CELLULAR AND NUCLEAR SIZING IN NORMAL AND ABNORMAL SOFT

涉及皮肤和皮下软组织的疾病很多。这些组织涉及广泛的先天性、炎性、感染性、创伤性和肿瘤过程。除了体格检查，大多数这些病变的诊断是侵入性的、耗时的和昂贵的，通常需要手术切除或活检，然后进行病理学检查。评估皮肤和软组织细胞变化的非侵入性、有效、低成本的技术在许多状况的管理中将是极其有价值的。已知正常细胞核具有4 - 7 μ m的特征直径，而发育异常细胞核通常增大并可大至20 μ m。因此，在大多数人体组织中，增大的细胞核是癌症、发育异常和细胞再生的主要细胞指标。目前用于测定组织中细胞核大小的方法如图像分析和流式细胞术需要将细胞样品分散在液体培养基或组织切片中，这不能用于细胞的体内分析。该R 03应用旨在开发一种新的光学方法，使我们能够直接从组织中的多重散射光或光子迁移测量中提取体内核尺寸分布。我们的想法核大小是使用基于模型的计算算法来重建核的大小分布中所包含的组织散射光谱。我们的核尺寸分布测量可以实时提供。我们的方法也将使我们能够恢复其他散射体，如血细胞的大小分布。有了这种能力，我们将能够检测到由于这些其他子结构的大小变化而导致的异常。因此，本申请的具体目的是：（1）继续开发用于提取核尺寸分布的正则化逆算法，（2）使用组织模型、培养细胞和人类皮肤等效实验来评估和优化计算机算法和基于CD的连续波光谱系统的集成功能，以及（3）基于在人类受试者试验期间获得的初步数据来评估我们的集成光学系统。

Diseases involving the skin and underlying soft tissues are legions. These tissues are involved by a wide range of congenital, inflammatory, infectious, traumatic, and neoplastic processes. Aside from physical examination, the diagnosis of most of these lesions is invasive, time- consuming, and costly, often requiring surgical excision or biopsy followed by pathological investigations. A non-invasive, efficient, low cost technique of evaluating skin and soft tissue cellular changes would be extremely valuable in the management of numerous conditions. It is known that normal nuclei have a characteristic diameter between 4 and 7mum, while dysplastic nuclei generally enlarge and can be as large as 20 mum. Thus, enlarged nuclei are primary cellular indicators of cancer, dysplasia and cell regeneration in most human tissues. Current methods such as image analysis and flow cytometry used to size nuclei in tissue require dispersed cellular samples in liquid media or tissue sections which cannot be used for in vivo analysis of cells. This R03 application is aimed to develop a novel optical method that will allow us to extract in vivo nuclear size distribution directly from multiply scattered light or photon migration measurement in tissue. Our idea for nuclear sizing is to use model-based computational algorithms to reconstruct the nuclear size distribution that is contained in the tissue scattering spectral. Our measurement of nuclear size distribution can be provided in real-time. Our approach will also allow us to recover the size distributions of other scatterers such as blood cells. With this capability, we will be able to detect abnormalities due to the size changes of these other substructures. Thus, the specific aims of this application are: (1) to continue the development of regularized inverse algorithms for the extraction of nuclear size distribution, (2) to evaluate and optimize the integrated functioning of the computer algorithms and a CD-based continuous-wave optical spectral system using tissue phantom, cultured cell and the human skin equivalent experiments, and (3) to evaluate our integrated optical system based on preliminary data obtained during a human subjects trial.