Lab-on-a-chip Flow Cytometer Using COlor-Space-Time (COST) Coding Method

描述（由申请人提供）：“Lab-on-a-chip flow cytometer using color-space-time（CoST）coding method”NanoSort，LLC RESEARCH & RELATED Other Project Information 7.流式细胞术（FACS）或流式细胞术使临床医生和研究人员能够定量表征细胞的物理（细胞大小，形状，粒度）和生化（DNA含量，细胞周期分布，细胞表面标志物和活力）特性。除了在基础研究（如免疫学、细胞和分子生物学）中的应用外，该仪器还使临床医生能够检测和监测急性骨髓性白血病（AML）和艾滋病毒/艾滋病等疾病的进展。凭借高通量分选以富集生物样本和提取稀有细胞类型的能力，最先进的流式细胞仪使得进行稀有事件研究（诸如细菌细胞、干细胞或肿瘤细胞的鉴定或分离）成为可能。然而，当今的流式细胞仪面临两个挑战，这限制了大幅降低其成本并将其日常使用扩展到临床环境的能力。第一个限制是系统的架构在利用越来越多的可用荧光颜色方面效率非常低。在当今的流式细胞仪设计中，每种荧光颜色都需要专用的PMT和光学器件;并且成本、复杂性和故障风险随着检测参数的数量而增加。其次，在流式细胞仪（计数细胞）和流式细胞仪（计数和分类细胞）之间存在巨大的价格差距（2- 3倍的价格差异）。外地资产管制系统大多设在共用的核心设施内，由训练有素的博士级专家操作。如果细胞分选变得比现在更容易获得和负担得起，那么对FACS的需求将会增加。基于罗教授在加州大学圣地亚哥分校的研究小组近10年的研究，我们将把芯片实验室技术开发成可以解决上述两个挑战的产品。为了促进转型，我们将应用我们的专利技术：1）颜色空间时间（COST）编码方法，使用单个PMT检测多个参数。COST技术从根本上改变了所有现有流式细胞仪中系统性能与系统复杂性之间的关系。我们还提出了一种高效和具有成本效益的芯片上压电细胞分选技术，具有低剪切和高细胞活力。我们提出的研究包括对芯片实验室FACS系统分选后细胞活力的系统研究，这是大多数研究实验室忽略的成功商业化的限制性问题。拟议的第一阶段研究使用创新方法将实验室技术转化为商业产品，这些产品的市场超过10亿美元，并且在拟议的成本降低和功能改进后有可能扩大市场。这些技术和商业目标将对基础研究和临床应用产生直接影响，以提高整个人口的健康和福祉。 公共卫生相关性：“Lab-on-a-chip flow cytometer using color-space-time（CoST）coding method”NanoSort，LLC RESEARCH & RELATED Other Project Information 8.项目叙述拟议的项目旨在开发高性能、易于操作和维护的芯片实验室流式细胞仪，并且比目前可用的任何流式细胞仪或荧光激活细胞分选仪（FACS）的成本显著更低，从而为研究和临床实践中的新设置提供这种广泛适用的技术。这项研究将把卢教授实验室开发的芯片实验室技术转化为产品，以满足先进生物医学研究和即时护理诊所的市场需求。所提出的系统实质上扩展了原有的大学技术，发明了COlor-空时（COST）编码技术，该技术允许使用单个光电倍增管（PMT）检测器进行多参数检测。

DESCRIPTION (provided by applicant): "Lab-on-a-chip flow cytometer using color-space-time (CoST) coding method" NanoSort, LLC RESEARCH & RELATED Other Project Information 7. PROJECT SUMMARY Fluorescence-activated-cell-sorting (FACS) or flow cytometry enables clinicians and researchers to quantitatively characterize the physical (cell size, shape, granularity) and biochemical (DNA content, cell cycle distribution, cell surface markers, and viability) properties of cels. Besides its applications in basic research (e.g. immunology, cell and molecular biology), the instrument has allowed clinicians to detect and monitor the progression of diseases such as acute myeloid leukemia (AML) and HIV/AIDS. With the capability of high- throughput sorting to enrich biospecimens and extract rare cell types, a state-of-the-art flow cytometer makes it possible to conduct rare-event studies such as the identification or isolation of bacterial cells, stem cells, or tumor cells. However, today's flow cytometers face two chalenges that limit the ability to drastically reduce their cost and extend their day-to-day utilization to clinical settings. The first limit is that the system's architecture is highly inefficient in utilizing the increasing number of available fluorescent colors. In today's flow cytometer design, each fluorescent color requires a dedicated PMT and optics; and the cost, complexity, and risk of failure grow with the number of detection parameters. Secondly, there exists a huge price gap (2-3X price difference) between flow cytometers (that count cells) and FACS (that count and assort cells). FACS are mostly located in shared core facilities and operated by well-trained, PhD level specialists. There is a large demand for FACS that would increase if cell sorting were to become more accessible and affordable than it is now. Based on nearly 10 years of research of Professor Lo's group at UCSD, we will develop the lab-on-a-chip technology into products that can address the above two challenges. To facilitate the transformation, we will apply our patented game-changing technologies: 1) COlor-Space-Time (COST) coding method to detect multiple parameters using a single PMT. The COST technique fundamentaly changes the relationship between the system performance and the system complexity in all existing flow cytometers. We also propose a highly efficient and cost effective on-chip piezoelectric cell sorting technique with low shearing and high cell viability. Our proposed research includes the systematic study of post-sorting cell viability for lab-on-a-chip FACS system, a limiting problem for successful commercialization that is overloked by most research laboratories. The proposed Phase I research uses innovative approaches to transform a laboratory technology into commercial products that have a market of over $1B and the potential for an expanded market following the proposed cost reductions and functionality improvements. These technology and business goals will have a direct impact on basic research and clinical applications to enhance the health and wellbeing of the entire population. PUBLIC HEALTH RELEVANCE: "Lab-on-a-chip flow cytometer using color-space-time (CoST) coding method" NanoSort, LLC RESEARCH & RELATED Other Project Information 8. PROJECT NARRATIVE The proposed project aims to develop lab-on-a-chip flow cytometers that are high performance, easy to operate and maintain, and significantly lower cost than any flow cytometers or fluorescence-activated-cell- sorters (FACS) available today, thus providing this broadly applicable technique to new settings in research and clinical practice. The research will transform the lab-on-a-chip technologies developed in Professor Lo's laboratory into products that meet the market needs in advanced biomedical research and point-of-care clinics. The proposed system substantially extends the original university technology with the invention of the COlor- Space-Time (COST) coding technique which allows multi-parameter detection using a single photo multiplier tube (PMT) detector.