Ex vivo reactivation upon post-mortem harvesting of TG is also observed, and virus reactivation in vivo can be induced through different means. The goal of this study was to achieve multimodal high spatial resolution imaging deep within an HSV-1-infected TG using signals intrinsically generated by, and generated within the tissue. TG contain many myelinated neurons of various sizes, whose myelin sheathes can generate a strong and specific CARS signal by virtue of their richness in symmetric CH2 vibrational modes. The neuron soma generates very little CARS signal, but is strongly autofluorescent and emits in the green spectrum close to that of the commonly used green fluorescent protein. Infection of cells can be detected using a virus expressing a fluorescent reporter gene. To date, the combination of CARS and multiphoton fluorescence microscopy in the context of a viral infection has been limited to the analysis of single monolayers of cells in culture. Herein, we describe for the first time the combined use of CARS microscopy and two-photon fluorescence microscopy to visualize infected cells and components of the surrounding tissue in an unfixed and unsectioned virus-infected tissue. For our pathogenesis studies of HSV-1, we produced and characterized a recombinant strain of HSV-1 that behaves like the wild-type parental strain KOS, and that stably expresses the second generation red fluorescent protein mCherry. This RFP was chosen due to its stability and the fact that it functions as a true monomer. Furthermore, due to the inherent autofluorescence of neural tissue in the green spectrum, the use of an RFP minimized background signal from uninfected cells in the TG. The cloning strategy chosen was designed so as not to disrupt any viral open Simetryn reading frames or regulatory regions of the surrounding genes. An mCherry expression cassette under control of a mammalian Ofloxacin promoter was inserted into the intergenic region between the ORFs of the HSV-1 Us7 and Us8 genes. To avoid disrupting the polyadenylation of transcripts coterminal with Us7, as well as to avoid interrupting the promoter of Us8, the Us7 and Us8 intergenic region was duplicated in tandem.