Velocimetric third-harmonic generation microscopy: micrometer-scale quantification of morphogenetic movements in unstained embryos.
We demonstrate the association of third-harmonic generation (THG) microscopy and particle image velocimetry (PIV) analysis as a novel functional imaging technique for automated micrometer-scale characterization of morphogenetic movements in developing embryos. Using a combined two-photon-excited fluorescence and THG microscope, we characterize the optical properties of Drosophila embryos and show that sustained THG imaging does not perturb sensitive developmental dynamics. Velocimetric THG imaging provides a quantitative description of the dynamics of internal structures in unstained wild-type and mutant embryos.
Débarre
Delphine
D
Laboratory for Optics and Biosciences, Centre National de la Recherche Scientifique, Institut National de la Santé et de la Recherche Médicale, Ecole Polytechnique, F-91128 Palaiseau, France.
Supatto
Willy
W
Farge
Emmanuel
E
Moulia
Bruno
B
Schanne-Klein
Marie-Claire
MC
Beaurepaire
Emmanuel
E
D000465:Algorithms; D000818:Animals; D004330:Drosophila / Q000166:cytology / Q000196:embryology / Q000502:physiology; D004625:Embryo, Nonmammalian; D007089:Image Enhancement / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods*; D009068:Movement / Q000502:physiology*; D015203:Reproducibility of Results; D012212:Rheology / Q000379:methods*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling