Three-Dimensional (3D) Acoustofluidic Scanning Nanoscope with Super Resolution and Large Field of View

具有超分辨率和大视场的三维 (3D) 声流控扫描纳米镜

• 批准号: 10278520
• 负责人: Chenglong Zhao
• 金额: $ 38.12万
• 依托单位: UNIVERSITY OF DAYTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10278520
• 关键词: 3-Dimensional; Acoustics; Address; Architecture; Area; Autophagocytosis; Benchmarking; Biodistribution; Biological; Biological Models; Biological Phenomena; Biological Process; Biology; Biomedical Research; Cell physiology; Cells; Cellular Structures; Chemistry; Collaborations; Complex; Confocal Microscopy; Development; Devices; Endocytosis; Engineering; Fluorescence Microscopy; Gold; Hela Cells; Image; Imaging Techniques; Imaging technology; Industry Standard; Joints; Laboratories; Laboratory Research; Lateral; Light; Lysosomes; Medicine; Microscope; Microscopy; Microspheres; Modification; Morphology; Optics; Organelles; Pathologic; Pattern; Peer Review; Performance; Physiological; Play; Positioning Attribute; Research; Research Personnel; Resolution; Role; Sampling; Scanning; Shapes; Signal Pathway; Solid; Speed; Structure; Subcellular structure; Surface; System; Techniques; Technology; Three-Dimensional Imaging; Time; Universities; base; cancer cell; cell behavior; cost; fluorophore; imaging modality; improved; insight; journal article; lens; nanoimaging; nanoparticle; nanorod; nanoscale; nanoscope; operation; optical imaging; particle; prototype; relating to nervous system; three-dimensional visualization; tool

PROJECT SUMMARY Over the past two decades, a number of “super-resolution” 3D imaging technologies have been developed, enabling researchers to observe nanoscale biological structures that were previously invisible to traditional, diffraction-limited imaging techniques. The ability to visualize cellular and subcellular structures at the nanoscale has revealed key insights into a variety of biological processes. Although impressive progress has been made in the development of 3D super-resolution imaging techniques, researchers are often forced to accept a tradeoff in terms of the resolution, field-of-view, speed, and ease of use of their 3D imaging technique. Recently, we have developed an acoustofluidic scanning nanoscope that can simultaneously achieve both super-resolution and large field-of-view imaging in 2D. In this R01 project, we will develop and validate a 3D acoustofluidic scanning nanoscope with the following features: (1) Super-resolution imaging with lateral and axial resolutions of ~50 nm and ~120 nm, respectively: The proposed 3D imaging method will achieve a resolution that is four times better than that from a confocal microscope, which makes the optical imaging of more detailed inner architecture of many subcellular structures possible; (2) Large field-of-view (~1,100×1,100 µm2): Conventional optical imaging methods achieve high-throughput imaging at the cost of reduced resolution and vice versa. By utilizing acoustics to simultaneously manipulate multiple microsphere lenses, the proposed imaging method will solve this long-standing technical barrier for large field-of-view imaging while maintaining superior lateral and axial resolution; (3) Imaging speed 10 times faster than that from a confocal microscope: Rapid z-stacking at a speed 10 times faster than that of a confocal microscope can be achieved by using surface acoustic waves to scan an array of microspheres across the sample volume in a precise, controllable manner; (4) Seamless connection to a conventional optical microscope for ease of use: Our device can be seamlessly connected to a conventional optical microscope without modification of the optical setup, which can significantly reduce the cost and the complexity of operation. With the aforementioned advantages, the proposed 3D acoustofluidic scanning nanoscope technology has the potential to significantly exceed current standards in the field and address many unmet needs. We will validate its performance by imaging 3D nanorod samples and the organelles of live HeLa cells. In this regard, we aim to demonstrate the far-reaching potential of our 3D acoustofluidic scanning nanoscope technology to enable improved research in areas ranging from subcellular imaging to the visualization of 3D neural activity.

项目摘要 在过去的二十年中，已经开发了许多“超分辨率” 3D成像技术 使研究人员能够观察纳米级生物结构，这些结构以前是传统的， 衍射受限的成像技术。能够可视化纳米级的细胞和亚细胞结构的能力 已经揭示了对各种生物过程的关键见解。尽管取得了令人印象深刻的进展 在开发3D超分辨率成像技术时，研究人员经常被迫接受权衡 就其3D成像技术的分辨率，视野，速度和易用性而言。最近，我们有 开发了一种声液扫描纳米镜，可以轻松实现超分辨率和 2D中的大型视野成像。在这个R01项目中，我们将开发和验证3D声学扫描 纳米镜具有以下特征：（1）横向和轴向分辨率约50的超分辨率成像 NM和〜120 nm：提出的3D成像方法将达到四倍的分辨率 比共聚焦显微镜更好，这使得更详细的内部体系结构的光学成像 许多亚细胞结构可能； （2）大型视野（〜1,100×1,100 µm2）：常规光学 成像方法以减少分辨率成本实现高通量成像，反之亦然。通过使用 声学要简单地操纵多个微球镜头，提出的成像方法将解决 这种长期存在的技术障碍，用于大型视野成像，同时保持横向和轴向优势 解决; （3）成像速度比共聚焦显微镜快10倍：快速z堆叠 速度比共聚焦显微镜快10倍，可以通过使用表面声波到 以精确的，受控的方式扫描样品体积上的微球阵列； （4）无缝 连接到常规光学显微镜以易于使用：我们的设备可以无缝连接 进行常规的光学显微镜，而无需修改光学设置，这可以显着降低 成本和操作的复杂性。具有近似优点，提出的3D声流体 扫描纳米镜技术有可能显着超过该领域的当前标准 满足许多未满足的需求。我们将通过成像3D纳米棒样品和细胞器来验证其性能 活的HeLa细胞。在这方面，我们旨在展示3D声流体的深远潜力 扫描纳米镜技术，以改善从亚细胞成像到 3D神经活动的可视化。