Development of high-throughput, commercially viable, Cell-CT instrument with init

描述（由申请人提供）：VisionGate开发了Cell-CT平台，最初用于以下三个市场：1）胃食管反流病（GERD）的检测和管理，2）肺癌风险评估（其他癌症的检测，在管道中），3）学术和制药研究（非FDA应用）。Cell-CT是一种台式设备，设计用于临床和研究实验室的体外使用。它是一个独特的强大的3D细胞分析平台，能够定量分析细胞内染色质、生物标记物、特定染色剂和其他吸收和/或荧光结构的3D形态，并进一步能够为任何具有细胞内容物的样品提供自动化早期疾病检测，这些细胞内容物可以在流体悬浮液中呈现（例如，血液、痰、宫颈刮片、乳房抽吸物、尿液、结肠刷、细针活组织检查）。与流式细胞术类似，细胞被注入毛细管中。但是Cell-CT中的试管是独一无二的，因为当细胞流过它时，它会轴向旋转。当试管旋转时，Cell-CT会在每个细胞旋转3600圈时创建数百个显微镜伪投影图像（具有扩展的焦深）。这些2D图像被馈送到反投影图像重建算法中以生成具有各向同性分辨率的细胞的3D图像。Cell-CT目前处于原型形式，并被证明能够以高分辨率和高保真度提供3D细胞图像。此外，Cell-CT测量和分类3D细胞特征，以提供单细胞分析和样本分类。这一点也得到了证明。然而，商业上可行的产品的最终障碍是提高产量。今天的Cell-CT需要15分钟来收集数据和处理单个细胞。一个典型的样本可能包含数千个细胞，因此目前的通量在临床实验室环境中将是无用的，在临床实验室环境中将执行Cell-CT的许多重要应用。根据该资助申请支持的开发工作将实现20分钟的目标速度，以在Cell-CT上完全处理细胞标本，这是临床实验室行业可接受的吞吐量。已经提出了几种仪器修改，以增加流体流动速度，采用1D和2D细胞仪来最佳地分类靶细胞，以获得更耗时的3D图像，并最终确定产品包装的仪器设计。这项工作的成功成果将是商业级的Cell-CT仪器。 公共卫生相关性：细胞定量显微分析的主要障碍是传统光学显微镜在解释被限制在载玻片上的三维细胞时的二维性质。Cell-CT以独特的方式将传统显微镜的图像质量与流式细胞术的流体学相结合，允许断层图像重建以计算细胞的真实3D结构。这种3D成像能力将应用于痰液中，以评估GERD和肺癌的早期检测，但在Cell-CT可以商业化使用之前，其目前的低通量必须得到显着改善，以允许在几分钟内而不是几小时内进行标本分析。

DESCRIPTION (provided by applicant): VisionGate has developed the Cell-CT platform for initial use in the three following markets: 1) Detection and management of gastroesophageal reflux disease (GERD), 2) Lung cancer risk assessment (detection of other cancers, in the pipeline), 3) Academic and pharma research (non-FDA applications). The Cell-CT is a bench-top device designed for in-vitro use in clinical and research laboratories. It is a uniquely powerful, 3D cell-analysis platform that enables the quantitative analysis of the 3D morphometry of chromatin, bio-markers, specific stains and other absorbing and/or fluorescing structures within cells, with the further capability of providing automated early disease detection for any sample with cellular content that can be presented in fluid suspension (e.g., blood, sputum, cervical scrape, breast aspirate, urine, colon brushing, fine needle biopsy). Similar to flow cytometry, cells are injected into a capillary tube. But the tube in the Cell-CT is unique, because it rotates axially as cells flow through it. As the tube rotates, the Cell-CT creates hundreds of microscopic pseudo-projection images (with extended depth of focus) around 3600 rotation of each cell. These 2D images are fed into a back-projection image reconstruction algorithm to generate the 3D image of a cell with isotropic resolution. The Cell-CT is currently in prototype form and is proven to deliver the 3D cell images at high- resolution and high-fidelity. Moreover, the Cell-CT measures and classifies 3D cell features to provide a single cell analysis and a specimen classification. This also has been demonstrated. However, the final hurdle for a commercially viable product is improved throughput. Today's Cell-CT requires 15 minutes just to gather data and process a single cell. A typical specimen might contain thousands of cells, so the current throughput would not be useful in a clinical lab setting where many of the important applications of the Cell-CT would be performed. The development effort to be supported under this grant application will achieve a target speed of 20 minutes to completely process a cell specimen on the Cell-CT, and this is an acceptable throughput for the clinical lab industry. Several instrument modifications have been proposed to increase fluid flow velocity, employ 1D and 2D cytometry to optimally triage target cells for more time consuming 3D imagery, and finalize the instrument design for product packaging. The successful outcome of this work will be a commercial-grade Cell-CT instrument. PUBLIC HEALTH RELEVANCE: A major impediment to quantitative microscopic analysis of cells is the two-dimensional nature of conventional optical microscopy in interpreting three-dimensional cells that are constrained to lie on a glass slide. The Cell- CT combines the image quality of conventional microscopy with the fluidics of flow cytometry in a unique manner that permits tomographic image reconstruction to compute the true 3D structure of cells. This 3D imaging capability will be applied to sputum for the assessment of GERD and for the early detection of lung cancer, but before the Cell-CT can be utilized commercially, its current low throughput must be improved significantly to allow specimen analysis in minutes rather than hours.