Visualization of a Mouse Brain Choroid Plexus Using Scanning Electron Microscopy

Take the choroid plexus from a mouse brain in a specimen basket.

The tissue, located within the brain ventricles, contains a layer of epithelial cells featuring microvilli on their apical surface.

Incubate the tissue in a fixative solution to preserve its cellular architecture.

Wash with a buffer to remove excess fixative and stabilize the pH.

Incubate the tissue with osmium tetroxide that binds to cell membrane lipids, preserving membrane stability, then rinse off excess reagent.

Dehydrate the tissue using increasing alcohol concentrations to prevent tissue distortion during imaging.

Dry the tissue, secure it onto a carbon-coated specimen mount, then coat it with a conductive platinum layer.

Using scanning electron microscopy, focus an electron beam onto the surface. The platinum layer dissipates electrons, preventing charge buildup and minimizing imaging artifacts.

A detector captures the electrons backscattered from the surface, producing an image of the tissue exhibiting epithelial cells with microvilli.

To perform the sample preparation for SEM, transfer the freshly isolated choroid plexus into freshly made fixation solution, and incubate overnight at 4 degrees Celsius. Once done, was