Structure determination of frozen, hydrated, crystalline biological specimens. 1978

K A Taylor

Low temperature transmission electron microscopy can be used to study the structure of biological materials in the hydrated state. Spatial averaging techniques are necessary to overcome the radiation damage problems, and for this reason the techniques described are most applicable to crystalline objects. However, with thin, crystalline biological specimens there are no difficulties with preserving periodicity during the freezing process. Improved specimen preparation methods are described which achieve the production of very thin aqueous films with the hydrated specimen embedded in them. Defocused bright field images of frozen, hydrated protein crystals possess a surprisingly high contrast, presumably due to the difference in density of the protein and the aqueous phase. The techniques described have been used to study the crystal structure of hydrated catalase and the outermost cell wall of Spirillum serpens.

UI MeSH Term Description Entries
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
D002374 Catalase An oxidoreductase that catalyzes the conversion of HYDROGEN PEROXIDE to water and oxygen. It is present in many animal cells. A deficiency of this enzyme results in ACATALASIA. Catalase A,Catalase T,Manganese Catalase,Mn Catalase
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
D003461 Crystallography The branch of science that deals with the geometric description of crystals and their internal arrangement. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Crystallographies
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
D013139 Spirillum A genus of gram-negative, curved and spiral-shaped bacteria found in stagnant, freshwater environments. These organisms are motile by bipolar tufts of flagella having a long wavelength and about one helical turn. Some species of Spirillum cause a form of RAT-BITE FEVER.
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