High resolution phase contrast endoscopy

摘要 相差显微镜是应用最广泛的生物成像技术之一，因为它 提供精美的高分辨率样本形态图像，无需使用样本标签。然而， 标准的相位对比技术，如差分干涉对比(DIC)，仅在传输中起作用 因此，在对厚样本进行成像时不能使用。因此，标准相位对比度 在活体生物医学或临床成像中，这些技术并没有产生太大的影响。 我们开发了一种新的相位对比度技术，称为斜背照明显微镜(OBM)，它 工作在反射光配置中，因此适用于体内内窥镜应用。OBM 不需要贴标签，并提供亚表面样品形态的高分辨率DIC类图像。到目前为止 正如我们所知，在表观检测配置中的DIC样成像以前从未被演示过。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，不需要物理活检。 我们将把我们的研究局限于评估OBM内窥镜在临床前癌症诊断中的作用。特别是， 我们将集中在啮齿动物体内和体外的结肠成像。使用小动物成像 内窥镜带来了与探头小型化相关的技术挑战，这些挑战比 人体成像。然而，我们认为这样的成像是必要的第一步，必须在 临床评估。因此，我们的具体目标是：1)构建一个小型化的OBM内窥镜 适用于大鼠在体结直肠成像的探针，2)对化学大鼠进行体内纵向成像 肿瘤模型，以及3)对遗传性小鼠癌症模型进行体外成像以与标准的 H&E组织病理学。 开发新技术的一个困难是，尽管我们已经建立了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起到什么作用 在临床应用中发挥作用(正是因为之前还没有人能够进行这样的成像)。我们的 该项目的目标是通过将OBM成像结果与已知的年表相关联来确定这一角色 在具有良好特征的动物模型中发现癌症，从而使我们能够定义与 这项新技术。最终，我们希望这项临床前研究将导致开发一种简单、 安全、通用、低成本的内窥镜技术，可与标准 内窥镜在临床原位结直肠癌诊断中的应用。

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.