2-photon laser scanning microscopy on native human cartilage

Joerg Martini; Katja Toensing; Michael Dickob; Dario Anselmetti

doi:10.1117/12.632940

7 October 2005 2-photon laser scanning microscopy on native human cartilage

Joerg Martini, Katja Toensing, Michael Dickob, Dario Anselmetti

Proceedings Volume 5860, Confocal, Multiphoton, and Nonlinear Microscopic Imaging II; 586003 (2005) https://doi.org/10.1117/12.632940
Event: European Conferences on Biomedical Optics 2005, 2005, Munich, Germany

Abstract

Native hyaline cartilage from a human knee joint was directly investigated with laser scanning microscopy via 2-photon autofluorescence excitation with no additional staining or labelling protocols in a nondestructive and sterile manner. Using a femtosecond, near-infrared (NIR) Ti:Sa laser for 2-photon excitation and a dedicated NIR long distance objective, autofluorescence imaging and measurements of the extracellular matrix (ECM) tissue with incorporated chondrocytes were possible with a penetration depth of up to 460 μm inside the sample. Via spectral autofluorescence separation these experiments allowed the discrimination of chondrocytes from the ECM and therefore an estimate of chondrocytic cell density within the cartilage tissue to approximately 0.2-2•10⁷cm³. Furthermore, a comparison of the relative autofluorescence signals between nonarthritic and arthritic cartilage tissue exhibited distinct differences in tissue morphology. As these morphological findings are in keeping with the macroscopic diagnosis, our measurement has the potential of being used in future diagnostic applications.

Citation Download Citation

Joerg Martini, Katja Toensing, Michael Dickob, and Dario Anselmetti "2-photon laser scanning microscopy on native human cartilage", Proc. SPIE 5860, Confocal, Multiphoton, and Nonlinear Microscopic Imaging II, 586003 (7 October 2005); https://doi.org/10.1117/12.632940

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