Correlative Light Electron Microscopy (CLEM) for Tracking and Imaging Viral Protein Associated Structures in Cryo-immobilized Cells

作者： Rachel Santarella-Mellwig1, Uta Haselmann2, Nicole L. Schieber1, Paul Walther3, Yannick Schwab1, Claude Antony1, Ralf Bartenschlager2,4, Inés Romero-Brey2 1European Molecular Biology Laboratory, 2Department of Infectious Diseases, Molecular Virology,Heidelberg University, 3Central Facility for Electron Microscopy,Ulm University, 4Heidelberg Partner Site,German Center for Infection Research

简介：

Overview

This article presents a correlative light electron microscopy (CLEM) method to visualize virus-induced intracellular structures. By integrating light microscopy (LM) and electron microscopy (EM), researchers can gain insights into virus-host interactions.

Key Study Components

Area of Science

• Cell Biology
• Microscopy Techniques
• Virus-Host Interactions

Background

• The method combines live cell imaging with electron microscopy.
• High pressure freezing is a critical component of the technique.
• It has been successfully applied to study virus interactions.
• The method can be generalized to various applications in cell biology.

Purpose of Study

• To visualize virus-induced structures in cells.
• To enhance understanding of bio-protein interactions in cellular membranes.
• To provide a practical guide for researchers using this method.

Methods Used

• Preparation of patterned sapphire discs for imaging.
• Use of inverted microscopy for pattern confirmation.
• Sterilization of discs using ultraviolet crosslinking.
• Cell culture techniques for seeding cells onto sapphire discs.

Main Results

• Successful visualization of virus-induced intracellular structures.
• Demonstration of the effectiveness of CLEM in studying virus-host interactions.
• Practical steps outlined for implementing the method.
• Potential applications in broader cell biology contexts.

Conclusions

• The CLEM method provides valuable insights into virus biology.
• It bridges the gap between light and electron microscopy.
• Further applications in cell biology are encouraged.

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