Axial Slice Light Sheet Microscopy

轴向切片光片显微镜

• 批准号: 8772761
• 负责人: Zhiwen Liu
• 金额: $ 20.83万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8772761
• 关键词: Address; Biological; Biological Process; Cells; Complex; Confocal Microscopy; Cultured Tumor Cells; Detection; Devices; Diagnosis; Dimensions; Dose; Electron Microscopy; Goals; Image; Imaging Techniques; Lasers; Lateral; Light; Lighting; Maps; Measures; Mechanics; Medical; Methods; Microscopy; Microspheres; Molds; Optics; Performance; Positioning Attribute; Process; Property; Resolution; Sampling; Scanning; Scheme; Signal Transduction; Silicon; Slice; Spatial Distribution; Specimen; Speed; Surface; System; Techniques; Tissue Sample; Tissues; Width; design; electron beam lithography; fluorescence imaging; imaging modality; in vivo imaging; lens; multi-photon; neoplastic cell; new technology; optical imaging; programs; prototype; public health relevance; research study; response; simulation; tool; two-photon

DESCRIPTION (provided by applicant): Three-dimensional optical imaging of biological cells and tissues has become increasingly critical for biomedical studies and medical diagnosis. Although images in the lateral direction (i.e., parallel to the sample surface) can be routinely acquires in parallel (e.g., using light-sheet microscopy) or at high speed (e.g., using high-speed scanning microscopy), no current imaging modality can image an axial slice (i.e., perpendicular to the sample surface) in parallel, limiting our ability to visualize fast biological processes happening across different depths. In this proposed study, we aim to overcome this limitation by developing a new technology capable of imaging axial slices of a sample in parallel. The key concept is the proposed use of an array of 45¿- tilted high-aspect-ratio micro mirrors arranged along the axial direction, which can demultiplex image signals emitted from different depth positions by exploiting their different wavefronts. Each micro mirror has a dimension comparable to the diffraction limited beam width. Consequently it behaves effectively as a confocal pinhole or slit, providing optical sectioning capability. Building upon our preliminary study, in the proposed program we will first design the proposed high-aspect-ratio micro mirror array and fabricate it by using micromolding. We will then develop the proposed axial slice light-sheet imaging system by incorporating the high-aspect-ratio micro mirror array device. The prototype system will be characterized and optimized. High- resolution axial slice light sheet imaging of fluorescent microspheres and 3D spheroid tumor cell culture complex systems will be demonstrated.

描述（由申请人提供）：生物细胞和组织的三维光学成像对于生物医学研究和医学诊断变得越来越重要。虽然横向方向上的图像（即，平行于样品表面）可以常规地并行获取（例如，使用光片显微术）或高速（例如，使用高速扫描显微镜），目前没有成像模式可以对轴向切片成像（即，垂直于样品表面），限制了我们可视化不同深度发生的快速生物过程的能力。在这项拟议的研究中，我们的目标是克服这一限制，开发一种新的技术，能够成像轴向切片的样品平行。关键概念是提出使用沿轴向方向沿着布置的45倾斜高纵横比微镜阵列，其可以通过利用从不同深度位置发射的图像信号的不同波前来解复用图像信号。每个微镜具有与衍射极限光束宽度相当的尺寸。因此，它有效地表现为共焦针孔或狭缝，提供光学切片能力。基于我们的初步研究，在拟议的计划中，我们将首先设计建议的高纵横比微镜阵列，并通过使用微成型制造它。然后，我们将开发建议的轴向切片光片成像系统，结合高纵横比微镜阵列器件。将对原型系统进行表征和优化。高分辨率轴向切片光片成像的荧光微球和三维球体肿瘤细胞培养复合系统将被证明。