Seeking H&E stains procedure for Ground Plastic section

From:

Bernard Ian R SSgt 59 CRES/MSROP

Hello again, to all fellow, Histo Plastic Researchers,

My Pathologist is adamant in me, finding a H&E procedure that will work, to stain some plastic processed (Methyl Methacrylate) pig & rabbit eye implants. I mentioned to him about other stains out there, that work similarly to the H&E and stains better. However, he insists that the H&E will give him a familiar and better contrast.

The one method I have, worked (acceptable to him) for another plastic procedure( A Butyl Methyl Methacrylate), but not this one. The stain came out to light. I even restained the slides following the same procedure again. And it still turned out light.

My adopted Plastic H&E Method abbreviated to key reagents:

1. I surface etched the plastic slides before staining to allow for stain penetration in 1 % Formic Acid ( 2 mins), 50/50 ETOH and rinse in Distilled H20.

2. Stain 6 min in Harris Hematoxylin

3. Differentiate in 0.5 % Acid Alcohol ( HCL in 70 % Alcohol)

4. Blue in Ammonia water (12 drops Ammonium Hydroxide in 250 mls Distilled water)

5. Stain 1 min in Eosin Y solution

6. Dehydrate to Xylene substitute.

These sections are less than 100 microns thick, no less than 40 microns

I need help with a good procedure.

Thanks in advance

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