Hello,

I am afraid there is too little information here to help you.

Could you provide some images of your negative and secondary only control as well as the final stain so we see which step exactly introduces the background?

This would be a good start to search for a solution.

Best

JGH

Hi Zahraa Fakih

When performing immunofluorescence on mouse skin tissue, a high background signal can be caused by several factors, including non-specific antibody binding, autofluorescence, and inadequate blocking of non-specific binding sites.

Here are some suggestions to help reduce background and improve the signal-to-noise ratio:

It is important to note that the optimization of immunofluorescence protocols can be highly specific to the target and tissue being studied, so it may be necessary to perform several rounds of optimization experiments to achieve the desired results.