High Resolution 3D Optics Coherence Phase Microscopy

高分辨率 3D 光学相干相位显微镜

• 批准号: 7929461
• 负责人: JOHANNES F DE BOER
• 金额: $ 13.18万
• 依托单位: VRIJE UNIVERSITEIT
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-10 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7929461
• 关键词: 3-Dimensional; Action Potentials; Astacoidea; Automobile Driving; Axon; B-Lymphocytes; Behavior; Binding; CD3 Antigens; Calibration; Cell Volumes; Cell physiology; Cells; Cellular Structures; Cellular biology; Detection; Development; Dyes; Electrodes; Environment; Fluorescence Microscopy; Foundations; Funding; Goals; Image; Imagery; Imaging Device; Imaging Techniques; Individual; Lateral; Lead; Life; Light; Lobster; Measures; Medicine; Methods; Microscope; Microscopy; Monitor; Nerve; Neurobiology; Neurons; Optical Coherence Tomography; Optics; Performance; Phase; Phase-Contrast Microscopy; Photons; Process; Research; Resolution; Specimen; Staining method; Stains; Stimulus; Structure; Swelling; System; T-Cell Receptor; T-Lymphocyte; Techniques; Technology; Three-Dimensional Image; Three-Dimensional Imaging; Tissues; base; cell behavior; cellular imaging; design; fluorescence imaging; imaging modality; insight; light microscopy; nanometer; novel; optical imaging; phase change; programs; public health relevance; relating to nervous system; response; submicron; three dimensional structure; transmission process; two-photon; voltage

DESCRIPTION (provided by applicant): Three-dimensional imaging of cells and sub cellular structures has enormous potential in studies on cell physiology and medicine, since it enables the observation of cell behavior in a variety of environments. Many three-dimensional imaging techniques to date require cells to be stained or dyed to produce images of structures that are normally not observable. We propose to develop a three-dimensional optical coherence phase-contrast microscope (OCPM) based on Spectral Domain Optical Coherence Tomography (SD-OCT) technology that can reveal three-dimensional structures and dynamics of cells in their natural state by measuring depth resolved phase differences with high sensitivity. The OCPM will be integrated with Two Photon Microscopy (TPM) into a standard inverted microscope. Phase sensitivity, lateral resolution, and structural imaging capability will be evaluated on calibration objects and cells, where nanometer axial and sub-micron lateral resolution are targeted. Interpretation of OCPM images will be investigated by simultaneous two-photon fluorescence imaging of the same specimen. The potential of OCPM will be studied by imaging cell responses to external stimuli. The ability for deep tissue imaging (250 ¿m) and the exploitation of phase sensitivity will be investigated by monitoring action potentials in axons embedded in larger nerve bundles. Neural activity has been demonstrated to induce rapid changes in nerves (swelling) on the order of 1-3 nanometers. The spatio-temporal resolution of OCPM is capable of detecting these rapid changes during action potential propagation. OCPM will serve as a powerful imaging tool for processes in cell biology and will enhance the understanding of the structure, function, and the behavior of living cells. The overall goal of this research is A) to develop an OCPM system for 3 dimensional high resolution phase contrast imaging of cells, B) to determine the phase resolution and sectioning capability of OCPM, C) to investigate the potential of OCPM for the study of cell dynamics in response to external stimuli, D) to non- invasively measure action potentials of axons within nerve bundles. PUBLIC HEALTH RELEVANCE: Three-dimensional imaging of cells and sub cellular structures has enormous potential in studies on cell physiology and medicine, since it enables the observation of cell behavior in a variety of environments. Many three-dimensional imaging techniques to date require cells to be stained or dyed to produce images of structures that are normally not observable. We will develop a new form of microscopy (OCPM) that is able to detect minute changes in cells on the order of nanometers. OCPM will serve as a powerful imaging tool for processes in cell biology and will enhance the understanding of the structure, function, and the behavior of living cells.

描述（通过应用提供）：细胞和亚细胞结构的三维成像具有增强的细胞生理和医学研究潜力，因为它可以在各种环境中观察细胞行为。迄今为止，许多三维成像技术要求细胞染色或染色，以产生通常无法观察到的结构图像。我们建议开发基于光谱结构域光学相干断层扫描（SD-OCT）技术的三维光学相互对比显微镜（OCPM）技术，该技术可以通过高敏感性来测量深度差异来揭示其自然状态的三维结构和细胞动态。 OCPM将与两个光子显微镜（TPM）集成到标准的倒置显微镜中。相位灵敏度，横向分辨率和结构成像能力将在校准对象和细胞上进行评估，其中纳米轴向和亚微米侧侧分辨率是针对的。 OCPM图像的解释将通过同一标本的简单两光子荧光成像进行研究。 OCPM的潜力将通过成像对外部刺激的细胞反应进行研究。深层组织成像（250€）的能力和相位灵敏度的利用将通过监测嵌入较大神经束中的轴突的作用电位来研究。已经证明神经活动可以诱导1-3纳米阶的神经（肿胀）的快速变化。 OCPM的时空分辨率能够在动作电位传播过程中检测这些快速变化。 OCPM将作为细胞生物学过程的强大成像工具，并将增强对活细胞的结构，功能和行为的理解。这项研究的总体目标是a）为3维高分辨率的对比成像开发OCPM系统，b）确定OCPM，c）确定OCPM的相位分辨率和切片能力，以研究OCPM对外部刺激的细胞动力学研究的潜力，d）对外部刺激的响应，d），d）在神经外侧的轴突中轴突的轴突的动作电位。 公共卫生相关性：细胞和亚细胞结构的三维成像增强了细胞生理和医学研究的潜力，因为它可以在各种环境中观察到细胞行为。迄今为止，许多三维成像技术要求细胞染色或染色，以产生通常无法观察到的结构图像。我们将开发一种新形式的显微镜（OCPM），该显微镜能够检测纳米顺序的细胞变化。 OCPM将作为细胞生物学过程的强大成像工具，并将增强对活细胞的结构，功能和行为的理解。