Confocal line-scanning versus point-scanning for imaging human skin in vivo

描述(申请人提供)：本项目的目标是将两台共焦线扫描显微镜与标准共焦点扫描显微镜进行比较，以便在活体内对人体皮肤进行反射成像。一种行扫描仪基于分割的物镜光瞳，另一种基于完全的光瞳配置。每种配置都有各自的优势。长期目标是创造一种从根本上更简单的共聚焦反射扫描显微镜，用于为临床和外科皮肤病成像人类皮肤癌。每年，仅在美国就有120万新的皮肤癌病例被诊断出来。在进行的550万例活组织检查中，80%被证明是正常的，因此可以避免，每年花费美国医疗保健16.5亿美元。共聚焦反射显微镜可以实现对这些皮肤癌的非侵入性和实时的筛查和诊断，而对活检的需求最小。我们的假设是，无论是分光还是完全光瞳的共焦线扫描仪都将是简单的，并且将提供非常高的分辨率和光学切片。这种线扫描显微镜可能会与当今最先进的点扫描显微镜在活体人体皮肤成像、临床诊断和手术指导应用方面展开竞争。用原型共聚焦分瞳行扫描仪的初步结果显示了活体人体皮肤表皮中的核和细胞细节的成像。该样机比点扫描仪要简单得多。其具体目标是创造一种结合了分瞳和全瞳孔配置的共焦线扫描仪。混合的分光和全瞳将能够在相同的光学和皮肤条件下对它们的线扩展函数(LSF)和成像性能进行实验评估。我们将在两种情况下对分瞳和全瞳LSF的共焦散布函数(PSF)进行实验比较：衍射受限条件(即标称仪器)以及实际皮肤的散射和像差条件(即人体真皮深处，穿过全层表皮)。人体皮肤的分瞳和全瞳线扫描图像，无论是在体内还是体外，都将与组织学和通过共聚焦扫描仪获得的特征良好的图像进行比较。对线扫描和点扫描两种扫描方法的图像背景噪声和对比度进行了定量比较。与公共卫生相关的共聚焦反射显微镜可以对黑色素瘤、基底细胞癌和鳞状细胞癌等皮肤癌进行筛查和诊断，或者可以直接和实时地指导此类癌症的手术。诊断或手术指导可能是非侵入性的，需要最小的活检，最小的痛苦和最小的费用。

DESCRIPTION (provided by applicant): The goal of this project is to compare two confocal line-scanning microscopes to a standard confocal pointscanning microscope for reflectance imaging of human skin in vivo. One line-scanner is based on a divided objective lens pupil and the other based on a full pupil configuration. Each configuration has advantages. The long-term objective is to create a fundamentally simpler class of confocal reflectance scanning microscopes for imaging human skin cancers for clinical and surgical dermatology. Every year, in the USA alone, 1.2 million new cases of skin cancers are diagnosed. Of the 5.5 million biopsies performed, 80% turn out to be normal and thus avoidable, costing US healthcare $1.65 billion annually. Confocal reflectance microscopy may enable screening and diagnosis of these skin cancers noninvasively and in real-time, with minimal need for biopsy. The hypothesis is that a confocal line-scanner with either a divided-pupil or a full pupil will be simple, and will provide very high resolution and optical sectioning. Such line-scanning microscopes may compete with today's state-of-the-art point-scanning microscopes for imaging human skin in vivo, for clinical diagnosis and surgical guidance applications. Preliminary results with a prototype confocal divided-pupil line-scanner demonstrate imaging of nuclear and cellular detail in the epidermis of human skin in vivo. The prototype is considerably less complicated than a point-scanner. The specific aims are to create a confocal line-scanner that combines both divided-pupil and full-pupil configurations. The combined divided-and-full pupil will enable experimental evaluation of their line spread functions (LSFs) and imaging performance under identical optical and skin conditions. Experimental comparisons of divided pupil and full-pupil LSFs to the confocal point spread function (PSF) will be made under two conditions: diffraction-limited (i. e., nominal instrument) conditions as well as scattering and aberrating conditions of actual skin (i. e., deep within human dermis, through full-thickness epidermis). Divided-pupil and full-pupil line-scanned images of human skin, both ex vivo and in vivo, will be compared to histology and to the well characterized images obtained by confocal point-scanners. Background noise and contrast in the images will be quantitatively compared between the two line-scanning and the point-scanning methods. RELEVANCE TO PUBLIC HEALTH Confocal reflectance microscopy may enable screening and diagnosis of skin cancers such as melanomas and basal- and squamous-cell carcinomas, or may guide surgery of such cancers, directly and in real-time on the patient. The diagnosis or surgical guidance may be noninvasive, with minimal need for biopsy, minimal pain and minimal expense.