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中央设施和功能，实现了运营和研究效率以及规模经济，这些设施和功能为克里克的所有研究人员提供了尖端研究设备和实验室启用功能，如高通量筛查设施(HTS)。克里克冠状病毒监测单元(CCSU)是高通量筛查设施的一个分支，建立的目的是运行一条活动管道，提供高通量活病毒抗体中和分析，以测量患者血清同时防止数千个样本感染SARS-CoV-2的能力。这一独特的中和分析平台(在HTS设施中开发)对许多正在进行的研究至关重要，例如那些评估关注的变异对当前接种人群和临床易受感染人群的可能影响的研究。该管道能够在收到新的病毒样本后2-3周内产生针对新VOCs(如Delta和Omicron)的中和数据，并可在GCP级别每周处理数千个样本。来自这一管道的数据已经在Crick-Legacy研究的参与者(辉瑞和牛津-阿斯利康疫苗的接受者)中测量了针对Delta和Omicron变体的中和抗体滴度。这些数据与英国政府科学顾问委员会共享，有助于延长英国对大流行的限制，使更多人能够在2021年夏天接种第二剂疫苗，并成功开展加强免疫活动。报道这些发现的出版物已被引用近600次。这一渠道还使健康成年人的抗体效价与癌症患者(Crick-Capture研究)和肾透析患者(Crick-Naomi研究)的抗体效价进行了比较，为确定脆弱患者群体的优先顺序提供了重要数据。此外，关于合成单抗Sotrovimab的体外中和效果的数据与NHSE和首席医疗官分享，并支持对感染Omicron BA.1和BA.2的患者继续使用这种药物。在英国(可能在国际上)没有其他类似的管道能够进行这种规模的活病毒检测，并且由于近乎实时的报告，产生的数据仍然是英国和国际大流行准备和反应的重要工具。目前，中和管道通过使用多个HTS液体处理设备来加强，以确保输送的连续性。然而，初级成像(以及由此得出的测量结果)仅限于一台显微镜，即凤凰歌剧院。这种对一种成像设备的依赖是这条管道的弱点。这项提议是在这项检测中部署CELIGO扫描细胞仪，从而消除对需要保持运行的管道对一台机器的依赖。此外，在CELIGO上的采购速度快4-5倍，这意味着管道吞吐量和容量可以显著增加。在2020/21年度，Covid中和管道占HTS使用量的50%。让高端显微镜与一种类型的化验一起使用必然会限制其他用户的访问(目前衡量的是访问凤凰歌剧院的延迟数周)。通过将CCSU活动的主要成像平台转移到CELIGO，我们将极大地提高其他克里克研究人员使用高含量显微镜的可用性，从而改善超过20/21年使用HTS设施的100多名克里克研究人员的服务交付。CELIGO成像仪的提供还将有助于改进辐射生物学、干细胞生物学和一般细胞活力领域中需要分析(大规模)细胞群体的现有活动。此外，用户将有新的机会探索以前手动分析的自动化，例如流感空斑分析

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