Contamination from Aerosol Generation (Cell Culture (Contamination))

Question

Accepted Answer

Multiple cultures becoming contaminated simultaneously or in sequence. Contamination spreading despite apparently good technique. Cross-contamination between neighboring cultures. Common causes: 1) Creating bubbles in medium during pipetting 2) Splash-back from waste beaker contaminating work area 3) Operator talking, coughing, or sneezing over open cultures 4) Not using plugged pipettes when transferring medium 5) Drawing liquid into pipette plug creating contaminated aerosols 6) Room air drafts, eddies, and turbulence in culture area 7) Excessive traffic and extraneous activity near culture work Solutions: 1) Avoid creating bubbles in medium or pipette during all manipulations 2) Use plugged pipettes (cotton-plugged at top) especially when transferring medium 3) Never draw liquid into pipette plug; change filter immediately if plug becomes wet 4) Discard waste into beaker with funnel or use vacuum aspiration 5) Avoid working when sick with a cold, or wear a mask 6) Face away from work when talking is necessary 7) Use automatic pipette aids and change filters regularly 8) Dedicate appropriate space for tissue culture, reduce traffic and extraneous activity 9) Allow 20 minutes after airflow initiation before opening containers in biosafety cabinet