The ultrastructure of calcified tissues: Methods and technical problems

Part of the Electron Microscopy in Biology and Medicine book series (EMBM, volume 7)


The organic/inorganic interface within calcified tissue matrices is both intriguing and problematic. It is intriguing because these specialized matrices initiate and control mineral deposition while serving as a means for storage and retrieval of various essential ions. Also, these matrices, at the cellular level, provide protection against the lethal stores of concentrated calcium phosphate in the form of microcrystalline apatite, Ca10(PO4)6(OH)2. The problematic aspect of calcified tissues is due to the complexity of this physico-chemical organic/inorganic interface, i.e., what is the exact nature of this interface, what chemical factors are involved with the initiation and control of mineral deposition, what factors govern crystal size and orientation, and what are the different phases of calcium phosphate other than apatite.


Collagen Fibril Scanning Transmission Electron Microscopic Matrix Vesicle Apatite Crystal Calcify Cartilage 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Arsenault AL, Hunziker EB: Electron microscopic analysis of mineral deposits in the calcifying epiphyseal growth plate. Calcif Tissue Int 42: 119–126, 1988.PubMedCrossRefGoogle Scholar
  2. 2.
    Arsenault AL, Ottensmeyer FP, Heath IB: An electron microscopic and spectroscopic study of murine epiphyseal cartilage: Analysis of fine structure and matrix vesicles preserved by slam freezing and freeze substitution. J Ultrastruct Mol Struct Res 98: 32–47, 1988.PubMedCrossRefGoogle Scholar
  3. 3.
    Moor H, Berlin G: The influence of high pressure freezing on mammalian nerve tissue. Cell Tissue Res 209: 201–216, 1980.PubMedCrossRefGoogle Scholar
  4. 4.
    Hunziker EB, Herrmann W, Schenk RK, Mueller M, Moor H: Cartilage ultrastructure after high pressure freezing, freeze substitution, and low temperature embedding. I. Chondrocyte ultrastructure — implications for theories of mineralization and vascular invasion. J Cell Biol 98: 267–276, 1984.PubMedCrossRefGoogle Scholar
  5. 5.
    Arsenault AL, Spritzer E, Simon GT: Improved preservation of cartilage extracellular matrix by freeze dried embedding. J Microsc 145: 357–360, 1987.PubMedGoogle Scholar
  6. 6.
    Ottensmeyer FP, Andrew JW: High-resolution microanalysis of biological specimens by electron energy loss spectroscopy and electron spectroscopic imaging. J Ultrastruct Res 72: 336–348, 1980.PubMedCrossRefGoogle Scholar
  7. 7.
    Arsenault AL, Ottensmeyer FP: Quantitative spatial distributions of calcium, phosphorus, and sulfur in calcifying epiphysis by high resolution electron spectroscopic imaging. Proc Natl Acad Sci USA 80: 1322–1326, 1983.PubMedCrossRefGoogle Scholar
  8. 8.
    Grove CA, Judd G, Ansell GS: Determination of hydroxyapatite crystallite size in human dental enamel by darkfield electron microscopy. J Dent Res 51: 22–29, 1972.PubMedCrossRefGoogle Scholar
  9. 9.
    Jackson SA, Cartwright AG, Lewis D: The morphology of bone mineral crystals. Calcif Tissue Res 25: 217–222, 1978.PubMedCrossRefGoogle Scholar
  10. 10.
    Arsenault AL, Grynpas MD: Crystals in calcified epiphyseal cartilage and cortical bone of the rat. Calcif Tissue Int 43: 219–225, 1988.PubMedCrossRefGoogle Scholar
  11. 11.
    Arsenault AL: A comparative electron microscopic study of apatite crystals in collagen fibrils of rat bone, dentin and turkey leg tendons. Bone and Mineral 6: 165–177, 1989.PubMedCrossRefGoogle Scholar
  12. 12.
    Arsenault AL: Crystal-collagen relationships in the calcified turkey leg tendon visualized by dark field electron microscopy. Calcif Tissue Int 43: 202–212, 1988.PubMedCrossRefGoogle Scholar
  13. 13.
    Meek KM, Chapman JA, Hardcastle RA: The staining pattern of collagen fibrils. Improved correlation with sequence data. J Biol Chem 254: 10710–10714, 1979.PubMedGoogle Scholar
  14. 14.
    Engström A: Apatite-collagen organization in calcified tendon. Exp Cell Res 43: 241–245, 1966.PubMedCrossRefGoogle Scholar
  15. 15.
    Berthet-Columinas C, Miller A, White SW: Structural study of the calcifying collagen in turkey leg tendons. J Mol Biol 134: 431–445, 1979.CrossRefGoogle Scholar
  16. 16.
    White SW, Hulmes DJS, Miller A, Timmins PA: Collagen-mineral axial relationship in calcified turkey leg tendon by x-ray and neutron diffraction. Nature 266: 421–425, 1977.PubMedCrossRefGoogle Scholar
  17. 17.
    Robinson RA, Watson ML. Collagen-crystal relationships in bone as seen in the electron microscope. Anat Ree 114: 383–410, 1952.CrossRefGoogle Scholar
  18. 18.
    Steve-Bocciarelli D. Morphology of crystallites in bone. Calcif Tissue Res 25: 217–222, 1970.Google Scholar
  19. 19.
    Landis WJ, Glimcher MJ. Electron diffraction and electron probe microanalysis of the mineral phase of bone tissue prepared by anhydrous techniques. J Ultrastruct Res 63: 188–223, 1978.PubMedCrossRefGoogle Scholar
  20. 20.
    Weiner S, Price PA. Disaggregation of bone into crystals. Calcif Tissue Int 39: 365 - 375, 1986.PubMedCrossRefGoogle Scholar
  21. 21.
    Arsenault AL: Microvascular organization at the epiphyseal-metaphyseal junction of growing rats. J Bone Mineral Res 2: 143–149, 1987.CrossRefGoogle Scholar
  22. 22.
    Arsenault, AL, Hunter WL, Hodsman AB: Microvascular organization at the epiphyseal-metaphyseal junction of normal and rachitic rats. In: Behavior of the Growth Plate. HK Uhthoff, JJ Wiley (eds), New York: Raven Press p 309–316, 1988.Google Scholar
  23. 23.
    Zinkernagel R, Riede UN, Schenk RK: Ultrastrukturelle Untersuchungen der juxtaepiphysaren Kapillären nach Perfusionsfixation. Experientia 28: 1205–1206, 1972.PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1990

Authors and Affiliations

  1. 1.Electron Microscopic Facility Faculty of Health SciencesMcMaster UniversityOntarioCanada

Personalised recommendations