DIGITAL OPTICAL PHASE CONJUGATION ASSISTED 4Pi FLUORESCENCE MICROSCOPY

数字光学相位共轭辅助 4Pi 荧光显微镜

• 批准号: 7978493
• 负责人: CHANGHUEI YANG
• 金额: $ 18.87万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7978493
• 关键词: Biochemical; Complex; Development; Developmental Biology; Drosophila genus; Embryo; Financial compensation; Fluorescence; Fluorescence Microscopy; Image; Lasers; Lateral; Lead; Light; Maps; Microscope; Microscopy; Modeling; Neurosciences; Optics; Phase; Research; Resolution; Sampling; Source; Spottings; System; Thick; Time; Tissues; Work; base; digital; improved; light scattering; prototype; public health relevance; research study; time use; tissue phantom; two-photon

DESCRIPTION (provided by applicant): This Exploratory/Developmental Research (R21) application proposes to develop an optical-phase- conjugation-assisted (OPC) 4Pi microscope that is capable of high lateral (200 nm) and axial (120 nm) resolution imaging. A typical 4Pi microscope consists of two counter-propagating laser beams that are focused to the same spatial point. The mutual interference of the two beams allows an improvement in the axial resolution by a factor of 5-7 versus a standard confocal microscope. However, a 4Pi microscope performs poorly for optically inhomogeneous samples as extensive scattering degrades the focal spot quality and can lead to a misalignment of the two focal spots. Furthermore, 4Pi microscopes are generally difficult to implement and use well because the exact superposition of the laser beams' focal spots is critical but easily disrupted. Our proposed research will leverage our current expertise in optical phase conjugation (OPC), time-reversal turbidity suppression via OPC, and tissue optics to significantly improve the capability of 4Pi microscopy. Optical phase conjugation can be roughly interpreted as the time-reversed playback of a target light field. We have recently established that, because scattering is deterministic, it is possible undo tissue scattering by time- reversing a scattered light field through the target tissue. We aim to demonstrate that the incorporation of a specially adapted OPC arrangement in a 4Pi microscope will allow such a microscope to adaptively align its focus and to correct sample aberration which has been an obstacle for the conventional 4Pi system. The application range of the proposed OPC 4Pi microscope is potentially broad. The anticipated aberration compensation ability will allow this system to image thicker and more heterogeneous samples. We expect that such systems will be useful in neuroscience and developmental biology experiments where precise 3D mapping of complex samples, such as embryos, are important. PUBLIC HEALTH RELEVANCE: The successful completion of this proposed project will result in more robust 4Pi microscope systems that are simpler to use and capable of imaging through thicker and more heterogeneous samples. We expect that such systems will be useful in neuroscience and developmental biology experiments where precise 3D mapping of complex samples, such as embryos, are important. The proposed work will also further our current research effort on the use of time-reversed light field to induce tissue transparency - a research direction that can ultimately lead to accurate and non-invasive biochemical sensing, and light-based deep tissue imaging.

描述（由申请人提供）：本探索性/开发性研究（R21）申请旨在开发一种光学相位共轭辅助（OPC）4Pi显微镜，该显微镜能够进行高横向（200 nm）和轴向（120 nm）分辨率成像。典型的4Pi显微镜由两个反向传播的激光束组成，它们聚焦到同一个空间点。这两个光束的相互干涉允许与标准共焦显微镜相比轴向分辨率提高5-7倍。然而，4Pi显微镜对于光学不均匀的样品表现不佳，因为广泛的散射会降低焦斑质量，并可能导致两个焦斑的不对准。此外，4Pi显微镜通常很难实现和使用，因为激光束焦斑的精确叠加是关键的，但很容易被破坏。我们提出的研究将利用我们目前在光学相位共轭（OPC），通过OPC的时间反转浊度抑制和组织光学方面的专业知识，以显着提高4Pi显微镜的能力。光学相位共轭可以粗略地解释为目标光场的时间反演回放。我们最近已经确定，因为散射是确定性的，所以可以通过时间反转穿过目标组织的散射光场来撤销组织散射。我们的目的是证明，在一个4Pi显微镜中的一个特别适合的OPC安排的合并将允许这样的显微镜自适应地调整其焦点，并纠正样品像差，这一直是传统的4Pi系统的障碍。建议的OPC 4Pi显微镜的应用范围是潜在的广泛。预期的像差补偿能力将允许该系统成像更厚和更异质的样品。我们预计，这样的系统将在神经科学和发育生物学实验中非常有用，在这些实验中，复杂样本（如胚胎）的精确3D映射非常重要。 公共卫生关系：该项目的成功完成将带来更强大的4Pi显微镜系统，该系统使用更简单，能够通过更厚和更异质的样品进行成像。我们预计，这样的系统将在神经科学和发育生物学实验中非常有用，在这些实验中，复杂样本（如胚胎）的精确3D映射非常重要。拟议的工作还将进一步推动我们目前关于使用时间反转光场诱导组织透明性的研究工作-这一研究方向最终可以导致准确和非侵入性的生化传感以及基于光的深层组织成像。