WHATS THE CONTAMINATION???

Dear all

Can you identify the following contamination for me. I have left DMEM + 10%FBS, with commercial antibiotic solution containing penicillin and streptomycin along with amphotericin overnight in humidified co2 incubator and observed this. Its the first time in my entire experience of cell culture that I got contamination. what could be the probable ccause for the same?

TO add to query.... Do I need to change the combination of antibiotics as it could be a resistant variety because I already added the antibiotic.

Hi Satinder

couldn't find the image, can you post it again directly as image in the post. may be contamination could be a fungal growth. 

hi hill trekker

Image is included as rar attachment on ppt slide. you can download and open that.

I also suspected a fungal growth but Im not sure and secondly I added antifungal amphotericin b in the media.

Satinder, I cannot open the file as ppt.

Webmaster, what would be the best way to post jpeg/ppt et al.? Especially for this forum, we need to have a way to do this. There is a "insert image" button, but it doesn't work for me.

Thanks

Dear All

The image is too large in size to be uploaded so I have made a PPT slide ad reduced the file size.

Kindly download the Image and extract the .ppt file using win RAR software and then open it.

Regards

 Hi all

I did got the similar kind of contamination again today in the media after over night incubation.

Please help..........

Satinder,

I know how frustrating this can be. Have you grown media alone in the TC incubator? Do you think it comes from the cells or else?

I don't know what 'rar' is. Can you help? It would help to see a picture somehow.

Stefan

Dear Sgirgenrath

I have grown only media supplemented with 10%FBS. No cells were involved. Rar is a software. down load the attachment and open the same using Win rar by choosing right click ption EXTRACT HERE. and then open the extracted file normally.

Could you convert the image into a JPEG file? As a .jpg, you can reduce its size easily and then not have to depend on people having software that can handle "rar" files (I've never heard of these before).

Hi satinder, is there any chance you have added any solution having property of beta-lactamase? it neutrilizes the property of penicillin hence has a chance of getting microbial contamination.

Dear Parvoman

I did tried that but It didnot worked well as the size of the image that we can post over here is limited

Dear scienceshare

I have not put beta lactamase. I have been doing the cell cultures for over 5 years now and this is the first time I have got the contamination and that too repeatedly.

I am reattaching the image again as PDF. I hope this could be of help...

Thanks, I can open the pdf.

Looks bizarre. I suppose this is larger than cells? Does it increase in size or number over time? Can you 0.2uM filter the media to eliminate the source? My best guess is you are doing this anyways when you prepare the medium. When these things happen to us, we make fresh media and sanitize the incubator.

Note to Parvoman - the reason that you have probably not heard of .rar files is because they are a pretty commonly used PC compression type. 

Easy to spot a Mac user a mile off.... if you do want to open .rar's you can use Stuffit :)

Dear Sgrigernath

I have filetered the media using 0.2 Micron filters from millipore. We did fumigated the incubator too. Thats why In first place Im worried because this kept reappearing.