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Proceedings Paper

Second harmonic imaging of collagen in mammalian tissue
Author(s): Guy C. Cox; Frank Manconi; Eleanor Kable
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Paper Abstract

It has recently been demonstrated that collagen is a very effective upconverter of light by second harmonic generation (SHG) but hitherto the potential this offers for biomedical imaging has not been realized. We show that bright SHG images van be obtained over a wide excitation range at illumination levels comparable to or lower than those required for two-photon excitation of fluorescent labels, with no damage to the collagen structure. Both paraffin and cryostat sections have been used, and medically significant results have been obtained in several fields. We show that the signal is easily distinguished from single and two-photon excited fluorescence by its forward propagation and narrow spectral width; in principle it could also be distinguished by lifetime. Key microscope requisites are: immersion objectives and condensers, high-efficiency PMT detectors for transmitted light, suitable filters, and effective blocking of stray light, especially from the mercury lamp.

Paper Details

Date Published: 17 June 2002
PDF: 9 pages
Proc. SPIE 4620, Multiphoton Microscopy in the Biomedical Sciences II, (17 June 2002); doi: 10.1117/12.470689
Show Author Affiliations
Guy C. Cox, Univ. of Sydney (Australia)
Frank Manconi, Univ. of Sydney (Australia)
Eleanor Kable, Univ. of Sydney (Australia)

Published in SPIE Proceedings Vol. 4620:
Multiphoton Microscopy in the Biomedical Sciences II
Ammasi Periasamy; Peter T. C. So, Editor(s)

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