I was culturing vero cell in Roller Bottle.
After 2 days of incubation i noticed white / creamy patches attached to inside surface of the bottle.
Upon shaking the patch detach from the surface and drops into media.
From gram staining i found nothing suggesting its not a bacterial contaminatio.
If its from fungal or any other source, how can i determine the type of contamination and further characterize the single causitive agent?
There are pictures of the patches under microscope in different magnification.
It must be a fungal contamination. To identify should go for lactophenol cotton blue staining procedure. Each class of fungus has some unique characteristics like spores and mycelia which can be identified under light microscope after staining. You can take the help of any microbiology book to identify it.
@Jayant Dewangan
Thanks a lot...
Can u further assits me that what could be the posssible source, handling error or contaminated reagent?
Definitely fungal. Consider using amphotericin in your culture medium. A lot of fungal contamination in cell culture tends to come from environmental contamination. Are you working in a biosafety cabinet?
Are you using pipette tips that have ever been opened outside of the BSC?
No...
But i yhink i should check the integrity of the pipette packets.
@Jordan R. Yaron
Definitely its a fungal contamination. Gram staining is only suitable for bacteria but for fungus you have to go for microscopic observation. Every class of fungus have some unique characteristics under microscope. By observing those you will be able to characterize the fungus.