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The Neural Crest (NC)

Investigating migration and differentiation of NC cells in chicken embryo

Chicken-embryo cross section at the level of the midbrain showing neural crest cells and cadherin-6B molecules. Raw widefield data (top) and THUNDER image (bottom). Chicken-embryo_cross_section_midbrain_neural_crest_cells_cadherin-6B_molecules_widefield_THUNDER_teaser.jpg

This article discusses how the study of neural crest (NC) development in chicken embryos is aided with haze-free imaging  using a THUNDER Imager 3D Assay. Proper specification, migration, and differentiation of NC cells during development is essential to prevent craniofacial deformation and neurocristopathies. Here, the epithelial-to-mesenchymal transition of NC cells and migration of cadherin-6B adhesion molecules were sharply imaged  with the THUNDER Imager and computational clearing revealing more details  than conventional widefield microscopy.

Neural crest cells

The neural crest (NC) is an embryonic cell population with remarkable multipotency and migratory ability enabling it to contribute to the development of diverse organ systems, including the craniofacial skeleton. i.e., the bone and cartilage of the face and skull, and the peripheral nervous system [1,2]. The dysregulation of cranial NC development often results in unusual craniofacial development, so studying the NC can help lead to a better understanding of how neurocristopathies can be prevented [1,2]. Researchers in developmental biology and embryology are interested in understanding the mechanisms underlying the specification, migration, and differentiation of NC cells in multiple vertebrate embryos. It is also known that NC cells normally downregulate levels of the cadherin-6B cell adhesion molecule (CAM).

Challenges when imaging embryos

Thick embryo specimens for developmental biology studies can be challenging for widefield microscopy, because there is often an out-of-focus blur or “haze” due to light scattering present in the images [3]. By removing the haze, then structures deep inside the specimen are better revealed.


The specimen studied was a cross section of a chicken embryo at the level of the midbrain showing neural crest (NC) cells. NC cells appear as magenta and Cadherin-6B adhesion molecules appear as green. Fluorescence imaging was done with a THUNDER Imager 3D Assay using a 40x, 1.3 numerical aperture (NA), plan apochromat, oil-immersion objective. Small volume computational clearing (SVCC) was applied [3,4].

Results imaging neural crest cells

The images below are a raw widefield image and THUNDER image of a chicken embryo. The NC cells (magenta) undergo epithelial-to-mesenchymal transition and begin to migrate. Cadherin-6B (green) begins to migrate laterally to the left side of image. NC cells in which the epithelial-to-mesenchymal transition is experimentally blocked are on the right side of image. Cadherin-6B levels remain high and the NC fails to leave the neural tube epithelium.


The image results shown demonstrate that a THUNDER Imager 3D Assay is able to visualize more clearly the neural crest cells and fluorescence signals of cadherin molecules in the embryo specimen compared to a conventional widefield microscope system.

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