Cell lineage reconstruction of early zebrafish embryos using label-free nonlinear microscopy.
Quantifying cell behaviors in animal early embryogenesis remains a challenging issue requiring in toto imaging and automated image analysis. We designed a framework for imaging and reconstructing unstained whole zebrafish embryos for their first 10 cell division cycles and report measurements along the cell lineage with micrometer spatial resolution and minute temporal accuracy. Point-scanning multiphoton excitation optimized to preferentially probe the innermost regions of the embryo provided intrinsic signals highlighting all mitotic spindles and cell boundaries. Automated image analysis revealed the phenomenology of cell proliferation. Blastomeres continuously drift out of synchrony. After the 32-cell stage, the cell cycle lengthens according to cell radial position, leading to apparent division waves. Progressive amplification of this process is the rule, contrasting with classical descriptions of abrupt changes in the system dynamics.
Olivier
Nicolas
N
Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS, INSERM, Palaiseau, France.
Luengo-Oroz
Miguel A
MA
Duloquin
Louise
L
Faure
Emmanuel
E
Savy
Thierry
T
Veilleux
Israël
I
Solinas
Xavier
X
Débarre
Delphine
D
Bourgine
Paul
P
Santos
Andrés
A
Peyriéras
Nadine
N
Beaurepaire
Emmanuel
E
D000818:Animals; D036703:Blastula / Q000166:cytology; D002453:Cell Cycle; D019070:Cell Lineage; D004625:Embryo, Nonmammalian / Q000166:cytology*; D007091:Image Processing, Computer-Assisted; D008853:Microscopy / Q000379:methods*; D015027:Zebrafish / Q000196:embryology*