CELL CULTURE INCUBATORS CLEANING AND DECONTAMINATION METHODS Models: MCO-170AC-series MCO-170AIC-series MCO-170AICD-series MCO-230AIC-series MCO-170M-series MCO-50AIC-series MCO-50M-series Basic care for optimum performance and care in the event of contamination

Introduction to cell culture incubators 4 Important points for the installation of incubators 6 Important points for usage 8 Cleaning and decontamination methods 8 Basic care of cell culture incubators 9 Tips to minimize contamination 13 Care in the event of contamination - 24 hour UV decontamination 14 - Dual Heat sterilisation 20 Care in the event of rust 21 Best practices and good laboratory technique 22

Introduction to cell culture incubators A cell culture incubator is designed to artificially replicate in vitro conditions essential to in vivo physiology typical of human and animal models. Cell growth outside of a natural environment presents a multitude of challenges associated with exposure to microorganisms that are not present in the in vivo state. Depending on the type of cell cultures being managed, several operating parameters must be carefully controlled with accuracy, repeatability and flexibility in setpoint choices. These include temperature and gas control. • Cell culture incubators...

THE INCUBATOR BUBBLE Unlike closed systems, such as hollow-fiber substrates, stirred tank or airlift bioreactors, the typical cell culture incubator is a conditioned chamber with a door that closes against a soft gasket. When the door is closed, the incubator creates an ideal environment for the cell culture process based on user defined setpoint parameters for temperature, CO2 and O2. Humidification is naturally evaporative from the humidifying tray, and an elevated relative humidity is sufficient to eliminate desiccation, especially in microplates with small media volumes. Some larger cell culture...

Important points for installation There are many factors to consider when determining the permanent location of the cell culture incubator. It is desirable to locate the unit where there is minimal foot traffic and where air disturbance is of little consequence. This reduces the volatility of outside air entering the incubator during a door opening. Avoid installing the incubator near windows, air conditioners, ceiling or floor HVAC air diffusers and return air intakes, all of which are sources of airborne contamination. INSTALLATION, LOCATION AND CLEARANCES It is important to consider the function...

Locations where there is a lot of foot traffic are not suitable for the unit. • Locate the incubator in a dean room or location where few people enter • Choose a clean room that is safe or a place where there are as few people as possible. Set up the unit as high off the floor as possible • Since there are fewer airborne bacteria in the upper part of a room, the incubator should be placed on a laboratory table or a special stand. • If stacking two or three units on top of each other, use a special roller base for that purpose. Place in a location that is not directly affected by outside air •...

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...

Basic care of cell culture incubators Always puts on gloves before cleaning the unit. As a basic rule, do not clean the incubator with bare hands. Be sure to use rubber gloves. Necessary materials • Rubber gloves • 70% ethanol • Sterile non-woven cloth/paper STEP 1 Turn off the power Remove the interior components. Clean the interior components Remove the components in the correct order. Use the correct cleaning procedure. 1 Remove the shelves 2 Remove the humidifying tray cover, and take out the tray 3 Remove the back duct 4 Remove the fan 5 Remove the stopper for the access port 1 Wash with...

Basic care of cell culture incubators STEP 4 Spray disinfecting alcohol inside the unit and wipe (70% ethanol). Disinfect all interior surfaces, internal components, shelves and water tray with 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 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. Sensor holes If it is operated with the inner door gasket not in place, the humidified air will leak and...

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. 1 Replace the stopper for the access port 2 Replace the fan, check to see if the fan spins smoothly by turning it with your hand. 3 Replace the back duct 4 Replace the humidifying tray cover, and put in the tray 5 Replace the shelves 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...

Basic care of cell culture incubators ClEanIng PrECauTIon Get into the habit of wiping carefully • Be sure to wear gloves to avoid cutting your hands on the interior component • Do not use acid, alkali, or chlorine-based cleansers, disinfectants, or sanitizers IMPorTanT PoInT Do not wipe with the same portion of the sterile non-woven cloth more than once. If you wipe another area with same portion of the cloth, you will be spreading bacteria around. Don’t forget to wipe the gasket and the inside of the door. ClEanIng ThE huMIdIfyIng WaTEr When replacing the water, also clean the tray. Do this...