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Safety

General guidelines

• Know the locations of the eyewash stations and overhead showers, fire extinguishers, fire alarm pull-stations, first aid kits, spill kit, and building exits. Maps are posted near both lab exits.
• No eating, drinking, handling contact lenses, or applying cosmetics in the lab. No storing food in the lab.
• Wear a lab coat when working in the lab. Wear gloves when working with infectious agents, and protective glasses or goggles as required (persons wearing contact lenses should wear protective eyewear).
• Remove lab coat and gloves and wash your hands before leaving the laboratory. The sink in N318 is for hand-washing and washing glassware, and the sink in N320 is for biohazard waste disposal.
• Store chemicals according to the color-coded spot on the container (color-coding chart is posted in N319). Flammable chemicals and acids should be stored in the appropriate cabinet under the fume hood; store other chemicals in the appropriate cabinet in N319. The fridges and freezers in our lab are not safe for flammable storage.

Disposing of waste

Biohazardous waste

Solid biohazardous waste should be sterilized in the kill autoclaves in the Biochemistry Building at 121 °C for an hour. Close the plastic waste bag, leaving a small opening at the top so that steam can enter and escape. Place the bag in an autoclave-safe tray, and place heat-sensitive tape over the biohazard symbol. After autoclaving, allow the bag to cool, place it in a black garbage bag, and then dispose of it in the dumpster.

To dispose of BL-1 liquid biohazardous waste, add 10% bleach to the container, let it sit for a minimum of 20 minutes, and then pour it down the drain. BL-2 liquid biohazardous waste should be sterilized in a kill autoclave for an hour before being poured down the drain. After sterilization, dispose of glass culture tubes in the broken glass box and save the caps.

Chemical waste

If the chemical is solid and non-hazardous, dispose of it in the trash. If it is only hazardous at high concentrations, pour it down the sink with lots of cold water.

For toxic or hazardous chemicals, leave the container in the N318 fume hood until it is ready for pickup (i.e. until a large container is full, or a few smaller containers have accumulated). When ready for pickup, fill out a hazardous waste disposal tag (located in the drawer next to the fume hood). Attach the top portion of the tag to the container with twine and mail the lower portion to the Safety & Health Office (MS 4472).

Sharps and glass

Dispose of broken glass and used, sterilized culture tubes in the blue and white broken glass box in N318.

Razor blades, needles, and Pasteur pipets should be disposed of in a red sharps container. Do not re-cap needles prior to disposal. When a red sharps container is full, deface the biohazard symbol with heat-sensitive tape and sterilize in a kill autoclave for one hour. After the container cools down, make sure the lid is securely shut and then put it in a garbage bag and dispose of it in the dumpster.

Cleaning spills

Biohazardous spills

To clean up a biohazardous spill...

• Remove any clothing and wash any body parts that came into contact with biohazardous material
• If biohazardous material is in your eyes, use the nearest eyewash station
• Alert others to the existence of the spill and/or mark the area
• Put on a lab coat, gloves, and safety glasses
• Cover the spill with paper towels
• Pour undiluted bleach over the area and allow it to soak for one hour
• Inform a supervisor about the spill
• If there is broken glass, use a broom and dustpan, forceps, or two pieces of cardboard to pick up the glass and dispose of it in a red biohazard sharps container
• Dispose of paper towels in an autoclave bag
• Wipe the area again with disinfectant
• Soak contaminated clothing or shoes in a tray with disinfectant

Chemical spills

Clean up solids using a broom and dustpan.

Small liquid spills can be cleaned with paper towels or a mop; larger spills can be contained with cat litter. For strong acids, neutralize with acid neutralizing powder, wait until the color change occurs, and then clean up as above. Cat litter and acid neutralizing powder are located in the spill kit under the N318 sink.

After cleaning, follow the procedure for disposal of chemical waste.

For a large spill of flammable solvent, hit the STOP botton, clear the area, and inform a supervisor.

Fire and power failure

In case of a fire, hit the STOP button to turn off power and gas in the lab. A fire extinguisher is located in the hallway near the elevators.

In case of power failure, make sure all gas valves are turned off.

Equipment

The autoclave

• Use autoclave gloves when loading/unloading the autoclave. A face shield can also be used.

• Fill containers to under 75% capacity.

• Loosen all caps before autoclaving, and tape them on one side.

• Make sure the pressure is at zero before opening the autoclave.

• Do not change the temperature on the autoclaves in the Biochemistry Building.

Liquids

Autoclave for 30 minutes using the liquid cycle. After autoclaving, crack the door to release steam and then slowly open the door. To prevent explosion of superheated liquids, allow liquids to cool slightly before removing the tray.

Solids

Autoclave for 30 minutes using the gravity cycle, and leave in the dryer overnight.

Do not autoclave...

• Hazardous chemical waste such as bleach, ethidium bromide, HCl, and chloroform.
• Non-heat-resistant plastics such as polystyrene and polyethylene.
• Radioactive waste.

The pH meter

• Rinse the pH probe with water before use.
• If necessary, calibrate the pH meter according to the instructions posted on the wall. At least check the calibration using the pH 7 buffer solution.
• Slide the tubing near the top of the pH probe up or down to open the vent, and close again when finished.
  • If the vent gets covered with gunk, wipe it off with a paper towel, don't rinse with water.
  • Don't let the filling solution get below the coil. If it completely dries out, rinse the probe with water before refilling.
• When taking measurements, make sure that the bulb and junction of the probe are submerged. Make sure that the stir bar does not break the bulb.
• When finished, rinse with water and put the probe back in the storage solution.