Contamination control in plant tissue culture is one of the most difficult issues to manage, and the real secret to your defense against contamination is prevention.
Bacteria, mold, and mycoplasma are common contaminants that most lab personnel are familiar with throughout their work with tissue culture media. Bacterias are easily identified and can be dealt with swiftly before they have too much of a devastating effect.
Contaminants like mycoplasma and fungal pathogens usually leave fewer visual clues and can, therefore, have a more severe impact on your tissue culture if left unchecked. Whatever the contaminant, it is safe to say they can be a nightmare to deal with.
In the case of tissue culture contamination, prevention is better than scrambling to eliminate. Use these seven tips to keep your experiments clean, healthy and free of contamination.
1. Use PPE
Wear gloves, lab-coats, and use hoods so that there is a barrier between the non-sterile factors (yes, that includes yourself) and the cells. Use personal protective equipment such as lab coats and gloves. Have your hood cleaned and serviced regularly. This maintenance will help to ensure that your hood is working efficiently and that your culture media is getting all the protection necessary. There are different kinds of hoods available, and each type offers a different level of protection.
2. Use the Hood Properly
If you followed the advice in step one, then you have a hood to protect your culture samples. But just having a hood is not enough - you need to use your hood correctly. Yes, this may seem obvious to you, but merely having a hood is not enough to ensure a secure environment. The first thing to check is that there is proper airflow inside the hood. Make sure that nothing is interfering with this airflow and that nothing is covering air inlets or outlets.
Cleanse the hood after each working session by wiping it down with sterilizing materials or using your ethanol solution. Although some people like to have a waste disposal basket in the hood area, it could compromise the environment, so it is best to keep your waste disposal outside the hood. And at the end of the day, be sure to switch on the UV light inside the hood to enable overnight sterilization.
3. Clean your Incubator
Some older model incubators do not have self-cleaning mechanisms. If this is one of yours, then make sure that you manually clean the incubator according to its specific protocol.
4. Sanitize, Sanitize, Sanitize
Use 70% ethanol mixed with water to kill bacteria. Spray everything, including your gloves and all equipment before you bring it into the hood. If you are right hand dominant, keep your equipment to your right, your hood directly in front, and the ethanol disinfectant on your left so that you can pick up the equipment with your right hand, spray it with your left hand, and then place it inside the hood. Simple, streamlined, and efficient systems like these will prevent careless contamination. Wash your hands before you interact with the tissue culture media, and make sure that anything you are wearing is clean and sanitized.
5. Use Plant Preservative Mixture
PPM™ not only targets and kills contaminants (including fungi), it can also prevent plant contamination from ever occurring. Use PPM™ with a high-quality culture medium to ensure healthy and protected experiments. Plant preservative mixture is a liquid concentrate that is effective against human contact, endogenous, waterborne, and airborne microbial contamination.
6. Keep Cell Exposure to a Minimum
Maintaining a sterile environment inside the hood and incubator is meaningless if you leave the cells exposed in an unsterilized environment. Yes, sometimes it is necessary when transferring between the incubator and hood or viewing under a microscope but plan so that they spend the least amount of time outside as possible.
7. Keep Organized
Even if your personal life is in shambles, make sure that your tissue culture lab is not. Following a dogmatic organizational approach will be your greatest asset in the lab. Label everything, and follow procedures down to the very last detail.
The only thing between your tissue cells and contamination is a set of strictly followed techniques and procedures to maintain aseptic environmental conditions. If this sounds complicated, just remember to wear the appropriate protective equipment, sterilize your equipment and use plant preservative mixture to prevent and treat contaminants.
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