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）：尽管可以常规地并行获取横向（即平行于样品表面）的图像，但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。例如，使用光片显微镜）或在高速（例如，使用高速扫描显微镜）下，当前的成像方式都无法对轴向切片（即垂直切片）进行成像。在这项拟议的研究中，我们的目标是通过开发一种能够并行成像样品轴向切片的新技术来克服这一限制。建议使用 45¿ - 沿轴向排列的倾斜高纵横比微镜，可以通过利用其不同的波前来解复用从不同深度位置发出的图像信号，每个微镜的尺寸与所测试的衍射限制光束宽度相当。共焦针孔或狭缝，提供光学切片能力，在我们的初步研究的基础上，在拟议的计划中，我们将首先设计所提出的高纵横比微镜阵列，并使用微成型来制造它。然后，我们将通过结合高纵横比微镜阵列装置来开发所提出的轴向切片光片成像系统，并对荧光微球和 3D 球状肿瘤的高分辨率轴向切片光片成像进行表征和优化。将展示细胞培养复杂系统。