Mitochondrial Ultrastructure Analysis in Astroglia

Transmission electron microscopy (TEM) was used to study the fine structure of the mitochondria and to count the number of mitochondria, as described previously [60 (link)]. Briefly, astroglia derived from 3xTg mice were treated with/without nilotinib (100 nM) for 24 hours. After incubation, cells were washed with PBS and fixed 3 hours in 3% v/v glutaraldehyde in 0.1 M Sorensen’s buffer. Cells were again fixed in 1% osmium tetroxide in 0.1 M Sorensen’s phosphate buffer for two hours, and dehydrated in a 30% to 100% ethanol series and embedded with Epon resin and infiltrated for 4 days. Later, sections were polymerized at 60 °C for 30 hours; Images were collected with Morgagni 268D electron microscope (Philips, Netherlands).