Microbiology Laboratory
RULES OF CONDUCT AND CLEAN UP PROCEDURES
The following rules have been established to protect you and your lab neighbors. It is wise and good technique to treat all microorganisms as potential pathogens and all lab equipment as potential accidents waiting to happen. Failure to observe the rules jeopardizes your safety and that of your classmates. Gross negligence will be reprimanded.
Rules of Conduct
1. ATTIRE: Lab coats and closed toe shoes are required and must be worn at all times in lab. Open-toed shoes are not permitted. Anyone without a lab coat, or wearing open shoes cannot attend lab that day.
a. Fold lab coats and keep them in their storage nook when not in use.
b. Before removing your lab coat from the lab for laundering or at the end of the
semester, it must be sterilized. Bring a paper grocery bag to place the coat in.
Clearly label the bag with your name and lab section and turn it in to the lab
technician for autoclaving. Speak with the lab technician before leaving your coat to determine when they will be sterilizing and can include you coat. This will ensure that you will have time to wash the coat and return it in time for your next lab meeting. Coats not collected within one week of the close of the semester or upon leaving the class will be confiscated.
c. Do not wear your coat outside of the lab. If you should need to leave the
lab during class, remove your coat and hang it over the back of your
seat. Upon returning to lab, put the coat back on.
2. SMOKING, EATING AND DRINKING are not allowed in the lab, including water, gum, and mints. During lab you should not put anything in your mouth i.e. pencils, fingers.
3. PIPETTE with pipette aids only. “In the old days…” it was a common practice to pipette by mouth. Today it is against lab safety protocol to use anything but various types of pipette aids. Take care not to draw anything into the pipette aids. If any cultures or reagents do get drawn into a pipette aid, that pipette aid must be withdrawn from use and cleaned appropriately by the lab technician before it can be put back into use. This is to prevent contamination of cultures and personnel.
4. LABEL everything clearly (tubes, petri dishes, slides) with the name of the organism, type of media or stain, date, your name (no initials), lab period and any other pertinent information that you may need to identify and retrieve your materials. There will be labs when each student will have several cultures. To prevent loss of cultures or confusion between them, it is necessary to label. Label petri dishes on the bottom and along the periphery so as not to obscure examining the plate. Label tubes along the length of the tube. DO NOT write on the white paint on the test tubes, as the writing cannot be removed and we reuse our tubes. Do not put tape on tube caps; the adhesive is not easily removed and makes it difficult for cleaning. When you use a marker or label on glass flasks and tubes, make sure all labels are removed, and all grease marks are erased before you leave. Put equipment back where you got it.
5. KEEP LAB COUNTERTOPS CLEAR AND CLEAN. Store packs, purses, etc. out of the way, under the counter (knee hole). Your lab counter area must be wiped down with disinfectant when you come in and when you leave. Throw the paper towels into the regular trash afterwards, NOT in the red biohazard bag. The biohazard bag is for any and all hazardous materials, including a toothpick you put in your mouth, gloves, disposable Petri dishes, etc. Use the regular trash for everything else. Before you leave, I need to check that your area has been disinfected, left clean, and that the microscope has been put away properly.
6. GAS LINES: Make sure the gas lines are completely shut off when not in use. Check again before leaving the lab for the day. Report any gas leaks to the instructor or the lab technician.
7. WASH YOUR HANDS with soap and water before leaving the lab. Use the sinks in front of Deck 1 and 3. Each sink has a soap dispenser and paper towel dispenser.
8. TURN OFF all phone/pagers while in lab.
Clean up Procedures
1. Biohazard Containers:
Biohazard containers are designed to receive biohazardous matter. This is largely due to the lining. which is a special red bag that can withstand autoclaving. All biohazard materials are to be disposed into these red containers.
a. Remove the bag when it is ¾ full. Tie the bag end into a knot.
b. Take the full bag to the prep room and give it to the lab technician for further
processing.
c. Line the container with a new red bag. New bags are located in drawers in the
front of the room marked with red tape.
2. Test tubes:
a. Remove all markings from test tubes and caps, using kimwipes or cotton balls and
ink remover or alcohol.
b. Boil all culture tubes in a boiling water bath with caps loosened for 30 minutes.
Begin counting time after water begins to boil.
c. Remove caps and dump tube contents, except Durham tubes into a biohazard
container – NOT THE SINK. Durham tubes can be “caught” by pouring the
suspension through a strainer over the biohazard container. A strainer is
located at each sink.
d. Rinse out all traces of media from culture tubes with water and a test tube brush.
