Protocol for quantification of intercellular connection transmission in ebolavirus infections using ImageJ

Area of Science:

• Virology
• Cell Biology
• Microscopy

Background:

• Ebolaviruses are known to spread between adjacent cells via intercellular connections.
• Quantifying this intercellular spread is crucial for understanding viral pathogenesis and developing interventions.
• Previous methods faced challenges due to high ebolavirus accumulation at the plasma membrane, hindering accurate quantification.

Purpose of the Study:

• To develop and present a reliable protocol for quantifying ebolavirus intercellular spread.
• To address limitations of previous quantification methods caused by intense viral signals.

Main Methods:

• A detailed protocol involving cell plating and Bundibugyo virus infection.
• Application of a neutralizing antibody during the infection process.
• Quantitative microscopy assay utilizing ebolavirus immunodetection.
• Specific procedures to minimize bias from strong plasma membrane viral accumulation.

Main Results:

• The developed protocol effectively quantifies ebolavirus intercellular spread.
• The method minimizes the confounding effect of high fluorescence signals from viral accumulation.
• Successful application of immunofluorescence for ebolavirus detection and spread assessment.

Conclusions:

• This protocol offers a refined approach for measuring ebolavirus intercellular transmission.
• The method enhances the accuracy of viral spread quantification in cell-based assays.
• Provides a valuable tool for ebolavirus research, aiding in the study of viral dynamics and control.