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已经证明了光学成像提供具有亚细胞分辨率的分子图像的潜力。在这个建议中，我们扩展了我们的工作在高分辨率共聚焦显微镜，基于一个更简单的替代方案，基于我们的观察，高分辨率的光学图像的顶部1-2层的细胞可以获得，而不需要一个共聚焦成像门，如果细胞被放置在直接接触一个平面的光学窗口。在这里，我们提出了一类新的光学成像设备-集成光学针（ION）-它可以被插入到一个小规格的针和先进的通过组织，以获取图像的组织在针的远端与亚细胞分辨率在真实的时间;使用有针对性的造影剂或活体染色可以产生额外的对比度以及功能和/或分子成像能力。在目标1中描述的光学针的第一个版本是基于耦合到宏观成像光学器件和CCD图像传感器的柔性相干光纤束。与目标1中使用的光纤束的结构相关联的欠采样限制了空间分辨率。第二种方法是在目标2和3中描述的集成光学针（ION），其使用LIGA技术将使用灰度光刻制造的微型透镜与光源和图像传感器集成在一起，以产生可以通过针插入的微型显微镜。该ION由NA = 0.4的显微镜组成，该显微镜可以从250 μ m的视场以1微米的横向分辨率成像;它被设计成适合于内径为1 mm的针。我们将使用细胞培养、组织培养、动物模型和试点临床试验进行各种临床前测试，以评估这些系统的相对性能。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，以了解临床所需的细胞细节，并确定是否离焦和散射光子降低对比度太大的程度。我们的初步结果表明，它是可能的，以获得良好的图像细胞的细节在人类受试者使用造影剂在常规临床使用。然而，为了判断各种器械和造影剂的图像质量，我们将使用图像分辨率、图像对比度和图像SNR的定量测量。本文提出的研究将开发一种新的工具，用于真实的实时图像引导组织评估，并在图像引导位置进行确认活检。ION可以对各种造影剂成像，从而获得特定部位的分子信息。集成光学针的成本即使在小批量（低于2，300美元）下也将是低的，但是当大量制造时，有很大的潜力进一步降低该成本。

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.