Share Email Print

Journal of Biomedical Optics • Open Access

Third-harmonic generation for the study of Caenorhabditis elegans embryogenesis
Author(s): Rodrigo Amador Aviles-Espinosa; Susana I. C. O. Santos; Andreas Brodschelm; Wilhelm G. Kaenders; Cesar Alonso-Ortega; David Artigas-García; Pablo Loza-Alvarez

Paper Abstract

Live microscopy techniques (i.e., differential interference contrast, confocal microscopy, etc.) have enabled the understanding of the mechanisms involved in cells and tissue formation. In long-term studies, special care must be taken in order to avoid sample damage, restricting the applicability of the different microscopy techniques. We demonstrate the potential of using third-harmonic generation (THG) microscopy for morphogenesis/embryogenesis studies in living Caenorhabditis elegans (C. elegans). Moreover, we show that the THG signal is obtained in all the embryo development stages, showing different tissue/structure information. For this research, we employ a 1550-nm femtosecond fiber laser and demonstrate that the expected water absorption at this wavelength does not severely compromise sample viability. Additionally, this has the important advantage that the THG signal is emitted at visible wavelengths (516 nm). Therefore, standard collection optics and detectors operating near maximum efficiency enable an optimal signal reconstruction. All this, to the best of our knowledge, demonstrates for the first time the noninvasiveness and strong potential of this particular wavelength to be used for high-resolution four-dimensional imaging of embryogenesis using unstained C. elegans in vivo samples.

Paper Details

Date Published: 1 July 2010
PDF: 7 pages
J. Biomed. Opt. 15(4) 046020 doi: 10.1117/1.3477535
Published in: Journal of Biomedical Optics Volume 15, Issue 4
Show Author Affiliations
Rodrigo Amador Aviles-Espinosa, ICFO - Instituto de Ciencias Fotónicas (Spain)
Susana I. C. O. Santos, ICFO - Instituto de Ciencias Fotónicas (Spain)
Andreas Brodschelm, TOPTICA Photonics AG (Germany)
Wilhelm G. Kaenders, TOPTICA Photonics AG (Germany)
Cesar Alonso-Ortega, ICFO - Instituto de Ciencias Fotónicas (Spain)
David Artigas-García, Univ. Politècnica de Catalunya (Spain)
Pablo Loza-Alvarez, ICFO - Instituto de Ciencias Fotónicas (Spain)

© SPIE. Terms of Use
Back to Top