Annals of Biomedical Engineering

, Volume 25, Issue 4, pp 678–689 | Cite as

A small angle light scattering device for planar connective tissue microstructural analysis

  • Michael S. Sacks
  • David B. Smith
  • Erik D. Hiester
Research Articles


The planar fibrous connective tissues of the body are composed of a dense extracellular network of collagen and elastin fibers embedded in a ground matrix, and thus can be thought of as biocomposites. Thus, the quantification of fiber architecture is an important step in developing an understanding of the mechanics of planar tissues in health and disease. We have used small angle light scattering (SALS) to map the gross fiber orientation of several soft membrane connective tissues. However, the device and analysis methods used in these studies required extensive manual intervention and were unsuitable for largescale fiber architectural mapping studies. We have developed an improved SALS device that allows for rapid data acquisition, automated high spatial resolution specimen positioning, and new analysis methods suitable for large-scale mapping studies. Extensive validation experiments revealed that the SALS device can accurately measure fiber orientation for up to a tissue thickness of at least 500 μm to an angular resolution of∼1o and a spatial resolution of±254 μm. To demonstrate the new device’s capabilities, structural measurements from porcine aortic valve leaflets are presented. Results indicate that the new SALS device provides an accurate method for rapid quantification of the gross fiber structure of planar connective tissues.


