these concerns are common to all bioresearch laboratories, and you should find that the same, or similar, procedures will be followed there. It is beyond the capacity of this module to provide comprehensive instruction on all lab safety procedures. The objective is, rather, to give you a basic working knowledge of lab safety, so that you are alert to such problems and will be able to ask appropriate questions as you enter into original research work.

The Techniques Facility and its exercises have been designed with reduction of potential hazards in mind. However, lab safety and proper procedure should always be observed in the Facility, as in any laboratory. Proper procedure means the safe use of all basic lab equipment such as gloves, safety glasses, tabletop centrifuge, beam balance, pipetman, electronic balance, pH meter, and spectrophotometer, as well as approved methods for the handling of any toxic chemicals, corrosive liquids, etc. When working in the laboratory, keep in mind that ignorance of safety procedures or misuse of equipment endangers not only you but those around you.

You must complete this lab safety module before proceeding to any others.

EXERCISE #1: LAB SAFETY VIDEOTAPE.

View the videotape on lab safety. Obtain the tape labeled "SAFETY" from the "VIDEO TAPES" file drawer. Use the Tech Facility VCR and monitor (ask for assistance). As you watch the videotape, take notes in your Tech Facility lab notebook. If you think you may have missed something, stop the tape, rewind it a bit, and watch that segment again. When you are finished, eject the tape and rewind it in the separate tape rewinder (ask for assistance). Return it to the "VIDEO TAPES" file drawer.

SAFETY REFERENCE WORKS

There are several reference works that are particularly useful with respect to safety concerns and research, such as the Handbook of Laboratory Safety and the Handbook of Chemistry and Physics. Both of these are on the book shelf in the Tech Facility.

EXERCISE #2: HANDBOOKS

Find the Handbook of Laboratory Safety and the Handbook of Chemistry and Physics (in the Tech Facility) and browse through them. Use them as aids whenever you feel unclear about a safety procedure or chemical reactivity. Feel free, in the future, to return to the Techniques Facility to consult the Handbooks. After browsing be sure to put the books back where you found them.

A. OSHA

The Occupational Safety and Health Administration, otherwise known as OSHA, is a U.S. Government agency that has set certain safety guidelines for investigators working in a research laboratory. Many of the guidelines presented in this module are derived from the OSHA regulations.

EXERCISE #3: OSHA GUIDELINES

Find and review a copy of the OSHA guidelines available to you in the Tech Facility Lab Safety Resource Book. This book is located on the Tech Facility book shelf.

B. PREPARATION, PLANNING, AND SAFETY

PLAN AHEAD

First and foremost, before going into a research lab you should know exactly what you are going to be working on. That includes the procedures, equipment, and materials you will be working with, as well as the handling and disposal of chemicals. Planning ahead is the key to a safely run experiment. Work out as many details as possible, and either check with others or do some research on any that you are not clear about before starting the work. Lack of planning not only jeopardizes successful completion of an experiment, but can make it dangerous.

Keep your lab notebook up to date. Get organized by making an outline of anticipated steps before starting the actual work. Examine equipment to be used and, if possible, do "trial runs" in advance. Trying to figure out what to do next or how to do it during your experiment wastes time and increases the likelihood of error.

Cleaning Up

Always clean up after yourself in the lab. Dispose of used materials in the proper receptacles. You don't have to wait until your experiment is done; when the opportunity presents itself, clear away any materials that you will no longer need such as test tubes, pipettes, etc. This will help to avoid dangerous clutter as you proceed. When you have completed an experiment, return reagent bottles to their proper place, make sure all equipment used is turned off, and clean up your work area so that it is ready for the next person.

EXERCISE #4: SAFETY SUMMARY

As you work through these exercises, write a summary of the key safety information sources and procedures in your Tech Facility notebook. Later on, add to it relevant information from other modules and use it as a ready reference. As a start, make a brief list of the sources of lab safety information that you have seen thus far.

C. PERSONAL PROTECTIVE EQUIPMENT

Safety glasses

Protective eyewear (goggles or safety glasses) and gloves are articles that you must always have available when in the lab.

Protective eyewear should be worn at all times in the lab! Even though you may not be doing anything dangerous, someone nearby may be.

