Worm like contamination (see photo) - serious problems (Jun/28/2007 )
Dear All
Could you please give us some suggestions about the conatmination problem we have been facing very seriously overhere.
Description:
1. cells seems ok untill the next day of sub-culture
2. after 24 hrs of seeding, there are very few bacillus like dark structures
3. after 48 hrs, there are more such structures both clumps and in individual (still cells grows well enough)
4. 72 hrs culture contains uncountable such worm like structures (see attached photo)
5. contamination is very serious and spreaded to many other cell lines in the same incubator.
(interesting thing is they look like moving worms under the microscope). 
I discarded all media and washing solution and then started new stock, the problem continued...
Whats these, possible routes of contamination, how to get rid of these----I cant imagine, how to solve these things.
Yours views, suggestions are highly appreciated.
Thank you very much.
Thapa
It seems like E. coli.
anyone doing c.elegans work using your lab?
c.elegans are much larger.
Noh sanjiun.
As far as I know, C. elegans is quite larger in size like 1 mm and almost visible through naked eyes. But here in my cell culture flask they are not visible (little turbid like bacterial growth). And you can see the 400X microspic image.
Could you please give us some suggestions about the conatmination problem we have been facing very seriously overhere.
Description:
1. cells seems ok untill the next day of sub-culture
2. after 24 hrs of seeding, there are very few bacillus like dark structures
3. after 48 hrs, there are more such structures both clumps and in individual (still cells grows well enough)
4. 72 hrs culture contains uncountable such worm like structures (see attached photo)
5. contamination is very serious and spreaded to many other cell lines in the same incubator.
(interesting thing is they look like moving worms under the microscope).
I discarded all media and washing solution and then started new stock, the problem continued...
Whats these, possible routes of contamination, how to get rid of these----I cant imagine, how to solve these things.
Yours views, suggestions are highly appreciated.
Thank you very much.
Thapa
very likely bacteria (not C. elegans
), f.i. Lactobacillus; with penicillin and G418 it is away within 24 h; G418 may harm your eucaryotes but it is the minor thread compared to bacteria invasion... Rapid growth make us to think about bacteria. But you know WHR, we use penicillin/streptomycin.
what X magnification is this?
you can always see bactrial contamination like this... very fast moving...
but its looking little bigger.. i don't know why..
anways.. no easy route to avoid contamination... but its better to..
1. clean your hood n check its pressure .. if it is out.. u'll never be able to grow any cell line..
2. test your media also for contamination..
3. grow new stock from liquid nitrogen in other hood if possible.. (use fresh media)
4. clean your fridge..
i did all when all my 6 cell lines got contaminated last year in june.. lol.. wasted whole month..
anywyas.. good luck.
you can always see bactrial contamination like this... very fast moving...
but its looking little bigger.. i don't know why..
anways.. no easy route to avoid contamination... but its better to..
1. clean your hood n check its pressure .. if it is out.. u'll never be able to grow any cell line..
2. test your media also for contamination..
3. grow new stock from liquid nitrogen in other hood if possible.. (use fresh media)
4. clean your fridge..
i did all when all my 6 cell lines got contaminated last year in june.. lol.. wasted whole month..
anywyas.. good luck.
Thanks Lab helpdesk for your guidance.
Yes, the magnification is 400X and culture is HT29 cells.
Most interesting part is I have started new stock from liq. N2 yesterday, still there is very few such creatures ( I am pretty sure the stock is well enough). Furthermore, all the precautions were taken like cleaning of hood and cabinet, changing media and all.
Do I use another hood and to further clarify??
Have you sterilised your hand help pipet, as it could be the thing which transfers the bacteria.