Araldite 502

One-millimeter cubes of each liver specimen were fixed for a minimum of 24 hours in 2% glutaraldehyde, 2% paraformaldehyde in 0.2 mol/L sodium cacodylate buffer, pH 7.3. The tissues were then washed in 0.2 mol/L sodium cacodylate buffer overnight and placed in a working solution of 2.5% potassium dichromate and 0.5% osmium tetroxide in reverse osmosis/deionized water for 6 hours at room temperature (RT), followed by washing in running water for 1 hour. The tissues were then stored overnight in 30% ethanol at 4°C, followed by dehydration in acetones (70%, 100% acetones for 30 min each). After dehydration, the tissues were infiltrated in 70% resin (15 mL DDSA, 5 mL LX-112, 5 mL Araldite 502, and 0.5 mL DMP-30 [Ladd Research Industries, Williston, VT]) in acetone for 1 hour. Tissue was then infiltrated in 2 changes of 100% resin, 1 at 3 hours and 1 overnight. Embedding was performed in “00” Beem capsule with 100% resin and polymerized at 60°C for a minimum of 48 hours. Sections of 1 µm thickness were cut using a Leica EM UC6 microtome (Leica Microsystems) with an ultradiamond knife (Diatome, Hatfield, PA). The sections were collected onto Superfrost Plus Slides and dried overnight on a 40°C slide warmer.