Low-Cost, Handheld Light Sheet Microscope for Guiding Anal Cancer Diagnosis

项目摘要/摘要 肛门癌的发病率一直在上升，在过去的三十年里增加了四倍。发病率为 艾滋病毒阳性男同性恋者的比例高得令人无法接受，每10万人中有131人。肛门癌的主要挑战 诊断和预防缺乏标准化的筛查程序。活体光学显微镜 共聚焦显微镜和光学相干层析成像等技术具有可视化的潜力 与肛门恶性肿瘤相关的细胞形态变化，但它们的临床适应性已被 很有挑战性。在这个开拓者R21项目中，我们建议开发一种低成本的、基于散射的光片 显微镜(SLSM)设备，可以帮助准确诊断肛门癌，目标是最终减少 与肛门癌相关的发病率和死亡率。我们的sLSM设备将独一无二地满足 现有的活体显微镜技术(例如CM、OCT)，同时实现了i)高横向分辨率(1- 2微米)用于检查上皮细胞的核特征，ii)大视野(~2.5 mm)，iii)可视化 单个图像中的全部上皮厚度，以及iv)低设备成本(约2,000美元)。我们预计sLSM设备 将揭示体内关键的诊断细胞特征，这将有助于早期肛门的检测 恶性程度高，及时启动适当治疗。SLSM设备还将引导活检 恶性病变具有较高的敏感性和特异性，可提高活检产量，降低发病率和死亡率 良性病变不必要的活检引起的并发症。作为迈向这一目标的第一步，我们将 研制手持式sLSM样机并评价其对肛门恶性肿瘤诊断的准确性。 目标1：研制用于肛门黏膜成像的手持式sLSM探头。SLSM设备 将在上皮细胞中提供细胞分辨率，并在单个 图像。我们将对照明和探测光学进行优化设计，以实现1.4微米的横向分辨率 超过2.5 mm视野，上皮轴向分辨率为6.5±0.9微米，轴向分辨率为18±8微米 固有层浅层。在目标2中，我们将进行一项对人类肛门组织进行成像的临床研究 手持sLSM设备。100例与肛门癌有关的肛门活组织检查将首先使用 SLSM设备。SLSM图像将与相应的组织学图像进行比较，以识别细胞和 在sLSM图像中可视化的架构特征，并制定sLSM诊断标准。敏感度 并评价sLSM诊断肛门恶性肿瘤的特异性。自动图像分析 将开发算法来定量评估蜂窝和建筑特征。除了……之外 提供一种低成本的显微镜工具，帮助准确诊断肛门恶性肿瘤，sLSM技术 可用于其他癌症(如宫颈癌、口腔癌、皮肤癌)的活体诊断、术中 核心针活检时手术切缘的评估和质量保证。

Project Summary/Abstract Anal cancer incidence has been increasing, by four times over the last three decades. The incidence rate is unacceptably high among HIV-positive homosexual men, 131 per 100,000. The main challenge in anal cancer diagnosis and prevention is the lack of a standardized screening program. In vivo optical microscopy technologies such as confocal microscopy and optical coherence tomography have the potential to visualize cellular morphologic changes associated with the anal malignancy, but their clinical adaptation has been challenging. In this Trailblazer R21 project, we propose to develop a low-cost, scattering-based light sheet microscopy (sLSM) device that can aid accurate diagnosis of anal cancer, with the goal of ultimately reducing the anal cancer-associated morbidity and mortality. Our sLSM device will uniquely address the unmet needs in existing in vivo microscopy technologies (e.g., CM, OCT), simultaneously achieving i) high lateral resolution (1- 2 µm) for examining cellular nuclear features in the epithelium, ii) large FOV (~2.5 mm), iii) visualization of the entire epithelial thickness in a single image, and iv) low device cost (~$2,000). We expect that the sLSM device will reveal the key diagnostic cellular features in vivo, which will facilitate detection of early-stage anal malignancy and timely initiation of adequate treatment. The sLSM device will also guide the biopsy to malignant lesions with high sensitivity and specificity to increase the biopsy yield and reduce the morbidity and complication caused by the unnecessary biopsy of benign lesions. As the first step towards this goal, we will develop the prototype handheld sLSM device and evaluate the accuracy of diagnosing anal malignancy in the following aims: In Aim 1, we will develop a handheld sLSM probe for imaging anal mucosa. The sLSM device will provide cellular resolution in the epithelium and reveal architectural details in deeper regions in a single image. We will optimally design the illumination and detection optics to achieve the 1.4 µm lateral resolution over a 2.5-mm FOV, 6.5 ± 0.9 µm axial resolution for the epithelium, and 18 ± 8 µm axial resolution for the superficial lamina propria. In Aim 2, we will conduct a clinical study of imaging human anal tissues with the handheld sLSM device. One hundred anal biopsies concerning for anal cancer will be first imaged with the sLSM device. sLSM images will be compared with the corresponding histologic images to identify cellular and architectural features visualized in sLSM images and to develop the sLSM diagnostic criteria. The sensitivity and specificity of the sLSM for diagnosing anal malignancy will be evaluated. Automated image analysis algorithms will be developed to quantitatively evaluate the cellular and architectural features. In addition to providing a low-cost microscopy tool that aids accurate diagnosis of anal malignancy, the sLSM technology can be used for in vivo diagnosis of other cancers (e.g., cervical, oral, skin cancers), intra-operative assessment of the surgical margin, and quality assurance during the core needle biopsy.