Advertisement

Epoxy Resins for Light and Transmission Electron Microscopy

  • Michael John Sumner
Chapter

Abstract

Compared to other epoxy resins, low viscosity Spurr’s resin infiltrates plant tissues more readily and has over the last 45 years become the resin of choice for embedding plant tissues for transmission electron microscopy. This chapter details the six fundamental steps in preparing plant tissues for correlative anatomical studies involving the use of both light and electron microscopy: fixation, dehydration, infiltration, embedding, sectioning and staining.

Keywords

Diamond knife Lead citrate Transmission electron microscope Spurr’s epoxy resin Toluidine blue O Ultramicrotome 

References

  1. 1.
    Glauert AM, Rogers GE, Glauert RH (1956) A new embedding medium for electron microscopy. Nature 178:803CrossRefPubMedGoogle Scholar
  2. 2.
    Richardson KC, Jarett L, Finke EH (1960) Embedding in epoxy resins for ultrathin sectioning in electron microscopy. Stain Technol 35:313–323PubMedGoogle Scholar
  3. 3.
    Luft JH (1961) Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol 9:409–414PubMedCentralCrossRefPubMedGoogle Scholar
  4. 4.
    Hayat MA (2000) Principles and techniques of electron microscopy: biological applications, 4th edn. Cambridge University Press, CambridgeGoogle Scholar
  5. 5.
    Ellis AE (2014) No more Epon 812: this product does not exist today. Microscopy Today 22:50–53CrossRefGoogle Scholar
  6. 6.
    Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43CrossRefPubMedGoogle Scholar
  7. 7.
    Glauert AM, Lewis PR (1998) Biological specimen preparation for transmission electron microscopy. Princeton University Press, PrincetonCrossRefGoogle Scholar
  8. 8.
    Bozzola JJ, Russell LD (1999) Electron microscopy, 2nd edn. Jones and Bartlett, SudburyGoogle Scholar
  9. 9.
    Russell SD, Preston L, Strout G (2014) Principles and techniques of transmission electron microscopy. Samuel Roberts Noble Microscopy Laboratory, University of Oklahoma online resource. http://www.ou.edu/research/electron/bmz5364/. Accessed 13 July 2015
  10. 10.
    O’Brien TP, McCully ME (1981) The study of plant structure: principles and selected methods. Termarcarphi, MelbourneGoogle Scholar
  11. 11.
    Hayat MA (1986) Basic techniques for transmission electron microscopy. Academic, LondonGoogle Scholar
  12. 12.
    Chang RFE, Klomparens KL (eds) (1988) Artifacts in biological electron microscopy. ­Plenum, New YorkGoogle Scholar
  13. 13.
    Gephart P, Murray RGE, Costilow RN, Wood WA et al (1981) Manual of methods for general bacteriology (Amer Soc Microbiol). ASM Press, Washington, D.C.Google Scholar
  14. 14.
    Fisher DB (1968) Protein staining of ribboned epon sections for light microscopy. Histochemie 16:92–96CrossRefPubMedGoogle Scholar
  15. 15.
    Fulcher RG, Wong SI (1980) Inside cereals—a fluorescence microchemical view. In: Inglett GE, Munch L (eds) Cereals for food and beverages—recent progress in cereal chemistry. Academic, New York, pp 1–26Google Scholar
  16. 16.
    Venable JH, Coggeshall R (1965) A simplified lead citrate stain for use in electron microscopy. J Cell Biol 25:407–408PubMedCentralCrossRefPubMedGoogle Scholar
  17. 17.
    Reynolds ES (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol 17:208–212PubMedCentralCrossRefPubMedGoogle Scholar
  18. 18.
    Hanaichi T, Sato T, Hoshin M, Mizuno N (1986) A stable lead stain by modification of Sato’s method. Proc XIth Int Cong Electron Microscopy. Kyoto, pp 2181–2182 (English corrected)Google Scholar
  19. 19.
    Thiery JP (1967) Mise en évidence des polysaccharides sur coupes fines en microscopie électronique. J Microsc 6:987–1017Google Scholar
  20. 20.
    Hepler PK (1981) The structure of the endoplasmic reticulum revealed by osmium tetroxide-potassium ferricyanide staining. Eur J Cell Biol 26:102–110PubMedGoogle Scholar
  21. 21.
    Hayat MA (1981) Fixation for electron microscopy. Academic, New YorkGoogle Scholar
  22. 22.
    Jensen WA (1962) Botanical histochemistry: principles and practice. Freeman, San FranciscoGoogle Scholar
  23. 23.
    Pappas PW (1971) The use of a chrome alum-gelatin (subbing) solution as a general adhesive for paraffin sections. Stain Technol 46:121–124PubMedGoogle Scholar
  24. 24.
    O’Brien TP, Feder N, McCully ME (1964) Polychromatic staining of plant cell walls by toluidine blue O. Protoplasma 59:368–373CrossRefGoogle Scholar
  25. 25.
    Hale AJ (1957) The histochemistry of polysaccharides. Int Rev Cytol 6:193–263CrossRefGoogle Scholar
  26. 26.
    Roland JC (1978) General preparation and staining of thin sections. In: Hall JL (ed) Electron microscopy and cytochemistry of plant cells. Elsevier/North-Holland Biomedical Press, Amsterdam, pp 1–62Google Scholar
  27. 27.
    Lane BP, Europa DL (1965) Differential staining of ultrathin sections of epon-embedded tissues for light microscopy. J Histochem Cytochem 13:579–582CrossRefPubMedGoogle Scholar
  28. 28.
    Hughes J, McCully ME (1975) The use of an optical brightener in the study of plant structure. Stain Technol 50:319–329PubMedGoogle Scholar
  29. 29.
    Schnepf E, Deichgraber G, Herth W (1982) Development of cell wall appendages in Acanthosphaera zachariasi (Chlorococcales): kinetics, site of cellulose synthesis and microfibril assembly, and barb formation. Protoplasma 110:203–214CrossRefGoogle Scholar
  30. 30.
    Wood PJ, Fulcher RG (1978) Interactions of some dyes with cereal beta glucans. Cereal Chem 55:952–966Google Scholar
  31. 31.
    Herth W, Schnepf E (1980) The fluorochrome, calcofluor white, binds oriented structural polysaccharide fibrils. Protoplasma 105:129–133CrossRefGoogle Scholar
  32. 32.
    Smith MM, McCully ME (1978) A critical evaluation of the specificity of aniline blue induced fluorescence. Protoplasma 95:229–254CrossRefGoogle Scholar
  33. 33.
    White DL, Mazurkeiwicz JE, Barrnett RJ (1979) A chemical mechanism for tissue staining by osmium tetroxide-ferrocyanide mixtures. J Histochem Cytochem 27:1084–1091CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  1. 1.Department of Biological SciencesUniversity of ManitobaWinnipegCanada

Personalised recommendations