Immunofluorescence Assays to Visualize Tick-Borne Flaviviruses in Mouse Skin or Brain

Rodrigo I Santos; Meghan E Hermance

doi:10.1007/978-1-0716-4587-1_10

Immunofluorescence Assays to Visualize Tick-Borne Flaviviruses in Mouse Skin or Brain

Methods Mol Biol. 2025:2936:85-97. doi: 10.1007/978-1-0716-4587-1_10.

Authors

Rodrigo I Santos¹, Meghan E Hermance²

Affiliations

¹ Jerry M. Wallace School of Osteopathic Medicine, Campbell University, Buies Creek, NC, USA.
² Frederick P. Whiddon College of Medicine, University of South Alabama, Mobile, AL, USA. mhermance@southalabama.edu.

PMID: 40512312
DOI: 10.1007/978-1-0716-4587-1_10

Abstract

Multiplex immunofluorescence (IF) utilizes a combination of different fluorophore-linked antibodies to facilitate the simultaneous observation of multiple antigenic epitopes within a tissue section. In this chapter, we describe how tick-borne flaviviruses can be visualized in mouse skin biopsies harvested from the tick feeding site or in mouse brain tissue. Using antibodies specific to the North American tick-borne flavivirus, Powassan virus (POWV), and to Mus musculus cell markers, IF multiplex labeling experiments allow us to visualize how POWV antigen co-localizes with host cell markers. Such IF multiplexing in mouse skin and brain tissue suggests which host cells are targets of viral infection.

Keywords: Autofluorescence; Brain; Immunofluorescence; Indirect immunofluorescence; Skin biopsy; Tick-borne flavivirus; Tick–host cutaneous interface.

MeSH terms

Animals
Brain* / metabolism
Brain* / virology
Encephalitis Viruses, Tick-Borne* / immunology
Encephalitis Viruses, Tick-Borne* / isolation & purification
Encephalitis, Tick-Borne* / virology
Flavivirus*
Fluorescent Antibody Technique* / methods
Mice
Skin* / pathology
Skin* / virology