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fungal contamination - fungal in cell culture (Mar/04/2007 )

hi,

Iam working on cho cell line, some of my flask get contaminented by filamenteous fungi and even confused by cell clumping some times. What would be the good method for conclusion of fungal contamination in cell culture as the it takes full swing of appearences of contamination took 8 days in the culture. If we make conclusion of initial stages of fungal contamination would not only save the time, butalso spreading of contamination during expansion of seed. So I request, if anyone has a good method to make conclusion of fungal contamination in early stages in cell culture?.

anyone has a good method to make conclusion of bacterial contamination in early stages in cell culture?.


thanks in advances.

-Gsanjay-

you can see fungal contamination by microscope inspection.

I don't know of any early detection techniques for bacteria. let alone mycoplasma infections which messes up experiment results, difficult to detect and harder to kill.

However I firmly believe that one should first make sure the working hood is sterile. Then work hard at perfecting one's sterile technique. Else contamination problems will hound you to death.

-perneseblue-

bacterial detection is easy; usually the doubling time is quite short. from a few hours to overnight, at most a couple days, you will have pretty substantial growth if bacteria are in your cultures. I think Perneseblue's point about good technique is excellent...but I would add that if you ever really need to know if you have bacteria, do a gram stain. it's cheap and easy and answers that question very quickly

-aimikins-

thanks for giving ur valuabe advices...

all looking the same in near future... of query also...

byeee

-Gsanjay-

As said before better aseptic techniques. Eliminate fungy is harder than bacteria. First clean well the incubator, use a new bottle of media, use a laminar flow hood if possible (make sure is clean!!), clean the outside of bottles with 70% alcohol, when transfering the media and or cells use a bunsen burner near and flame the top of bottles (beware to not kill cells or damage plastics!!!)and If possible add an antimicotic to the media, the majority of antibiotics that are in lab are for bacteria only so make sure you have an antimicotic also. Hope this help!!and Good luck

-merlav-