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

Year: 2024
Type: article
Abstract: Previous work demonstrates that ebolaviruses can spread to neighboring cells through intercellular connections. Here, we present a protocol to quantify the intercellular spread of ebolaviruses via immunofluorescence. We describe steps for cell plating, Bundibugyo virus infection, and adding a neutralizing antibody. We detail procedures for quantitative microscopy assay using ebolavirus immunodetection. Strong virus accumulation around the plasma membrane leads to high fluorescence signal preventing quantification of viral spread based on signal intensity. This protocol minimizes the impact of this bias. For complete details on the use and execution of this protocol, please refer to Santos et al. 1 • Ebolavirus infection and neutralizing antibody treatment in Huh7 cells • Immunofluorescence staining to quantify virus cell-to-cell transmission • Quantifying ebolavirus cell-to-cell transmission using ImageJ software Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Previous work demonstrates that ebolaviruses can spread to neighboring cells through intercellular connections. Here, we present a protocol to quantify the intercellular spread of ebolaviruses via immunofluorescence. We describe steps for cell plating, Bundibugyo virus infection, and adding a neutralizing antibody. We detail procedures for quantitative microscopy assay using ebolavirus immunodetection. Strong virus accumulation around the plasma membrane leads to high fluorescence signal preventing quantification of viral spread based on signal intensity. This protocol minimizes the impact of this bias.
DOI: 10.1016/j.xpro.2024.103363
Language: English
Field: Medicine
Open Access status: gold
Estimated APC paid: $1,500
OpenAlex ID: W4403148381
FWCI: 0.316
Cites: 12
Cited by: 1
Related to: 10