Image pro plus version 7.0 imaging software

After 34 months of postmenopausal diet, monkeys were euthanized using sodium pentobarbital (100mg/kg, IV), a method consistent with the recommendations of the panel on euthanasia of the American Veterinary Medical Association. The circulatory system was flushed with lactated Ringers. The heart was removed and perfusion fixed for 1 h at 100 mm/Hg pressure using 10% neutral buffered formalin. The right common iliac artery (RCI) (postmenopausal outcome [POC]) and the coronary arteries were collected for further histopathologic evaluation. The RCI was opened longitudinally, laid flat on cardboard, and a portion was immersion fixed in 10% neutral buffered formalin. Each coronary artery (left anterior descending [LAD], left circumflex artery [LCA], right coronary artery [RCA]) was sectioned in 5 blocks (3 mm each in length) and cut perpendicular to the long axis of the arteries. Five-μm sections were made from each block and stained with Verhoeff-van Gieson’s stain. Each section was captured digitally using a Nikon DS Fi1 camera mounted on an Olympus BH-2 microscope equipped with a mechanical stage. Morphometric measurements were made using Image-Pro Plus version 7.0 imaging software (Media Cybernetics, Inc., Bethesda, MD).

At necropsy, animals were anesthetized deeply with pentobarbital (60 mg/kg), the cardiovascular system was flushed with normal saline, and the heart removed and perfusion fixed with 10% neutral buffered formalin under a pressure of 100 mm Hg. After fixation, five standard serial blocks, 3 mm in length, were taken from each of the left anterior descending (LAD), left circumflex (LCX), and right coronary arteries (RCA). Each of the 15 blocks was embedded in paraffin, and 5-μm sections were cut and stained with Verhoeff-van Gieson’s stain. Images of each section were captured digitally using a Nikon DS Fi1 camera mounted on an Olympus BH-2 microscope equipped with a mechanical stage. Morphometric measurements were made using Image-Pro Plus version 7.0 imaging software (Media Cybernetics Inc., Bethesda, MD). Plaque extent is expressed as the cross-sectional area (mm²) of the area between the internal elastic lamina and the lumen (55 (link),56 ).