Integrated Optical Needle (ION) for Cellular and Molecular Imaging of Cancer

描述（由申请人提供）：共聚焦显微镜的最新进展证明了光学成像在提供亚细胞分辨率的分子图像方面的潜力。在本提案中，我们扩展了我们在高分辨率共聚焦显微镜方面的工作，基于一个更简单的替代方案，基于我们的观察，如果将细胞直接放置在平面光学窗口上，则可以在不需要共聚焦成像门的情况下获得顶部1-2层细胞的高分辨率光学图像。本文提出了一种新型的光学成像装置——集成光学针（ION），它可以插入小尺寸的针中，穿过组织，实时获取针远端亚细胞分辨率的组织图像；使用靶向造影剂或生命染色剂可以产生额外的对比度以及功能和/或分子成像能力。第一个版本的光学针，在目标1中描述，是基于一个灵活的，相干光纤束耦合到宏观成像光学器件和CCD图像传感器。与Aim 1中使用的光纤束结构相关的欠采样限制了空间分辨率。第二种方法是在目标2和目标3中描述的集成光学针（ION），它使用LIGA技术将使用灰度光刻技术制造的微型透镜与光源和图像传感器集成在一起，从而产生可以通过针插入的微型显微镜。该离子包括一个NA = 0.4的显微镜，可以在250微米的视野内以1微米的横向分辨率成像；它被设计成适合于内径为1毫米的针。我们将开展多种临床前试验，包括细胞培养、组织培养、动物模型和临床试验，以评估这些系统的相对性能。需要解决的问题是，没有针孔或其他深度切片技术的成像是否产生足够质量的图像，以欣赏临床所需的细胞细节，并确定失焦和散射光子是否会使对比度降低得太大。我们的初步结果表明，在常规临床应用中使用造影剂可以获得人类受试者细胞细节的良好图像。然而，为了通过各种设备和造影剂来判断图像质量，我们将使用图像分辨率、图像对比度和图像信噪比等定量指标。这里提出的研究将开发一种新的工具，用于实时图像指导组织评估，以及在图像指导位置进行确认性活检。离子可以成像范围广泛的造影剂，从特定部位获得分子信息。即使在小批量生产（低于2,300美元）时，集成光学针的成本也很低，但在批量生产时，进一步降低成本的潜力很大。

DESCRIPTION (provided by applicant): Recent advances in confocal microscopy have demonstrated the potential of optical imaging to provide molecular images with sub-cellular resolution. In this proposal, we extend our work in high resolution confocal microscopy, based on a much simpler alternative, based on our observation that high resolution optical images of the top 1-2 layers of cells can be obtained without the need for a confocal imaging gate if the cells are placed in direct contact with a flat optical window. Here we propose a new class of optical imaging device - the integrated optical needle (ION) - which can be inserted into a small gauge needle and advanced through tissue to acquire images of tissue at the distal tip of the needle with sub-cellular resolution in real time; the use of targeted contrast agents or vital stains can yield additional contrast as well as functional and/or molecular imaging capability. The first version of the optical needle, described in Aim 1, is based on a flexible, coherent fiber-optic bundle coupled to macroscopic imaging optics and a CCD image sensor. The under-sampling associated with the structure of the fiber bundle used in Aim 1 limits the spatial resolution. The second approach, an integrated optical needle (ION) described in Aims 2 and 3, uses LIGA technology to integrate miniature lenses fabricated using grayscale lithography, together with the light source and image sensor to yield a miniature microscope which can be inserted through a needle. This ION consists of a NA = 0.4 microscope that can image with 1 micron lateral resolution from a 250 um field of view; it is designed to fit within a needle that has an inner diameter of 1 mm. We will carry out a variety of preclinical tests using cell culture, tissue culture, animal models and pilot clinical trials to evaluate the relative performance of these systems. The question to be addressed is whether imaging without a pinhole or other depth-sectioning technique yields images of sufficient quality to appreciate cellular detail required clinically, and to determine whether out-of-focus and scattered photons reduce contrast to too large a degree. Our preliminary results indicate that it is possible to obtain good images of cellular detail in human subjects using contrast agents in routine clinical use. However, to judge image quality with the various devices and contrast agents, we will use quantitative measures of image resolution, image contrast and image SNR. Research proposed here will develop a new tool for image guided tissue assessment in real time, and confirmatory biopsy at image directed locations. The ION can image a broad range of contrast agents enabling molecular information from specific sites. The cost of the integrated optical needle will be low even in small quantities (below $2,300) but there is great potential to further reduce this cost when manufactured in quantity.