Stereological method for objectively quantifying myelin sheaths in the rat hippocampus

来源 :Neural Regeneration Research | 被引量 : 0次 | 上传用户:GoldGhost
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In the present study,tissue blocks were randomly sampled from the entire hippocampus of 6-week-old Long-Evans rats.Isotropic,uniform and random sections,60 nm thick,were prepared by isector.Fifteen fields of view were randomly selected for each section and photographed using a transmission electron microscope.The mean internal and external diameters of the myelin sheaths were obtained by measuring the longest profile diameter perpendicular to its longest axis.The inner and outer perimeters of the myelin sheaths were estimated using the equidistant parallel test lines.The thickness of the myelin sheaths was estimated by direct orthogonal measurements in uniform,random locations.These stereological methods should permit an unbiased quantitative assessment of changes in the myelin sheaths of myelinated fibers in the hippocam-pus. In the present study, tissue blocks were optionally sampled from the entire hippocampus of 6-week-old Long-Evans rats. Isotropic, uniform and random sections, 60 nm thick, were prepared by isector. Fifteen fields of view were randomly selected for each section and photographed using a transmission electron microscope. The mean internal and external diameters of the myelin sheaths were obtained by measuring the longest profile diameter perpendicular to its longest axis. The inner and outer perimeters of the myelin sheaths were estimated using the equidistant parallel test lines The thickness of the myelin sheaths was estimated by direct orthogonal measurements in uniform, random locations. These stereological methods should permit an unbiased quantitative assessment of changes in the myelin sheaths of myelinated fibers in the hippocam-pus.
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