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Correlative Light Electron Microscopy CLEM for Tracking and Imaging Viral Protein Associated Structures in Cryo-immobilized Cells
Published on: September 7, 2018
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Correlative light and electron microscopy enables viral replication studies at the ultrastructural level
Kirsi Hellström1, Helena Vihinen2, Katri Kallio1
1Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland.
Methods (San Diego, Calif.)
|April 29, 2015
Summary
Correlative light and electron microscopy (CLEM) identifies virus-infected cells and their replication sites. This technique reveals nanoscale membrane modifications crucial for positive-strand RNA virus replication.
Area of Science:
- Cell biology
- Virology
- Microscopy
Background:
- Electron microscopy (EM) studies cellular changes from infections or genetic modifications.
- Correlative light and electron microscopy (CLEM) aids in identifying specific cells for detailed EM analysis.
- Positive-strand RNA virus replication occurs at specific, nanoscale membrane sites.
Purpose of the Study:
- To apply CLEM for studying Semliki Forest virus (SFV) replication in cells.
- To develop methods for identifying infected cells and their replication sites using CLEM.
- To investigate virus-induced membrane modifications at the ultrastructural level.
Main Methods:
- Utilized CLEM to analyze cells transfected with viral components or infected with SFV.
- Developed a detectable RNA template for identifying cells with viral replication, even with lethal mutations.
- Employed fluorescently labeled viral proteins to locate replication-positive cells in mammalian and mosquito cells.
Main Results:
- Successfully identified SFV replication-positive cells using CLEM, even at low infection efficiencies.
- Localized viral replication to specific cellular regions characterized by extensive membrane modifications.
- Demonstrated the utility of a novel RNA template for studying viral protein function and replication.
Conclusions:
- CLEM is an effective technique for studying virus replication dynamics.
- CLEM enables visualization of virus-induced ultrastructural changes, particularly membrane alterations.
- The developed methods facilitate research on viral replication mechanisms and host-pathogen interactions.
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