Basic cell culture instructions

Aseptic Technique

For best results in tissue culture, we want to work to keep microbial (bacteria, yeast and molds) contamination to a minimum. To do this, there are certain things you must be aware of and guidelines to follow.

• Work in a culture hood set-aside for tissue culture purposes. Most have filtered air that blows across the surface to keep microbes from settling in the hood. Turn off the UV/antimicrobial light and turn on the hood 30 minutes prior to entering the hood.

• Wear short sleeves or roll your sleeves up. Turn your baseball caps back if you MUST wear them, tie long hair back and remove rings and watches.

• Wash hands with specialize soap (like Bacdown) and water before beginning the procedure and rewash if you touch anything that is not sterile or within the hood.

• Spray down your hands, work surface, and anything that will go into the hood with 70% ethanol. Rewipe at intervals if you are working for a long time in the hood. This will reduce the numbers of bacteria and mold considerably.

• Do not breathe directly into your cultures, bottles of media, etc. This also means to keep talking to a minimum. No singing or chewing gum.

• Work as quickly as you can within limits of your coordination. Also, keep bottles and flasks closed when you are not working with them. Avoid passing your arm or hand over an open bottle.

• Use only sterilized pipets, plates, flasks and bottles in the hood for procedures.

• Take special precautions with the sterile pipets. Remove them from the package just before use. Make certain to set up the numbers on the pipet so that they face you. Never mouth-pipet, use the pipetting aid. Change pipets for each manipulation. If the tip of the pipet touches something outside of the flask or bottle, replace with a new one. Never use a pipet twice.

• When handling flasks and plates, take care to not allow media to slosh near the rims or openings. This is one common way for microbial contamination to get into the dish.