Chapter 10
Live Imaging of the Neural Crest Cell
Epithelial-to-Mesenchymal Transition in the Chick Embryo
Mary Cathleen McKinney and Paul M. Kulesa
Abstract
Live embryo imaging may provide a wealth of information on intact cell and tissue dynamics, but can be
technically challenging to sustain embryo orientation and health for long periods under a microscope. In
this protocol, we describe an in vivo method to mount and image cell movements during the epithelial-tomesenchymal transition (EMT) of neural crest cells within the chick dorsal neural tube. We focus on
describing the collection of images and data preparation for image analysis throughout the developmental
stages HH15-21 in the chick trunk. Trunk neural crest cell EMT is crucial to development of the peripheral
nervous system and pigment cell patterning. The methods we describe may also be applied to other cell and
tissue phenomena at various chick developmental stages with some modifications.
Key words Chick, In vivo, Whole embryo culture, Time-lapse imaging, Neural crest, EMT
1 Introduction
Static imaging limits our understanding of the mechanisms underlying in vivo cell and tissue dynamics. However, to resolve individual cell behaviors within intact embryos present several technical
challenges. Early-stage embryos are often fragile and difficult to
fluorescently mark and manipulate. Later-stage embryos are often
difficult to keep oriented and in the field of view of the microscope
objective due to vibrations of the heart, tissue growth of normal
morphogenesis, thermal drift due to a heated chamber, or shrinkage due to dehydration. One of the crucial phenomena of early
development is the epithelial-to-mesenchymal transition of neural
crest cells enabling them to migrate and form peripheral structures.
Neural crest cells exit from the dorsal neural tube in a rostral-tocaudal manner starting near the midbrain [1]. Following single cell
behaviors in the dorsal neural tube and neural crest cell emigration
has proved difficult due to the challenges described above. Following this protocol, we were able to observe behaviors within the
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_10,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
107
Live Imaging of the Neural Crest Cell
Epithelial-to-Mesenchymal Transition in the Chick Embryo
Mary Cathleen McKinney and Paul M. Kulesa
Abstract
Live embryo imaging may provide a wealth of information on intact cell and tissue dynamics, but can be
technically challenging to sustain embryo orientation and health for long periods under a microscope. In
this protocol, we describe an in vivo method to mount and image cell movements during the epithelial-tomesenchymal transition (EMT) of neural crest cells within the chick dorsal neural tube. We focus on
describing the collection of images and data preparation for image analysis throughout the developmental
stages HH15-21 in the chick trunk. Trunk neural crest cell EMT is crucial to development of the peripheral
nervous system and pigment cell patterning. The methods we describe may also be applied to other cell and
tissue phenomena at various chick developmental stages with some modifications.
Key words Chick, In vivo, Whole embryo culture, Time-lapse imaging, Neural crest, EMT
1 Introduction
Static imaging limits our understanding of the mechanisms underlying in vivo cell and tissue dynamics. However, to resolve individual cell behaviors within intact embryos present several technical
challenges. Early-stage embryos are often fragile and difficult to
fluorescently mark and manipulate. Later-stage embryos are often
difficult to keep oriented and in the field of view of the microscope
objective due to vibrations of the heart, tissue growth of normal
morphogenesis, thermal drift due to a heated chamber, or shrinkage due to dehydration. One of the crucial phenomena of early
development is the epithelial-to-mesenchymal transition of neural
crest cells enabling them to migrate and form peripheral structures.
Neural crest cells exit from the dorsal neural tube in a rostral-tocaudal manner starting near the midbrain [1]. Following single cell
behaviors in the dorsal neural tube and neural crest cell emigration
has proved difficult due to the challenges described above. Following this protocol, we were able to observe behaviors within the
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_10,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
107