Autor/a:
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Aviles Espinosa, Rodrigo; Santos, Susana I.C.O.; Brodschelm, Andreas; Kaenders, Wilhelm G.; Alonso Ortega, Cesar; Artigas García, David; Loza Álvarez, Pablo
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Abstract:
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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 highresolution
four-dimensional imaging of embryogenesis using unstained
C. elegans in vivo samples. |