Team:METU HS Ankara/Safety
We used Escherichia coli (risk group 1), and cancer cells for our experiments (risk group 2) under the control of our advisors. We didn’t use organisms of risk group 3 or 4. We engineered the E. coli to cure colon cancer. Cancer cells were used for our experiments. Our work area for E. coli was open bench. However while working with cancer cells, we used biosafety cabinet / laminar flow hood, to prevent even the smallest contaminations.
We did our experiments in the Biology Department of METU and we used organisms of safety level 1 and safety level 2 in the labs for our experiments.
We always considered the importance of safety while we were doing our experiments and any type of lab work. At the beginning of the year, before we started the experiments in the lab, our advisors informed us about lab safety, and provided us with all the necessary information to maintain safe working conditions for all of our team members [1] .
We didn’t face any unusual conditions in the lab and in our experiments. *While doing our experiments or any lab work, all of our team members wear their lab coats to protect themselves from body contact with hazardous chemicals [1]. For sterilization we use %70 ethyl alcohol solution to sterilize our working area, bench, and our hands before starting experiments. Also, we sterilize our equipments by autoclave. We were trained to note the potential burns or cuts that can occur from handling or sorting hot sterilized items or sharp instruments when removing them from autoclaves/ sterilizers or from steam lines that service the autoclaves, so we did these jobs really carefully [2].
When we worked by the fire, we acted extremely careful, and worked accordingly. To prevent any risk of contamination, we put our hair into ponytails and didn’t talk. We labelled the chemicals and our equipments, to prevent chemicals from getting mixed and to keep the lab organized. We kept the chemicals in the places where they should be kept. Also, we stored biological wastes in different bags and removed them by following the disposal procedures. We first autoclaved them in order to decontaminate them.
Centrifuges, due to the high speed at which they operate, have great potential for injuring users if not operated properly. Unbalanced centrifuge rotors can result in injury. Sample container breakage can generate aerosols that may be harmful if inhaled [2]. We watched our steps so our experiments did not result with accidents. We did not uncap the centrifuge machine before the time run off, or we did not power it before attaching the cap.
As METU HS iGEM team, we all regard the importance of safety in the lab and care for our team members’ safety and protection. We continue our experiments while implementing all the lab rules. We do not use organisms belonging to risk group 3 or 4 and we follow all the lab rules so we didn’t face any dangerous, unusual conditions so far.
BioSafety
The application of knowledge, techniques and equipment to prevent personal, laboratory and environmental exposure to potentially infectious agents or biohazards is called as biosafety. Before starting the project some topics were discussed with our advisors such as safety equipment, personal equipment, safety measures, waste disposal and biosafety rules.In Turkey there are biosafety regulations [3], that were followed in the lab.
KILL SWITCH
Our kill switch is based on the expression of holin and antiholin based on the presence of arabinose.As long as our arabinose promoter is functioning our construct will be able to produce enough antiholin to form dimers with holin and prevent it from forming pores in the cell membrane. But it’s absence will cause the holin levels to increase as the amount of antiholin present in the cell decreases. Therefore, there will not be anything to counter holin from causing cell lysis.
GENERAL RULES FOR LAB SAFETY
1. SAFETY FIRST - USE COMMON SENSE to avoid accidents.
2. Shoes (no sandals) must be worn at all times. Bare feet or open shoes (front or back) are not permitted in the laboratory.
3. All students must wear lab coats at all times.
4. No eating, drinking, or smoking is permitted in the laboratory.
5. Only authorized experiments may be performed. Equipment should be performed only for its intended purpose.
6. No chemicals or equipment may be removed from the laboratory.
7. Do not invite anyone into the lab.
8. If you are or a student near you injured or if any type of accident or fire occurs, IMMEDIATELY call your assistant for assistance.
9. The working space should be as uncluttered as possible to allow work space and avoid accidents. Also, keep the aisles clear to
prevent tripping over your gear. Place jackets, coats, book bags, pocketbooks, etc. at/on the designated area. Ask your assistant
for help if needed.
10. Keep your work space clean and tidy. The working space, desk drawers, cabinets, instruments must be kept neat and clean at all times.
11. When lab work is completed, all materials must be returned to their proper places and used benches, instruments and glassware
must be cleaned up.
12. Any student who has, or who develops a medical condition (epilepsy, asthma, allergies, diabetes, etc.) should immediately notify
the lab instructor.
WASTE
13. ONLY PURE WATER CAN GO DOWN THE SINK.
14. Follow the instruction of waste disposals. Never discard chemicals through the sink or to a regular trash.
15. Only discard the proper waste that is identified by the label on the waste container/collector.
16. Never mix the waste. Mixing may result in explosions and serious injuries.
17. Never overfill a waste container. If it is full, request a new one from your lab instructor.
18. Laboratory specific wastes:
a. Solid waste must be discarded to solid waste container.
b. Silica waste must be dried in the student hood before poured into silica waste container in the hood.
19. Never place regular trash into the waste containers. Normal trash can be thrown into trash bins or to bins labeled as trash.
20. Rinse all disposable or broken glassware with water before discarding it in the broken glass waste container. Rinse all
regular glassware thoroughly with water before returning.
