Second harmonic imaging of collagen in mammalian tissue

Guy C. Cox; Frank Manconi; Eleanor Kable

doi:10.1117/12.470689

17 June 2002 Second harmonic imaging of collagen in mammalian tissue

Guy C. Cox, Frank Manconi, Eleanor Kable

Proceedings Volume 4620, Multiphoton Microscopy in the Biomedical Sciences II; (2002) https://doi.org/10.1117/12.470689
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

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.

Citation Download Citation

Guy C. Cox, Frank Manconi, and Eleanor Kable "Second harmonic imaging of collagen in mammalian tissue", Proc. SPIE 4620, Multiphoton Microscopy in the Biomedical Sciences II, (17 June 2002); https://doi.org/10.1117/12.470689

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