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

描述（由申请人提供）：“使用颜色-时空（成本）编码方法的片上实验室流式细胞仪”NanoSort， LLC荧光活化细胞分选（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特性（细胞大小、形状、粒度）和生化特性（DNA含量、细胞周期分布、细胞表面标记物和活力）。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诸如急性髓性白血病（AML）和艾滋病毒/艾滋病等疾病的进展。流式细胞仪具有丰富生物标本和提取稀有细胞类型的高通量分选能力，可以进行罕见事件研究，如细菌细胞、干细胞或肿瘤细胞的鉴定或分离。然而，今天的流式细胞仪面临着两个挑战，限制了其大幅降低成本和将其日常应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。在今天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；成本、复杂性和故障风险随着检测参数的增加而增加。其次，流式细胞仪（用于细胞计数）和流式细胞仪（用于细胞计数和分选）之间存在巨大的价格差距（2-3倍的价格差异）。FACS大多位于共享的核心设施中，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，对流式细胞仪的需求将会增加。基于加州大学圣地亚哥分校罗教授团队近10年的研究，我们将把芯片实验室技术发展成能够解决上述两个挑战的产品。为了便于转换，我们将应用我们的专利技术：1)颜色-时空（COST）编码方法，使用单个PMT检测多个参数。成本技术从根本上改变了现有所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且具有成本效益的片上压电细胞分选技术，具有低剪切和高细胞存活率。我们提出的研究包括系统地研究实验室芯片上FACS系统的分选后细胞活力，这是大多数研究实验室忽视的成功商业化的限制问题。拟议的第一阶段研究使用创新方法将实验室技术转化为具有超过10亿美元市场的商业产品，并且在拟议的成本降低和功能改进之后具有扩大市场的潜力。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。

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.