Aseptic technique prevents contamination of cell cultures and reagents by microorganisms. Watch our aseptic technique video protocol that shows you how to sterilize work areas and use appropriate sterile handling techniques, personal protective equipment and good hygiene (2:34 minutes).
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Sterile work areas
Cell culture should be carried out in a laminar flow safety cabinet located in a room that is specific to cell culture.
The interior of safety cabinets should be wiped with Trigene disinfectant followed by 70% ethanol before and after use. Ultraviolet (UV) light can also be used to sterilize safety cabinets between uses.
Areas of work should be uncluttered and contain only the items required for the current procedure.
Sterile handling techniques
Spray gloved hands with 70% ethanol before commencing work in the safety cabinet.
When bringing equipment or reagents into safety cabinets, spray them beforehand with 70% ethanol.
Never pour cell culture media or other reagents. Instead use sterile glass or disposable plastic pipettes to manipulate liquids and use each one only once.
Many cell culture reagents are certified sterile from the manufacturer; however, they can become contaminated during handling. Sterilize reagents either by filtration (0.2 µm filters) or autoclaving if necessary.
Never leave bottles of reagents, flasks or dishes open in the hood longer than necessary. Only open vessels immediately before use and if a lid has to be placed on the surface of the safety cabinet, ensure the opening is facing down. Replace lids as soon as possible.
Never open multiple flasks, dishes or bottles at the same time, this may result in cross contamination.
Personal protective equipment and good hygiene
Always wash hands thoroughly before and after working with cell cultures.
Wear a lab coat, gloves and safety glasses to protect yourself from any hazardous materials and to prevent contamination of the cell cultures from microbes present on skin and clothes.
Aseptic technique prevents contamination of cell culture and reagents from microorganisms in the environment. This video will outline the procedures necessary to work in a sterile manner within a cell culture hood.
Before commencing work ensure that you have wash your hands and are wearing the correct personal protective equipment, this includes gloves a lab coat and safety glasses. Thoroughly spray the inside of the cell culture hood with disinfectant and wipe clean with tissue.
Dispose of used tissue in the appropriate waste bin. Next spray the inside of the hood with 70% ethanol and wipe clean with tissue. The hood is now ready to use. Take the cell culture medium and supplements from the fridge and place in a 37oC incubator to warm. To prepare the medium aseptic spray each item with ethanol. Aim to work quickly and efficiently to minimize the risk of contamination and place each bottle in easy reach. Add each supplement to the bottle of medium using a new sterile tip or pipette. Only open bottles immediately before use and if the lid is put down insure the opening is facing upwards. Never sweep your hands and sleeves over the top of an opened bottled as this could cause contamination. To minimize the risk of this happening replace the lid as soon as you are finished with the bottle.
Label the medium bottle with your initials the date and details of the contents.
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