Summary
Highlights
Successful cell culture relies on preventing microorganism contamination through proper sterile techniques. This includes wearing personal protective equipment (PPE), reviewing MSDS information, and wearing appropriate clothing.
Washing hands before handling cells removes bacteria and dead skin particles, which are potential contaminants. A 70% ethanol wash kills microorganisms. Crucially, clean the hood before and after use, and wipe down all items placed in the hood with ethanol. Avoid using spray alcohol near flames.
Arrange items in the hood logically to avoid crossing hands over sterile areas and to maintain airflow. Do not overcrowd the work surface or disrupt the airflow. Keep the front sash in the lower position to maintain the sterile field. Only touch media and reagents with sterile objects.
When opening pipette packages, ensure the pipette does not touch non-sterile surfaces. Grasp the pipette high on the neck and insert it into the pipette aid. Only open media and reagents inside the cell culture hood. When holding bottle caps, avoid touching the inside edge. When pipetting, prevent the pipette from touching non-sterile surfaces. Use each pipette only once.
For large volume transfers, use a sterile pipette or pour aseptically, quickly and deliberately, guiding the liquid with the bottle's front corner. Wipe any spills immediately with ethanol. When finished, close all containers tightly before removing them from the hood. Wipe down the work surface with ethanol and organize the hood.