Laboratory Investigations

Calcified Tissue International

, Volume 44, Issue 1, pp 20-24

FT-IR microscopy of endochondral ossification at 20μ spatial resolution

  • Richard MendelsohnAffiliated withDepartment of Chemistry, Rutgers Universit, Newark College of Arts and Science
  • , Alborz HassankhaniAffiliated withDepartment of Chemistry, Rutgers Universit, Newark College of Arts and Science
  • , Edward DiCarloAffiliated withDepartment of Orthopaedic Pathology, Cornell University Medical College and The Hospital for Special Surgery
  • , Adele BoskeyAffiliated withDepartment of Ultrastructural Biochemistry, Cornell University Medical College and the Hospital for Special Surgery

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Summary

A Fourier transform infrared spectrometer has been coupled with an optical microscope to study the distribution and characteristics of the mineral phase in calcifying tissues at 20μ spatial resolution. This represents the first biophysical application of this technique. High quality spectra were obtained in a relatively short scan time (1–2 minutes) from thin longitudinal sections of normal and rachitic rat femurs. Substantial spatial variations in the extent and structure of the mineral phase were observed as a function of spatial position both within and beyond the growth plates, as judged by the phosphate vibrations in the 900–1200 cm−1 spectral region. The current experiments reveal the utility of FT-IR micrscopy in identification of sites where mineralization has occurred. In addition to vibrations from the inorganic components, the Amide I and Amide II motions of the protein constituents are readily observed and may be useful as a probe of protein/mineral interactions.

Key words

FT-IR FT-IR microscopy Endochondral ossification Infrared spectroscopy Hydroxyapatite