Towards In Situ Optical Pathology of Nonmelanoma Skin Cancers

描述(申请人提供)：非黑色素瘤皮肤癌是人类最常见的癌症。因此，迫切需要对这些肿瘤进行高分辨率的检测和显微控制的切除。目前，病理学家使用低倍显微镜观察肿瘤边缘，高倍显微镜检查细胞特征，以诊断和治疗非黑色素瘤皮肤癌。我们建议开发一种多模式设备，使其能够在活体和实时进行宽视场和高分辨率成像。该设备将结合CCD宏观成像和共焦显微镜的功能，前者可以快速检查大表面上的浅层组织层，后者可以在混浊介质中以与组织学相当的分辨率进行成像。该仪器将在这些模式之间进行缩放，允许更宽的视野引导高分辨率的视野。这两种模式都将获得反射、荧光和荧光偏振图像。为了增强图像的对比度，我们将使用FDA批准的传统组织学染色，在低浓度下无毒，即亚甲基蓝，以模拟H&E组织病理学的染色模式。这项技术的可行性将分别通过切除的人类非黑色素瘤皮肤癌标本和小动物的体外和体内试点试验来探索。在技术开发阶段，将进行三项试点试验。对切除的非黑色素瘤皮肤癌标本进行的第一次试验旨在通过与组织学的比较来确定光学图像中的诊断特征。在第二次试验过程中，第一阶段确定的诊断标准将用于临床试验，届时该系统将用于莫氏显微切除人类非黑色素瘤皮肤癌期间开放手术野的成像。最后，开发的技术的诊断能力将在小动物身上进行测试，方法是对可疑鳞状细胞癌的病变进行成像，并在组织病理学分析的基础上将结果与诊断进行比较。在该项目过程中开发的非侵入性设备和方法将能够在原位识别肿瘤边缘和小的肿瘤巢，从而改善癌症治疗的结果。这项技术的诊断应用具有取代活组织检查的潜力，从而减少了与侵入性手术相关的发病率和时间。

DESCRIPTION (provided by applicant): Nonmelanoma skin cancer is the most common form of human cancer. Therefore, there is a strong need for high-resolution detection and microscopically controlled removal of these neoplasms. Currently pathologists use microscopes at low power to view tumor margins and at high power to inspect cell features for diagnosis and management of nonmelanoma skin cancers. We propose to develop a multimodal device that will enable both wide-field and high-resolution imaging in vivo and in real time. The device will combine the capabilities of the CCD macro-imaging that enables rapid inspection of a superficial tissue layer over large surfaces, and confocal microscopy that allows imaging within turbid media with resolution comparable to that of histology. The instrument will zoom between these modes, allowing the wider view to direct the high resolution view. Both modes will acquire images in reflectance, fluorescence, and fluorescence polarization. To enhance the contrast of the acquired images, we will use an FDA-approved conventional histological stain, non-toxic at low concentrations, i.e. Methylene Blue, to mimic the staining pattern of H&E histopathology. The feasibility of the technology will be explored by in vitro and in vivo pilot trials with excised human nonmelanoma skin cancer specimens and in small animals, respectively. During the technology development stage three pilot trials will be conducted. The first trial with excised nonmelanoma skin cancer specimens is designed for determining diagnostic features in optical images by comparison to histology. In the course of the second trial, the diagnostic criteria defined by the first stage will be used in a clinical trial, when the system will be employed for imaging open surgical fields during Mohs micrographic removal of nonmelanoma skin cancers in humans. Finally, the diagnostic capability of the developed technology will be tested in small animals by imaging lesions suspicious for squamous cell carcinoma and comparing the results to the diagnosis, based on the analysis of histopathology. The noninvasive device and methods developed in the course of this project will enable identification of tumor margins and small tumor nests in situ, leading to improved outcomes of cancer treatments. The diagnostic application of this technology holds the potential to replace biopsy, thus reducing morbidity and time associated with that invasive procedure.