Hello Ohoud,

Are you pre-treating your microscope slides before initial tissue mount? How much time and at what temperature are you fixing with acetone?

In my experience the pre-treatment of microscope slides is a crucial step before initial tissue mount, either for general histological staining or immunohistochemistry on frozen tissue sections. If you are not pre-treating your slides this is a very helpful protocol for the preparation of gelatin-coated slides: https://www.rndsystems.com/resources/protocols/protocol-preparation-gelatin-coated-slides-histological-tissue-sections

Also, this is a good source to review some aspects of IHC fixation protocols like time and temperature of acetone fixation: (https://www.abcam.com/protocols/ihc-fixation-protocol)

I hope any of this information is useful for you.

-Angel

Do not worry, Tissue can be frozen without fixing.

You can make sections of tissues using cryostat. Fix tissue sections in 4%PFA ( few drops, freshly diluted, I buy capsule from EM sciences) for 30 minutes, wash with PBS, 0.2% glutaraldehyde for 10 minutes and ready to go for IHC

Thank you Angel and subhash

I will try those porptocol and see what will work