Rapid parallel 3D confocal/fluorescence cell imaging with varifocal lens

使用变焦镜头进行快速并行 3D 共焦/荧光细胞成像

• 批准号: 7880578
• 负责人: Guoqiang Li
• 金额: $ 1.06万
• 依托单位: UNIVERSITY OF MISSOURI-ST. LOUIS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7880578
• 关键词: Antiviral Agents; Automobile Driving; Behavior; Biological Sciences; Cell Line; Cell physiology; Cells; Chinese Hamster Ovary Cell; Coupled; Detection; Devices; Disease; Early Diagnosis; Electronics; Emerging Technologies; Feedback; Fluorescence; Fluorescence Microscopy; Frequencies; Goals; Human; Image; Incubated; Knowledge; Label; Lasers; Lead; Length; Life; Ligands; Light; Lighting; Mechanics; Medical; Methods; Microscope; Microscopic; Morphologic artifacts; Morphology; Opioid Receptor; Optics; Organism; Performance; Phase; Quantum Dots; Reaction Time; Research Personnel; Resolution; Sampling; Scanning; Speed; Surface; System; Technology; Three-Dimensional Imaging; Time; Tissues; Translations; bioimaging; cellular imaging; charge coupled device camera; cost; data acquisition; design; digital; disease diagnosis; disorder control; drug candidate; flexibility; fluorescence imaging; improved; keratinocyte; lens; light microscopy; liquid crystal; millisecond; novel; optical imaging; physical science; public health relevance; receptor; time use; tissue phantom; tool; uptake; vibration; voltage

DESCRIPTION (provided by applicant): Acquisition speed of light microscopy is very critical for live cell imaging, which greatly enhance our ability to understand the dynamic cellular processes. The overall goal of this study is to develop a novel parallel 3D confocal/fluorescence optical imaging system equipped with an electro-optic varifocal lens for rapid depth scanning and digital micromirror device for transverse confocal scanning and hence fast image acquisition. The system provides a new tool to assess tissue and cell function and morphology in real time. This is the first demonstration system using varifocal optical lens for high-resolution imaging of biomedical tissues. Both longitudinal and transverse scanning are performed electro-optically without translational components and the moving effect of the sample due to mechanic vibration of the conventional imaging system can be avoided. The response time of the varifocal lens can be in the order of millisecond. With correct matching CCD camera and electronics, it is feasible to achieve an acquisition speed of a few hundred frames per second. The pixel dwell time is three orders of magnitude higher than the conventional raster scanning confocal imaging. It allows lower laser power and high sensitivity. The proposed system has potential advantages such as high resolution, high sensitivity, large dynamic range, minimized photo bleaching, cost-effective, and flexibility of imaging at different wavelengths. The system has versatile functions, including widefield, confocal, and fluorescence imaging. In the proposed exploratory phase of this technology-driven project, we will evaluate the performance of our novel imaging system on tissue phantoms and live cells. Specifically, we will study live cells (CHO cells expressing specific surface receptors; e.g.,-opioid receptor) that are incubated with quantum dots coated ligand and we will also study uptake and intracellular distribution of antiviral compounds into human keratinocytes and related cell lines. The high performance of the system will be attractive for live cell imaging, tissue imaging, and diagnosis of diseases. PUBLIC HEALTH RELEVANCE: The proposed parallel confocal/fluorescence imaging system using varifocal lens and reconfigurable DMD allows high frame rate, high-resolution 3D live cell imaging without any mechanical translation components. The system has versatile functions, including wide field, confocal, and fluorescence imaging. It has many advantages in comparison with the conventional 3D microscope imaging systems. The pixel dwell time is significantly increased. It allows lower excitation laser power, higher sensitivity, and less thermal damage. Such a system is very promising for live cell imaging and it will significantly improve our ability to understand the dynamic interactions inside the cells and control diseases. Applications of the varifocal lens can be extended to broad fields where adaptive change of focusing power with large aperture, low voltage and low power dissipation is desirable.

描述（由申请人提供）：光学显微镜的采集速度对于活细胞成像至关重要，这极大地增强了我们了解动态细胞过程的能力。这项研究的总体目的是开发一种具有电动型变斑透镜的新型平行3D共聚焦/荧光光学成像系统，用于快速深度扫描和数字微骨器设备，用于横向共聚焦扫描，从而快速的图像采集。该系统提供了一种新工具，可以实时评估组织，细胞功能和形态。这是使用Varifocal光学镜头进行生物医学组织的高分辨率成像的第一个示范系统。纵向扫描和横向扫描都是在没有翻译成分的情况下进行的，并且由于常规成像系统的机械振动而引起的样品的移动效果。 Varifocal镜头的响应时间可以按毫秒的顺序。使用正确匹配的CCD摄像头和电子设备，可以每秒获得几百帧的采集速度。像素停留时间比传统的栅格扫描共聚焦成像高三个数量级。它允许较低的激光功率和高灵敏度。所提出的系统具有潜在的优势，例如高分辨率，高灵敏度，大动态范围，最小化的照片漂白，成本效益和在不同波长下成像的灵活性。该系统具有多功能功能，包括广场，共焦和荧光成像。在该技术驱动的项目的拟议探索阶段，我们将评估新型成像系统在组织幻像和活细胞上的性能。具体而言，我们将研究与量子点涂层配体一起孵育的活细胞（表达特定表面受体的CHO细胞；例如 - 阿片类受体），我们还将研究抗病毒药物化合物对人角膜素类和相关细胞系的摄取和细胞内分布。该系统的高性能将对活细胞成像，组织成像和疾病的诊断具有吸引力。公共卫生相关性：使用Varifocal镜头和可重构DMD的拟议的平行共聚焦/荧光成像系统允许高框架速率，高分辨率3D Live Cell成像，而无需任何机械翻译组件。该系统具有多功能功能，包括广阔的场，共聚焦和荧光成像。与常规3D显微镜成像系统相比，它具有许多优势。像素停留时间显着增加。它允许降低激发激光功率，更高的灵敏度和较小的热损伤。这种系统对于活细胞成像非常有前途，它将显着提高我们了解细胞内部动态相互作用和控制疾病的能力。值得期望的是，可以扩展到宽阔的镜头的应用，在宽阔的田间中，需要大量孔径，低压和低功率耗散。