Protocol for Extraction and Electron Microscopy Visualization of Lipids in Viburnum tinusFruit Using Cryo-Ultramicrotomy

We recently reported that Viburnum tinusfruit generates its metallic blue color using globular lipid inclusions embedded in its epicarpal cell walls. This protocol describes steps to visualize the lipidic nature of the nanostructure using cryo-ultramicrotomy, chloroform extraction, and transmission electron microscopy (TEM) imaging. This method is useful to localize and characterize novel lipidic nanostructures embedded in both plant and animal tissues at the TEM resolution.