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Journal of Materials Science

, Volume 44, Issue 14, pp 3813–3818 | Cite as

High-throughput SEM preparation of proteinaceaous extracellular matrix

  • Maja WiegemannEmail author
  • Frank Lehmann
Article

Abstract

Adhesives of aquatic organisms are of high scientific interest with a view to biomimicry. The visualization of their fine-structure, however, is difficult due to the fragility of the extracellular matrix (ECM). Care must be taken in its preparation for high-resolution SEM. Relating matrix structures to substrate properties demands a high throughput of samples for reliable comparisons. In order to acquaint ourselves with a suitable method we found an easy way to manage the critical steps of preparation for SEM. We show that thin layers of proteinaceous matrix can be satisfactorily prepared by combining freeze-drying and sputtering in a conventional sputter coater after one-step fixation (2.5% glutaraldehyde). The results were superior to CPD.

Keywords

PDMS Chemical Fixation Glutaraldehyde Fixation Fast Freezing Substrate Piece 

Notes

Acknowledgement

We thank Prof. M. Kern—head of the Clinic of Prosthodontics, Propaedeutics and Dental Materials at the University Kiel, Germany—for supporting this study. The publication is part of a project funded by the German Research Foundation—Deutsche Forschungsgemeinschaft (DFG).

References

  1. 1.
    Graham LD (2005) Encycl Biomater Biomed Eng. doi: https://doi.org/10.1081/E-EBBE-120041680
  2. 2.
    Páez J, Jorge-Herrero E (2006) In: Edwards JV, Buschle-Diller G, Coheen SC (eds) Modified fibers with medical and specialty applications, 1st edn. Springer, NetherlandsGoogle Scholar
  3. 3.
    Smith AM, Callow JA (2006) Biological adhesives, 1st edn. Springer, Berlin, HeidelbergCrossRefGoogle Scholar
  4. 4.
    Stevens MJ, Steren RE, Hlady V, Stewart RJ (2007) Langmuir 23(9):5045CrossRefGoogle Scholar
  5. 5.
    Hennebert E, Viville P, Lazzaroni R, Flammang P (2008) J Struct Biol 164(1):108CrossRefGoogle Scholar
  6. 6.
    Sullan RMA, Gunari N, Tanur AE, Yuri C, Dickinson GH, Orihuela B, Rittschof D, Walker GC (2009) Biofouling 25(3):263CrossRefGoogle Scholar
  7. 7.
    Walker G (1999) In: Harrison FW, Humes AG (eds) Microscopic anatomy of invertebrates, vol 9, 1st edn. Wiley, New York, ChichesterGoogle Scholar
  8. 8.
    Wiegemann M, Watermann B (2003) J Adhes Sci Technol 17:1957CrossRefGoogle Scholar
  9. 9.
    Walker G (1987) Crit Rep Appl Chem 16:112Google Scholar
  10. 10.
    Mitchell R, Kirchmann D (1984) In: Costlow JD, Tipper RC (eds) Marine biodeterioration: an interdisciplinary study. Naval Institute Press, Annapolis, MDGoogle Scholar
  11. 11.
    Callow JA, Osborne MP, Callow ME, Baker F, Donald AM (2003) Colloids Surf B Biointerfaces 27:315CrossRefGoogle Scholar
  12. 12.
    Danilatos GD (1988) Adv Electron Electron Phys 71:109CrossRefGoogle Scholar
  13. 13.
    Bergmans L, Moisiadis P, Van Meerbeek B, Quirynen M, Lambrechts P (2005) Int Endod J 38:775CrossRefGoogle Scholar
  14. 14.
    Muscariello L, Rosso F, Marino G, Giordano A, Barbarisi M, Cafiero G, Barbarisi A (2005) J Cell Physiol 205:328–334CrossRefGoogle Scholar
  15. 15.
    Kirk SE, Skepper JN, Donald AM (2009) J Microsc 233:205–224CrossRefGoogle Scholar
  16. 16.
    Walther P (2007) In: Schatten H, Pawley J (eds) Biological low-voltage scanning electron microscopy, 1st edn. Springer, BerlinGoogle Scholar
  17. 17.
    Stokes DJ, Mugnier JY, Clarke CJ (2004) J Microsc 213:198–204CrossRefGoogle Scholar
  18. 18.
    Peters T, Ashley CA (1967) J Cell Biol 33:53–60CrossRefGoogle Scholar
  19. 19.
    Geyer G (1977) Elektronenmikroskopische Histochemie Teil 1: Nachweis- und Kontrastierungsmethoden für Kohlenhydrate, Proteine und Aminosäuren, Nucleinsäuren, Lipide und Mineralstoffe. Gustav Fischer Verlag, Stuttgart, New YorkGoogle Scholar
  20. 20.
    Pawley JB (2007) In: Schatten H, Pawley J (eds) Biological low-voltage scanning electron microscopy, 1st edn. Springer, BerlinGoogle Scholar
  21. 21.
    Nei T (1973) J Microsc 99:227CrossRefGoogle Scholar
  22. 22.
    Porter KR, Anderson KL (1982) Eur J Cell Biol 29(1):83Google Scholar
  23. 23.
    Lindroth M, Frederiksson BA, Bell PB (1991) J Microsc 161:229CrossRefGoogle Scholar
  24. 24.
    Lubig R, Kusch P, Röper K, Zahn H (1981) Monatsh Chem 112(11):1313CrossRefGoogle Scholar
  25. 25.
    Erlandsen SL (2007) In: Schatten H, Pawley J (eds) Biological low-voltage scanning electron microscopy, 1st edn. Springer, BerlinGoogle Scholar
  26. 26.
    Bancroft JD, Stevens A (1975) Histopathological stains and their diagnostic uses. Churchill Livingstone, EdinburghGoogle Scholar
  27. 27.
    Billings-Gagliardi S, Pockwinse SM, Schneider GB (1978) Am J Anat 152(3):383CrossRefGoogle Scholar
  28. 28.
    Lindroth M, Bell PB, Frederiksson BA (1988) J Microsc 151:103CrossRefGoogle Scholar
  29. 29.
    Nordestgaard BG, Rostgaard J (1985) J Microsc 137:189CrossRefGoogle Scholar
  30. 30.
    Boyde A, Franc F (1981) J Microsc 122(1):75CrossRefGoogle Scholar
  31. 31.
    Warley A, Skepper JN (2000) J Microsc 198:116CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.BAH, Alfred Wegener InstituteHelgolandGermany
  2. 2.Department of Prosthodontics, Propaedeutics and Dental MaterialsChristian-Albrechts UniversityKielGermany

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