Low resolution scanning electron microscopy of cerebellar neurons and neuroglial cells of the granular layer

Teleost fishes, Arius Spixii and Salmo trout and adult Swiss albino mice have been processed with the freeze-fracture technique for SEM to explore the inner cytoplasmic and nuclear surface details of neurons and neuroglial cells. The specimens were fixed by vascular perfusion with Karnovsky fixative and 2-3 mm thick cerebellar slices were subsequently fixed by immersion in the same fixative. They were postfixed in osmium tetroxide, dehydrated in ethanol, frozen in Freon 22, cooled by liquid nitrogen and fractured. After thawing in ethanol, they were critically point dried, coated with gold-palladium and viewed by SEM. The surface features of perikaryon were examined at low resolution and magnifications. The image of endoplasmic reticulum, GERL complex and chromatin were described in fractured cerebellar neurons (granule and Golgi cells). The fractured protoplasmic astrocytes displayed a characteristic glass surface appearance of cytoplasmic body and processes, which facilitated their recognition at the neuropile and perivascular region. The oligodendrocytes appeared as fusiform cells depicting a thin rim of perinuclear cytoplasm. The surface view of endoplasmic reticulum was well studied at the nuclear poles. Fine cytoplasmic beaded canaliculi appeared connecting the outer surface of nuclear envelope with the plasma membrane inner surface. The nucleus exhibited well developed peripheral heterochromatin masses forming anastomotic bands separated by vacuolar spaces. The SEM nerve and neuroglial cell fractographs were compared with similar images obtained by conventional transmission electron microscopy and freeze etching technique.