Stroboscopic opto-acoustic scattering (SOAS) flow cytometer for pre-cancerous detection

用于癌前检测的频闪光声散射 (SOAS) 流式细胞仪

• 批准号: BB/X003620/1
• 负责人: Brian Huntly
• 金额: $ 23.17万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX003620%2F1
• 关键词: Stroboscopic; opto; acoustic; scattering; SOAS

ProblemIt is now well described that most malignancies develop along a time-continuum that takes decades, acquiring the malignant phenotype and clonal dominance in a stepwise-manner due to the accumulation of multiple mutations. Early diagnosis and treatment of cancer invariably leads to better outcomes. Identification of these premalignant phases offers improved risk stratification, better deployment of early detection techniques, and possibly even prevention. However, early cellular changes are subtle, and occur in only a small subpopulation of cells, making detection of these targets challenging. We hypothesise that alterations in the physical characteristics of these pre- malignant cells will allow their detection through the use of advanced sensing techniques, and propose to develop this for blood cancers as an exemplar. "Liquid tumours" have the benefits of simple sample acquisition and a predefined premalignant state, so-called clonal haematopoiesis of indeterminate potential (CHIP). Currently, symptomatic patients are "screened" for cellular and biochemical abnormalities in the blood. Automated cell counters assess physical size through light scatter in addition to protein content properties across large numbers of cells. This allows quantitation of cell types/frequencies in comparison to population normal ranges. However, they derive no qualitative measures and are therefore unlikely to find our target premalignant cells. At the other end of the diagnostic scale, overt haematological malignancies are diagnosed using a complicated combination of techniques (Histology, FACS, NGS) that are costly (in time and equipment) and are not feasible for wider screening. Moreover, regarding FACS analysis, prior knowledge of differential protein expression between normal and premalignant cells would be required that is currently lacking. Due to the predicted infrequency of premalignant cells, analysis of sufficient sized cellular populations using these techniques would encounter issues of throughput that our proposed techniques will overcome. SolutionWe propose an innovative approach using fast and efficient quantitative identification and real-time multi-parametric characterisation of biophysical properties of suspended cell components: shape, density, elasticity and compressibility. The scalability of this approach allows the existing throughput limitation to be cost-effectively overcome for the first time. We propose combining acoustic standing gradient forces with novel stroboscopic opto-acoustic scattering tomography integrated onto a Lab-on-chip device. The new multi-parametric stroboscopic opto-acoustic sensors will be used to compare biophysical properties of different cell populations or sub-populations within heterogeneous samples. We have high confidence that we can achieve analysis rates of up to 60,000 cells/second. Such a rate would allow analysis of a 500 ul sample in under 2 minutes (2.25-5.5M cells). The unique ability that our proposed optical technique with acoustic fields offers to measure multiple cells in parallel over several length scales is the route to ultra-fast throughput. Providing information on the mechanical properties of the cellular population at single-cell levels lays the foundation of a new generation of opto-acoustic sensing. More importantly it immediately opens up new clinical opportunities for reducing cancer rates and could have impacts in other areas such as airborne particle analysis, water/soil microbial sensing and production-line industrial particle sensing.

问题现在已经很好地描述了，大多数恶性肿瘤的发展需要数十年的时间连续性，由于多种突变的积累，以逐步的方式获得恶性表型和克隆优势。癌症的早期诊断和治疗总是会带来更好的结果。这些癌前阶段的识别可以改善风险分层，更好地部署早期检测技术，甚至可能进行预防。然而，早期的细胞变化是微妙的，并且仅发生在一小部分细胞中，这使得检测这些目标具有挑战性。我们假设这些癌前细胞的物理特征的改变将允许通过使用先进的传感技术来检测它们，并建议将其作为血癌的范例。 “液体肿瘤”具有样本采集简单和预定义的癌前状态的优点，即所谓的不确定潜力克隆造血（CHIP）。 目前，对有症状的患者进行血液细胞和生化异常的“筛查”。除了大量细胞的蛋白质含量特性之外，自动细胞计数器还通过光散射评估物理尺寸。这允许与群体正常范围相比对细胞类型/频率进行定量。然而，他们没有得出定性测量结果，因此不太可能找到我们的目标癌前细胞。在诊断范围的另一端，明显的血液恶性肿瘤使用复杂的技术组合（组织学、FACS、NGS）进行诊断，这些技术成本高昂（时间和设备），并且不适合更广泛的筛查。此外，关于 FACS 分析，需要对正常细胞和癌前细胞之间差异蛋白表达的先验知识，而目前尚缺乏这些知识。由于预计癌前细胞的频率较低，使用这些技术对足够大小的细胞群进行分析将遇到我们提出的技术将克服的吞吐量问题。 解决方案我们提出了一种创新方法，利用快速有效的定量识别和实时多参数表征悬浮细胞成分的生物物理特性：形状、密度、弹性和可压缩性。这种方法的可扩展性使得现有的吞吐量限制首次得以经济高效地克服。我们建议将声学驻梯度力与集成到芯片实验室设备上的新型频闪光声散射断层扫描相结合。新型多参数频闪光声传感器将用于比较异质样品中不同细胞群或亚群的生物物理特性。我们非常有信心能够实现高达 60,000 个细胞/秒的分析速率。这样的速率可以在 2 分钟内分析 500 ul 样品（2.25-5.5M 细胞）。 我们提出的声场光学技术提供了在多个长度尺度上并行测量多个细胞的独特能力，这是实现超快吞吐量的途径。在单细胞水平上提供有关细胞群机械特性的信息，为新一代光声传感奠定了基础。更重要的是，它立即为降低癌症发病率开辟了新的临床机会，并可能对空气颗粒分析、水/土壤微生物传感和生产线工业颗粒传感等其他领域产生影响。