Whether in a professional tissue culture lab or DIY home tissue culture lab, strict hygiene protocols should be observed if you want to steer clear of contamination.
Contamination within the tissue cell culture process is difficult to avoid altogether due to the highly susceptible nature of your media. There are certain precautions you can take, such as using biocides or a preservative mixture like PPM™ to reduce existing contaminants and prevent new contamination from occurring, but in general, contamination is one of the greatest issues facing tissue culture cultivators.
Different contaminants require different methods of treatment. The following list of common contaminants will help you to know what to look out for, and what you are dealing with:
What are the Most Common Types of Contamination?
Knowing how to treat and prevent lab contamination is an essential skill for anyone using tissue culture media. Keeping the environment clean by using aseptic techniques is critical to maintaining contaminant-free lab equipment.
Whether the contamination is airborne microbial contamination, contamination that occurs due to contaminated lab equipment, or even diseases arising from contaminated water, unwanted invaders must be eliminated so that no cell line contamination can take place.
When you have identified the source of your contamination, you need to take control as soon as possible. Bacterial and fungal contaminants are usually easy to identify. Mycoplasma and other microbial infections have a subtle visual appearance, and are difficult to identify with the naked eye. Viruses can be even more challenging to identify, as the signs can be even more subtle.
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Types of Contamination
- Microbial Contaminants
- Fungal, bacterial, and yeast contaminations are among the easiest to identify. But while they are easy to identify, they can be vicious unwelcome guests. If you are using agar or another medium with added phenol red, then be on alert if your medium changes color as this indicates microbial contamination.
- Microscopy can also be used in the identification of microbial cells. The best way to keep your plants free of microbial contamination is to perform daily observations and administer the proper dosage of PPM™ as a sustainable treatment. Be regimental in your daily culture observations and you will be able to detect contamination early. This early detection will become one of your best defenses. Prompt elimination is essential if you want to contain the infection and prevent the contamination from spreading throughout the plants and the lab environment.
- If you are wondering which contaminant should cause you the most concern, you’ve just found it. Mycoplasmas can be a severe contaminant and are known as one of the most widespread contaminants faced in tissue culture.
- While there are two types, fungal and bacterial, these biological contaminants are reasonably simple to spot. Mycoplasmas and viruses are more difficult to identify, and therefore present more of a risk if they are left to grow and multiply.
- Mycoplasmas are difficult to detect, and they can have a profoundly negative effect on cells. While they are classified as bacterias, mycoplasmas contain characteristics that separate them from common bacterias. Lacking a cell wall and being smaller than other bacterias, they can easily permeate filter membranes that are utilized during sterilization processes. Antibiotics used to be the common solution to bacterial and fungal contaminations, but researchers have begun to realize the long term harm these treatments have on plant cultures and instead, are now advising the use of preservative mixtures such as PPM™. Mycoplasmas are resistant to antibiotics since antibiotics target bacterial cell walls. PPM™, on the other hand, can eliminate both bacterial and mycoplasma contamination.
- Bacteria and Fungi
- Molds and yeasts are common problems that can affect your cultures. Bacterias and fungi tend to flourish in cultures and are quick to multiply, making them the most common contaminants in tissue culture labs. Yeasts cause the medium to become cloudy as the cells begin to die, and molds will cause branched mycelium, which will be look like furry clumps that float on the surface of the culture medium.
- As we have mentioned, viruses are difficult to identify. Unlike other common contaminates, they do not leave a trail of visual cues behind them. There is no cloudy medium, nor are there any changes in the culture medium’s pH.
- Although the viruses can cause severe damage to the host, they are usually self-limiting. Therefore, the main danger that viruses pose is to the laboratory personnel, people like you and me. Always follow safety precautions, especially if your lab contains animal or human cells.
- If you are working with tissue culture, you should be prepared for contamination at every stage of the process. Therefore, follow safety and hygiene precautions with rigidity, and use the appropriate preventative and curative measures, such as high-quality plant preservative mixture.
While many people used to turn exclusively to antibiotics, researchers now understand the drawbacks of such treatments. Antibiotics lead to the spawn of resistant contaminant strains, are not sustainable for long term use, and are largely ineffective against fungal infections. PPM™, on the other hand, is rapidly becoming the favorite amongst lab personal because of its ability to prevent and eliminate fungal contaminations, as well as secure a resolution for all existing microbial contaminants.