I immunostained PDGFRb which should be mainly positive on peicytes of vessel walls in glioma xenografts from mice. However, non-specific signals are frequently seen in vessel lumens. For tissue processing, I used PBS for perfusion, then fixed the tumor in 4% PFA in PBS for one day and immersed it in 30% glucose in PBS for 3 days. Could anyone tell me the reason for the non-specific signals in vessel lumen? Thanks!
They look like erythrocytes into the lumen, if you change filter do you still see them very intense ? Anyway as far as I know the autofluorescence of the erythrocytes is quite normal and you can do pretty nothing to avoid it, just learn to differenziate from a truly positive cells, but is easy when you change filter and you still see them very intensely fluorescent.
I hope that can help
Hello,
if you inject the tumor cells in the brain directly you damage the blood brain barrier and macrophages can also invade the damaged area.
These macrophages are also autofluorescent (visible with all fluorescence filters) and moreover also autofluorescent calcium particles can be observed after damaging the brain.
But especially in case of your image I also agree with Federico that erythrocytes are shown.
Kind regards
Stefan
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