title: Visualization of mouse neuronal ganglia infected by Herpes Simplex Virus 1 (HSV-1) using multimodal non-linear optical microscopy.
abstract: Herpes simplex virus 1 (HSV-1) is a neurotropic virus that causes skin lesions and goes on to enter a latent state in neurons of the trigeminal ganglia. Following stress, the virus may reactivate from latency leading to recurrent lesions. The in situ study of neuronal infections by HSV-1 is critical to understanding the mechanisms involved in the biology of this virus and how it causes disease; however, this normally requires fixation and sectioning of the target tissues followed by treatment with contrast agents to visualize key structures, which can lead to artifacts. To further our ability to study HSV-1 neuropathogenesis, we have generated a recombinant virus expressing a second generation red fluorescent protein (mCherry), which behaves like the parental virus in vivo. By optimizing the application of a multimodal non-linear optical microscopy platform, we have successfully visualized in unsectioned trigeminal ganglia of mice both infected cells by two-photon fluorescence microscopy, and myelinated axons of uninfected surrounding cells by coherent anti-Stokes Raman scattering (CARS) microscopy. These results represent the first report of CARS microscopy being combined with 2-photon fluorescence microscopy to visualize virus-infected cells deep within unsectioned explanted tissue, and demonstrate the application of multimodal non-linear optical microscopy for high spatial resolution biological imaging of tissues without the use of stains or fixatives.
authors.affiliation: INRS - Institut Armand-Frappier, Laval, Québec, Canada.
authors.forename: Pierre-Alexandre
authors.initials: PA
authors.lastname: Rochette
authors.affiliation: INRS - Centre Énergie Matériaux Télécommunications, Varennes, Québec, Canada.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Laliberté
authors.affiliation: INRS - Centre Énergie Matériaux Télécommunications, Varennes, Québec, Canada.
authors.forename: Antony
authors.initials: A
authors.lastname: Bertrand-Grenier
authors.affiliation: INRS - Centre Énergie Matériaux Télécommunications, Varennes, Québec, Canada.
authors.forename: Marie-Andrée
authors.initials: MA
authors.lastname: Houle
authors.affiliation: INRS - Centre Énergie Matériaux Télécommunications, Varennes, Québec, Canada.
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: Blache
authors.affiliation: INRS - Centre Énergie Matériaux Télécommunications, Varennes, Québec, Canada.
authors.forename: François
authors.initials: F
authors.lastname: Légaré
authors.affiliation: INRS - Institut Armand-Frappier, Laval, Québec, Canada.
authors.forename: Angela
authors.initials: A
authors.lastname: Pearson
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002522:Chlorocebus aethiops; D006561:Herpes Simplex / Q000473:pathology*; D018259:Herpesvirus 1, Human / Q000502:physiology*; D051379:Mice; D008853:Microscopy; D008856:Microscopy, Fluorescence; D012668:Trigeminal Ganglion / Q000821:virology*; D014709:Vero Cells; D014779:Virus Replication / Q000502:physiology