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

描述（申请人提供）：光学显微镜的采集速度对活细胞成像至关重要，这大大提高了我们了解细胞动态过程的能力。本研究的总体目标是开发一种新型的平行三维共焦/荧光光学成像系统，该系统配备了用于快速深度扫描的电光变焦透镜和用于横向共焦扫描的数字微镜装置，从而实现快速图像采集。该系统为实时评估组织和细胞的功能和形态提供了一种新的工具。这是第一个使用变焦光学透镜对生物医学组织进行高分辨率成像的演示系统。纵向和横向扫描均采用光电扫描，无需平移元件，避免了传统成像系统因机械振动引起的样品移动效应。变焦透镜的响应时间可以达到毫秒级。通过正确匹配CCD相机和电子器件，可以实现每秒几百帧的采集速度。像素停留时间比传统的光栅扫描共聚焦成像提高了3个数量级。它允许较低的激光功率和高灵敏度。该系统具有高分辨率、高灵敏度、大动态范围、最小化光漂、低成本和不同波长成像灵活性等潜在优势。该系统具有多种功能，包括广角、共聚焦和荧光成像。在这个技术驱动项目的拟议探索阶段，我们将评估我们的新型成像系统在组织幻影和活细胞上的性能。具体来说，我们将研究与量子点包被配体孵育的活细胞（表达特定表面受体的CHO细胞，例如-阿片受体），我们还将研究抗病毒化合物在人角质形成细胞和相关细胞系中的摄取和细胞内分布。该系统的高性能将对活细胞成像、组织成像和疾病诊断具有吸引力。公共卫生相关性：建议的平行共聚焦/荧光成像系统使用变焦透镜和可重构DMD，允许高帧率，高分辨率3D活细胞成像，而无需任何机械平移组件。该系统具有多种功能，包括宽视场、共聚焦和荧光成像。与传统的三维显微镜成像系统相比，它具有许多优点。像素停留时间显著增加。它允许更低的激发激光功率，更高的灵敏度，和更少的热损伤。这样的系统对于活细胞成像非常有前景，它将显著提高我们了解细胞内部动态相互作用和控制疾病的能力。变焦透镜的应用可以扩展到广泛的领域，在大光圈、低电压和低功耗的要求下自适应改变聚焦功率。

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.