Scoring of collagen organization in healthy and diseased human dermis by multiphoton microscopy

J Biophotonics. 2010 Jan;3(1-2):34-43. doi: 10.1002/jbio.200910062.


We have used nonlinear imaging to evaluate collagen organization in connective tissue ex-vivo samples. Image analysis methods were tested on healthy dermis, normal scars, and keloids. The evaluation of the second harmonic to autofluorescence aging index of dermis (SAAID) has allowed a first characterization of tissues by scoring the collagen/elastin content. Further analyses on collagen morphology in healthy dermis and keloids were performed by image-pattern analysis of SHG images. The gray-level co-occurrence matrix (GLCM) analysis method has allowed classification of different tissues based on the evaluation of geometrical arrangement of collagen fibrillar bundles, whereas a pattern analysis of the FFT images has allowed the discrimination of different tissues based on the anisotropy of collagen fibers distribution. This multiple scoring method represents a promising tool to be extended to other collagen disorders, as well as to be used in in-vivo skin-imaging applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Anisotropy
  • Cicatrix / pathology*
  • Collagen*
  • Dermis / anatomy & histology*
  • Dermis / pathology*
  • Humans
  • Image Processing, Computer-Assisted / methods
  • In Vitro Techniques
  • Keloid / pathology*
  • Microscopy / instrumentation
  • Microscopy / methods
  • Microscopy, Fluorescence, Multiphoton / instrumentation
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Nonlinear Dynamics


  • Collagen