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This grid is then placed between two low mass copper platelets where it serves as a spacer, and hence limits the thickness of the aqueous layer to be frozen (Miiller and Moor, 1984a). Adequately manufactured copper platelets are now commercially available (Balzers Union, Balzers FL). The frozen sandwiches are fractured under liquid nitrogen for further processing by freeze-substitution or in a freeze-fracture apparatus for freeze-drying or replication. Sandwiches always split between either side of the spacer grid and a copper platelet.
Ultra-thin sections are cut in ultramicrotomes with either glass or diamond knives. A detailed description of the principle and function of the microtomes has been given by Reid (1974) and Sitte and Neumann (1983). E. Post-staining Thin sections (c100 nm) generally do not possess enough contrast to show fine structural details. However, the contrast of biological samples can be enhanced by post-staining of the sections with solutions of heavy metal salts like uranyl or lead salts. 0 (Reynolds, 1963).
15,469-479. Zierold, K. Cryotechniques in Biological Electron Microscopy (R. A. Steinbrecht and K. Zierold, eds), pp. 132-148. Springer-Verlag, Berlin. etching. Metal Shadowing. Negative Staining and Ultrathin Sectioning UWE B. SLEYTR. PAUL MESSNER and DIETMAR PUM Zentrum f i r Ultrastrukturforschung und Ludwig Boltzmann-lnstitut fur Ultrastrukturforschung. Universitat fur Bodenkultur. A-I180 Wien. Austria I . Introduction . . . . 11. Freeze-etching . . . A . General principles . . . B.