High resolution phase contrast endoscopy

摘要 相差显微镜是用于生物成像的最广泛使用的技术之一，因为它 提供样品形态的精美高分辨率图像，无需使用样品标记。然而 标准的相位对比技术，如微分干涉对比（DIC），仅在传输中起作用 方向，因此不能在成像厚样品时使用。因此，标准相位差 这些技术在体内生物医学或临床成像中没有产生很大的影响。 我们已经开发了一种新的相衬技术，称为斜背照明显微镜（OBM）， 以反射光结构工作，因此适合于体内显微内镜应用。OBM 不需要标记，并提供亚表面样品形态的高分辨率DIC样图像。远 如我们所知，以前从未展示过落射探测配置中的类似DIC的成像。我们的目标 在这个项目中，是为了证明OBM可以使临床医生进行组织病理学和组织诊断 在原位，而不需要物理活检。 我们将把我们的研究限制在OBM显微内镜用于临床前癌症诊断的评价上。特别是， 我们将集中于啮齿动物体内和体外的结肠直肠成像。小动物成像， 内窥镜提出了与探针小型化相关的技术挑战， 人体成像尽管如此，我们认为这种成像是必要的第一步，必须在 临床评价。因此，我们的具体目标是：1）构建一个小型化的OBM显微内镜 适用于大鼠体内结肠直肠成像的探针，2）进行化学大鼠的纵向体内成像 癌症模型，和3）进行遗传小鼠癌症模型的离体成像以与标准的 H & E组织病理学。 开发新技术的困难在于，虽然我们已经建立了OBM， 在厚组织中产生高分辨率图像，我们还没有一个路线图来说明这些图像可以发挥什么作用， 在临床应用中发挥作用（正是因为以前没有人能够进行这种成像）。我们 该项目的目标是通过将OBM成像结果与已知的 癌症在良好表征的动物模型中，从而使我们能够定义与以下相关的分类标准： 这项新技术。最终，我们希望这项临床前研究将导致一个简单的， 安全、多功能、低成本的显微内窥镜技术，可与标准技术结合使用 内镜在大肠癌原位诊断中的临床应用

ABSTRACT Phase contrast microscopy is one of the most widely used techniques for biological imaging because it provides exquisite high-resolution images of sample morphology, without the use of sample labeling. However standard phase contrast techniques, like differential interference contrast (DIC), only work in the transmission direction and thus cannot be used when imaging thick samples. For this reason, standard phase contrast techniques have not made a great impact in in-vivo biomedical or clinical imaging. We have developed a new phase contrast technique, called Oblique Back-illumination Microscopy (OBM), that works in a reflected light configuration, and is thus amenable to in-vivo endomicroscopy applications. OBM requires no labeling and provides high resolution DIC-like images of sub-surface sample morphology. As far as we know, DIC-like imaging in an epi-detection configuration has never been demonstrated before. Our goal in this project is to demonstrate that OBM can enable clinicians to perform histopathology and tissue diagnosis in situ, without the need for a physical biopsy. We will limit our study to the evaluation of OBM endomicroscopy for preclinical cancer diagnosis. In particular, we will concentrate on colorectal imaging in rodents, in-vivo and ex-vivo. Small animal imaging with endomicroscopes presents technical challenges related to probe miniaturization that are more restrictive than human imaging. Nevertheless, we believe such imaging is a necessary first step that must be taken prior to clinical evaluations. Accordingly, our specific aims are to 1) construct a miniaturized OBM endomicroscope probe suitable for in-vivo colorectal imaging in rats, 2) perform longitudinal in-vivo imaging of a chemical rat cancer model, and 3) perform ex-vivo imaging of a genetic mouse cancer model to compare with standard H&E histopathology. A difficulty with the development of a new technique is that, while we have already established that OBM produces high resolution images in thick tissue, we do not yet have a roadmap for what role these images can play in clinical applications (precisely because no-one has been able to perform such imaging before). Our goal for this project is to establish this role by correlating OBM imaging results with the known chronology of cancer in well characterized animal models, thus enabling us to define classification criteria associated with this new technology. Ultimately, we hope this preclinical study will lead toward the development of a simple, safe, versatile, low-cost endomicroscopy technique that can be operated in conjunction with standard endoscopy for in-situ colorectal cancer diagnosis in the clinic.