Robust High-throughput Mechanical Phenotyping Platform for Clinical Diagnostics

用于临床诊断的强大高通量机械表型平台

• 批准号: 8592948
• 负责人: Henry Tse
• 金额: $ 20万
• 依托单位: CYTOVALE, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8592948
• 关键词: Acute; Address; Area; Benign; Biological Assay; Biological Markers; Biology; Biopsy; Body Fluids; Cell Line; Cell Shape; Cells; Characteristics; Chest; Chronic; Clinical; Clinical Research; Complement; Complex; Computer Analysis; Coupled; Cytology; Cytometry; Data; Data Set; Decision Analysis; Diagnosis; Diagnostic; Diagnostic Sensitivity; Disease; Engineering; Event; Flow Cytometry; Future; Goals; Gold; Health Care Costs; Healthcare; Heterogeneity; Image; Image Analysis; Imaging Techniques; Immune; Immunohistochemistry; In Vitro; Individual; Inflammation; Inflammatory; Label; Lead; Leukocytes; Liquid substance; Low Prevalence; Lung; Malignant - descriptor; Malignant Neoplasms; Malignant Pleural Effusion; Masks; Measurement; Measures; Mechanics; Medicine; Mesothelial Cell; Methods; Microfluidics; Modality; Molecular; Monitor; Neoplasm Metastasis; Outcome; Output; Patients; Performance; Phase; Phenotype; Pleural; Pleural cavity; Pleural effusion disorder; Population; Population Heterogeneity; Positioning Attribute; Preparation; Procedures; Process; Property; Recording of previous events; Research; Risk; Sampling; Side; Sorting - Cell Movement; Source; Specificity; Speed; Staging; Stress; System; Techniques; Thoracentesis; Time; Work; Workload; analog; base; cancer cell; care burden; cell type; clinical application; clinically relevant; computing resources; cost; cost effective; data acquisition; detector; diagnostic accuracy; granulocyte; high risk; improved; instrument; interest; nephelometry; neutrophil; novel diagnostics; pressure; prototype; public health relevance; screening; sensor; success; tool

DESCRIPTION (provided by applicant): There has been increased interest in using mechanical properties of cells as label-free biomarkers for applications in basic biology and clinical diagnostics. Increased single-cell deformability has been correlated with malignancy in cell lines and patient samples for example. In contrast to conventional molecular biomarkers a mechanical property is intrinsic to the cells of interest and does not require extensive sample preparation or costly labels. Still, clinical applications in this field have been limited to the research setting as previous tools have lacked the throughput to examine the heterogeneity of complex clinical samples effectively. The proposed work builds upon the deformability cytometry platform, which employs inertial microfluidic principles to create a fluid shearing extensional flo junction that can analyze mechanical properties of over 1000 cells per second. At the research platform stage, the deformability cytometry system has shown very promising clinical diagnostic utility in identifying malignant cells in pleural fluid with much higher sensitivity than the gold standard cytological analysis. Here we build off this initial success and address two major challenges to create a commercially viable high- throughput mechanical phenotyping platform. We aim to reduce costs associated with high-speed camera readout, and decrease computational resources required to achieve near-real-time-rapid-decision analysis, faster data turnaround time and downstream sorting capabilities. Specifically, the aims of this proposal are to explore the feasibility of lower cost single-point interrogation modalities and high-speed imaging alternatives while retaining similar diagnostic performance. Single-point interrogation methods include transit-time of a pre- deformed cell prior to mechanically applied stress compared to post-deformed cells. Alternatively, single-point measurements of scattered light during the deformation events, similar to side-scatter flow cytometry measurements will be correlated to deformed cell shape. Both these approaches are expected to reduce computational resources as output data is 1D. Three alternative high-speed imaging techniques will also be evaluated 1) Position sensitive detectors (PSDs) are one axis imaging sensors - by placing two PSDs orthogonally 2D spatial data can be collected during deformation events, 2) Commercially available OEM high- speed image sensors. are customizable where onboard FPGA integration can be used to prescreen data in order to reduce offline image analysis workload, 3) adapted lower frame rate camera (10,000 fps) coupled to 500ns strobe source. Any of the proposed alternative acquisition methods will reduce the cost of the instrument and lower analysis time to create a clinically relevant label-free mechanical biomarker-based diagnostic product.

描述(由申请人提供)：在基础生物学和临床诊断中应用细胞的机械性能作为无标记生物标记物的兴趣日益增加。例如，单细胞变形性的增加与细胞系和患者样本中的恶性肿瘤有关。与传统的分子生物标记物相比，机械特性是感兴趣的细胞固有的，不需要大量的样品制备或昂贵的标记。然而，由于以前的工具缺乏有效地检查复杂临床样本的异质性的吞吐量，该领域的临床应用一直局限于研究环境。这项拟议的工作建立在可变形细胞仪平台的基础上，该平台利用惯性微流控原理创建了一个流体剪切延伸型Flo结，可以每秒分析超过1000个细胞的力学性质。在研究平台阶段，变形性细胞学系统在胸水中识别恶性细胞方面显示出非常有前途的临床诊断实用价值，其灵敏度远远高于金标准细胞学分析。在这里，我们在这一初步成功的基础上，解决两个主要挑战，以创建一个商业上可行的高通量机械表型鉴定平台。我们的目标是降低与高速摄像机读数相关的成本，并减少实现近乎实时的快速决策分析、更快的数据周转时间和下游分类能力所需的计算资源。具体地说，这项提议的目的是探索在保持类似诊断性能的同时，采用成本较低的单点审讯模式和高速成像替代方案的可行性。单点询问方法包括与变形后单元相比，预变形单元在机械施加应力之前的渡越时间。或者，变形事件期间的散射光的单点测量，类似于侧向散射流式细胞术测量，将与变形的细胞形状相关联。由于输出数据是一维的，因此这两种方法都有望减少计算资源。还将评估三种替代的高速成像技术1)位置敏感探测器(PSD)是一轴成像传感器-通过将两个PSD垂直放置，可以在变形事件期间收集2D空间数据，2)商业OEM高速图像传感器。都是可定制的，其中可以使用板载FPGA集成来预筛选数据，以减少离线图像分析工作量，3)采用耦合到500 ns频闪源的较低帧速率摄像头(10,000 fps)。任何拟议的替代采集方法都将降低仪器的成本和分析时间，以创建一种临床相关的基于机械生物标记物的诊断产品。