A ball microscope for viewing the entire surface of amphibian embryos.

Waves on the surface of developing eggs/embryos need to be viewed from all sides of their 3D tissue. The ball microscope will enable tracking of cellular waves and determine their interactions with the cells on the surface. Nine microscopes are arrayed in a spherical formation around an imaging stage to create whole surface images of objects anywhere from 0.5 mm3 to 60 mm3 in size. The 3D printed ball-based microscope is made using nine, Opti-Tekscope OT-HD Digital USB Microscope Camera Magnifiers. Eight of the microscope cameras fit into the ball at 90° angles to each other and one bottom microscope is used for a base to hold the stage. The base will support a customised cuvette to hold the embryo in water. The microscopes are the size of a pen (13 cm long and 1 cm in diameter) and each have a ring light around their diameter for self illumination. The nine microscopes can be attached to a microcontroller for time-lapse automated imaging. This microscope will be compared to other microscopes developed for the same purpose. The microscope can be used for time lapse imaging of the surface of small 3D objects and can be used to view Axolotl salamander embryo development as the Axolotl embryos are 2 mm in diameter. Other amphibian eggs can also be imaged using this technique.

Acquisition and reconstruction of 4D surfaces of axolotl embryos with the flipping stage robotic microscope.

We have designed and constructed a Flipping Stage for a light microscope that can view the whole exterior surface of a 2 mm diameter developing axolotl salamander embryo. It works by rapidly inverting the bottom-heavy embryo, imaging it as it rights itself. The images are then montaged to reconstruct the whole 3D surface versus time, for a full 4D record of the surface. Imaging early stage axolotl development will help discover how cell differentiation and movement takes place in the early embryo. For example, the switch from ectodermal to neural plate cells takes place on the top, animal surface portion the egg/embryo and can be observed using the flipping stage microscope. Detailed pictures of the whole surface need to be obtained so that cell tracking and event histories, such as cell divisions and participation in differentiation waves, of individual cells can be recorded. Imaging the whole exterior of the eggs/embryos will allow for the analysis of cell behavior and the forces the cells experience in their natural setting in the intact or manipulated embryo. This will give insights into embryogenesis, development, developmental disruptions, birth defects, cell differentiation and tissue engineering.

Effects of microgravity on cell cytoskeleton and embryogenesis.

The aim of this review is to compile, summarize and discuss the effects of microgravity on embryos, cell structure and function that have been demonstrated from data obtained during experiments performed in space or in altered gravity induced by clinostats. In cells and tissues cellular structure and genetic expression may be changed in microgravity and this has a variety of effects on embryogenesis which include death of the embryo, failure of neural tube closure, or final deformities to the overall morphology of the newborn or hatchling. Many species and protocols have been used for microgravity space experiments making it difficult to compare results. Experiments on the ways in which embryonic development and cell interactions occur in microgravity could also be performed. Experiments that have been done with cells in microgravity show changes in morphology, cytoskeleton and function. Changes in cytoskeleton have been noted and studies on microtubules in gravity have shown that they are gravity sensitive. Further study of basic chemical reactions that occur in cells should be done to shed some light on the underling processes leading to the changes that are observed in cells and embryos in microgravity.