The brain was removed for histological examination and placed in fixative formalin for 24 h. Next, during the dehydration and clarification steps, the samples were embedded into molten paraffin. Using a microtome (Cambridge Medical Instruments, United Kingdom), 6-micron-thick sections were prepared. The samples were placed on a slide, and then half of the samples were stained with hematoxylin-eosin and the other half with thioflavin S.
The main goal of preparing sections of the hippocampus and staining them was to investigate possible changes in brain tissue and to quantitatively examine the amyloid fibrils formed in the absence and presence of silibinin and trans-chalcone in each section. Five tissue sections in each group were randomly examined microscopically. Since we observed that silibinin performed better than trans-chalcone in the initial tests, both DG and CA1 regions were checked for the former and DG for the latter.
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