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Visualizing Intracellular Sialylation with Click Chemistry and Expansion Microscopy
Published on: February 7, 2025
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Click-ExM enables expansion microscopy for all biomolecules.
De-En Sun1,2, Xinqi Fan1,2, Yujie Shi1,2,3
1College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
Nature Methods
|December 8, 2020
Summary
Expansion microscopy (ExM) now images more than proteins. Click-ExM uses click chemistry to enable nanoscale imaging of diverse biomolecules, including lipids and nucleic acids, on standard microscopes.
Area of Science:
- Biochemistry
- Microscopy
- Molecular Biology
Background:
- Expansion microscopy (ExM) offers super-resolution imaging on conventional microscopes.
- Current ExM methods are primarily limited to proteins and nucleic acids.
Purpose of the Study:
- To develop a versatile method for nanoscale imaging of diverse biomolecules using ExM.
- To integrate click chemistry into ExM for broader biomolecule compatibility.
Main Methods:
- Developed click-ExM by integrating click labeling into the ExM protocol.
- Utilized click labeling with biotin and fluorescently labeled streptavidin for detection.
- Applied click-ExM to various biomolecules including lipids, glycans, proteins, DNA, RNA, and small molecules.
Main Results:
- Demonstrated click-ExM's applicability to a wide range of biomolecules (18 clickable labels).
- Showcased click-ExM's utility in both cell culture and tissue imaging.
- Confirmed compatibility of click-ExM with signal amplification and two-color imaging.
Conclusions:
- Click-ExM provides a convenient and versatile 'one-stop-shop' for nanoscale imaging.
- This method expands the scope of ExM to diverse biomolecules.
- Click-ExM is suitable for routine use in cell and tissue sample analysis.
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