Novel Nanoscale Single-Cell Analysis of Exfoliative Cytology

剥脱细胞学的新型纳米级单细胞分析

• 批准号: 9026609
• 负责人: Yang Liu
• 金额: $ 34.65万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9026609
• 关键词: Algorithms; Attention; Benign; Cancerous; Cell Cycle; Cells; Cellular Structures; Cervical; Characteristics; Clinic; Clinical; Clinical Management; Cytology; Detection; Diagnosis; Diagnostic Neoplasm Staging; Disease; Disease model; Electron Microscopy; Ensure; Evaluation; Fluorescence Microscopy; Frequencies; Future; Gold; Health; Image; In Vitro; Individual; Intraobserver Variability; Label; Length; Lesion by Stage; Light Microscope; Liquid substance; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of urinary bladder; Methods; Microscopy; Morphology; Nanostructures; Nuclear; Optics; Patients; Performance; Phase; Photophobia; Play; Procedures; Property; Research; Resolution; Role; Sampling; Screening for cancer; Specimen; Speed; Staging; Structure; System; Systems Analysis; Techniques; Technology; Testing; Time; Training; Unnecessary Procedures; Urine; Validation; cancer cell; carcinogenesis; clinical Diagnosis; clinical application; cohort; contrast imaging; cost effective; diagnostic accuracy; exfoliative cytology; improved; instrument; light microscopy; microscopic imaging; nano; nanoscale; nanostructured; novel; prevent; prototype; reconstruction; research study; screening; single cell analysis; validation studies

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.

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起着重要作用。准确的细胞学诊断在很大程度上取决于检测单个单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受限分辨率限制了对癌细胞主要微米级特征的检测，这会导致观察者内部的差异，错过早期癌症(假阴性)，以及经常导致在没有癌症的情况下进行不必要的侵入性手术的不确定病例(假阳性)。迫切需要进行高速、准确、定量和经济有效的细胞学分析并适合常规临床使用的技术。细胞纳米结构 改变代表了一种很有前途的识别癌前细胞或癌细胞的策略。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到常规光学显微镜遗漏的癌前细胞的结构变化。我们建议开发一种新的先进显微镜，它集成了这两种技术的互补属性，可以对自然状态下的无标记单细胞进行实时定量结构成像和高速、全面的纳米级结构分析，用于研究和临床应用。这种显微镜系统的技术性能将得到表征，其分析纳米级结构的能力将得到严格验证。通过开发细胞分割算法和多尺度结构分析，分析未染色的脱落细胞学标本将优化为常规临床使用。我们将专注于两个具有临床挑战性的大容量细胞学样本作为我们的疾病模型：宫颈细胞学和尿细胞学。该系统的准确性将通过培训和验证集使用相对较大规模的宫颈和尿液细胞学样本进行测试。如果成功，这项技术可用于支持传统细胞学进行准确、定量、高速、全面和经济有效的细胞学分析，以提高诊断准确性，避免昂贵的不必要的程序。

项目成果

Stochastic Optical Reconstruction Microscopy (STORM).

随机光学重建显微镜（STORM）。

• DOI: 10.1002/cpcy.23
• 发表时间: 2017
• 期刊: Current protocols in cytometry
• 作者: Xu,Jianquan;Ma,Hongqiang;Liu,Yang
• 通讯作者: Liu,Yang

Super-Resolution Imaging of Higher-Order Chromatin Structures at Different Epigenomic States in Single Mammalian Cells.

• DOI: 10.1016/j.celrep.2018.06.085
• 发表时间: 2018-07-24
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Xu J;Ma H;Jin J;Uttam S;Fu R;Huang Y;Liu Y
• 通讯作者: Liu Y

Targeted DNA damage at individual telomeres disrupts their integrity and triggers cell death.

• DOI: 10.1093/nar/gkv598
• 发表时间: 2015-07-27
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Sun L;Tan R;Xu J;LaFace J;Gao Y;Xiao Y;Attar M;Neumann C;Li GM;Su B;Liu Y;Nakajima S;Levine AS;Lan L
• 通讯作者: Lan L

Nuclear Nano-architecture Markers of Gastric Cardia and Upper Squamous Esophagus Detect Esophageal Cancer "Field Effect".

• DOI: 10.7150/jca.6990
• 发表时间: 2013
• 期刊: Journal of Cancer
• 影响因子: 3.9
• 作者: Fasanella KE;Bista RK;Staton K;Rizvi S;Uttam S;Zhao C;Sepulveda A;Brand RE;McGrath K;Liu Y
• 通讯作者: Liu Y

Fast and Precise 3D Fluorophore Localization based on Gradient Fitting.

• DOI: 10.1038/srep14335
• 发表时间: 2015-09-22
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Ma H;Xu J;Jin J;Gao Y;Lan L;Liu Y
• 通讯作者: Liu Y