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Imaging VIPER-labeled Cellular Proteins by Correlative Light and Electron Microscopy
Julia K Doh1, Young Hwan Chang1,2, Caroline A Enns3
1Department of Biomedical Engineering, Oregon Health & Science University, Portland, Oregon 97239, USA.
Bio-Protocol
|March 3, 2021
Summary
Researchers developed Versatile Interacting Peptide (VIP) tags, including VIPER, to overcome limitations in imaging cellular proteins. These genetically-encoded tags enable simultaneous fluorescence and electron microscopy for advanced cell biology research.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Molecular Imaging
Background:
- Fluorescence microscopy (FM), electron microscopy (EM), and correlative light and EM (CLEM) enable detailed study of cellular proteins and nanostructures.
- Current limitations exist in tagging and tracking proteins across scales, particularly for EM compatibility.
- A scarcity of suitable genetic tags hinders comprehensive protein visualization and quantification.
Purpose of the Study:
- To introduce Versatile Interacting Peptide (VIP) tags as a solution for multi-modal protein imaging.
- To present VIPER, a specific VIP tag, for labeling proteins for both FM and EM.
- To provide a protocol for using VIPER in CLEM for cell-surface receptor imaging.
Main Methods:
- Development of genetically-encoded Versatile Interacting Peptide (VIP) tags.
- Implementation of VIPER for labeling cellular proteins with fluorophores (FM) or electron-dense nanoparticles (EM).
- Application of VIPER in a correlative light and electron microscopy (CLEM) protocol for cell-surface receptors.
Main Results:
- VIPER enables robust labeling of target proteins for high-resolution imaging.
- Successful application of VIPER for imaging a cell-surface receptor using CLEM.
- Demonstration of VIP tags' versatility for bridging fluorescence and electron microscopy.
Conclusions:
- VIPER tags overcome existing limitations in genetic tagging for multi-modal microscopy.
- This protocol facilitates advanced imaging of protein spatial organization in cell biology.
- VIP tags represent a significant advancement for correlative imaging studies.
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