EXERCISE #5: PROTECTIVE EYEWEAR

Go to the drawer in which the protective eyewear is kept (look for a label ) and select a pair for yourself. Try them on. These will be your glasses for all work performed in the Tech Facility. You can label them with initials and keep them in an area or drawer designated for your use. Take them with you upon completion of all modules (A Tech Facility "souvenir"!).

Protective gloves

Gloves will protect you from hazardous materials and will minimize contamination of others within or outside of the lab. There is a proper way to put on and remove gloves to avoid contamination. The removal procedure is especially important, doing this improperly can be just as bad as not wearing gloves at all!

EXERCISE #6: GLOVE SAFETY VIDEO

View the video presentation on glove safety. Obtain the tape labeled "GLOVES" from the "VIDEO TAPES" file drawer. When finished, eject the tape, rewind it (in rewinder), and return it to "VIDEO TAPES" file drawer.

EXERCISE #7: PUTTING GLOVES ON

Procedure:

1. Get a pair of latex gloves from the supply box (choose appropriate size: small, medium or large).

2. Remove any rings or jewelry that might tear the gloves (keep these in a secure place!).

3. Put on the gloves as you would an ordinary pair of gloves, but be careful that, as you do so, you do not perforate them with your fingernails.

EXERCISE #8: TAKING GLOVES OFF

When removing the gloves you must follow a specific procedure to avoid contamination.

Procedure:

1. Pinch and grasp the area about one inch from the open end of the right glove with your left index and forefinger.

2. Pull the glove down and off (turning it inside out).

3. Using your gloved hand, ball it up and then grasp the "ball" in your gloved hand.

4. Put your right index finger under the top of your left glove, towards the inner wrist; push it down, grasping the inside as you do so.

5. Turn the left glove inside out, wrapping it around the balledup right glove.

6. Repeat exercises #7 and #8 until you feel comfortable with them (use the same pair of gloves!).

EXERCISE #9: TESTING YOUR GLOVE TECHNIQUE

When you feel you can do exercises #7 and #8 adequately, put your gloves on (#7), go to the sink, and run water over them until they are wet. Now, repeat the removal procedure (#8). Does any water get on your hands? If so, repeat the glove removal procedure (#8) until no water gets on your hands.

Proper use of gloves in a research laboratory is extremely important, as it will minimize the risk of contamination to you and the rest of the laboratory. Improper use can result in contamination within and outside of the lab, and in possible injury to yourself or someone you encounter.

ADDITIONAL NOTES:

Gloves

1. Wash your hands with soap and warm water before leaving the laboratory even if you have used gloves!

2. Always remember to remove your safety glasses after you have removed gloves and washed your hands. Otherwise, you may contaminate your glasses.

3. Once you have dealt with a potentially hazardous substance change your gloves and discard them to minimize the chance of contamination.

Lab coats and other clothing

Although lab coats will not be used in the Techniques Facility, keep in mind that when you do begin working with more hazardous materials in a research laboratory, a lab coat will not only provide you an extra layer of protection but will also help keep your own clothes from being damaged or stained. Neither open toed shoes nor shorts should be worn in the lab at any time.

D. HANDLING CHEMICALS

Before using any chemical, find out what potential hazards it may pose. This information can be found in the Handbook of Chemistry and Physics or in the Handbook of Laboratory Safety both of which are on the Tech Facility book shelf. Safety information is usually present to some extent on the chemical bottle or container. Read the "MSDS" for the particular chemical (see below), and consult with others in the laboratory who may be experienced in handling that chemical.

MSDS

A "Material Safety Data Sheet" or MSDS should be available for all the chemicals or reagents that you will be using in the laboratory. These can be found in the Tech Facility Lab Safety Resource Book. Always refer to this sheet when first using a chemical with hazards you may not be aware of! It contains information provided by the manufacturer/supplier of the chemical to the purchaser, as required by law. The labels of bottles are also sources of information that should be attended to regarding chemical safety. Some chemical manufacturers such as Fisher Chemicals (most of the chemicals used in the Tech Facility are manufactured by Fisher Chemicals) have a labeling system which gives information on the hazards of that packaged chemical.

EXERCISE #10: THE "MSDS"

Find the MSDS for NaCl in the Tech Facility Lab Safety Resource Book. Make note of all of the potential hazards of NaCl and the protective equipment you will need when using it.

