Laboratory Safety: The RAMP Acronym

• Laboratory safety is key to reducing workplace injuries and illnesses
• It involves recognizing, assessing, minimizing, and preparing for hazards, creating a safe environment for lab workers and the community
• The four principles of safety are encapsulated by the acronym RAMP, which serves as a guide;
  • Recognize hazards
  • Assess risks of hazards
  • Minimize risks of hazards
  • Prepare for emergencies

Historical Background on Laboratory Hazards

• A laboratory hazard is any agent with the potential to cause harm to a vulnerable target.
• Past incidents highlight the importance of laboratory safety.
• Marburg virus outbreak (1967):
  • In 1967, workers at the Marburg Laboratory in Germany experienced symptoms of fever, diarrhea, vomiting, and internal bleeding
  • 7 workers died.
  • The outbreak involved a previously unknown virus carried by monkeys imported from Uganda for polio research
  • The Marburg virus was named after the city where it was discovered
• Chernobyl Disaster (1986):
  • In 1986, a safety test at the Chernobyl nuclear power plant in Ukraine led to a catastrophic explosion and meltdown.
  • It released massive amounts of radioactive material into the environment.
  • The disaster caused immediate deaths, long-term health effects, and severe environmental contamination, marking it as the worst nuclear accident in history.
• Karen Wetterhahn Mercury Poisoning (1997):
  • In 1997, chemistry professor Karen Wetterhahn at Dartmouth College, USA, encountered exposure during an experiment with dimethylmercury, a highly toxic compound.
  • A few drops spilled on her latex gloves, penetrated them, and reached her skin.
  • She suffered severe mercury poisoning, leading to neurological damage and death within months,
  • This incident indicated that latex gloves are inadequate protection against penetrating toxic chemicals
• University of Minnesota Glass Vessel Explosion (2008):
  • In 2008, a researcher at the University of Minnesota, USA, lost an eye when a glass vessel exploded under pressure
  • The accident resulted from a lack of proper shielding and safety protocols for pressurized experiments
• University of California Electrical Arc Flash (2012):
  • In 2012, a graduate student at the University of California, USA, suffered severe burns from an electrical arc flash while working on high-voltage equipment
  • The incident occurred due to a lack of proper training and safety protocols for electrical work
• Gold King Mine Wastewater Spill (2015):
  • In 2015, a cleanup operation at the Gold King Mine in the USA accidentally released 3 million gallons of toxic wastewater into a river
  • Poor planning and a lack of environmental safety protocols led to this environmental disaster.

Hazards in Cell Culture Laboratories

• Cell culture laboratories present hazards from handling human or animal cells, tissues, and toxic or mutagenic reagents.
• Common hazards include:
  • Accidental punctures from contaminated sharps, e.g., syringe needles
  • Spills and splashes onto skin or mucous membranes
  • Ingestion through mouth pipetting
  • Inhalation of infectious aerosols

Types of Hazards

• Chemical hazards:
  • Chemical hazards include toxic, corrosive, or irritating substances such as medications, solutions, gases, vapors, aerosols, and particulate matter
  • Some chemical reactions generate heat, leading to thermal burns
  • Inhalation of toxic solvents and ingestion of chemicals (e.g., through contaminated hands or food) are significant risks.
  • Safe storage of food and drinks away from chemical exposure is critical
• Physical hazards:
  • Physical hazards involve risks that can cause physical harm due to factors such as improper handling of equipment, exposure to extreme temperatures, noise, or vibration
  • Common injuries include cuts from glassware or sharp objects and burns from hot apparatus
  • Safe handling practices are essential to prevent injuries
• Electrical hazards:
  • Electrical hazards arise from unsafe electrical systems, such as faulty wiring or equipment near liquids
  • Electrical fires can occur due to unsafe cords or plugs
  • Ground-fault circuit interrupters should be used to prevent electrical shocks
• Radiological hazards:
  • Radiological hazards involve the uncontrolled release of radioactive materials, which can harm people or the environment
  • Proper shielding and handling protocols are necessary to prevent exposure
• Biological hazards:
  • Biological hazards include bacteria, viruses, blood, tissues, and bodily fluids that can carry diseases or allergens
  • These hazards can be transmitted to laboratory workers, who may then become carriers, posing risks to others outside the lab.

Why is Laboratory Safety Important?

• Laboratory safety is crucial for reducing injuries and illnesses.
• By minimizing or eliminating exposure to hazards, laboratory workers can protect their health and lives.
• Everyday precautions like seat belts show the importance of proactive safety measures.

