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Related Experiment Video

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Jacky G Goetz1, Fabien Monduc, Yannick Schwab

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Methods in Molecular Biology (Clifton, N.J.)
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Summary
This summary is machine-generated.

This study presents a new method combining live imaging and electron microscopy to visualize cellular structures. This correlative light and electron microscopy (CLEM) approach enhances understanding of cellular dynamics in vivo.

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Developmental Biology

Background:

  • Live imaging offers dynamic cellular insights but lacks spatial resolution.
  • Electron microscopy provides high-resolution ultrastructure but typically requires fixed samples.
  • Correlative Light and Electron Microscopy (CLEM) bridges this gap by integrating live and fixed imaging modalities.

Purpose of the Study:

  • To develop and describe a protocol for high-resolution ultrastructural analysis of endothelial cells.
  • To enable the study of cellular dynamics in vivo using live confocal microscopy.
  • To correlate live cellular events with detailed ultrastructure using electron microscopy.

Main Methods:

  • Live confocal microscopy was used to image endothelial cells during zebrafish embryonic development.
  • A novel protocol was established for sample preparation allowing subsequent electron microscopy.
  • The same cells were imaged first live and then their ultrastructure was extracted and analyzed.

Main Results:

  • The protocol successfully allowed the extraction of endothelial cell ultrastructure after live imaging.
  • High-resolution electron microscopy images were obtained from previously live-imaged cells.
  • The method preserves cellular context for detailed ultrastructural examination.

Conclusions:

  • This CLEM protocol enables detailed ultrastructural analysis of live-imaged cells in their native context.
  • The technique is valuable for studying dynamic cellular processes with high spatial resolution.
  • It advances the understanding of endothelial cell behavior during embryonic development.