Travel Grant for Adaptive Optics in CARS Microscopy

CARS 显微镜中的自适应光学旅行补助金

• 批准号: EP/F020155/1
• 负责人: John Girkin
• 金额: $ 0.72万
• 依托单位: University of Strathclyde
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FF020155%2F1
• 关键词: Travel; Grant; Adaptive; Optics; CARS

Optical microscopy has provided scientists, and clinicians, with the ability to image living tissue with sub-cellular resolution for around four hundred years. This ability has lead to a number of scientific breakthroughs both in the life and physical sciences. Significant advances in general optics have frequently been achieved by the desire to image samples with increasing resolution so that the finest structures can be resolved. Indeed just over 130 years ago Abbe advanced the study of optics with his work on improved optical microscopy with the introduction of diffraction theory and the study of optical aberrations. However, crucial to all forms of microscopy is the ability to have contrast between the different structures present within the sample. For most of the last fifty years this has been provided by the addition of fluorescent labels to the sample culminating in the use of genetically modified samples so that they naturally express a fluorescent label. Even in this case the sample is not in its natural state and thus the method is not practical for a number of samples, and makes the application of microscopy to living tissue difficult, and impossible for many clinical applications.Coherent Anti-Stokes Raman Scattering (CARS) microscopy is a method by which optical microscopy can be undertaken using the spectral fingerprint of a specific molecule as the contrast agent. By tuning two laser sources to the correct wavelengths specific molecular spectral features can be excited enabling highly specific microscopy. The process is non-linear and hence the imaging volume is restricted in three dimensions offering the ability of the microscope to naturally generate high resolution optical slices which can be reconstructed to produce three dimensional images.However, as one wishes to image more deeply into a sample the laser light has to pass through the sample and is aberrated leading to a loss of signal and spatial resolution at depths much beyond 100microns. This problem is similar to that faced by ground based astronomers examining space through the atmosphere. Through the use of active optical elements (adaptive optics) they have demonstrated that the atmospheric aberrations can be removed, or at least minimised. Previously we have adopted the same technology to multiphoton and confocal microscopy with considerable success; however, both of these imaging modalities require the addition of fluorescent compounds to the sample in some form thus perturbing the natural state of the tissue. The aim of this project is to explore the use of adaptive optics (AO) in CARS microscopy by linking, for two weeks, world leading expertise in both the application of adaptive optics to microscopy and CARS imaging. By fitting the Institute of Photonics AO system to the Harvard CARS microscope we will rapidly be able to determine if AO has a role to play in CARS microscopy as is strongly suspected. This proposal thus offers a rapid, low cost route to the evaluation of AO for in-depth CARS microscopy.

光学显微镜为科学家和临床医生提供了大约四百年的亚细胞分辨率成像活组织的能力。这种能力导致了生命科学和物理科学中的许多科学突破。一般光学的显著进步常常是通过期望以增加的分辨率对样品进行成像以使得可以分辨最精细的结构来实现的。事实上，就在130多年前阿贝先进的光学研究与他的工作，改善光学显微镜引进衍射理论和研究光学像差。然而，对于所有形式的显微镜来说，至关重要的是能够在样品中存在的不同结构之间进行对比。在过去50年的大部分时间里，这是通过向样品中添加荧光标记来提供的，最终使用遗传修饰的样品，使得它们天然地表达荧光标记。相干反斯托克斯拉曼散射（汽车）显微术是一种使用特定分子的光谱指纹作为对比剂进行光学显微术的方法，它可以在显微镜下观察到特定分子的光谱指纹。通过将两个激光源调谐到正确的波长，可以激发特定的分子光谱特征，从而实现高度特异性的显微镜检查。该过程是非线性的，因此成像体积被限制在三维中，从而提供显微镜自然生成高分辨率光学切片的能力，所述高分辨率光学切片可以被重构以产生三维图像。当人们希望更深入地成像到样品中时100微米。这个问题类似于地面天文学家通过大气层检查太空所面临的问题。通过使用主动光学元件（自适应光学），他们已经证明可以消除或至少最小化大气像差。以前，我们已经采用了相同的技术，多光子和共聚焦显微镜相当成功，然而，这两种成像方式需要添加荧光化合物的样品以某种形式，从而扰乱组织的自然状态。该项目的目的是探索自适应光学（AO）在汽车显微镜中的使用，方法是在两周内将自适应光学应用于显微镜和汽车成像方面的世界领先专业知识结合起来。通过将光子学研究所的AO系统安装到哈佛的汽车显微镜上，我们将能够迅速确定AO是否像人们强烈怀疑的那样在汽车显微镜中发挥作用。因此，这一建议提供了一个快速，低成本的路线，以评估AO深入汽车显微镜。

项目成果

Application of Adaptive Optics in CARS Microscopy

自适应光学在CARS显微镜中的应用

• 发表时间: 2008
• 作者: S Poland

Adaptive optics applied to coherent anti-Stokes Raman scattering microscopy

自适应光学应用于相干反斯托克斯拉曼散射显微镜

• DOI: 10.1117/12.764445
• 发表时间: 2008
• 作者: Girkin J