Biosafety in Cell Culture Laboratory

• The goal of any biosafety program is to minimize/eliminate exposure to harmful biological agents, protecting lab workers and the environment.
• The most critical element of safety in a cell culture laboratory is strict adherence to standard microbiological practices and techniques
• Safety equipment includes primary barriers (biosafety cabinets) and personal protective equipment (PPE) along with Material Safety Data Sheets (MSDS) for hazard communication

Biosafety Cabinets

• Biosafety cabinets are designed to eliminate or minimize exposure to hazardous materials
• They provide an aseptic work area while containing infectious splashes or aerosols generated during microbiological procedures
• Three Classes of biosafety cabinets (Class I, II, and III) are available to meet varying research and clinical needs:
  • Class I biosafety cabinets: Provide protection for lab personnel and the environment, but not cultures, similar to chemical fume hoods
  • Class II biosafety cabinets: Designed for work involving BSL-1, 2, and 3 materials, they provide the aseptic environment needed for cell culture while protecting personnel and the environment, used for handling potentially hazardous materials
  • Class III biosafety cabinets: Gas-tight and provide the highest level of protection for personnel and the environment, required for work involving known human pathogens and BSL-4 materials, used in high-containment laboratories where the risk of exposure to life-threatening agents is significant

Air-Flow Characteristics of Cell Culture Hoods

• Biosafety cabinets protect the working environment from dust and other airborne contaminants via constant, unidirectional HEPA-filtered air flow.
• HEPA (High-Efficiency Particulate Air) filters can remove 99.97% of airborne particles as small as 0.3 microns, including dust, pollen, mold, and bacteria
• Airflow can be horizontal (parallel to the work surface) or vertical (from the top of the cabinet onto the work surface)

Personal Protective Equipment (PPE)

• PPE is an immediate barrier between personnel and hazardous agents, including gloves, lab coats, gowns, shoe covers, boots, respirators, face shields, safety glasses, and goggles
• PPE is often used with biosafety cabinets and other containment devices

Material Safety Data Sheets (MSDS)

• An MSDS is a document containing detailed information about the properties of a specific substance.
• It includes:
  • Physical data (melting point, boiling point, flash point)
  • Hazard information (toxicity, reactivity, health effects)
  • Safety guidelines (storage, disposal, recommended protective equipment, spill procedures)

Biosafety Levels (BSL)

• Biosafety Levels (BSL) are biocontainment precautions designed to protect lab personnel, the environment, and the community from infectious agents.
• Levels range from BSL-1 (minimal risk) to BSL-4 (highest risk).
  • Biosafety Level 1 (BSL-1):: Minimal risk to personnel and the environment. It is appropriate for non-pathogenic agents, like E. coli, in normal, healthy humans, following standard microbiological practices like handwashing/no eating in the lab, using open bench tops and basic PPE such as lab coats, gloves, and safety glasses, commonly found in teaching laboratories
  • Biosafety Level 2 (BSL-2):: Involves moderate risk where agents pose a hazard to personnel but are unlikely to spread to the community. Pathogens that cause mild to moderate disease include Salmonella spp. and hepatitis A/B/C viruses. Access to the lab is restricted, aerosols are handled in biosafety cabinets (BSCs), and PPE (lab coats, gloves, face shields) is required, as well as Class I or II biosafety cabinets for aerosol-generating procedures. Additionally, an autoclave for waste decontamination and self-closing doors are required. Typically exists in clinical labs handling human blood or tissues
  • Biosafety Level 3 (BSL-3):: Involves high-risk agents causing serious/lethal disease through inhalation, such as Mycobacterium tuberculosis and SARS-CoV-1. Includes strict access control and medical surveillance, mandatory work in biosafety cabinets/containment devices. Required safety equipment includes enhanced PPE (respirators, full-body suits) and Class II or III biosafety cabinets. Facility requirements include double-door entry (airlock) and HEPA filtration for exhaust air, commonly found in research laboratories
  • Biosafety Level 4 (BSL-4): Involves maximum-risk, life-threatening agents without vaccines. Examples include exotic pathogens like Ebola/Marburg virus. Requires strict access control and extensive training. Personnel wear full-body, air-supplied positive pressure suits, all work is in Class III biosafety cabinets/isolated rooms. Safety equipment includes full-body, air-supplied suits with independent breathing apparatus and Class III biosafety cabinets, includes an isolated building or zone with double-HEPA filtration for exhaust air. Laboratories study Ebola or other hemorrhagic fever viruses.