解析病毒的复制周期是病毒学研究中一个重要目标和难点，具体包括从病毒入侵到病毒释放这一复杂过程中，病毒在时间和空间上与宿主细胞的相互作用。而传统静止的研究方法很难满足对病毒感染过程中与宿主相互作用的时空效应问题研究需要。为了解决这一难点问题，本研究将使用微流控芯片技术以及Halotag标记的方法，利用荧光标记动态成像技术，实现PRV感染神经元细胞时，从最初的膜融合阶段开始，一直到病毒基因组从核衣壳内释放阶段的单颗粒病毒的全程动态示踪。由于Halotag能高效特异性结合不同荧光的配体，我们能区分亲代与子代病毒，完成完整病毒生命周期的研究，从时空效应上阐明PRV入侵细胞的动态规律与致病机制。我们还将使用微流控芯片研究PRV在轴突顺向或逆向转运过程中与宿主细胞亚结构和细胞器的相互作用。本研究不仅能为α疱疹病毒侵染神经细胞的分子机制提供理论依据，也为在动态水平研究其他病毒致病机制提供新的技术平台。

One of the most pivotal and difficult fields in virology research is deciphering the complex process of virus replication cycle from viral entry to the egress of viral progeny particles.In a word, viral infections include many dynamic steps, featuring spatio-temporal interactions of virus particles with cellular structures, which make it impossible to dissect the detailed molecular mechanisms using traditional and stationary approaches. To this end, combined with dynamic fluorescent labeling visualization technique, this study will apply Halotag to label PRV and realize single particle tracking of PRV infection cycle in neurons, elucidating dynamics from steps of virus-host membrane fusion to viral DNA genome releasing from viral capsid.In addtion, the divisity of halotag ligands enable us to label parental and progeny virus with fluorescent dyes of diffrent emission wavelengths and study the complete life cycle of PRV in neurons. We will also focus on the temporal and spacial interations between PRV particle and host sub-cellular structures and organelles during the process of retrograde and anterograde axonal transport, which will shed light on fundamental issues in viral entry and pathogenesis mechanisms of PRV infection. This study will not only provide theoretical basis for the molecular mechanism of the herpes virus infection on neuron cells, but also provide a new platform for the study of other viruses pathogenesis in the dynamic level.