Protocol 25 Electron microscopy

Two additional rats were exposed to 0, 50, 150 or 1500 mg kg-1 BW-1 APAP and deeply anesthetized with Pentobarbital at 6 h after dosing. The vessels superior and inferior of the liver were clamped. The posterior vena cava was canulated and the portal vessels severed. The retrograde perfusion was via the posterior vena cave first with RPMI media at 37°C to clean the liver followed by cold 3% glutaraldehyde (in 0.1 M sodium cacodylate buffer pH 7.2) for 30 min. The left lateral lobe was minced into 1-mm cubes and incubated overnight in 3% glutaraldehyde solution, and post fixed in OsO4. Centrilobular areas were identified on 0.5-^m-thick sections stained with toluidine blue. Thin sections, approximately 80 nm, were examined on a Philips EM 400 electron microscope after staining with uranyl acetate and lead citrate.

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