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Updated: Jan 23, 2026
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Correlative Super-resolution and Electron Microscopy to Resolve Protein Localization in Zebrafish Retina
Published on: November 10, 2017
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Super-Resolution Microscopy of Phloem Proteins
Ryan C Stanfield1, Alexander Schulz2
1Department of Renewable Resources, University of Alberta, Edmonton, AB, Canada.
Methods in Molecular Biology (Clifton, N.J.)
|June 15, 2019
Summary
Super-resolution microscopy enhances phloem protein studies by offering higher resolution than confocal or electron microscopy. This protocol details advanced imaging for sieve element and companion cell plasma membrane proteins using 3D-SIM.
Area of Science:
- Plant cell biology
- Microscopy techniques
- Molecular imaging
Background:
- Super-resolution microscopy (SRM) offers significant resolution improvements over traditional light microscopy.
- SRM techniques, based on fluorescence, enable detailed visualization of cellular structures.
- Phloem research has been limited by the resolution of existing microscopy methods.
Purpose of the Study:
- To present a protocol for investigating phloem proteins using SRM.
- To demonstrate the advantages of SRM for visualizing plasma membrane proteins in sieve elements and companion cells.
- To provide insights into phloem function through high-resolution imaging.
Main Methods:
- Utilizing fluorescence-based SRM techniques, specifically three-dimensional structured illumination microscopy (3D-SIM).
- Developing a protocol for immunolocalization of phloem proteins.
- Adapting sample preparation for high-resolution imaging of delicate phloem tissue.
Main Results:
- SRM provides higher resolution (2.5-10x) than confocal microscopy for phloem imaging.
- Precise co-localization of membranes, plasmodesmata, and structural proteins is achievable.
- SRM reveals details not visible with confocal or electron microscopy, overcoming limitations of fixed and sectioned tissues.
Conclusions:
- SRM, particularly 3D-SIM, is a powerful tool for studying phloem protein localization and function.
- The presented protocol facilitates high-resolution imaging of immunolocalized phloem proteins.
- SRM opens new avenues for understanding phloem biology at the molecular level.
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