Biosafety Levels One and Two

Lab Safety

Introduction

There are four biosafety levels: BSL levels 1 – 4. This protocol provides information for biosafety level 1 (BSL-1) and biosafety level 2 (BSL-2). Our labs do not permit us to work with BSL-3 or BSL-4 organisms. We can work only with BSL-1 organisms in the Teaching Lab (Rm 359); we can work with BSL-1 and BSL-2 organisms in the Research Lab (Rm 363).

The purpose of the four levels is to distinguish between different types of hazards and to provide protective measures for each level, as each level has different safety requirements. BSL-1 is designated for those working with microbes that don’t cause disease in healthy humans, for example, non-pathogenic E. coli. BSL-2 is for labs that work with pathogens including organisms such as Staphylococcus aureus or Vibrio cholerae. BSL-2 includes all of the precautions needed in BSL-1, along with additional precautions to prevent injuries, ingestion, and exposures to hazardous materials.

Equipment

  • Lab coat
  • Close-toed shoes
  • Gloves
  • Eye protection or face shields (can include glasses)
  • Eye wash
  • Safety shower
  • Sharps container
  • Biohazard waste container
  • Fire extinguisher

Guidelines

BSL-1 Guidelines

Before You Work

  1. Right after entering the lab, put on the proper personal protective equipment (PPE) and wear it the whole time you are working in the lab.
  2. Do not eat, drink, chew gum, or apply makeup in the laboratory.
  3. Before you start your experiment, make sure your workspace is clean and uncluttered. Disinfect your workspace with 70% ethanol solution provided in the lab. Make sure you have enough space to work at your lab bench.
  4. It’s possible that while working, you may accidentally come into contact with hazardous materials. A sink, eyewash station, safety shower, and extinguisher are also required to be present in the room. Make sure that you know where these are located before you start.
    • Use the eyewash station if unwanted or biohazardous materials are splashed onto your face and/or get into your eyes.
    • Wash your hands before and after working in the lab.
  5. Ensure that a designated chemical waste accumulation site is present, and that you understand how to dispose of chemicals and biological waste before starting the work. Before working with chemicals, first review their material safety data sheets (MSDS). MSDS are avaible for every chemical we stock in the lab (see Chemicals on this site.)

While You Work

  1. Dispose of sharps in a designated sharps container. Sharps include anything that can pierce your skin, including, but not limited to broken glass or needles. Sharps containers are thick heavy plastic, impenetrable by a needle, and can be closed securely.
  2. Biohazard signs should be placed in all areas containing biohazardous materials, including biohazardous waste. Dispose of waste that may be contaminated with pathogens or chemicals that present a danger to people and the environment in biohazard containers. Biohazardous materials are decontaminated prior to disposal, commonly done with an autoclave.
  3. Only mechanical pipetting should be done in the laboratory. Never pipette by mouth!
  4. There may be some times when you may be working with a protocol that requires shaking or mixing, which may produce aerosols or splashes. These procedures should be done inside a chemical hood.
  5. Clean any spills immediately, and decontaminate with 70% ethanol as necessary. For large spills that cannot be easily cleaned, evacuate and close off the area and alert the chemical hygiene officer.

BSL-2 Guidelines

Remember, the BSL-1 laboratory guidelines above are expected to be followed in addition to BSL-2 guidelines below, including PPE protocols.

  1. Working in a BSL-2 laboratory requires laboratory glasses in addition to coat and gloves. This lab coat should not be worn outside of the BSL-2 area.
  2. BSL-2 laboratories must be clearly marked as “BSL-2.” The names and contact information of the laboratory manager should be clearly visible in the room and on the door.
  3. BSL-2 centrifugation steps require an aerosol-tight lid and rotors should be loaded and unloaded in a chemical hood.
  4. BSL-2 safety protocols require bloodborne pathogens training. It is strongly recommended that anyone participating in BSL-2 work receives a hepatitis B vaccination or titer prior to starting work in the laboratory.
  5. For some biohazardous waste, an autoclave or other method for decontaminating must be used for proper disposal. Liquid BSL-2 waste can be decontaminated in a final concentration of 10% bleach for 30 minutes before pouring down the drain. Solid BSL-2 waste can be collected in designated biohazardous waste containers that can be autoclaved.

Conclusion

Although simple, following appropriate BSL-1 and BSL-2 protocol goes a long way. It is the easiest way to ensure that you’re doing your work in the safest way possible. Protecting yourself and working smart in the lab is important!

Procedure