[Histonet] brain tissue sections

[Ryan Dominique Salazar]

Hi,

Please help me with my brain tissue sections, I'm having difficulties in staining them because they disintegrate during H&E staining, resulting into folded and incomplete sections into the slide.

I processed the tissues 2 days after fixation using Leica ASP300S (all new reagents). I have no problem in cutting 5 u thickness during microtomy.
I used adhesive pre-treated slides and Milli-Q water during orientation and fishing out in the floatation bath. I use flattening table as hot plate and heat the freshly cut slides at 62C, for not less than 10 minutes. During staining, I use 3 changes of xylene and abs. ethanol followed by 80% then 70% ethanol, all of them for 3 minutes. I used milli-Q water for hydration and washing during staining. For differentiation and bluing, I use 1% acid alcohol and ammonia water.

Can you also provide me some techniques and difficulties for Perl's prussian blue and amyloid staining.

Thank you very much for your help and your website which changes the lives of ordinary histologist like me.

More power and God bless!

       
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