Biomaterial Database

Protocol for Extraction and Electron Microscopy Visualization of Lipids in Viburnum tinus Fruit Using Cryo-Ultramicrotomy. 2020

Miranda Sinnott-Armstrong, and Silvia Vignolini, and Yu Ogawa

Department of Ecology and Evolutionary Biology, Yale University, New Haven, CT 06511, USA.

We recently reported that Viburnum tinus fruit 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. For complete details on the use and execution of this protocol, please refer to Middleton et al. (2020).

UI	MeSH Term	Description	Entries
D008055	Lipids	A generic term for fats and lipoids, the alcohol-ether-soluble constituents of protoplasm, which are insoluble in water. They comprise the fats, fatty oils, essential oils, waxes, phospholipids, glycolipids, sulfolipids, aminolipids, chromolipids (lipochromes), and fatty acids. (Grant & Hackh's Chemical Dictionary, 5th ed)	Lipid
D008854	Microscopy, Electron	Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.	Electron Microscopy
D002473	Cell Wall	The outermost layer of a cell in most PLANTS; BACTERIA; FUNGI; and ALGAE. The cell wall is usually a rigid structure that lies external to the CELL MEMBRANE, and provides a protective barrier against physical or chemical agents.	Cell Walls,Wall, Cell,Walls, Cell
D005638	Fruit	The fleshy or dry ripened ovary of a plant, enclosing the seed or seeds.	Berries,Legume Pod,Plant Aril,Plant Capsule,Aril, Plant,Arils, Plant,Berry,Capsule, Plant,Capsules, Plant,Fruits,Legume Pods,Plant Arils,Plant Capsules,Pod, Legume,Pods, Legume
D016616	Cryoultramicrotomy	The technique of using a cryostat or freezing microtome, in which the temperature is regulated to -20 degrees Celsius, to cut ultrathin frozen sections for microscopic (usually, electron microscopic) examination.	Cryosectioning,Freeze Sectioning,Sectioning, Freeze
D046529	Microscopy, Electron, Transmission	Electron microscopy in which the ELECTRONS or their reaction products that pass down through the specimen are imaged below the plane of the specimen.	Electron Diffraction Microscopy,Electron Microscopy, Transmission,Microscopy, Electron Diffraction,Transmission Electron Microscopy,Diffraction Microscopy, Electron,Microscopy, Transmission Electron
D049329	Nanostructures	Materials which have structured components with at least one dimension in the range of 1 to 100 nanometers. These include NANOCOMPOSITES; NANOPARTICLES; NANOTUBES; and NANOWIRES.	Nanomaterials,Nanostructured Materials,Material, Nanostructured,Materials, Nanostructured,Nanomaterial,Nanostructure,Nanostructured Material
D020285	Cryoelectron Microscopy	Electron microscopy involving rapid freezing of the samples. The imaging of frozen-hydrated molecules and organelles permits the best possible resolution closest to the living state, free of chemical fixatives or stains.	Electron Cryomicroscopy,Cryo-electron Microscopy,Cryo electron Microscopy,Cryo-electron Microscopies,Cryoelectron Microscopies,Cryomicroscopies, Electron,Cryomicroscopy, Electron,Electron Cryomicroscopies,Microscopies, Cryo-electron,Microscopies, Cryoelectron,Microscopy, Cryo-electron,Microscopy, Cryoelectron
D027925	Viburnum	A plant genus in the family CAPRIFOLIACEAE. The common name derives from its traditional use for menstrual cramps. It is a source of viburnine, valerianic acid, vibsanin, and ursolic acid. Note that true cranberry is VACCINIUM MACROCARPON.	Cramp Bark,Cranberry, Highbush,Highbush Cranberry,Bark, Cramp,Barks, Cramp,Cramp Barks,Cranberries, Highbush,Highbush Cranberries,Viburnums