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Neuroscience JoVE (Open Access) Citable · DOI

High-resolution Live Imaging of Cell Behavior in the Developing Neuroepithelium

DOI: 10.3791/3920-v
What you'll learn
  • ✓ Perform embryo electroporation to deliver fluorescent markers to developing neural tissue
  • ✓ Prepare chick spinal cord slices and culture medium for long-term imaging
  • ✓ Acquire high-resolution time-lapse imaging of live neuroepithelial cell behavior
  • ✓ Analyze cellular and subcellular dynamics in developing nervous system tissue
Protocol

Imaging embryonic tissue in real-time is challenging over long periods of time. Here we present an assay for monitoring cellular and sub-cellular changes in chick spinal cord for long periods with high spatial and temporal resolution. This technique can be adapted for other regions of the nervous system and developing embryo.

Difficulty
advanced
Total time
~4–6 hours per embryo (electroporation + slice preparation + imaging acquisition)
Model organism
Chick embryo (Gallus gallus domesticus), spinal cord
Biosafety
BSL-1

Steps

1
Perform embryo electroporation with fluorescent markers

Introduce plasmid DNA into developing chick spinal cord using electrical pulses to enable cell tracking and visualization of progenitor cell dynamics.

▶ 01:33
2
Prepare collagen matrix and culture medium

Prepare collagen solution and imaging-compatible culture medium required for slice culture viability and optical clarity during live imaging.

▶ 03:04
3
Generate and mount spinal cord tissue slices

Dissect electroporated embryonic spinal cord, generate thin sections, and mount in collagen matrix within imaging chambers for sustained culture and observation.

▶ 04:22
4
Acquire high-resolution time-lapse image sequences

Configure microscopy system for long-term imaging with appropriate spatial and temporal resolution to capture neuroepithelial cell morphology and movement over extended periods.

▶ 07:52
5
Analyze time-lapse sequences of progenitor cells

Review and quantify cellular behaviors including migration, division, and morphological changes in spinal cord progenitor cells from recorded image series.

▶ 08:58
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