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High Resolution Metal Replication of Freeze-Dried Specimens

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Abstract

Freeze-drying followed by heavy-metal shadowing is an established and straightforward approach to routine structural studies of dehydrated biological specimens (Nermut and Frank 1971; Kistler et al. 1977; Gross et al. 1985). Very thin specimens such as isolated membranes or isolated macromolecules can be directly adsorbed on carbon-coated grids. After rapid freezing, the grids are transferred into suitable high-vacuum equipment for freeze-drying and shadowing. After stabilizing the heavy-metal coat with carbon (C-backing), the specimens are warmed to room temperature and transferred via atmospheric conditions into the transmission electron microscope (TEM). In other words, the entire sandwich consisting of C-supporting film, biological specimen, metal coat and C-backing film is examined in the microscope.

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References

  1. Amrein M (1985) Der Einfluß von Restwasser nach Gefriertrocknen auf die Strukturerhaltung eines zweidimensionalen Proteingitters — untersucht mittels Schwermetallbeschattung bei extrem tiefer Objekttemperatur. Diplomarbeit, Eidgen Tech Hochschule Zürich.

    Google Scholar 

  2. Baumeister W, Bart M, Hegerl R, Guckenberger R, Hahn M, Saxton WO (1986) Three-dimensional structure of the regular surface layer (HPI) of Deinococcus radiodurans. J Mol Biol 187:241–253.

    Article  PubMed  CAS  Google Scholar 

  3. Crowther RA, Klug A (1975) Structural analysis of macromolecular assemblies by image reconstruction from electron micrographs. Annu Rev Biochem 44:161–182.

    Article  PubMed  CAS  Google Scholar 

  4. Frank J (1980) The role of correlation methods in computer image processing. In: Hawkes PW (ed) Topics in physics, vol 13. Springer, Berlin Heidelberg New York, pp 187–222.

    Google Scholar 

  5. Gross H, Kuebler O, Bas E, Moor H (1978) Decoration of specific sites on freeze-fractured membranes. J Cell Biol 79:646–656.

    Article  PubMed  CAS  Google Scholar 

  6. Gross H, Müller T, Wildhaber I, Winkler H, Moor H (1984) Freeze-fracturing and replication at-260°C. In: Proc 42nd Annu EMSA Meet, Detroit MI. San Francisco Press, San Francisco, pp 12–15.

    Google Scholar 

  7. Gross H, Müller T, Wildhaber I, Winkler H (1985) High resolution metal replication, quantified by image processing of periodic test specimens. Ultramicroscopy 16:287–304.

    Article  Google Scholar 

  8. Guckenberger R (1985) Surface reliefs derived from heavy-metal-shadowed specimens — fourier space techniques applied to periodic objects. Ultramicroscopy 16:357–370.

    Article  Google Scholar 

  9. Kistler J, Aebi Y, Kellenberger E (1977) Freeze-drying and shadowing a two-dimensional periodic specimen. J Ultrastruct Res 59:76–85.

    Article  PubMed  CAS  Google Scholar 

  10. Kühlbrandt W (1984) Three-dimensional structure of the light-harvesting chlorophyll a/b-prote-in complex. Nature (London) 307:478–480.

    Article  Google Scholar 

  11. Kühlbrandt W, Thaler Th, Wehrli E (1983) The structure of membrane crystals of the light-harvesting chlorophyll a/b protein complex. J Cell Biol 96:1414–1424.

    Article  PubMed  Google Scholar 

  12. Maissel LI, Glang R (eds) (1970) Handbook of thin film technology. McGraw-Hill, New York

    Google Scholar 

  13. Nermut MV, Frank H (1971) Fine structure of influenza A2 (Singapore) as revealed by negative staining, freeze-drying and freeze-etching. J Gen Virol 10:37–51.

    Article  PubMed  CAS  Google Scholar 

  14. Rachel R, Jakubowski U, Baumeister W (1986) Electron microscopy of unstained, freeze-dried macromolecular assemblies. J Microsc (Oxford) 141:179–191.

    Article  Google Scholar 

  15. Robards AW, Sleytr UB (1985) Low temperature methods in biological electron microscopy. In: Glauert AM (ed) Practical methods in electron microscopy, vol 10. Elsevier, Amsterdam.

    Google Scholar 

  16. Saxton WO, Baumeister W (1982) The correlation averaging of a regularly arranged bacterial cell envelope protein. J Microsc (Oxford) 127:127–138.

    Article  CAS  Google Scholar 

  17. Saxton WO, Pitt TJ, Horner M (1979) Digital image processing: the Semper system. Ultramicroscopy 4:343–354.

    Article  Google Scholar 

  18. Sleytr UB, Messner P (1983) Crystalline surface layers on bacteria. Annu Rev Microbiol 37:311–339.

    Article  PubMed  CAS  Google Scholar 

  19. Smith PR, Kistler J (1977) Surface reliefs computed from electron micrographs of heavy metal shadowed specimens. J Ultrastruct Res 61:124–133.

    Article  PubMed  CAS  Google Scholar 

  20. Unwin PNT, Henderson R (1975) Molecular structure determination by electron microscopy of unstained crystalline specimens. J Mol Biol 94:425–440.

    Article  PubMed  CAS  Google Scholar 

  21. Venables JA, Spiller GDT, Hanbücken M (1984) Nucleation and growth of thin films. Rep Prog Phys 47:399–459.

    Article  Google Scholar 

  22. Wildhaber I, Gross H, Moor H (1982) The control of freeze-drying with deuterium oxide. J Ultrastruct Res 80:367–373.

    Article  PubMed  CAS  Google Scholar 

  23. Wildhaber I, Hegerl R, Barth M, Gross H, Baumeister W (1986) Three dimensional reconstruction of a freeze-dried and metal shadowed bacterial surface layer. Ultramicroscopy 19:57–68.

    Article  Google Scholar 

  24. Winkler H (1986) Computerunterstützte Interpretation elektronenmikroskopischer Bilder von dekorierten und beschatteten biologischen Oberflächen. PhD, Diss (Nr 8186) Eidgen Tech Hochschule Zürich.

    Google Scholar 

  25. Winkler H, Wildhaber I, Gross H (1985) Decoration effects on the surface of a regular layer protein. Ultramicroscopy 16:331–339.

    Article  CAS  Google Scholar 

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© 1987 Springer-Verlag Berlin Heidelberg

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Gross, H. (1987). High Resolution Metal Replication of Freeze-Dried Specimens. In: Steinbrecht, R.A., Zierold, K. (eds) Cryotechniques in Biological Electron Microscopy. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-72815-0_10

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  • DOI: https://doi.org/10.1007/978-3-642-72815-0_10

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-72817-4

  • Online ISBN: 978-3-642-72815-0

  • eBook Packages: Springer Book Archive

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