A Celigo Whole Well Imaging Cytometer for the Crick Covid neutralisation pipeline

克里克通过被称为科学技术平台的集中设施和功能实现了运营和研究效率和规模经济，这些设施和功能为克里克的所有研究人员提供了最先进的研究设备和实验室功能，如高通量筛选设施（HTS）。克里克Covid监测单位（CCSU）是HTS设施的一个分支，旨在运行一个活动管道，提供高通量活病毒抗体中和试验，以测量患者血清同时在数千个样本中预防SARS-CoV-2细胞感染的能力。这种独特的中和试验平台（在HTS设施中开发）对许多正在进行的研究至关重要，例如评估关注变异体对当前接种疫苗人群和临床易感人群的可能影响。该管道能够在收到新病毒样本后2-3周内生成针对新挥发性有机化合物（例如Delta和Omicron）的中和数据，并且每周可以处理数千个GCP水平的样本。来自该管道的数据测量了克里克- legacy研究参与者（辉瑞和牛津-阿斯利康疫苗的接受者）针对Delta和Omicron变体的中和抗体滴度。这些数据与英国政府科学咨询委员会共享，有助于延长英国的大流行限制，使更多的人能够在2021年夏季接种第二剂疫苗，并成功开展加强运动。报道这些发现的出版物被引用了近600次。该管道还可以将健康成人的抗体滴度与癌症患者（克里克- capture研究）和肾透析患者（克里克- naomi研究）的抗体滴度进行比较，为弱势患者群体的优先级提供重要数据。此外，合成单克隆抗体Sotrovimab体外中和效果的数据已与国家卫生和社会保障局和首席医疗官共享，并支持继续使用该药物治疗感染了Omicron BA.1和BA.2的患者。在英国（可能是国际上），没有其他类似的管道能够进行这种规模的活病毒分析，并且具有近乎实时的报告，所产生的数据仍然是英国和国际大流行防范和应对的重要工具。目前，中和管道通过使用多个HTS液体处理装置来确保输送的连续性，从而使管道变得坚固。然而，主要成像（以及由此产生的测量）仅限于一种显微镜，即Opera Phenix。这种对单一成像设备的依赖是该管道的一个弱点。该建议是在该试验中部署Celigo扫描细胞仪，从而消除对一台机器的依赖，以维持流水线的运行。此外，Celigo的采集速度快了4-5倍，这意味着管道的吞吐量和产能可以显著提高。2020/21年期间，Covid中和管道占HTS使用量的50%。使用高端显微镜进行一种类型的检测必然会限制其他用户的访问（目前使用Opera Phenix需要延迟数周）。通过将CCSU活动的主要成像平台转移到Celigo，我们将大大增加其他克里克研究人员使用高含量显微镜的可用性，从而改善为超过100名克里克研究人员提供的服务，这些研究人员在2011 /21年使用了HTS设备。Celigo成像仪的可用性还将改进辐射生物学、干细胞生物学和一般细胞活力领域中需要分析细胞群（大规模）的现有活动。此外，用户将有新的机会探索以前手工分析的自动化，例如流感斑块分析

The Crick achieves operational and research efficiencies and economies-of-scale through centralised facilities and functions, known as Science Technology Platforms, that provide all researchers at the Crick, irrespective of affiliation, with access to cutting-edge equipment for research and laboratory enabling functions such as the High Throughput Screening facility (HTS).The Crick Covid Surveillance unit (CCSU) is an offshoot of the HTS facility and was established to run a pipeline of activity delivering a high throughput live-virus antibody neutralisation assay to measure the ability of patient serum to prevent cell infection by SARS-CoV-2 across thousands of samples simultaneously. This unique neutralisation assay platform (developed in the HTS facility) is essential to many ongoing research studies, for example those assessing the likely impact of Variants of Concern on current vaccinated populations and in clinically vulnerable cohorts. The pipeline is able to generate neutralization data against new VOCs (e.g. Delta and Omicron) within 2-3 weeks upon receipt of a novel virus sample and can process thousands of samples per week at GCP level. Data from this pipeline has measured neutralising antibody titres against Delta and Omicron variants in participants in the Crick-Legacy study (recipients of both the Pfizer and Oxford-AstraZeneca vaccines). These data are shared with UK government scientific advisory boards, contributing to the extension of UK pandemic restrictions to allow more people to receive a second vaccine dose in summer 2021 and the successful booster campaign. Publications reporting these findings have been cited nearly 600 times. This pipeline also enabled the comparison of antibody titres in healthy adults with those of both cancer patients (Crick-CAPTURE study) and kidney dialysis patients (Crick-NAOMI study), providing important data on prioritisation of vulnerable patient groups. In addition, data on the in vitro neutralising efficacy of the synthetic monoclonal antibody, Sotrovimab was shared with NHSE and the Chief Medical Officer and supported the ongoing use of this drug for patients infected with Omicron BA.1 and BA.2. There are no other similar pipelines capable of this scale of live virus assay in the UK (possibly internationally) and with near real time reporting, the data produced remains a vital tool in the UK and international pandemic preparedness and response.Currently, the neutralisation pipeline is made robust by using multiple HTS liquid handling devices to ensure continuity of delivery. However, the primary imaging (and measurements derived from them) is restricted to one microscope, the Opera Phenix. This reliance on one imaging device is a weak point for this pipeline. This proposal is to deploy the Celigo scanning cytometer in this assay and thereby remove the dependence on one machine for a pipeline that needs to remain operational. Moreover, acquisition on the Celigo is 4-5 times faster meaning that pipeline throughput and capacity can be significantly increased.The Covid neutralisation pipeline accounted for 50% of HTS usage over 2020/21. Having a high-end microscope occupied with one type of assay necessarily restricts access by other users (currently measured in weeks of delay for access to the Opera Phenix). By moving the primary imaging platform for the CCSU activity to the Celigo, we would greatly increase the availability of high content microscopy to other Crick researchers and thereby improve service delivery for more than 100 Crick researchers who used the HTS facility over 20/21. The availability of a Celigo imager will also enable improvements to existing activities requiring analysis of cell populations (at scale) in the areas of radiation biology, stem cell biology and general cell viability. In addition, there will be new opportunities for users to explore automation of previously manual assays e.g. Influenza plaque assays