Analysis of cytokinesis by electron microscopy

Area of Science:

• Cell Biology
• Developmental Biology
• Microscopy

Background:

• Correlative Light and Electron Microscopy (CLEM) is crucial for understanding cellular processes.
• Previous work established protocols for early Caenorhabditis elegans embryos in mitosis.

Purpose of the Study:

• To adapt and extend an established protocol for ultrastructural analysis of permeabilized or injected embryonic systems.
• To focus on initial preparation steps for high-pressure freezing and electron microscopy of postmitotic embryos.

Main Methods:

• Adaptation of an established protocol for ultrastructural analysis.
• Preparation of drug-treated C. elegans embryos and fluorescently labeled Lytechinus pictus embryos.
• High-pressure freezing and subsequent electron microscopy, with an emphasis on electron tomography.

Main Results:

• The study details initial preparation steps for high-pressure freezing and electron microscopy.
• The adapted protocol is suitable for both C. elegans and sea urchin embryos.
• Focus on ultrastructural analysis of animal cytokinesis in early embryonic stages.

Conclusions:

• The extended protocol facilitates ultrastructural studies on animal cytokinesis.
• The method allows for detailed analysis of permeabilized or injected embryonic systems.
• Advantages and limitations of the protocol for electron tomography will be discussed.