I received this gastric tumor biopsis and processed using standard protocol to establish gastric cancer organoids. Contamination looks like this. I have never seen this before and the culture medium used for the organoid contains primocin. There were no obvious change in the medium pH.
Hi Ran
It is very difficult to conclude the type of contamination or whether the culture is contaminated in the first place (given the pH is fixed, primocin is sufficiently added, samples were disinfected prior to culture as much as possible).
From here it appears like fungal/ contamination (black circular structures may be sporangia). It may also be the dispersed tissues and debris (less likely). One way to figure out the contamination is by imaging at a higher magnification and look for yeast/fungal morphologies (buds). OR if possible take a small amount of the sample (20 ul) and test it out on an adherent plate and see if the contamination persist. But this may expose your incubator to contamination. In worst case I believe the sample may be heavily contaminated to begin with and hence either more primocin and pre-washes are required when you work with it next time.
Can you use cotton blue to visualise possible cells? Could be only crystallization, but first, it is necessary confirm it it isn´t fungi.
Looks like a precipitate (primocin or else). Avoid repeated freeze thaw cycles.
Check medium in microscope and optically. In case, pass through filter or replace primocin by new batch which has been kept at room temperature without any freezing step.
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