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-空间-时间(成本)编码方法的芯片上流式细胞仪”NanoSort，LLC研究及相关其他项目信息7.项目摘要荧光激活细胞分选(FACS)或流式细胞术使临床医生和研究人员能够定量表征CEL的物理(细胞大小、形状、粒度)和生化(DNA含量、细胞周期分布、细胞表面标记和活力)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髓细胞白血病(AML)和艾滋病毒/艾滋病等疾病的进展。通过高通量分选来丰富生物标本和提取稀有细胞类型，最先进的流式细胞仪使进行罕见事件研究成为可能，例如鉴定或分离细菌细胞、干细胞或肿瘤细胞。然而，今天的流式细胞仪面临着两个挑战，这两个挑战限制了大幅降低成本和将其日常使用扩展到临床环境的能力。第一个限制是，该系统的体系结构在利用不断增加的可用荧光颜色方面效率极低。在当今的流式细胞仪设计中，每种荧光颜色都需要专用的光电倍增管和光学元件；随着检测参数的增加，成本、复杂性和故障风险都会增加。其次，流式细胞仪(对细胞进行计数)和流式细胞仪(对细胞进行计数和分类)之间存在巨大的价格差距(2-3倍的价格差异)。FAC大多位于共享的核心设施中，由训练有素的博士级别的专家运营。如果细胞分选变得比现在更容易获得和负担得起，对FAC的需求将会增加。基于加州大学圣迭戈分校罗教授团队近10年的研究，我们将把芯片实验室技术开发成能够应对上述两个挑战的产品。为了促进这一转变，我们将应用我们的专利改变游戏规则的技术：1)颜色-空间-时间(成本)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的所有流量计的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率和高成本效益的低剪切率和高细胞存活率的芯片上压电细胞分选技术。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问题。拟议的第一阶段研究使用创新的方法将实验室技术转化为市场价值超过10亿美元的商业产品，并在拟议的成本降低和功能改进后扩大市场潜力。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，以增强整个人口的健康和福祉。 与公共健康相关：“使用颜色-空间-时间(成本)编码方法的芯片上实验室流式细胞仪”NanoSort，LLC研究和相关的其他项目信息8.项目简介拟议的项目旨在开发高性能、易于操作和维护的芯片上实验室流式细胞仪，并且比目前可用的任何流式细胞仪或荧光激活细胞分类器(FAC)的成本低得多，从而在研究和临床实践的新环境中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满足高级生物医学研究和护理点诊所市场需求的产品。提出的系统大大扩展了原有的大学技术，发明了颜色-空间-时间(成本)编码技术，允许使用单个光电倍增管(PMT)探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.