Dear All,
Hope everyone doing well,
I am trying to prepare 2X EMEM and 2X DMEM stock for my experiment. After prepration and filtration when I aliqout some part of my medium for contamination testing I could see there are some strange looking particles are floating inside my medium. However they are not moving or growing even if I incubate my flask for few days.
I have troubleshoot and changed a lot of stuffs for medium preparation (eg: Medium filter, Source of MiliQ water, bottles, magnetic stirrer, pH meter and even different brand of Medium Hiclone and Gibco) and these things still exist in my 2X medium.
I attached the microscopic photo here. is anybody experience same issue before here?
Looking forward to hear your comments
Thank you so much
It could be protein aggregates from the FBS you are using. try sterile filtering it at room temperature before you use it.
hi
I agree with victoria , after thawing of FBS ,remove fibrin precipitate with centrifuge ( 10.000RPM / 5min /RT) .
good luck
Thanks for your answers. But I am facing this in my 2X medium which i havent add any FBS, L- Glu and NEAA inside the only thing I add inside is Sodium bicarbonate. In our lab we prepare 2x stock then from there I prepare my 10% and 2% medium. Since I encounter this problem I dont dare to waste my DBS and other supliments to prepare my growth media
Ok, this is strange. What filter size have you used? Judging by the size it shold be removed by a 0,2 µm filter.
Have you checked your components under the microscope individually?
I use 0.2 um filter to filter my medium. I haven't check the other component under microscope yet. the only component left for me to check its Sodium Bicarbonate which Im going to check it later
as far as I know, preparation of 2X medium does not allow complete solubilization of all the solutes. so it is possible that what you were looking at under the microscope was insoluble solutes precipitating or forming sediments. Have you noticed when you allow the 2X medium standing for a moment that a sediment is formed at the bottom of bottle/tube?
- Badges
- Science topic