High-throughput digital microplate microscopy reader for the study of cellular responses to infection and stress

病毒需要活细胞才能繁殖。当这样做时，病毒会引起被感染细胞的完全重组，总体上允许产生数千个新的病毒颗粒，但也会抑制宿主抵抗感染的能力。对这种复杂相互作用的关键见解可以通过成像活细胞和病毒获得，这只有使用显微镜才能实现。然而，大多数显微镜一次只能对几个细胞成像，这使得从具有代表性的细胞数量中获取信息变得非常费时费力。此外，细胞有自己的生命周期，即使在同一个培养皿中，细胞之间也存在差异。克服这个问题的一种方法是使用高通量显微镜，能够对数千个细胞进行成像，并对单个细胞的特性进行分析。这一提议涉及为萨里大学病毒学部门购买一套这样的成像系统，该部门是由来自6个不同小组的150名研究人员组成的，他们研究人类病毒及其与人类细胞的相互作用。我们打算获得的仪器将使我们能够追踪病毒在细胞内的位置以及它们如何操纵细胞内环境。我们将能够可视化病毒的特征，如基因表达和复制工厂，以及细胞对感染的反应，以压力颗粒的形式，线粒体重组和细胞核形态。更重要的是，我们将能够在特定条件下在数百个单个细胞中定性和定量地测量这些事件，在短时间内生成高质量的可重复数据。例如，我们最近发现痘病毒（包括新出现的猴痘病毒）中表达的一种病毒蛋白可诱导线粒体（细胞的能量工厂）聚集。组装和拆卸的动力学以及这些簇在不同条件下的大小和形态仍然未知，但可以用高通量多模式显微镜单元来阐明。同样，我们最近发现，感染单纯疱疹病毒的细胞不能从细胞核中输出它们的mrna，从而导致细胞功能的封锁，从而允许有效的病毒感染。这一过程背后的机制尚不清楚，但可以通过配备点位计数模块的成像系统进行研究。总的来说，所要求的仪器不仅将提高我们进行突破性研究的能力，而且还将提供目前其他设备无法提供的关于病毒感染的新视角。

Viruses require living cells to multiply. When doing so, viruses cause a complete re-organisation of the infected cell, overall allowing the production of thousands of new viral particles but also suppressing the ability of the host to fight infection. Crucial insights into this complex interplay can be obtained imaging living cells and viruses, which is only possible using microscopes. Most microscopes are, however, able to image a few cells at a time making it very lengthy and laborious to obtain information from a representative number of cells. In addition, cells undergo their own life cycle and variations exist between them even in the same petri dish. A way to overcome this problem is the use of high-throughput microscopes able to image thousands of cells and analyse their properties on an individual basis. This proposal concerns the acquisition of one of such imaging systems for the Section of Virology at the University of Surrey, a unit of research formed by >20 investigators from 6 different groups studying human viruses and their interplay with human cells. The instrument we propose to acquire will enable us to track where viruses go inside a cell and how they manipulate the intracellular environment. We will be able to visualise viral features such as gene expression and replication factories, as well as the cell's response to infection in the form of stress granules, mitochondrial reorganisation and cell and nucleus morphology. More importantly, we will be able to measure these events qualitatively and quantitatively in hundreds of individual cells in specified conditions, generating high quality reproducible data in a short period of time. As examples, we have recently discovered a viral protein expressed in poxviruses (including the emerging monkeypox virus) that induces the clustering of mitochondria (the energy factories of the cell). The kinetics of assembly and disassembly as well as the size and morphology of these clusters in different conditions remains unknown but can be elucidated with a high-throughput multimode microscopy unit. Similarly, we have recently discovered that cells infected with herpes simplex virus fail to export their mRNAs from the nucleus, causing a blockade of cellular functions that allows efficient virus infection. The mechanisms behind this process are unclear but can be researched with an imaging system equipped with a spot counting module. Altogether the requested instrument will not only increase our capacity to deliver ground-breaking research, but also provide novel perspectives on viral infection that are at present not possible with other equipment.