Spring 2012

Richland College LABORATORY SAFETY BIOL. 2420 & 2421 Microbiology

Experiments in laboratories involve equipment and materials that can pose safety consideration if used incorrectly. The proper handling of the tools of scientific investigation will be demonstrated by your lab instructor. Be sure to follow all suggestions and cautions as indicated by the instructor for safe behaviors in lab. As to the chemicals and microorganisms that we will encounter, normal handling and adequate ventilation will minimize exposure. Many of the chemicals that you will work with are found in products that you use on a daily basis. Our lab use of these chemicals represents a minimal exposure to you. Acetone Acid alcohol (HCl + ethanol) Amyl alcohol Calcium chloride Coverage Plus disinfectant p-Dimethylaminobenzaldehyde (Kovacs reagent) Ethyl alcohol Ferric chloride Hydrochloric acid Hydrogen peroxide Hydroxylamine hydrochloride Iodine Isopropyl alcohol Lysol Methyl red alpha-Naphthol (Barritts A) alpha-naphthylamine (nitriate B) Ninhydrin Petroleum jelly Potassium chloride Potassium hydroxide (Barritts B) Sodium chloride Sulfanilic acid (nitrate A) Dimethyl p-phenylenediamine (oxidase) dihydrochloride Xylene Zinc powder STAINS: carbol fuschin crystal violet safrinin lactophenol cotton blue malachite green methylene blue LIVING MICROORGANISMS: bacteria protozoa algae fungi bacterial viruses

It is strongly suggested that a student who is pregnant or has a medical condition that may be affected or aggravated by chemicals or microorganisms used in the lab inform your instructor. In such cases, the student should meet with the instructor to discuss ways to safety use or to avoid certain chemicals or microorganisms. A doctor should be consulted if more information is wanted, or if one needs an opinion about the safety of the lab. Be very aware that we are using LIVE, POTENTIALLY DANGEROUS MICROORGANISMS throughout the semester. You need to take the upmost care to NOT contaminate yourself or others around you. The use of good aseptic technique in lab will reduce this possibility.

NATIONAL FIRE PROTECTION AGENCY LABEL (NFPA) HAZARD RATING SYMBOL KEY
For more information, go to http://chemlabs.uoregon.edu/Safety/NFPA.html

Each chemical has a Material Safety Data Sheet (MSDS) which contains more information about hazards and safety precautions.

MSDS SHEETS ARE AVAILABLE FOR STUDENTS TO READ UPON REQUEST.

YOUR INSTRUCTOR WILL BE SHOWING YOU A SAFETY POWERPOINT AT THE BEGINNING OF LAB, to go over main points about lab safety and lab orientation.

MICROBIOLOGY LABORATORY PROCEDURES AND SAFETY RULES

It is extremely important that you follow the safety rules and lab procedures carefully. 1. DO NOT EAT OR DRINK IN THE LAB! Although you can go out into the hallway to drink something or eat something, no food or drink will be opened inside of the lab. Any drink containers must be closed and off the tables. Smoking is prohibited in all campus buildings. 2. DRESSING FOR THE LAB: MANDATORY FOR ALL STUDENTS IN ALL LAB SECTIONS! You are expected to wear appropriate clothing in the lab, including lab coat and safety glasses. o closed-toed shoes (NO flip-flops or sandals) o safety glasses or goggles used (required when using bacteria and/or staining) o long hair tied back o lab coat/smock (paper coats should be in bookstore on campus) o gloves YOU MUST PROVIDE THE GLOVES. Realize that some people are allergic to latex, so you might consider non-latex gloves, such as nitrile. Lab coats, goggles, and gloves can be kept in the lab cabinets around the room perimeter. Each instructor/lab section has an assigned cabinet for storing students materials. DO NOT GET INTO A CABINET OF ANOTHER SECTION. Do NOT take lab coats home until AFTER it has been sterilized at the end of the semester. WASH YOUR HANDS BEFORE BEGINNING THE LAB AND WHEN FINISHING, or whenever leaving the lab to go to the restroom. Alcohol gel pumps are on all tables and handwash soap at all sink (in dispensers on wall). Please inform your instructor when the paper towels are low or when the hand foam cleaner at the sink dispensers is empty. 3. KEEP ALL AISLES AND WALKWAYS IN THE LAB CLEAR FOR PROVIDE A SAFE WALKING SURFACE. Backpacks and miscellaneous items are to be placed in the coat rack shelves on either side of the lab. Backpack straps are dangerous when moving around the lab. Purses/bags are to be placed under the table by your seat. 4. KEEP THE LABORATORY CLEAN AND ORGANIZED. Each side of the lab table has a slideout tray with common materials needed for the labslides, cover slips, inoculating tools, lens paper, bibulous paper, etc. Replace items when finished. There should also be clothespins for holding slides during stain procedures. One slide box with preserved slides will be found on either side of the table. YOU BRING YOUR OWN MARKER AND MASKING TAPE. 5. READ EXPERIMENTS BEFORE COMING TO LAB. 6. USE THE DISINFECTANT TO CLEAN YOUR LAB TABLES. There is a large spray container of disinfectant on each table. Wipe the entire table, at the beginning of lab and then again at the end of lab. There is a large stock bottle of the disinfectant on the lab bench, along with a funnel to fill the bottles. Students are responsible for filling disinfectant spray bottles when empty. 7. DISCARD: a. PAPER PRODUCTS: Paper towels go into trashcansNOT into pipet jars or biohazard bags. The paper wrappers around pipets and cotton swabs are thrown into garbage cans, NOT THE PIPET JARS. Do NOT replace pipets, loops, or swabs back into wrappers. b. MICROBIAL CULTURES: All tubes will be placed in ALUMINUM BASKETS with instructor name.

