Imaging Flow Cytometry and Confocal Immunofluorescence Microscopy of Virus-Host Cell Interactions

Abstract

Viruses are diverse pathogens that use host factors to enter cells and cause disease. Imaging the entry and replication phases of viruses and their interactions with host factors is key to fully understanding viral infections. This review will discuss how confocal microscopy and imaging flow cytometry are used to investigate virus entry and replication mechanisms in fixed and live cells. Quantification of viral images and the use of cryo-electron microscopy to gather structural information of viruses is also explored. Using imaging to understand how viruses replicate and interact with host factors, we gain insight into cellular processes and identify novel targets to develop antiviral therapeutics and vaccines.

Opportunities in optical and electrical single-cell technologies to study microbial ecosystems

“…The main limitation of image flow cytometry is that large amounts of data are generated in minimal amounts of time and the cell images produced by image flow cytometry are much more complex to analyze. Successful applications of this technique can be found in the field of virology and more specific in the study towards virus-host interactions (Han et al, 2016;McClelland et al, 2021). Sorting realtime deformability cytometry is a microfluidics technique that uses brightfield images to sort out cells with clear morphological differences.…”

Viral-Host Dependency Factors as Therapeutic Targets to Overcome Antiviral Drug-Resistance: A Focus on Innate Immune Modulation

“…Advanced imaging methodologies, including improved highresolution light microscopy, confocal microscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM) or cryo-EM, among others, take advantage of new optical imaging tools to perform spatio-temporal assessment of the viral replication cycle, for a better understanding of viral structures and interactions within the cell between host proteins and viral counterparts. These techniques have been extensively reviewed elsewhere (56)(57)(58)(59)(60)(61).…”

“…Among them, it is worth to mention Imaging Flow cytometry (IFC), a technique which merges flow cytometry and fluorescence microscopy, enabling a high-throughput analysis of cell morphology besides to a multi-channel fluorescence imaging of single cell (see (57) for review). Applied to virology, ICF is useful to study subtle virus-host interactions, such as viral entry interactions, tracking intracellular distribution of viral proteins or nucleic acids, with a high resolution.…”