In-vivo third-harmonic generation microscopy at 1550nm three-dimensional long-term time-lapse studies in living C. elegans embryos
2011
In-vivo microscopic long term time-lapse studies require controlled imaging conditions to preserve sample viability.
Therefore it is crucial to meet specific exposure conditions as these may limit the applicability of established techniques.
In this work we demonstrate the use of third harmonic generation (THG) microscopy for long term time-lapse three-dimensional
studies (4D) in living Caenorhabditis elegans embryos employing a 1550 nm femtosecond fiber laser. We
take advantage of the fact that THG only requires the existence of interfaces to generate signal or a change in the
refractive index or in the χ 3 nonlinear coefficient, therefore no markers are required. In addition, by using this
wavelength the emitted THG signal is generated at visible wavelengths (516 nm) enabling the use of standard collection
optics and detectors operating near their maximum efficiency. This enables the reduction of the incident light intensity at
the sample plane allowing to image the sample for several hours.
THG signal is obtained through all embryo development stages, providing different tissue/structure information. By
means of control samples, we demonstrate that the expected water absorption at this wavelength does not severely
compromise sample viability.
Certainly, this technique reduces the complexity of sample preparation (i.e. genetic modification) required by established
linear and nonlinear fluorescence based techniques. We demonstrate the non-invasiveness, reduced specimen
interference, and strong potential of this particular wavelength to be used to perform long-term 4D recordings.
Keywords:
- Correction
- Source
- Cite
- Save
- Machine Reading By IdeaReader
0
References
0
Citations
NaN
KQI