SAFETY RULES AND REGULATIONS SPECIFIC TO THE GENERAL CHEMISTRY LABORATORY
- Students should come to the lab on time.
- No student is permitted in the laboratory without the instructor being present.
- No one is permitted in the laboratory with bare feet, sandals, or shoes (with exposed toes) which do not provide adequate foot coverage.
- Students with hair shoulder length or longer should tie with a hairclip. You cannot use beret, hat, turban, etc
- No unauthorized experiments are to be performed in the laboratory.
- Do not eat food, drink beverages, or chew gum in the laboratory.
- Report all injuries (cuts, burns) to the laboratory instructor immediately.
- Know the location of fire extinguishers and fire blankets.
- Know the emergency evacuation route.
- Leave glassware clean and dry at the close of each laboratory period. Wash and wipe desktop with paper towels. Be sure that gas and water are off. Each student is responsible for cleaning up spilled chemicals or broken glassware.
- Read the label twice before taking anything from a container.
- Do not take the reagent bottles away from their places. Carry liquids to your bench in clean test tubes or beakers and carry solids in clean beakers or on weighing paper.
- Take the exact amount of reagent indicated. Larger amounts will not be more effective and may lead to uncontrollable reactions.
- Never return unused chemicals to stock bottles. Dispose properly.
- Never use one pipette for different chemicals. Do not insert your pipette or dropper into the reagent bottles. Use the one that is designated (labeled) for that reagent.
- Discard all water-insoluble solids and organic chemicals in the special containers that are provided.
- Never throw matches, litmus paper or any solid waste into the sink
- Never discard any liquid containing chemicals into the sink without your assistant’s permission.
- Never pick up hot objects with your hands. After heating glassware or crucibles place the item on a wire gauze to cool.
- To insert glass tubing through a rubber stopper, lubricate the tube and stopper with water or vaseline. Use a cloth or paper towel to protect your hand and hold the tubing near the end to be inserted.
- Any chemical spilled on your eye should be washed off with plenty of water for at least 15 minutes at the eye wash. Notify an instructor immediately.
- Any chemicals spilled on the skin should be washed off immediately and the skin should be flooded with water for several minutes. Notify an instructor immediately.
- Never taste any laboratory chemicals.
- Never inhale gaseous fumes or position your noise directly above the sample. If you need to determine the odor of a gas, gently fan a small amount of the vapor toward your nose with your hand.
- In case of a skin burn, notify the instructor. Minor skin burns should immediately be placed under cold running tap water for 5-10 minutes.
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Be familiar with the following terms and their effects.
- FLAMMABLE They burn.
- IRRITANTS They irritate eyes, lungs and skin.
- TOXIC They are poisonous, effective either the short or long term.
- CARCINOGENIC They cause cancer.
- MUTAGENIC They cause genetic mutations.
- EXPLOSIVE They explode, usually on being mixed with air.
- CORROSIVE They burn the eyes, lungs and skin.
ACCIDENTS
Make sure to be in a safe position/place before helping others. If you do not know how to help to others STAY AWAY from the accident area and move to a safer place.
FIRE
If a person’s clothing catches on fire, he/she needs help. Prevent him/her from running. If he/she is close enough, put him/her under the safety shower because it is more effective than a blanket. If not, make him/her lie down and smother the flames by rolling, wrapping with lab coats, blankets, towels, etc. There are blankets located in the middle of the laboratories. Never turn a carbon dioxide extinguisher on a person.
If a fire breaks out, (if time allows) turn off all burners and remove solvents, place the chemical and equipment safely to the nearest possible table/bench, exit the building calmly. If you do not use the fire extinguisher, leave the room immediately to a safer place possibly outside. There are carbon dioxide extinguishers in the laboratory and the positions and operation of these should be known.
Point the extinguisher at the base of the flames. Very small fires can be put out with a damp towel by smothering. Only after the safety of all is assured should the matter of extinguishing the fire be considered.Because a few seconds delay can result in very serious injury, laboratory personal (assistants and technicians) will guide you on what to do and how to exit during the case of such an emergency.
CHEMICALS
If corrosive chemicals are spilled on the clothing, immediate showering (with clothing on) is the best remedy. When no safety shower is available remove the affected clothing immediately, and wash the area for 15 full minutes either in the sink or with the eye wash. Notify an assistant immediately and ask for help. If chemicals are spilled on the skin, wash them off with large volumes of water. Bromine should be washed off with water and the skin then massaged with ethanol or glycerine. Do not apply a burn ointment. If the chemical is spilled in the eye, it should immediately be washed out thoroughly with water using the eyewash. If acid was involved, a weak solution of sodium bicarbonate in an eyecup should then be used. If a base, boric acid is effective.
If corrosive chemicals are spilled on the desk, dilute them with a large volume of water and then neutralize with sodium bicarbonate if an acid, or dilute acetic acid if a base.
References
1. Lab Safety Handbook- New York University
2. Laboratory Safety Guidance Occupational Safety and Health Administration U.S. Department of Labor- 2011
3. The Biosafety Clearing-House of the Cartagena Protocol on Biosafety
4. Laboratory Safety Rules and Regulations Middle East Technical University Chemistry Department