Collagen Fiber architecture Lasers Light scattering Optical methods Heart valves 


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  1. 1.
    Baer, E., J. J. Cassidy, and A. Hiltner. Hierarchical structure of collagen and its relationship to the physical properties of tendon. In: Collagen, vol. 2, edited by M. E. Nimni. Boca Raton, FL: CRC Press, 1988, pp. 177–199.Google Scholar
  2. 2.
    Borch, J., P. R. Sundarajan, and R. H. Marchessault. Light scattering by cellulose. III. Morphology of wood.J. Polym. Sci. 9:313–329, 1971.Google Scholar
  3. 3.
    Bortolotti, U., A. Milano, and A. Mazzucco. Results of re-operation for primary tissue failure of porcine bioprostheses.J. Thorac. Cardiovasc. Surg. 90:564–569, 1985.PubMedGoogle Scholar
  4. 4.
    Chaudhuri, S., H. Nguyen, R. M. Rangayyan, S. Walsh, and C. B. Frank. A Fourier domain directional filtering method for analysis of collagen alignment in ligaments.IEEE Trans. Biomed. Eng. BME-34:509–518, 1987.PubMedCrossRefGoogle Scholar
  5. 5.
    Chien, J. C. W., and E. P. Chang. Small-angle light scattering of reconstituted collagen.Macromolecules 5:610–617, 1972.CrossRefGoogle Scholar
  6. 6.
    Chuong, C. J., M. S. Sacks, R. L. Johnson, and R. C. Reynolds. On the anisotropy of the diaphragmatic central tendon.J. Biomech. 24:563–576, 1991.PubMedCrossRefGoogle Scholar
  7. 7.
    Cowley, J. M. Principles of image formation. In: Introduction to analytical electron microscopy, chap 1, edited by J. J. Hren, J. I. Goldstein, and D. C. Joy. New York: Plenum Press, 1979, pp. 1–42.Google Scholar
  8. 8.
    Ferrans, V. J., S. L. Hilbert, T. Tomita, M. Jones, and W. C. Robert. Morphology of collagen in bioprosthetic heart valves. In: Collagen, vol. 3, edited by M. E. Nimni. Boca Raton, FL: CRC Press, 1988, pp. 145–189.Google Scholar
  9. 9.
    Frank, C., B. MacFarlane, P. Edwards, R. Rangayyan, Z. Q. Liu, S. Walsh, and R. Bray. A quantitative analysis of matrix alignment in ligament scars: a comparison of movement versus immobilization in an immature rabbit model.J. Orthoped. Res. 9:219–227, 1991.CrossRefGoogle Scholar
  10. 10.
    Fung, Y. C. Biomechanics: Mechanical Properties of Living Tissues New York: Springer Verlag, 1993, pp. 1–568.Google Scholar
  11. 11.
    Gabbay, S., P. Kadam, S. Factor, and T. K. Cheung. Do heart valves bioprostheses degenerate for metabolic or mechanical reasons?J. Thorac. Cardiovasc. Surg. 55:208–215, 1988.Google Scholar
  12. 12.
    Guinier, A. X-Ray Diffraction. San Francisco: W. H. Freeman and Company, 1963, pp. 1–378.Google Scholar
  13. 13.
    Halliday, D., and R. Resnick. Physics, New York: John Wiley and Sons, 1960, pp. 1–1214.Google Scholar
  14. 14.
    Hilbert, S. L., V. J. Ferrans, and W. M. Swanson. Optical methods for the nondestructive evaluation of collagen morphology in bioprosthetic heart valves.J. Biomed. Mater. Res. 20:1411–1421, 1986.PubMedCrossRefGoogle Scholar
  15. 15.
    Hukins, D. W. L. Collagen orientation. In: Connective tissue matrix, edited by D. W. L. Hukins. Munich: Verlag, Chemie, 1984, pp. 211–240.Google Scholar
  16. 16.
    Kastelic, J., and E. Baer. Deformation of tendon collagen. In: The mechanical properties of biological materials, edited by J. F. Vincient, and J. D. Currey. Weinheim, U.K.: Society for Experimental Biology Symposium XXXIV, 1980, pp. 397–433.Google Scholar
  17. 17.
    Kronick, P. L., and P. R. Buechler. Fiber orientation in calfskin by laser light scattering or X-ray diffraction and quantitative relation to mechanical properties.J. Am. Leather Chem. Assoc. 81:221–229, 1986.Google Scholar
  18. 18.
    Kronick, P. L., M. S. Sacks, and M. Dahms. Vertical fiber defect quantified by small angle light scattering.Connect. Tiss. Res. 27:1–13, 1991.Google Scholar
  19. 19.
    Liu, Z. Q., R. M. Rangayyan, and C. B. Frank. Statistical analysis of collagen alignment in ligaments by scale-space analysis.IEEE Trans. Biomed. Eng. 38:580–587, 1991.PubMedCrossRefGoogle Scholar
  20. 20.
    Marshall, G. E. Gaussian laser beam diameters and divergence. In: Optical scanning, edited by G. E. Marshall. New York: Marcel Dekker, 1991, pp. 1–11.Google Scholar
  21. 21.
    Milano, A., U. Bortolotti, and E. Talenti. Calcific degeneration as the main cause of porcine bioprostheses.Am J. Cardiol. 53:1066–1070, 1984.PubMedCrossRefGoogle Scholar
  22. 22.
    Moritani, M., N. Hayashi, A. Utsuo, and H. Kawai. Light-scattering patterns from collagen films in relation to the texture of a random assembly of anisotropic rods in three dimensions.Polym. J. 2:74–87, 1971.CrossRefGoogle Scholar
  23. 23.
    Muggli, R., and R. Marton. Light scattering by cellulose. V. Anisotropy scattering by wood fibers.J. Polym. Sci. 36:121–139, 1971.Google Scholar
  24. 24.
    Otano, S. E., M. S. Sacks, and T. I. Malinin. Mechanical Behavior of Human Dura Mater, vol. 29.Proceedings in the 1995 Bioengineering Conference, Beaver Creek, CO, 1995, pp. 329–330.Google Scholar
  25. 25.
    Purslow, P. P., A. Bigi, A. Ripamonti, and N. Roveri. Collagen fibre reorientation around a crack in biaxially stretched materials.Int. J. Macromol. 6:21–25, 1984.CrossRefGoogle Scholar
  26. 26.
    Raman, C. V., and M. R. Bhat. The structure and optical behavior of some natural and synthetic fibers.Proc. Indian Acad. Sci. A40:109–116, 1954.Google Scholar
  27. 27.
    Sacks, M. S., Focus on materials with scattered light.Res. Dev. 30:73–78, 1988.Google Scholar
  28. 28.
    Sacks, M. S., and C. J. Chuong. Characterization of collagen fiber architecture in the canine central tendon.J. Biomech. Eng. 114:183–190, 1992.PubMedGoogle Scholar
  29. 29.
    Sacks, M. S., C. J. Chuong, and R. More. Collagen fiber architecture of bovine pericardium.ASAIO 40:M632-M637, 1994.CrossRefGoogle Scholar
  30. 30.
    Sacks, M. S., M. S. Chuong, W. M. Petroll, M. Kwan, and C. Halberstatd. Collagen fiber architecture of a cultured tissue.J. Biomech. Eng. 119:124–127, 1997.PubMedGoogle Scholar
  31. 31.
    Sasaki, N., and S. Odajima. Stress-strain curve and Young’s modulus of a collagen molecule as determined by the X-ray diffraction technique.J. Biomech. 29:655–658, 1996.PubMedCrossRefGoogle Scholar
  32. 32.
    Schoen, F. J., Cardiac valve prostheses: review of clinical status and contemporary biomaterial issuesJ. Biomed. Mater. Res. 21:91–117, 1987.PubMedGoogle Scholar
  33. 33.
    Stein, R. S., P. Erhardt, J. J. van Aartsen, and S. Clough. Theory of light scattering from oriented and fiber structures.J. Polym. Sci. 13:1–35, 1966.Google Scholar
  34. 34.
    Stein, R. S., and P. R. Wilson. Scattering of light by polymer films possessing correlated orientation fluctuation.J. Appl. Phys. 33:1914–1922, 1962.CrossRefGoogle Scholar
  35. 35.
    Whittaker, P., and P. B. Canham. Demonstration of quantitative fabric analysis of tendon collagen using two-dimensional polarized light microscopy.Matrix 11:56–62, 1991.PubMedGoogle Scholar

Copyright information

© Biomedical Engineering Society 1997

Authors and Affiliations

  • Michael S. Sacks
    • 1
  • David B. Smith
    • 1
  • Erik D. Hiester
    • 1
  1. 1.Tissue Mechanics Laboratory, Department of Biomedical EngineeringUniversity of MiamiCoral Gables

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