Multimodal nonlinear optical imaging of collagen arrays

We report multimodal nonlinear optical imaging of fascia, a rich collagen type I sheath around internal organs and muscle. We show that second harmonic generation (SHG), third harmonic generation (THG) and Coherent anti-Stokes Raman scattering (CARS) microscopy techniques provide complementary infor...

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Bibliographic Details
Published in:Journal of structural biology 2008-10, Vol.164 (1), p.140-145
Main Authors: Pfeffer, Christian P., Olsen, Bjorn R., Ganikhanov, Feruz, Légaré, François
Format: Article
Language:English
Subjects:
Animals
Atomic force microscopy
Collagen
Collagen Type I - chemistry
Collagen Type I - ultrastructure
Connective Tissue
Fascia - chemistry
Fascia - ultrastructure
Mice
Mice, Inbred C57BL
Microscopy
Microscopy, Atomic Force
Nonlinear optical microscopy
Spectrum Analysis, Raman
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Summary:We report multimodal nonlinear optical imaging of fascia, a rich collagen type I sheath around internal organs and muscle. We show that second harmonic generation (SHG), third harmonic generation (THG) and Coherent anti-Stokes Raman scattering (CARS) microscopy techniques provide complementary information about the sub-micron architecture of collagen arrays. Forward direction SHG microscopy reveals the fibrillar arrangement of collagen type I structures as the main matrix component of fascia. SHG images detected in the backward direction as well as images of forward direction CARS microscopy show that the longitudinal collagen fiber bundles are further arranged in sheet-like bands. Forward-THG microscopy reveals the optically homogeneous content of the collagen sheet on a spatial scale of the optical wavelength. This is supported by the fact that the third harmonic signal is observed only at the boundaries between the sheets as well as by the CARS data obtained in both directions. The observations made with THG and CARS microscopy are explained using atomic force microscopy images.
ISSN:1047-8477
1095-8657
DOI:10.1016/j.jsb.2008.07.002