hi i would like to know if sterility or accidental addition of microbes during immunofluorescence part must be observed.
The steps starting from taking out the culture plates from incubator, washing it with PBS and fixing it with paraformaldehyde. should i be using sterile PBS and paraformaldehyde etc?
Once tissue or cells is fixed, you do not have to use sterile solutions at all. But you may use solution which does not have any contamination of any kind like dust. It is good idea to filter after you make solution to get rid of undissolved particles of reagents. If you do not filter even and you feel solutions are clean, that will be ok.
Hi,
You can observe bacterial contamination of your slips.
Last month I noticed some rod spots in my IF samples, when I was detecting DAPI. I concluded that was bacterial contamination of one of my stock solutions that I didn't filter.
I generally work in non sterile conditions when I perform IF (I do not work under hood), but I use 0,22 um filtered solutions.
Andrea
No need to work under the hood but preferably the sterile solution, certainly not contaminated reagents.
Once tissue or cells is fixed, you do not have to use sterile solutions at all. But you may use solution which does not have any contamination of any kind like dust. It is good idea to filter after you make solution to get rid of undissolved particles of reagents. If you do not filter even and you feel solutions are clean, that will be ok.
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