Hello,
My collagues and I are trying to stain paraffin-embedded slices (mouse brain). After deparaffinization, hydration and dehydration ( as explained on https://pathologycenter.jp/method-e/nissl.html), the dye went away on the first step in alcohol. We have tried different steps but the slices didn't retain the staining. Can you help us to understand why this is not working? Which step is the problem? Thank you everybody
Dear
See the method in the following paper
A rapid method combining Golgi and Nissl staining to study neuronal morphology and cytoarchitecture
May be the differentiation step in 95% ETOH + Acetic acid is too strong. After CV-staining rinse the sections in several changings of distilled water and put them for 3 min each in 70% ETOH, 2x 100% (95%) , Isopropanol, 2x Xylene.
After the first 100% ETOH you can check the quality of the defferentiation under the microscope. I have add a protocol which we use for then 20 years and an images of stained rat cortex, paraffin 5µm thick
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