Automated Cell Preparation in Tubes for 3D Microscopy

用于 3D 显微镜的管内自动细胞制备

• 批准号: 7472827
• 负责人: Deirdre R. Meldrum
• 金额: $ 3.16万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-09 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7472827
• 关键词: bioimaging /biomedical imaging; biomedical automation; cell morphology; clinical research; cytodiagnosis; early diagnosis; flow cytometry; high throughput technology; human tissue; molecular probes; neoplasm /cancer diagnosis; technology /technique development; three dimensional imaging /topography; tissue /cell preparation

DESCRIPTION (provided by applicant): 3D microscopy represents a powerful new cell analysis tool for early detection and diagnosis of cancer, but its future use may be limited because methods for preparation of samples are cumbersome, inefficient, labor intensive and generally imprecise. Current methods for cytological sample collection are manual and distributed in nature through various physicians' office laboratories and local hospitals, with the actual analysis being centralized at regional clinical laboratories. Among the cytological specimens are sputum, gynecological and colorectal scrapes, fine needle aspirates, urinary tract, and gastrointestinal samples. We propose the development of a new automated system that will transform these difficult and messy clinical specimens into an optimal format for 3D microscopy morphological and molecular analysis. The model and method we propose is comprised of three sequential steps. First, at the distributed site, an automated sample processor dissociates and fixes cells and debris for shipment in an automation compatible canister. Second, after shipment to a centralized clinical laboratory, the specimen canisters are loaded into an automatic processor that performs cleanup (debris removal), specimen/assay specific staining (and counterstaining), and finally embedding of cells of interest in glass microcapillary tubes (about 50 mu m ID), with cells being spaced at regular intervals (about 200 mu m) within a tube. This preparation format is uniquely suited for integration with multiple 3D imaging platforms for true 3D volumetric assessment of cell morphology and molecular probe and/or stain density distribution. The proposed system also enables use of cytometric flow sorting for enrichment of cells of interest at an intermediate stage of the sample preparation process. The potential impact to improved human health through rapid diagnostic screening will be illustrated using a high impact emerging technology, optical tomography. In summary, the aim of the proposed project is to develop, design, and build a complete sample processing system that automates the process of sample cleanup, assay specific staining, and mounting of cells into glass microcapillary tubes, and a tube positioning and rotation scanner mechanism for 3D microscopy analysis of cell morphology for the early detection of cancer.

描述（由申请人提供）：3D 显微镜代表了一种用于癌症早期检测和诊断的强大的新型细胞分析工具，但其未来的使用可能受到限制，因为样品制备方法繁琐、低效、劳动密集型且通常不精确。目前的细胞学样本采集方法是手动的，并且本质上通过各个医生办公室实验室和当地医院进行分发，实际分析集中在区域临床实验室。细胞学标本包括痰、妇科和结直肠刮片、细针抽吸物、尿道和胃肠道样本。我们建议开发一种新的自动化系统，将这些困难且混乱的临床标本转化为 3D 显微镜形态学和分子分析的最佳格式。我们提出的模型和方法由三个连续的步骤组成。首先，在分布式站点，自动样品处理器分离并固定细胞和碎片，以便在自动化兼容罐中运输。其次，在运送到集中临床实验室后，样本罐被装入自动处理器中，该处理器执行清理（去除碎片）、样本/测定特定染色（和复染），最后将感兴趣的细胞嵌入玻璃微毛细管中（大约50μm ID），细胞在管内以规则的间隔（约200μm）间隔开。这种制备形式特别适合与多个 3D 成像平台集成，以对细胞形态和分子探针和/或染色密度分布进行真正的 3D 体积评估。所提出的系统还能够在样品制备过程的中间阶段使用细胞流式分选来富集感兴趣的细胞。将使用高影响力的新兴技术光学断层扫描来说明通过快速诊断筛查对改善人类健康的潜在影响。总之，拟议项目的目的是开发、设计和构建一个完整的样品处理系统，该系统可自动执行样品净化、测定特定染色、将细胞安装到玻璃微毛细管中的过程，以及管定位和旋转扫描仪3D 显微镜分析细胞形态以早期检测癌症的机制。