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Related Experiment Videos

Optimal methods for processing mineralized tissues for Fourier transform infrared microspectroscopy.

S Aparicio1, S B Doty, N P Camacho

  • 1Mineralized Tissue Laboratory, The Hospital for Special Surgery, 535 E 70th St., New York, NY 10021, USA.

Calcified Tissue International
|June 11, 2002
PubMed
Summary

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Fourier transform infrared microspectroscopy (FTIRM) and imaging (FTIRI) analyze bone properties. Non-aqueous fixatives and specific embedding media like LR White, Spurr, Araldite, and PMMA minimize spectral interference for accurate bone analysis.

Area of Science:

  • Biophysics
  • Materials Science
  • Biochemistry

Background:

  • Fourier transform infrared microspectroscopy (FTIRM) and infrared imaging (FTIRI) are valuable for analyzing bone mineral and matrix properties in health and disease.
  • These techniques preserve spatial arrangement, enabling quantitative data on bone mineral (hydroxyapatite) and matrix composition at specific locations.
  • Optimal fixation and embedding are crucial as most media spectrally overlap with mineralized tissues, potentially affecting spectral analysis.

Purpose of the Study:

  • To evaluate the impact of various fixatives and embedding media on bone spectral properties analyzed by FTIRM and FTIRI.
  • To identify fixation and embedding protocols that minimize spectral interference for accurate bone analysis.

Main Methods:

  • Mouse calvaria were subjected to seven different fixatives and six embedding media.
Keywords:
Non-programmatic

Related Experiment Videos

  • Fourier transform infrared microspectroscopy (FTIRM) and infrared imaging (FTIRI) were used to assess spectral changes.
  • Comparisons were made to unprocessed ground bone and cryosections of unfixed tissue.

    • Main Results:

    • Non-aqueous fixatives and embedding in LR White, Spurr, Araldite, and PMMA demonstrated the least effect on spectral parameters.
    • These parameters included mineral to matrix ratio, mineral crystallinity, and collagen maturity.
    • The selected methods showed minimal spectral interference compared to cryo-sectioned or non-fixed bone.

    Conclusions:

    • Non-aqueous fixatives and embedding in LR White, Spurr, Araldite, and PMMA are recommended for FTIRM/FTIRI analysis of bone.
    • These protocols preserve spectral integrity, ensuring reliable quantitative data on bone composition and structure.
    • This research provides critical guidance for optimizing sample preparation in bone research using infrared spectroscopy.