Jsm 6510lvs scanning electron microscope

The epithelial cells grown on filter inserts were fixed 30 min after the application of osmotic changes with 10% paraformaldehyde and 1% glutaraldehyde/0.1 M HEPES buffer for 2 h. Then the cells were post-fixed with 1% osmium tetroxide/0.1 M HEPES buffer for 60 min, followed by the dehydration with ethanol in a similar manner as described in the previous subsection. Next, the ethanol was replaced with t-butanol, and the epithelia were frozen at −20°C followed by the sublimation of t-butanol. The filters were mounted on aluminum planchets with carbon adhesive, and coated with platinum using an ion coater. The samples were then observed by a JSM-6510LVS scanning electron microscope (JEOL).

Epithelia grown on filters under the ‘Apical’ and ‘Basal’ conditions were fixed with 10% paraformaldehyde and 1% glutaraldehyde/0.1 M HEPES buffer for 2 h, post-fixed with 1% osmium tetroxide/0.1 M HEPES buffer for 60 min, and dehydrated with ethanol in a similar manner as described in Transmission electron microscopy. The ethanol was then replaced with t-butanol, and the epithelia were frozen at −20°C followed by the sublimation of t-butanol. The filters were mounted on aluminum planchets with carbon adhesive and coated with platinum using an ion coater, and the samples were observed by a JSM-6510LVS scanning electron microscope (JEOL).