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After 3 days of decalcification with formic acid, the tissue was embedded in paraffin, sectioned, and stained with hematoxylin and eosin.
Thereafter, the tissue was embedded in Yazulla embedding medium (30%% egg albumin and 3%% gelatin in distilled water) and 10 μm cryosections were cut in a cryostat (Thermo Scientific Microm HM560).
The tissue was embedded in paraffin, cut in 6 µm-thick sections.
The tissue was embedded in OCT (Sakura Finetek, Torrence, CA), frozen, and cut at 15 µm thickness on a cryostat.
The tissue was embedded by freezing in OCT (Sakura Finetek, Torrance, CA, USA) and 40 µm sections cut in either the sagittal or transverse plane.
For the histological analyses cross sections of the colon were fixed in 4% formalin and the tissue was embedded in paraffin, sliced in sections of 2 µm thickness and stained with haematoxylin-eosin and scored as described [36], [39].
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The hard solid block in which the tissue is embedded assists the microtomy process by preventing shrinkage, distortion, and loss of cellular constituents.
Following cryoprotection in 25% sucrose buffer overnight at room temperature, the tissues were embedded in Tissue Tek O.C.T compound and rapidly frozen in -80°C.
The tissues were embedded in a mixture of Araldite and Epon.
The tissues were embedded in paraffin blocks and sectioned, and the sections were mounted on glass slides.
The tissues were embedded in paraffin blocks, then sliced into 5 μm in thickness and placed onto glass slides.
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