Cleaning and decontamination methods
Most incubator manufacturers recommend a solution of 70% ethanol and manual cleaning prior to initial start-up and regularly thereafter. The 70% ethanol solution is intentionally diluted to give the ethanol time to kill the contaminant before the ethanol evaporates.
Why is 70% ethanol better than 100% ethanol in bacterial inhibition? 100% ethanol coagulates and dehydrates proteins so quickly that a layer of relatively impermeable denatured protein forms in the exterior parts of the bacterial cell (in and under the cell wall), and this prevents further diffusion of the alcohol into the cell. This protects the core of the cell from denaturation.
With 70% ethanol, the process is slower and the alcohol manages to diffuse throughout the cell denature proteins. In addition to conventional manual wipe down using 70% ethanol, the incubator may be equipped with a sterilisation cycle such as a high heat (180°C) system or a decontamination cycle such as hydrogen peroxide vapor H2O2 system. The cycle should be performed prior to first use. If commissioning and Current Good Manufacturing Practices (cGMP) criteria are in place, all contamination control efforts must be in compliance with previously approved best practices and facility protocol.
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Instrumentation and equipment interface?
Shakers, cell bottle rollers, magnetic stirrers and other devices are commonly used in the cell culture incubators. These must be free of contaminants before they are placed in the incubator.
Cell culture vessels usually include flasks with and without vent caps, petri dishes, roller bottles and multi-well plates. These are usually prepackaged and sterilized by gamma radiation prior to shipment. They should be opened only in a biosafety cabinet to preserve the integrity of the sterilisation.
Other labware returned from a central sterilisation room must be considered a source of contamination if exposed to ambient air during cart transit and shelf storage.
How to clean a CO2 Incubator?
Step 1 - Turn off the power
Turn off the incubator and unplug it from the power source.
Step 2 - Remove the interior components
Remove all the cultures, shelves, trays, and any other items from the incubator. Place them in a clean and safe location.
Step 3 - Clean the interior components
Wipe the interior surfaces of the incubator with a clean and damp cloth. Avoid using abrasive or harsh cleaners as they can damage the interior surfaces. Instead, use a mild detergent or disinfectant solution recommended by the manufacturer.
Step 4 - Spray disinfecting alcohol inside the unit and wipe (70% ethanol)
Do not spray 70% ethanol into the sensor holes directly! Just wipe off with non-woven cloth/paper which is sprayed with 70% ethanol.
Step 5 - Disinfect all interior surfaces, internal components, shelves and water tray with 70% ethanol
Step 6 - Please make sure to spread the ethanol for disinfection to all corners of the inner door gasket and sufficiently remove stains while wiping it clean
If it is operated with the inner door gasket not in place, the humidified air will leak and will cause condensation between the unit and outer door. After wiping, confirm that the inner door gasket is securely in place and doesn’t have any creases.
Adjust the shape of inner door gasket by sliding fingers from each corner in direction of arrows. Specifically, insert fingers behind fin of inner door gasket and slide.
The inner door gasket has an important role to maintain the chamber humidity. If it is operated with the inner door gasket not in place, the humidified air will leak and will cause condensation between the unit and outer door. After wiping, confirm that the inner door gasket is securely in place and doesn’t have any creases. If the inner door gasket is not in place, please refer to the back side and adjust the shape of the inner door gasket.
Step 7 - Replace the interior components
Replace the components in the reverse order of [STEP 2], and put sterilized distilled water in the humidifying tray.
- Replace the stopper for the access port
- Replace the fan, check to see if the fan spins smoothly by turning it with your hand.
- Replace the back duct
- Replace the humidifying tray cover, and put in the tray
- Replace the shelves
Step 8 - Let it dry with the door ajar
Before turning the power back on (restarting) let the inside dry out, check that there is no alcohol smell remaining. If you turn the power on while it is still damp inside, the O2 and CO2 sensors may be damaged.
Step 9 - Dual heat sterilisation function or H2O2 (EXTRA)
If using a unit that is equipped with dual heat sterilisation function or H2O2 (hydrogen peroxide) decontamination function, performing sterilisation/decontamination before use will make prevention of contamination (bacterial contamination) more effective.
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