Cell contamination - Small rapidly moving black rods (Feb/08/2008 )
Hi,
Our whole lab is having an absolute nightmare at the moment with regards a contamination that just seems to be getting worse despite our best efforts. Various cell types are being cultured (e.g. T47D, MDA-MB-231, HeLa, Kerationcytes etc) & we are all encountering the same problem - the cells go down due this contamination. Under the microscope they look like small black dots (rods on closer inspection) that appear to be moving quite rapidly... We have done a million different media tests with various components & cant seem to determine which is the culprit as the manner in which they go down is quite random. The contamination becomes apparent very rapidly - media is turbid after 12-24hrs. We have thrown out all suspected aliquots, frozen stocks etc but still cant figure out the cause. We then thought that the source was something in the room itself rather than a media component so we have sterilized both the hood & the incubator (¬ to mention the whole cell culture room), yet we are still experiencing the problem if anything it just seems to be getting worse. We have also determined that it is resistant to penstrep. It all began about a month ago when 3 or 4 of us were affected but now the whole lab is affected and all work has pretty much come to a standstill.
If anyone has any idea as to what it might be & how to eradicate it we would be eternally grateful.
Maybe there is something in your refrigerator or where you keep your media? But I assume that you will have cleaned this also....
very strange... or perhaps the filters in your sterile hoods are contaminated, so the airflow maybe the culprit?? I am just guessing
Wish you good luck to find it ! 
very strange... or perhaps the filters in your sterile hoods are contaminated, so the airflow maybe the culprit?? I am just guessing
Wish you good luck to find it !
Yeah we cleaned out the refrigerator. The filter in the hood is our main suspect at the moment. It really is mind boggling though... I'd be very curious to find out what exactly it is & how to get rid of it so as to prevent future problems. I've been set back quite a bit already - a month without any cell work so far!! Frustrating beyond belief!
It is also possible that the filter inside the incubator got contaminated, or it just needs to be changed.
Another possibility is the water bath.
Hi, I posted about the dots on another post.
It's your Serum. Check your serum. Someone messed
up the serum.
it could be anything - the growth media, the serum, some chemical you added (even your pen/strep)
throw it all away (stuff frozen before the contamination MAY be ok but its still a risk)
if you still have contamination with new media its probably in the source (the cells) so - throw them away.
the fact that its moved from cell type to cell type is not a good sign - make sure nobody in your lab is borrowing other peoples stuff (if they are - throw THEM away)
i'm afraid the best advice i can give is the only way of killing a contaminant is to put it in the bin
be quick before rhombus shows up and shouts at you
good luck
dom
I've thrown everything away... If in doubt & all that!!! As for the filters - the hoods arent even a year old so they SHOULD be ok but that doesnt mean that they aren't. I'm pushing to have them changed just to be sure!
I made up new media on Friday & did a media test over the weekend & thankfully all ok today so going to attempt to bring up my cells again. I noticed that a few other people had their cells go down again over the weekend so i'll keep you updated on what happens over the course of the next few days. Thanks for all your suggestions!
we had the same problem just last week. We tentatively labeled the problem as a mold contamination. Our culprit so far has been one particular media bottle and a heating filter in the room.
Since this has been happening for a little while, be sure to clean everything: hoods, incubators, etc. We've had some luck using Sporicidin--just be careful as it does contain phenol as the main active ingredient.
Also, make sure that the water baths and water pans for the incubator do not have any growth. Use sterilized water for those especially and supplement with some algicide.
It's a good thing that you did a media test. if that looks ok, then definitely go for it.
Good luck!
Since this has been happening for a little while, be sure to clean everything: hoods, incubators, etc. We've had some luck using Sporicidin--just be careful as it does contain phenol as the main active ingredient.
Also, make sure that the water baths and water pans for the incubator do not have any growth. Use sterilized water for those especially and supplement with some algicide.
It's a good thing that you did a media test. if that looks ok, then definitely go for it.
Good luck!
Mould contamination could be likely as we found some mouldy mildewy stuff by the window. (Has all been cleaned now). Everything has been sterilized - incubator, hood, waterbath, watertray. When waterbath & watertray were refilled there was algicide added to both. Fingers crossed this is the end of the problem!
With luck it should be!
Let me know how it turns out. 