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Proceedings Paper

Two-photon laser scanning microscopy on native cartilage and collagen membranes for tissue engineering
Author(s): Jörg Martini; Katja Tönsing; Michael Dickob; Ronald Schade; Klaus Liefeith; Dario Anselmetti
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Paper Abstract

In our experiments 2-Photon laser scanning microscopy (2PLSM) has been used to acquire 3-dimensional structural information on native unstained biological samples for tissue engineering purposes. Using near infrared (NIR) femtosecond laser pulses for 2-photon excitation and second harmonic generation (SHG) it was possible to achieve microscopic images at great depths in strongly (light) scattering collagen membranes (depth up to 300 μm) and cartilage samples (depth up to 460 μm). With the objective of optimizing the process of chondrocyte growth on collagen scaffolding materials for implantation into human knee joints, two types of samples have been investigated. (1) Both arthritic and non-arthritic bovine and human cartilage samples were examined in order to differentiate between these states and to estimate the density of chondrocytes. In particular, imaging depth, fluorescence intensity and surface topology appear promising as key information for discriminating between the non-arthritic and arthritic states. Human chondrocyte densities between 2-106/cm3 and 20-106/cm3, depending on the relative position of the sample under investigation within the cartilage, were measured using an automated procedure. (2) Chondrocytes which had been sown out on different types of I/III-collagen membranes, were discriminated from the scaffolding membranes on the basis of their native fluorescence emission spectra. With respect to the different membranes, either SHG signals from the collagen fibers of the membranes or differences in the emission spectra of the chondrocytes and the scaffolding collagenes were used to identify chondrocytes and membranes.

Paper Details

Date Published: 23 February 2006
PDF: 9 pages
Proc. SPIE 6089, Multiphoton Microscopy in the Biomedical Sciences VI, 60891N (23 February 2006); doi: 10.1117/12.641430
Show Author Affiliations
Jörg Martini, Bielefeld Univ. (Germany)
Katja Tönsing, Bielefeld Univ. (Germany)
Michael Dickob, Orthopedic Surgery (Germany)
Ronald Schade, IBA e.V. (Germany)
Klaus Liefeith, IBA e.V. (Germany)
Dario Anselmetti, Bielefeld Univ. (Germany)

Published in SPIE Proceedings Vol. 6089:
Multiphoton Microscopy in the Biomedical Sciences VI
Ammasi Periasamy; Peter T. C. So, Editor(s)

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