All petri dishes go into the RED biohazard bag on floor at discard area. c. PIPETS go into pipet jar on table. The jar will be emptied at the END OF LAB into the red biohazard bag. d. COTTON SWABS, WOODEN STICKS, PLASTIC DROPPERS, PLASTIC INOCULATING LOOPS (if used) go into the piet jar bags, and then the bag is discarded into biohazard bag. 8. SPILLS OF MICROORGANISMS: Flood the floor, table, etc. with the lab disinfectant: leave on for 5 minutes. After 5 minutes, pick up the glass with a broom/dustpan (ask instructor) and place in BROKEN GLASS CONTAINER (green and white or blue and white box at front of lab) in the lab. Use paper towels to absorb the disinfectant/microbial fluid. Place into garbage can. 9. HANDLING MICROORGANISMS: Properly label test tubes and petri plates for incubation your initials + lab table # section #, date name of microorganism Do NOT write on test tubes with a sharpie pen: use masking tape to mark on. However, you CAN write directly on the plastic petri dishes since they are autoclaved and tossed. 10. CHEMICAL INFORMATION: Read labels on chemical bottles for safety precautions. If chemicals come in contact with the skin, wash immediately with water. Make sure you know how to properly dispose of any chemicals used in lab. Do not return chemicals to the original containers. Make sure all chemical containers are closed tightly after use. There are eyewashes at EACH sink (and by 1 lab entrance door). There is an overhead shower by 1 lab entrance door. 11. DYE BOTTLES & TUBS: Dye bottles are located by each sink, and the bottles are color-coded. This means that that color is to stay right there. If a bottle is empty, DO NOT grab a bottle from another sink. REFILL THE BOTTLE WITH FRESH REAGENT. Stock solutions for reagents are located in a marked cabinet. There will be a dye tub + rack, a drain pad underneath the sink. Your group will be dyeing AT THE SINK, rather than at the tables. When done, place everything back correctly for the next group to use. IMPORTANT: There are stock bottles of dye and other reagents in a marked cabinet, for refilling the dye kits. 12. MICROSCOPE SLIDES/COVER SLIPS Each lab has 3 boxes of clean slides to be kept in the locked cabinet of each section. The slides will be used by the lab section, then cleaned by students as needed. The BON AMI cleanser solution for cleaning is by each sink. Broken slides are to be placed into broken glass container. Remove cover slips from wet mounts and place into red sharps containers---NOT DOWN THE SINK.

13. MICROSCOPE USE: Put the microscope back in correct cabinet location, with stage DOWN, the low power objective in place, the oil wiped off the lens, and the electric cord coiled properly around the holder (but NOT pulled tight). You are ASSIGNED a microscope for the semester, and it is your responsibility to take care for it. Arm of microscope is facing outward toward you. Please refer to the instructions and picture inside of each microscope cabinet at the student table. 14. FIRE INFORMATION: A fire extinguisher is mounted in the corner next to the emergency shower, at one door entrance.. A fire blanket is mounted by the white board. A gas valve OFF BUTTON is located on the side of all entrance doors into the lab. PLEASE TELL THE INSTRUCTOR IF YOU ACCIDENTALLY HIT THIS button so that it can be re-set. The exit doors from the building are at the west end of the building. 15. THE LAST PERSON AT TABLE MAKES SURE THAT ALL GAS JETS ARE TURNED OFF. 16. LAB EQUIPMENT: A glass pipet should never be forced into a pi-pump. Wrap the pipet in a paper towel when inserting it into a pi-pump and when removing it. Broken glassware must be disposed of in the glass disposal box located in the lab. This box is for glass only. A brush and dustpan are hanging on the wall by the glass box: ask the instructor. Use extreme caution around an open flame. Keep long hair and loose clothing well away from the flame. Make sure that gas jets are off completely when burner is not in use. Use extra care when working with inoculating equipmentloops and needlesand bunsen burners. They get VERY HOT! Handle hot glassware with appropriate clamps or tongs. Small beakers or flasks may be removed from heat using paper towels. Test tube holders are for lifting test tubes only. 17. If you are cut, burned, or injured in any way during the lab, you must inform the instructional associate or your instructor immediately. In addition to administering first aid if needed, the instructor must take special precautions when cleaning areas and objects contaminated with human blood. Do not attempt to clean blood-contaminated spills! 18. Know where to find emergency equipment such as the eyewash, fire extinguisher, fire blankets, electricity and gas shut off switch, and telephone (dial 911 for campus police). Report all accidents to your instructor immediately, as well as any hazardous condition. 19. Do not bring children to the lab. Unaccompanied children are not permitted on the campus. 20. YOU MUST SIGN A LABORATORY SAFETY ROSTER and a SAFETY AGREEMENT. By doing so, you are saying that you understand the safety rules and that you agree to follow them and all other instructions stated in the handouts. 167523821.doc