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#Asepsis technique how to#
#Asepsis technique skin#
#Asepsis technique free#

Never sweep your hands and sleeves over the top of an opened bottle as this could cause contamination. Only open bottles immediately before use and if the lid is put down ensure the opening is facing downwards. Add each supplement to the bottle of media using a new sterile tip or pipette. Aim to work quickly and efficiently to minimize the risk of contamination and place each bottle in easy reach. To prepare the media aseptically spray each item with ethanol. Take the cell culture media and supplements from the fridge and place in a 37 oC incubator to warm. Next, spray the inside of the hood with 70% ethanol and wipe clean with tissue.

Thoroughly spray the inside of the cell culture hood with disinfectant and wipe clean with tissue.ĭispose of used tissue in the appropriate waste bin. This video will outline the procedures necessary to work in a sterile manner within a cell culture hood.īefore commencing work ensure that you have washed your hands and are wearing the correct personal protective equipment, this includes gloves, a lab coat, and safety glasses. All cell lines from Abcam are supplied mycoplasma-free, so this is something you should not need to worry about.Īseptic technique prevents contamination of cell culture and reagents from microorganisms in the environment.

#Asepsis technique free#

Once a mycoplasma contamination has been detected the only way to ensure a mycoplasma free environment is through hydrogen peroxide fumigation, also known as “bombing”, of the lab. The most common source is through contamination of cultures with microorganisms such as bacteria, fungi and yeasts, which can usually be observed under the microscope.Īnother important contaminant is mycoplasma, which is resistant to traditional cell culture sterilisation techniques, isn’t visible and requires specific routine testing to detect.

#Asepsis technique how to#

Sources of contamination and how to address them View our cytopresevation of mammalian cell lines protocol.

#Asepsis technique skin#

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.

Always wash hands thoroughly before and after working with cell cultures.

Personal protective equipment and good hygiene

Never open multiple flasks, dishes or bottles at the same time, this may result in cross contamination.

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.

Never leave bottles of reagents, flasks or dishes open in the hood longer than necessary.

Sterilize reagents either by filtration (0.2 µm filters) or autoclaving if necessary.

Many cell culture reagents are certified sterile from the manufacturer however, they can become contaminated during handling.

Instead use sterile glass or disposable plastic pipettes to manipulate liquids and use each one only once. Never pour cell culture media or other reagents.When bringing equipment or reagents into safety cabinets, spray them beforehand with 70% ethanol.Spray gloved hands with 70% ethanol before commencing work in the safety cabinet.Areas of work should be uncluttered and contain only the items required for the current procedure.Ultraviolet (UV) light can also be used to sterilize safety cabinets between uses. The interior of safety cabinets should be wiped with Trigene disinfectant followed by 70% ethanol before and after use.Cell culture should be carried out in a laminar flow safety cabinet located in a room that is specific to cell culture.For other video protocols please visit our video protocols library here.