EXERCISE #11: HAZARD IDENTIFICATION SYSTEM

On the wall (near the sink) there is a Hazard Identification System chart. It is applicable to the labels of all the chemical compounds used in the Tech Facility. Go and look at it. Using the label for sucrose, determine what safety precautions you should take when working with that compound.

Liquids

Liquids should always be dealt with carefully. Splashing is a potential hazard, so take your time. Use pipettes, squeeze bottles, and funnels whenever possible. Never pipette by mouth! Use bulbs and other pipetting devices.

Solids

Solids should always be handled using the proper utensils (such as spatulas). Do not pour solids out of their containers, as you will not have as much control over their whereabouts. Some solids are actually fine or fluffy powders that will make a cloud in the air unless handled with spatulas, slowly and carefully.

LABORATORY WASTE DISPOSAL

When you work in a research laboratory, waste disposal is a critical concern. Are you using toxic substances? If so, how are they to be disposed of when experimental work is completed? If you do not know, you must ask your research sponsor before beginning any experiment.

Where does one put other types of laboratory waste? research labs have specific receptacles for paper, for biomedical waste (such as used gloves), and for glass, syringes or other sharp objects. You must know in advance where all of these receptacles are located, and what goes into them. Incorrect disposal of waste, such as putting used gloves or syringes in the paper waste bin, is likely to alarm those who pick up such waste and cause a shutdown of disposal service to your laboratory.

There are three types of disposal containers in the Tech Facility and in most bioresearch laboratories:

1) Normal waste

You can put regular non-contaminated paper, wrappings, and food in normal waste containers. These are large boxes with clear plastic liners, or typical wastebaskets.

2) Biomedical waste

Biomedical waste includes rubber gloves, animal bedding, any disposable labware, kimwipes, paper towels that have been used for any hazardous materials, etc. All such waste must be put into the biomedical waste container. These containers are large boxes with red plastic liners, and have a "biohazard sign" on them indicating the nature of their contents.

In the Tech Facility you will also find smaller red containers with red plastic liners. These too are for biomedical waste, and are used as a convenient local disposal station so that you do not have to go to the large container so often. When the smaller biomedical waste containers become full, the bag is closed up with tape and transferred into the large biomedical waste box.

3) Sharp waste ("Sharps")

You must put anything sharp, such as used razor blades, broken glass, used glass pipettes, syringes (see below), etc. into the sharps container. This is a hard, thick red cardboard or plastic box with a circular hole in the top. The sharps box is slightly smaller than a "hazardous waste" box, and fits inside the latter. Disposing of sharp objects in anything other than the sharps disposal container is extremely irresponsible, and can cause severe injury to anyone handling the container.

NEVER PUT SHARPS IN ANY OTHER DISPOSAL CONTAINER!

In the Tech Facility, the sharps container has already been placed inside a larger hazardous waste box. It is better if razor blades are first packed into sealed smaller thick-walled containers of some kind. These small containers are then sealed and dropped into the sharps box.

Syringes:

For proper disposal of syringes, do not break off the needle or break the barrel or plunger before disposing in the sharps container. Place the entire syringe and cap into the sharps container. The less you handle the syringe the less likely it is that you will injure yourself.

MATERIALS THAT CAN NOT BE POURED DOWN THE SINK

The following materials are banned from the New York City sewer system:

--all flammable liquids

--explosives

--water/air reactives

--acids with a pH below 5.0 (but NOTE: Many Hunter College labs have "acid waste sinks". These empty into neutralizing tanks before going into the sewer system. Check with your sponsor.)

--bases with pH above 9.5

--toxic substances that are harmful to humans, animals and

aquatic life

--noxious or malodorous substances

--radioactive material not in compliance with NYC Health Code Article 175 (your sponsor should be aware of the of material that you will be handling)

FINAL PACKAGING AND REMOVAL OF WASTE

Each biosciences laboratory (both research and teaching) at Hunter College is responsible for the proper packaging of its own biomedical waste and sharps, and for removal of the packages to a storage area from which they are eventually picked up by a commercial waste disposal company.

