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Third-harmonic generation for the study of Caenorhabditis elegans embryogenesis
Aviles Espinosa, Rodrigo; Santos, Susana I.C.O.; Brodschelm, Andreas; Kaenders, Wilhelm G.; Alonso Ortega, Cesar; Artigas García, David; Loza Álvarez, Pablo
Universitat Politècnica de Catalunya. Departament de Teoria del Senyal i Comunicacions
Live microscopy techniques i.e., differential interferencecontrast, confocal microscopy, etc. have enabled the understandingof the mechanisms involved in cells and tissue formation. In long-termstudies, special care must be taken in order to avoid sample damage,restricting the applicability of the different microscopy techniques.Wedemonstrate the potential of using third-harmonic generation THG microscopy for morphogenesis/embryogenesis studies in living Caenorhabditiselegans C. elegans . Moreover, we show that the THGsignal is obtained in all the embryo development stages, showing differenttissue/structure information. For this research, we employ a1550-nm femtosecond fiber laser and demonstrate that the expectedwater absorption at this wavelength does not severely compromisesample viability. Additionally, this has the important advantage thatthe THG signal is emitted at visible wavelengths 516 nm . Therefore,standard collection optics and detectors operating near maximum efficiencyenable an optimal signal reconstruction. All this, to the bestof our knowledge, demonstrates for the first time the noninvasivenessand strong potential of this particular wavelength to be used for highresolutionfour-dimensional imaging of embryogenesis using unstainedC. elegans in vivo samples.
Multiphoton processes
Fiber optic cables
Imaging systems in biology
Signal theory (Telecommunication)
Senyal, Teoria del (Telecomunicació)
Open Access

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