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

Year: 2010

Authors: Cicchi R., Kapsokalyvas D., De Giorgi V., Maio V., Van Wiechen A., Massi D., Lotti T., Pavone F.S.

Autors Affiliation: L.E.N.S. and Department of Physics, University of Florence, Via Nello Carrara 1, 50019, Sesto Fiorentino, Italy;
Department of Dermatology, University of Florence, Via della Pergola 58, 50121, Florence, Italy;
Department of Human Pathology and Oncology, University of Florence, Viale G.B. Morgagni 85, 50134, Florence, Italy;
Kavli Institute of Nanoscience, Delft University of Technology, Lorentzweg 1, 2628 CJ, Delft, The Netherlands;
C.N.I.S.M., Unity of Florence, Sesto Fiorentino, Italy

Abstract: 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.


Volume: 3 (1-2)      Pages from: 34  to: 43

More Information: We thank the European Laboratory for Non-linear Spectroscopy (Integrate Infrastructure Initiative \”LASERLAB EUROPE\” JRA-Optobio), Italian Space Agency (ASI, MOMA project), and Fondazione Ente Cassa di Risparmio di Firenze for financial support to this project. We thank R. Ballerini and A. Hajeb from the mechanical workshop of LENS, and M. Giuntini, M. De Pas and A. Montuori from the electronic workshop of LENS for their help in building the mechanics and electronics of our experimental setup.
KeyWords: Aging index; Analysis method; Autofluorescence; Collagen disorders; Collagen fiber; Collagen morphology; Connective tissues; Ex-vivo; Geometrical arrangements; Gray level co-occurrence matrix; Human dermis; Image analysis method; Imaging applications; In-vivo; Multi-photon microscopy; Nonlinear imaging; Pattern analysis; Scoring methods; Second harmonic generation; Second harmonics; Second-harmonic generation microscopy, Harmonic analysis; Harmonic generation; Histology; Image analysis; Musculoskeletal system, Collagen, collagen, algorithm; anisotropy; article; dermis; histology; human; image processing; in vitro study; instrumentation; keloid; methodology; microscopy; multiphoton microscopy; nonlinear system; pathology; scar, Algorithms; Anisotropy; Cicatrix; Collagen; Dermis; Humans; Image Processing, Computer-Assisted; Keloid; Microscopy; Microscopy, Fluorescence, Multiphoton; Nonlinear Dynamics
DOI: 10.1002/jbio.200910062

Citations: 196
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