The current packaging regulations are as follows: the loaded biomedical waste container must not exceed 30 lbs. When the container has been filled, the red bag is taped closed and the box is sealed completely with box tape, including all of the corners. If the full red sharps container is not already inside a larger cardboard hazardous waste box, it must be put into a larger box with its lid taped closed, and the larger box sealed as above.

The following information must be written clearly on all boxes:

1- Date

2- Name of lab (with Professor's name)

3- Room # of lab

4- Hunter College address

The sealed waste boxes (hazardous waste and sharps) are then transported to a holding area.

FOR FURTHER DETAILS, SEE CURRENT POSTED INSTRUCTIONS ON TECH FACILITY BULLETIN BOARD, MARKED WASTE DISPOSAL.

Dead animal disposal

Dead animals should be put into a red plastic biomedical waste bag and kept frozen in a lab freezer. They are transported (still frozen) to an animal waste freezer in the Animal Facility (15th Floor), where they are kept until pickup.

Preserved animal disposal

The manner in which preserved animals are disposed of depends upon the medium in which they are preserved. See the Tech Facility bulletin board instructions marked "Animal Disposal".

SPECIAL CHEMICAL: HANDLING AND DISPOSAL

Many chemicals, particularly potential carcinogens, neurotoxins, and radioactively labeled compounds, require specific handling and disposal procedures that are not covered in this Tech Facility workbook. Be aware that special training must be obtained before you are allowed to handle them. If you are in doubt about any chemical, do not hesitate to ask whether special handling and disposal are required for your particular research project.

A listing of all commonly used chemicals that are of particular concern with regard to health risks is provided in the OSHA guidelines. This can be found in the Tech Facility Lab Safety Resource Book located on the book shelf.

SPECIAL CHEMICALS: TWO EXAMPLES

(a) PHENOL

There are certain chemicals that give little warning when they make contact with the skin. Phenol, for example, acts as a local anesthetic but is also caustic. Phenol can cause damage without its effects being felt until it is too late.

(b) MERCURY

Mercury, a component of most laboratory thermometers, is highly toxic and affects the skin, respiratory system, CNS, kidneys, and eyes.

EXERCISE #12:

Locate all three types of disposal containers in the Techniques Facility (normal waste; biomedical/hazardous waste; sharps). Make at least a mental note of their location for future use in module exercises.

F. SAFETY WHEN USING SPECIFIC PIECES OF EQUIPMENT

There are safety concerns associated with each piece of laboratory equipment. Most of these are addressed in the modules covering equipment use. However, a few specific concerns are reinforced here.

Centrifuge

When using the table top centrifuge, or any centrifuge, opposing tubes with their holders must be balanced by weighing (including the tube holders) before doing a centrifugation "run". Unbalanced tubes frequently break, causing loss of experimental samples and production of dangerous broken glass or plastic fragments. Instructions on balancing are given in the module on "Using a Centrifuge". In addition, you should always wait until the centrifuge comes to a full stop before lifting the lid and removing your centrifuge tubes from it. For a more complete treatment of proper and safe centrifuge use refer to Module XII.

G. ACCIDENTS

By following safety procedures outlined here and provided by other sources, you can reduce chances that accidents will occur, and help to minimize their consequences. However, no matter how careful you are, there exists the possibility that an accident will happen. If it does, you should be prepared to take proper action. Make certain that the lab supervisor or professor is informed as quickly as possible. This is critical if it involves equipment or another student.

Example: One of your centrifuge tubes breaks in the centrifuge, but you fail to inform anyone. You investigate but can find no apparent cause. Later when someone uses that same centrifuge, one of their centrifuge tubes breaks and they are injured by the glass. You could have prevented this from happening!

If you become injured, notify the lab supervisor or professor immediately! Sometimes what doesn't seem to be serious at all, without timely attention, can become very serious indeed.

H. LAB SAFETY EQUIPMENT

One of the first things you should do upon beginning work in a research laboratory is to find out where all of the lab safety equipment is located, such as fir extinguishers eye wash stations, showers, first aid kits, blankets etc.

Fires

Laboratory fires are rare. However, if a fire does occur, immediately alert others in the lab. Lab fires can be extremely dangerous because of the potential for explosion and noxious fumes.

CLASSIFICATION OF FIRES

FIRES ARE CLASSIFIED ACCORDING TO THE MATERIAL THAT IS BURNING.

