Multiview robotic microscope reveals the in-plane kinematics of amphibian neurulation.

A new robotic microscope system, called the Frogatron 3000, was developed to collect time-lapse images from arbitrary viewing angles over the surface of live embryos. Embryos are mounted at the center of a horizontal, fluid-filled, cylindrical glass chamber around which a camera with special optics traverses. To hold them at the center of the chamber and revolve them about a vertical axis, the embryos are placed on the end of a small vertical glass tube that is rotated under computer control. To demonstrate operation of the system, it was used to capture time-lapse images of developing axolotl (amphibian) embryos from 63 viewing angles during the process of neurulation and the in-plane kinematics of the epithelia visible at the center of each view was calculated. The motions of points on the surface of the embryo were determined by digital tracking of their natural surface texture, and a least-squares algorithm was developed to calculate the deformation-rate tensor from the motions of these surface points. Principal strain rates and directions were extracted from this tensor using decomposition and eigenvector techniques. The highest observed principal true strain rate was 28 +/- 5% per hour, along the midline of the neural plate during developmental stage 14, while the greatest contractile true strain rate was--35 +/- 5% per hour, normal to the embryo midline during stage 15.

Mechanical effects of cell anisotropy on epithelia.

Theoretical, numerical and experimental methods are used to develop a comprehensive understanding of how cell shape affects the mechanical characteristics of two-dimensional aggregates such as epithelia. This is an important step in relating the mechanical properties of tissues to those of the cells of which they are composed. Statistical mechanics is used to derive formulas for the in-plane stresses generated by tensions gamma along cell-cell interfaces in sheets with anisotropic cellular fabric characterized by average cell aspect ratio kappa. These formulas are then used to investigate self-deformation (strain relaxation) of an anisotropic sheet composed of cells of thickness h and having effective viscosity mu. Finite element simulations of epithelia and of isolated cells and novel relaxation studies of specimens of embryonic epithelia reported herein are consistent with the predictions of the theory. In all cases, geometric factors cause the relaxation responses to be more complex than a single decaying exponential.