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: 8707576
• 负责人: Henry Tse
• 金额: $ 19.97万
• 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 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8707576
• 关键词: Acute; Address; Algorithmic Analysis; 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使用细胞的机械特性作为无标记生物标志物的兴趣越来越大。增加的单细胞变形性已经与例如细胞系和患者样品中的恶性肿瘤相关。与传统的分子生物标志物相比，机械性质是感兴趣的细胞固有的，并且不需要大量的样品制备或昂贵的标记。尽管如此，在这一领域的临床应用已被限制在研究设置为以前的工具缺乏有效地检查复杂的临床样本的异质性的吞吐量。拟议的工作建立在变形性细胞仪平台上，该平台采用惯性微流体原理来创建流体剪切延伸流动连接，该连接可以分析每秒超过1000个细胞的机械特性。在研究平台阶段，变形性细胞术系统在识别胸水中的恶性细胞方面显示出非常有前途的临床诊断实用性，其灵敏度比金标准细胞学分析高得多。在这里，我们建立了这一初步的成功，并解决两个主要的挑战，创造一个商业上可行的高通量机械表型平台。我们的目标是降低与高速相机读出相关的成本，并减少实现近实时快速决策分析、更快的数据周转时间和下游分选能力所需的计算资源。具体而言，本提案的目的是探索成本较低的单点询问模式和高速成像替代方案的可行性，同时保持类似的诊断性能。单点询问方法包括与变形后的细胞相比，在机械施加应力之前变形前的细胞的渡越时间。或者，类似于侧向散射流式细胞术测量，变形事件期间散射光的单点测量将与变形细胞形状相关。这两种方法都有望减少计算资源，因为输出数据是1D的。还将评估三种替代的高速成像技术：1）位置敏感检测器（PSD）是单轴成像传感器-通过正交放置两个PSD，可以在变形事件期间收集2D空间数据，2）市售OEM高速图像传感器。可定制，其中板载FPGA集成可用于预筛选数据，以减少离线图像分析工作量，3）适配的较低帧率相机（10，000 fps）耦合到500 ns频闪源。任何提出的替代采集方法都将降低仪器成本并缩短分析时间，以创建临床相关的无标签机械生物标记物诊断产品。