Brushes are located at each sink. DO NOT USE SOAP OR CLEANSER. Place
brushes into the containers designated to keep them free of soap that may collect
in the sink from hand and slide washing. These products can leave a residue that
kills our cultures when we reuse the tubes.
e. Place rinsed tubes with caps removed in the wash basin near the prep room door.
A separate container for Durham tubes is inside the washbasin.
3. Plastic petri dishes:
These are placed into the biohazard container when your work with them is completed.
It is not necessary to remove any writing.
4. Pipettes:
Pipettes are placed tip down into special container for pipettes – a pipette bin. The pipette bin should be lined with a biohazard red bag. Pipette sleeves can be disposed of into the trash unless you replace a used pipette into the sleeve. In this case the whole pipette and sleeve needs to be placed into the pipette bin.
5. Micropipette tips:
Micropipette tips are ejected into special containers for them. These will be labeled and placed on counter tops when experiments require micropipettes.
6. Prepared slides: (price range from $2.25 to $16.00)
a. Do not break slides due to careless handling or improper microscope technique.
b. Remove all traces of oil from each slide. Wipe oil away with a kimwipe. If
necessary, one or two drops of alcohol can be used to remove residual oil. Take
care not to remove permanent slide labels.
c. Dry the slide with kimwipes and return to the proper tray. Trays are labeled on
their ends with the name of the slides.
7. Student prepared heat fixed slides:
These are washed and reused. Each student will have a box of clean slides in their drawers for their use.
a. Remove immersion oil with kimwipe
b. Wash slides clean with a non-abrasive cleanser; do not use the brush used
for tubes.
c. Rinse thoroughly and dry slide with kimwipes, pass through bunsen burner
flame once and return to your slide storage box.
8. Wet mount preparation:
These are non-heat-fixed slides and include the hanging drop method (depression slide
$2.25).
a. Saturate slide with coverslip in place with disinfectant for 15 minutes; capillary
action will draw the disinfectant under the coverslip.
b. Wash slide and cover slip with non-abrasive cleanser and rinse thoroughly
with tap water. Take care in handling and cleaning coverslips. They break
very easily and can cause cuts.
c. Dry slide with kimwipes, pass through a Bunsen burner flame once and return
to your slide storage box.
9. Contaminated cotton swabs (applicators) should be placed back into their sleeve
immediately after use and then discarded into the biohazard container.
11. Culture spills:
a. Flood the area with disinfectant.
b. Cover with paper towels and let it sit for 15 minutes.
c. Notify the instructor or lab technician.
d. Alert students as to the location of the spill while paper towels are in place so as
to prevent anyone from slipping on it.
e. Wipe up the area with paper towels and place all towels in the biohazard
container, along with your contaminated gloves.
f. Using a hand broom and dustpan, sweep up any glass and dispose of it into a
biohazard container.
12. Broken glass:
a. Notify the instructor or lab technician.
b. Carefully sweep up all broken glass. A hand broom and dustpan are located at
each sink on either end of the lab.
c. Deposit broken glass into boxes and/or plastic container marked “BROKEN
GLASS”. Never put broken glass into the trash cans, as maintenance personnel
could be injured.
d. Broken glass as a result of a culture spills is disposed of as instructed above for
culture spills.
13. IN CASE OF INJURY
a. Notify the instructor or lab technician immediately.
b. In the event of a cut, instruct the injured person to apply pressure to control
bleeding.
c. In the event of a burn, have the injured person flood the burn with cold water.
d. If lab personnel are injured or absent, call the campus health services at X-4494
using your cell phone or the phone in the prep room if you have no cell phone.
In the event of a life threatening emergency call X-1222.
e. For other emergencies or if the health services does not answer, call the
campus police emergency number X-4492.
7. FIRE SAFETY: Fires in this lab are started from flammable items becoming ignited by Bunsen burners. Do not leave a lit Bunsen burner unattended. Keep alert to the hazards of open flames and boiling water. Keep flammable materials away from the flame and never reach over a burning flame. Remember, a lab coat can be taken off and used to put out a fire if someone’s hair catches fire, etc.
14. FIRE EVENT: Immediately notify staff
a. In the event of fire, use the fire extinguisher located on the wall next to the prep
room window.
b. Should a student’s clothing catch fire, assist student to the shower and pull lever
to release shower water. A fire blanket is also located next to the fire
extinguisher that can be draped over the student to help extinguish the fire.
c. Should a student’s eyes get splattered with chemicals, assist the student to the
eye wash and open wash basin; have student lower face into basin to rinse eyes
for several minutes.
15. At times there may be a need to specify additional procedures for special experiment. These will be addressed at that time.