Class A: Ordinary combustibles or fibrous materials such as wood, paper, cloth, rubber, and some plastics.

Class B: Flammable or combustible liquids such as gasoline, kerosene, paint, paint thinner, and propane gases.

Class C: Energized electrical equipment such as appliances, switches, panel boxes, and power tools.

EXERCISE #13: FIRE SAFETY

Find and read the fire safety booklet in the Tech Facility Lab Safety Resource Book. Pay close attention to the types of fires and the proper extinguisher type. What type of fire extinguisher is most appropriate for a research laboratory?

EXERCISE #14: FIRE EXTINGUISHER

Find the fire extinguisher in the Tech Facility. Read the directions for its use. What type of fires can it be used for? Is it appropriate for all types of fires that could occur in the Tech Facility?

EXPOSURE TO CHEMICAL OR OTHER IRRITANTS

THE EYE

If your eyes have received chemical exposure (such as by accidental splash) use the eye wash station IMMEDIATELY!! Flush eyes with water, keep eyelids open for at least 15 minutes, and roll your eyes around to flush as much of the eye as possible.

NOTE: Contact lenses should be avoided when working in the lab. They can make irrigating the eye very difficult and prolong exposure to a damaging substance. Some soft lenses actually absorb chemical vapors! There are also gases released in certain reactions that will damage the surface of your contacts.

EXERCISE #15: EYEWASH STATIONS

Look for the Tech Facility eyewash station near the sink. Make a mental note of its location. Remove one of the eyewash bottles from its holder, examine it, and read the directions for its use. When finished, put it back into the holder.

SHOWER STATIONS

Shower stations are to be used when chemical spills or splashes posing a potential threat to the body occur. Many bioresearch laboratories have an overhead shower that is activated by pulling on a large ring or loop. If such a spill occurred, you would go quickly to the shower station, activate it by pulling the large loop, and remove the contaminated clothing while in the shower. The shower station can also be used to bathe the face and eyes in a large spill accident or to extinguish clothing fires.

The Tech Facility does not have an overhead shower station, but there is a pressurized water sprayer attached to the sink. The water in this sprayer is always ON, so that you need only push the sprayer control.

EXERCISE #16:

Go to the sink and test the water sprayer (aim it into the sink!).

FIRST AID KIT

Every research lab should have a first aid kit. This kit should include basic materials needed for minor laboratory injuries.

EXERCISE # 17: FIRST AID KIT

Go to the area of the lab with the eyewash station, and find the first aid kit. Open the kit and take note of the contents. A first aid manual is included; browse through it, paying particular attention to the sections on chemical burns, cuts, and first aid for the eyes.

NOTES ON EQUIPMENT NOT IN THE TECH FACILITY

Bunsen burner

All other necessary equipment should be set up and the area checked before lighting a bunsen burner. Keep all papers, tissues, and flammable materials or chemicals far away from the bunsen burner, and keep the burner away from your own clothing and body parts! Long hanging hair can be a particular danger, and should be tied back.

Bunsen burner flames are not always easy to see. While the burner is in use, post a NON-FLAMMABLE sign near it that says "BURNER IS ON". However, do not leave it unattended while on. When the burner is not in use turn it off at the source; the knob on the burner itself should be used only to control the flame and not to turn it off. Keeping your work area free of clutter will minimize chances of accidents.

Fume hood

A fume hood must be used properly in order for it to be effective. Put your sample in the hood about four to six inches from the leading edge, and bring the protective glass down until the guide arrows on the sliding glass and on the hood frame are aligned. It is at this height that the ventilation system is designed to work properly, so it is very important that you follow this procedure. Make sure that there is, in fact, a draft, by checking the airflow meter (if there is one) or by observing the movement of a hanging strip of paper taped to the bottom inside of the hood's sliding glass. Try to keep storage of materials in the fume hood to a minimum so as to avoid clutter. (See pages 145-150 of the Handbook of Laboratory Safety for a good overview of different types of fume hoods.)

Chemical Spills

Each particular spill requires a specific cleanup procedure, depending upon what has been spilled. Do not proceed to clean up a spill until you are sure of the correct procedure FOR THAT CHEMICAL. General instructions on precautions to be taken in dealing with spills can be found in Tech Facility Lab Safety Resource Book.