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（由适用提供）：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已经变得越来越重要。尽管可以平行地（例如，使用灯表显微镜）或高速（例如，使用高速扫描显微镜）并行地沿侧向（即平行于样品表面）沿侧向（即平行于样品表面）进行图像，但没有当前的成像模态可以轴向切片（即依赖于样品表面）的轴向切片（即在平行的均衡范围内）。在这项拟议的研究中，我们旨在通过开发能够并行成像样品轴向切片的新技术来克服这一局限性。关键概念是提议使用45¿-沿轴向方向排列的倾斜的高光谱理由的微镜，通过利用其不同的波前，它可以从不同深度位置发出的图像信号拆除。每个微镜的尺寸与衍射有限的光束宽度相当。因此，它的表现为共聚焦针孔或缝隙，提供了光学切片能力。在我们的初步研究的基础上，在拟议的程序中，我们将首先设计提出的高光谱比率微镜阵列，并使用微镀料进行制造。然后，我们将通过转换高光谱的微镜阵列设备来开发提出的轴向切片灯表成像系统。原型系统将被表征和优化。将展示荧光微球和3D球状肿瘤细胞培养复合系统的高分辨率轴向切片光片成像。