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Updated: Jul 3, 2026
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4D Microscopy of Yeast
Published on: April 28, 2019
A protocol for isolation and visualization of yeast nuclei by scanning electron microscopy
Stephen Murray1, Elena Kiseleva
1TEM Service Facility, Paterson Institute for Cancer Research, University of Manchester, Manchester M20 4BX, United Kingdom.
Methods in Cell Biology
|July 12, 2008
Summary
This study presents a new protocol for isolating yeast nuclei for high-resolution imaging. It enables detailed visualization of nuclear structures like the nuclear envelope and nuclear pore complexes using Field Emission Scanning Electron Microscopy (FESEM).
Area of Science:
- Cell Biology
- Microscopy Techniques
- Molecular Biology
Background:
- Studying yeast nuclear structure requires advanced imaging methods.
- Existing protocols may not offer sufficient resolution or ease of use for visualizing nuclear components.
Purpose of the Study:
- To describe a detailed protocol for yeast nuclear isolation.
- To enable high-resolution visualization of the nuclear envelope (NE), nuclear pore complexes (NPCs), and associated cytoskeletal structures.
Main Methods:
- Enzymatic treatment of yeast cells (Saccharomyces cerevisiae and Schizosaccharomyces pombe) to create spheroplasts.
- Isolation of nuclei via centrifugation.
- Immunogold labeling of proteins.
- Visualization using Field Emission Scanning Electron Microscopy (FESEM).
Main Results:
- Successful isolation of yeast nuclei from both budding and fission yeast.
- Optimized protocol allows for clear visualization of the NE and NPCs.
- FESEM imaging reveals associated cytoskeletal structures.
Conclusions:
- The described protocol is effective for high-resolution imaging of yeast nuclear structures.
- This method provides a valuable tool for cell biology research.
- Samples can be stored under vacuum for extended periods, facilitating image acquisition.
