We work with lymphoblastoid cell lines derived from EBV transformed PBMCs. Recently, one of the lines I have been culturing has ceased growth, despite forming healthy-looking clumps. I have noticed a lot of smaller cells which appeared to be debris settling at the bottom of the plate. Recent observations at 20X magnification showed that those cells seem to be moving, almost in a vibrating sort of manner.
Have been looking at papers online which identify these as black dots, and using antibiotic combinations seems to help them get removed. What concentration of piperacillin/tazobactam can I use?
7/7/21
Dear Anushka,
From the photos, I can't tell for sure if you have contamination in your cell culture. You might try looking at the cells at with a 40x objective (total magnification 400x) or, better still, a 100x objective (oil immersion lens; total mag: 1000x). The vibration you see is not definitive, as it could be simple Brownian motion. You could try doing a simple stain (e.g., crystal violet, etc.) or a Gram stain and see if you can visualize bacteria.
If you want to be sure, transfer a small aliquot of your cell culture to a flask of sterile microbial broth designed for growing bacterial cells, (e.g., Nutrient Broth, etc.) and incubate it at a suitable temperature (e.g., 30-35C) with shaking. If you a have microbial contamination, the broth will become turbid in a few days.
I hope this information helps you.
William J. Colonna Ames, Iowa, USA [email protected]
You can have an option to identify. You need to collect the reference image so that you can compare them with this image using pattern matching algorithms.
Hope it works
I think it is fungal contamination, best is to discard yr culture, sterilise everything and start fresh. the best way to identify contamination is you will see hypae like structure at higher magnification (40x or 60x), in bacterial contamination, media becomes turbid.
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