Shannon John P, Kamenyeva Olena, Reynoso Glennys V, Hickman Heather D
Viral Immunity and Pathogenesis Unit, Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Biological Imaging Section, Research Technology Branch, NIAID, NIH, Bethesda, MD, USA.
Methods Mol Biol. 2019;2023:301-311. doi: 10.1007/978-1-4939-9593-6_19.
Intravital multiphoton microscopy (MPM) allows the direct visualization of viral infections in real time as they occur in living animals. Here we describe the routes and considerations for murine infection with vaccinia virus (VACV) for imaging, and the preparation of the skin and inner lip (labial mucosa) of infected animals for MPM. Using different recombinant VACVs expressing fluorescent proteins in combination with transgenic fluorescent reporter mice, MPM imaging can be used to examine the movements, interactions, and functions of virus-infected cells or selected immune cell populations after infection.
活体多光子显微镜(MPM)能够实时直接观察病毒感染在活体动物体内发生的情况。在此,我们描述用于成像的痘苗病毒(VACV)感染小鼠的途径和注意事项,以及用于MPM的感染动物皮肤和内唇(唇黏膜)的制备方法。使用表达荧光蛋白的不同重组VACV与转基因荧光报告小鼠相结合,MPM成像可用于检查感染后病毒感染细胞或选定免疫细胞群体的运动、相互作用和功能。