Page 4 - Cell Culture Training Hand Book - A4 Edited 20.1.2020

ASEPTIC TECHNIQUES

Work Area
 Only items required for intended cell culture procedure are allowed on the working surface.
 Routinely, disinfect the working surface and surrounding area thoroughly with 70% ethanol
before, during and after work.
 Use ultraviolet light to sterilise the air, work surface and items required for cell culture for at
least 30 minutes.

Personal Hygiene
 Wash your hands before and after working with cell cultures.
 Complete personal protective equipment (PPE) is required at all time to protect yourself from
hazardous materials, reduces probability of contamination from shed skin as well as dirt and
dust from your clothes.

Reagents and Media
 Wipe the outside of the bottles, flask and plates with 70% ethanol before placing them on
your working surface or cell culture hood.
 Make sure all of your plastic wares are stored in sterile re-sealable bags.
 Routinely inspect your media and reagents if they look cloudy, has floating particles, foul
smell or unusual colour.

Sterile Handling
 Make sure to wipe your hands with 70% ethanol.
 Wipe the surfaces of all items ie: pipettes, containers, flasks, plates, and dishes with 70%
ethanol before placing them in the cell culture hood.

 Place caps or covers with the opening face down on the work surface if you need to remove
it from the bottle/flask. Always cap the bottles after use and seal with parafilm.

 Always use sterilise pipettes when working with liquids. Use once, to avoid cross
contamination.

 Do not open the packaging of the steriled flask, bottle, petri dish, or pipettes outside the cell
culture hood. Do it in the hood, when you are ready to use. Never leave the plastic wares
packaging open to the environment. Seal the packaging as soon as you are finished.

 It is best not to talk, sing, or whistle when you are performing sterile procedures.

Cell Culture Training Hand Book | CMBL Page 4 of 12