Sometimes I see these small particles surrounding my cells in cell culture plates. Are these particles microbial contamination?
The first image is a 22RV1 cells used to be clear in the begining but after I split them into two plates I started to see those particles.
The second image is LNCaP treated with siRNA and I started to see those particles the next day after treatment.
Normally I add antibiotics to my medium (of course not while siRNA treatment).
How can I get rid of those small particles from my cells?
I think this is not bacterial but a fungal (mostly yeast) infection.
How to get rid of this infection?
1. Complete desinfection of your incubator. The warm moisty and dark environment is good for the fungi...
2. Start a new cell culture. According to my experience the infected culture can not be effectively "cleaned" without antifungal agents which might significantly modify the outcome of the planned experiment.
3. Use cell culture flasks equipped with filter cap instead of culture dish (if possible)
Good luck!
Gabor
I think this is not bacterial but a fungal (mostly yeast) infection.
How to get rid of this infection?
1. Complete desinfection of your incubator. The warm moisty and dark environment is good for the fungi...
2. Start a new cell culture. According to my experience the infected culture can not be effectively "cleaned" without antifungal agents which might significantly modify the outcome of the planned experiment.
3. Use cell culture flasks equipped with filter cap instead of culture dish (if possible)
Good luck!
Gabor
I agree with Gabor. It seems a yeast infection. Unfortunately your cultures are not good any more. You have to start all over again after your desinfect all your working place. Are you working under a laminar flow hood?
Yes, I always consider sterilized conditions, but my plates always get contaminated after splitting them. even though I filter the Trypsin
Hello
You have the option of adding an anti-mycotic. These are available from most cell culture companies, often in combination with an antibiotic also, it is advisable to routinely sterilize incubators and to do routine checks. .
Good Luck
Treatment of fungal infections of the skin
Treatment of fungal infections depends on several factors - age of the patient, type of fungus, the place where the infection occurred ... Since it is very important in the treatment and prevention of skin hygiene, it is recommended to use antimycotic in the form of shampoo and cream during treatment. A tablet or capsule therapy is added thereto, and laser is used more often in therapy.
It is important to emphasize that, if a fungal infection has affected the skin by hair or a nail plate, then the therapy and treatment are long lasting, that is, they need to last for several months.
It must be preventive
Prevent increased sweating and maintain hygiene in detail by appropriate means.
Do not wear anybody else's shoes.
In the summer period wear light and wide clothes, as well as open shoes.
In public places, such as sports clubs, pools and shared showers, wear shoes, or do not go barefoot.
If swelling or redness occurs on the skin, contact a dermatologist immediately. Do not use corticosteroid fat without prior examination because it mask infection, but it does not prevent its further spread.
When there is a decrease in pet hair, contact a veterinarian, as this may be a fungal infection.
Discard these plates as well as any stocks you have prepared during this period. Sterilize your incubator, water bath (if using for media warming) and add p/s as well as fungicides to your media. Keep your media for check before starting. Yeast contamination is notorious appears after 2-3 days of seeding.
You are most likely dealing with a yeast contamination. I am afraid that you have to discard your plates and start all over again. Good luck
I agree with all of you, it looks like yeast. You have probably to discard your cultures and try to start de novo
good luck
It is most likely yeast contamination. Do a contamination check for your media.
No confusion here. Its yeast.
Ditch your culture and media, and start again.
Dear Majid,
The fist image already suggests that your culture is contaminated by yeast cells.
Before thawing and putting in culture new cells, 1) clean your cell culture facility, 2) prepare new reagents, 3) put aliquots thereof at 37C/RT for 2 days at least and 4) subsequently test under the microscope for signs of contamination.
Thaw new cells only in the case that the new media, PBS, etc are not contaminated.
Keep in mind that the source contamination could be your frozen stocks. So, you may also defrost a different cell line in parallel, as a further control.
Hope this helps.
Best,
Metello
I agree with all answers it's yeast infection, I suggest you to change media, SVF, PBS, trypsin solutions and to clean verry well the incubator . take new cells also.
lamia
Thanks you guys for all the answers. I found the source of the contamination in my experiment. The siRNA i used was reconstituted by another student who didn't use a sterilized DEPC water for the reconstitution which explains why only my siRNA treated cells are getting contaminated while the non-treated cells are clean and healthy.
The question is now: Can I filter the reconstituted siRNA by 0.22um filters to lower the possibility of contaminating my cells?
I think that filtering should do the trick indeed. Yeast tends to be bigger than 0.22um.
Dear Majid,
I agree with all previous comments and would recommend all their answers.
Regarding your second question, I do not completely agree with Stephanie Hoekstra although it seems logical to do. But, be careful, because what the yeast has already produced in the reconstituted siRNA solution is going to affect your outcomes, in an unpredictable way.
I think you have to prepare a fresh siRNA solution and redo the experiment(s) with all precautions taken in consideration.
I would just filter it, for the sake of doing it, but not include this or any of its results in any research related publication.
My best,
Ahmed
You could try filtering your siRNA. It may work but you would be better off preparing fresh siRNA. You won't waste time working with something that may still contain contaminants.
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