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

项目概要/摘要 肛门癌的发病率一直在增加，在过去的三十年里增加了四倍。发病率 在艾滋病毒阳性的男同性恋者中，这一比例高得令人无法接受，每100 000人中有131人。肛门癌的主要挑战 诊断和预防是缺乏一个标准化的筛查计划。体内光学显微镜 技术，如共焦显微镜和光学相干断层扫描有潜力可视化 细胞形态学改变与肛门恶性肿瘤有关，但其临床适应性一直是 挑战性在这个开拓者R21项目中，我们建议开发一种低成本、基于散射的光片 显微镜（sLSM）设备，可以帮助准确诊断肛门癌，其目标是最终减少 肛门癌相关的发病率和死亡率。我们的sLSM设备将以独特的方式满足 现有的体内显微技术（例如，CM，OCT），同时实现i）高横向分辨率（1- 2 µm）用于检查上皮细胞核特征，ii）大FOV（~2.5 mm），iii） 单个图像中的整个上皮厚度，以及iv）低设备成本（~ 2，000美元）。我们希望sLSM设备 将揭示体内关键的诊断细胞特征，这将有助于检测早期肛门 恶性肿瘤和及时开始适当的治疗。sLSM器械还将引导活检， 恶性病变具有高敏感性和特异性，以提高活检率并降低发病率， 良性病变不必要活检引起的并发症。作为实现这一目标的第一步， 开发原型手持式sLSM设备，并评估诊断肛门恶性肿瘤的准确性， 以下目标：在目标1中，我们将开发一种用于肛门粘膜成像的手持式sLSM探头。sLSM设备 将在上皮中提供细胞分辨率，并在单个细胞中揭示更深区域的结构细节。 形象我们将优化设计照明和检测光学器件，以实现1.4 µm的横向分辨率 在2.5 mm FOV范围内，上皮细胞轴向分辨率为6.5 ± 0.9 µm， 浅层固有层在目标2中，我们将进行一项临床研究， 手持sLSM设备。100例与肛门癌有关的肛门活检将首先使用 sLSM设备。将sLSM图像与相应的组织学图像进行比较，以识别细胞和 在sLSM图像中可视化的结构特征，并开发sLSM诊断标准。灵敏度 并评价sLSM诊断肛门恶性肿瘤的特异性。自动图像分析 将开发算法以定量评估细胞和结构特征。除了 sLSM技术提供了一种低成本的显微镜工具，有助于准确诊断肛门恶性肿瘤， 可用于其它癌症的体内诊断（例如，宫颈癌、口腔癌、皮肤癌），术中 手术切缘的评估和芯针活检过程中的质量保证。

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.