EAGER: Mobile-phone based single molecule imaging of DNA and length quantification to analyze copy-number variations in genome

主要研究者：厄兹坎，艾多安加州大学洛杉矶分校提案编号：1444240标题：EAGER：基于手机的DNA单分子成像和长度量化，以分析基因组中的拷贝数变化本提案的目的是创建一个变革性的荧光显微镜系统，该系统与下一代手机集成，用于成像单DNA分子。这种在智能手机上运行的现场便携式成像接口将具有在大视场内对单分子DNA片段进行成像的灵敏度和对比度。在最先进的手机上展示DNA成像将成为下一代移动的微观分析、传感和诊断工具的垫脚石，并可能导致在智能手机上进行单分子DNA测序。拟议的设计将有能力广泛应用于各种临床应用，包括癌症的早期检测（例如胃和脑）、神经系统疾病和感染性疾病的耐药性。这个基于手机的单分子成像、DNA平台还可以帮助医疗保健专业人员、流行病学家和政策制定者跟踪新兴趋势，并揭示更多的因果关系。知识描述：单分子成像和DNA长度定量，这两项目前都不适合使用基于手机的成像系统;需要极高的检测灵敏度、信噪比（SNR）、空间分辨率和自动化样品处理和处理界面。为了为这些重要任务提供变革性的解决方案，PI将设计一种安装在智能手机上的多功能便携式成像设备，该设备将允许在同一光机械附件内进行样品制备和单分子成像。智能手机上的这种荧光显微镜将通过将激光二极管、一次性纳米通道芯片、外部透镜和基于薄膜的发射滤光片集成在由3D打印技术创建的坚固附件中来设计。高SNR荧光信号检测将通过实施高角度/倾斜照射来实现，使得直接激发光束将不会进入低NA收集透镜。他们还将开发一种基于压缩采样的DNA长度估计方法，该方法将利用（i）移动电话上荧光显微镜的测量点扩散函数;（ii）物体的空间稀疏性（荧光标记的DNA分子）;以及（iii）在视场内拉伸的DNA分子的线性，作为-先验约束来估计感兴趣的DNA片段的长度，其准确度显著优于其初始成像系统的分辨率。

PI: Ozcan, AydoganInstitution: University of California-Los AngelesProposal number: 1444240Title: EAGER: Mobile-phone based single molecule imaging of DNA and length quantification to analyze copy-number variations in genomeThe aim of this proposal is to create a transformative fluorescent microscopy system that is integrated with next generation mobile-phones for imaging single DNA molecules. This field-portable imaging interface running on a smartphone will have the sensitivity and contrast to image single molecule DNA fragments over a large field of view. Demonstrating DNA imaging on a state-of-the-art mobile-phone would serve as a stepping stone to next-generation mobile micro-analysis, sensing and diagnostic tools and could lead to single molecule DNA sequencing on a smartphone.The proposed design will have the capability to be broadly used in various clinical applications including early detection of cancers (e.g. stomach and brain), nervous system disorders and drug resistance in infectious diseases. This cellphone based single molecule imaging, DNA platform could also assist health-care professionals, epidemiologists and policy makers to track emerging trends and shed more light on cause-effect relationships.Intellectual Description:Single molecule imaging and DNA length quantification, both of which are currently not feasible using mobile-phone based imaging systems; require extreme detection sensitivity, signal-to-noise ratio (SNR), spatial resolution and automated sample handling and processing interfaces. To provide a transformative solution to these important tasks, the PI will design a multifunctional portable imaging device installed on a smartphone which will allow sample preparation and single molecule imaging within the same opto-mechanical attachment. This fluorescence microscope on a smartphone will be designed by integrating a laser diode, a disposable nano-channel chip, an external lens and a thin-film based emission filter in a robust attachment created by 3D printing techniques. High SNR fluorescence signal detection will be achieved by implementing high-angle/oblique illumination so that the direct excitation beam will not enter the low NA collection lens. They will also develop a compressive sampling based DNA length-estimation method which will utilize (i) the measured point spread function of the fluorescent microscope on the mobile-phone; (ii) the spatial sparsity of the objects (fluorescently labeled DNA molecules); and (iii) the linearity of the stretched DNA molecules within the field of view as a-priori constraints to estimate the length of the DNA fragment of interest with an accuracy that is significantly better than the resolution of their initial imaging system.