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Scanning Electron Microscopy

Abstract

High magnification studies of the fenestrated capillary endothelium in the zona fasciculata (ZF) of rat adrenal glands were performed using the objective lens stage of an analytical scanning electron microscope (SEM) equipped with a lanthanum hexaboride emitter (LaB6). Resolution of surface substructure of the luminal membrane obtained with specimens decorated with gold/palladium (Au/Pd) was compared with that observed in others sputter coated with tantalum (Ta). High magnification (50,000x) of the fenestrated endothelium demonstrates that tantalum coating of the cryofractured adrenals improves the substructural detail compared to that seen in Au/Pd decorated specimens.

The procedures used in specimen preparation, metal deposition and secondary electron imaging (SEI) are described. Quality imaging achieved using the objective lens stage is a result of the elimination of the SE-III component derived from backscattered electrons.

Rat adrenals exhibited uniformly patent capillaries. High magnification micrographs of capillary walls were randomly recorded in two morphometric studies of the fenestral content of capillaries in the rat adrenal cortex.

Adrenocorticotropic hormone (ACTH), when administered to rats following dexamethasone (DEX) treatment, significantly reduced the fenestrae/μm2 of endothelial surface and increased the mean size of fenestrae. After hypophysectomy, the number of fenestrae/μm2 declined over 48 h; within 2 h after ACTH was given to rats hypophysectomized 48 hours earlier, the fenestrae/μm2 had increased two-fold. These studies indicate that ACTH plays an important role in modulating fenestral content of the capillary endothelium in the adrenal cortex.

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