Novel Nanoscale Single-Cell Analysis of Exfoliative Cytology

描述（申请人提供）：脱落细胞的体外分析在疾病筛查和临床诊断中发挥着重要作用。准确的细胞学诊断在很大程度上取决于检测单个细胞的细胞和核结构特征变化的能力。传统光学显微镜的衍射极限分辨率限制了对癌细胞的主要微米级特征的检测，这导致观察者内的变化，错过早期癌症（假阴性），以及不确定的情况，这些情况通常导致在没有癌症的情况下进行不必要的侵入性手术（假阳性）。迫切需要进行高速、准确、定量和具有成本效益的细胞学分析并适合常规临床使用的技术。细胞纳米结构 这些变化代表了用于鉴定癌前细胞或癌细胞的有希望的策略。我们最近开发了两种互补的技术，这两种技术都可以在单细胞水平上评估纳米级的结构特征：定量相位显微镜和空间频率的光谱编码（SESF）。我们证明了它们能够检测传统光学显微镜错过的癌前细胞的结构变化。我们建议开发一种新的先进的显微镜，它集成了这两种技术的互补属性，以进行实时定量结构成像和高速和全面的纳米级结构分析的无标记的单细胞在其自然状态下用于研究和临床应用。该显微镜系统的技术性能将被表征，其分析纳米结构的能力将被严格验证。通过开发细胞分割算法和多尺度结构分析，将其优化用于常规临床使用，以分析未染色的脱落细胞学标本。我们将重点放在两个大体积和临床上具有挑战性的细胞学标本作为我们的疾病模型：宫颈和尿液细胞学。将通过训练集和验证集使用相对较大规模的宫颈和尿液细胞学标本来测试系统的准确性。如果成功，该技术可用于支持常规细胞学进行准确，定量，高速，全面和具有成本效益的细胞学分析，以提高诊断准确性并防止昂贵的不必要程序。

DESCRIPTION (provided by applicant): In vitro analysis of exfoliated cells plays an important role in disease screening and clinical diagnosis. An accurate cytological diagnosis depends largely on the ability to detect changes in cellular and nuclear structural characteristics of individual single cells. The diffraction-limited resolution of a conventional light microscope limis the detection of mostly micron-scale features of cancer cells, which result in intra-observer variation, missed early-stage cancers (false negatives), and indeterminate cases that often result in unnecessary invasive procedures in the absence of cancer (false positives). Technologies that perform high-speed, accurate, quantitative and cost-effective cytological analysis and are suitable for routine clinic use are urgently needed. Cellular nanoscale structural changes represent a promising strategy for identifying pre-cancerous or cancerous cells. We have recently developed two complementary techniques, both of which can assess nanoscale structural characteristics at the single-cell level: quantitative phase microscopy and spectral-encoding of spatial frequency (SESF). We demonstrated their ability to detect structural changes in pre-cancerous cells missed by conventional light microscopy. We propose to develop a new advanced microscopy that integrates the complementary attributes of these two techniques to perform real-time quantitative structural imaging and high-speed and comprehensive nanoscale structural analysis of label-free single cells in their natural state for use in both research and clinical applications. The technical performance of this microscopy system will be characterized and their ability to analyze nanoscale structures will be rigorously validated. It will be optimized for routine clinical use to analyze unstained exfoliative cytology specimens by developing cell segmentation algorithm and multi-metric structural analysis. We will focus on two large-volume and clinically challenging cytology specimens as our disease models: cervical and urine cytology. The accuracy of the system will be tested using a relatively large scale of cervical and urine cytology specimens via a training and a validation set. If successful, this technique could be used to support conventional cytology for accurate, quantitative, high-speed, comprehensive and cost-effective cytological analysis to improve the diagnostic accuracy and prevent costly unnecessary procedures.