Hello
Why I am getting a red dot( or false satining)when I stain my primary cultures only with DAPI?This spots looks red in DAPI filter and looks green in FITC filter and red in TRITC filter irrespective of the fact that I am not staining it with FITC or TRITC. .The red dots are not in my nuclear region but looks to be in surrounding region .It appears as false positive.This problem is only with tissues and primary cell culture staining.Secondary cultures do not give such false staining.
It will be good idea if you could upload one image, than someone can comment who is expert in these areas, I also see very fine red dot in the center of nucleus,
Sounds like you could have some sort of fluorescent debris in the sample if it is lighting up in all channels, or possibly the fluorescence cubes allow bleedthrough of the signal between fluorescence channels so that you detect the object in all channels.
Since it is missing in the secondary only control specimens, one possibility is that there are aggregates of the primary antibody in your solutions that for some reason like to stick to the nucleus. You might try centrifuging the primary antibody to see if you can remove any aggregates before applying the antibody to the sample.
Hi Samrina. Let me take a guess that you are staining endothelial cells.
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