Optoelectronic Tweezers on Sapphire for Compact High Throughput Fluorescence Activated Cell Sorter

主要研究者：邱佩瑜研究所：加州大学洛杉矶分校题目：用于紧凑型高通量荧光激活细胞分选仪的蓝宝石光电镊子智能优点：高通量单细胞分析是定量生物学的未来趋势。荧光激活细胞分选仪（FACS）是用于高速和多参数单细胞分析的最强大和金标准工具。它具有广泛的应用，从免疫学，细胞生物学，分子生物学的基础研究，到临床应用，如艾滋病和癌症的检测和分析。然而，目前的外地资产管制系统体积庞大，价格昂贵，需要训练有素的技术人员在中央设施中操作。在诊所和资源有限的地区应用的高性能和低成本的流式细胞仪是高度寻求，但尚未实现。在过去的十年中，已经提出了许多微流体FACS系统，但是以高分选纯度和高细胞活力高速分选活哺乳动物细胞仍然是一个主要挑战，并且集成系统并不紧凑。用于光学检测的庞大光学器件（例如PMT管）和用于流体输送的庞大外部流体泵送系统是FACS小型化的两个主要障碍。该方案旨在开发一种紧凑且高通量的FACS，其通过在透明蓝宝石衬底上制造的新型双灵敏度光电子镊子（OET）实现。该平台提供数百万个光致单细胞捕获威尔斯孔，以提供基于荧光信号的高通量单细胞分选。大规模并行OET使能的光学操作使O-FACS在智能手机大小的系统上的吞吐量为55，000个细胞/秒，相当于当前高端台式FACS系统的吞吐量。更广泛的影响：所提出的紧凑型、手持式和高通量O-FACS平台将在细胞分选领域提供范式转变的影响。从基础生物学研究到临床应用，它将有广泛的应用。 在该项目过程中取得的成果和积累的专门知识也将纳入方案执行指标？在本科和研究生两个层次的教学活动。少数民族研究生和本科生将通过独立的研究课程参与这些项目。参与该项目的学生将在加州大学洛杉矶分校工程学院获得优秀的多学科培训环境。PI还将参与加州大学洛杉矶分校的杰出外展计划（CEED），通过工程专业介绍（E 87）课程招募代表性不足的大学生进行短期项目。

PI: Chiou, Pei-YuInstitution: University of California-Los AngelesTitle: Optoelectronic Tweezers on Sapphire for Compact High Throughput Fluorescence Activated Cell SorterIntellectual Merit: High throughput single cell analysis is the future trend for quantitative biology. Florescence activated cell sorter (FACS) is the most powerful and gold standard tool for high speed and multi-parametric single cell analyses. It has broad applications from fundamental research in immunology, cell biology, molecular biology, to clinical applications, such as AIDS and cancer detection and analysis. However, the current FACS is bulky, expensive and requires well-trained technician to operate in a centralized facility. High performance and low cost FACS for applications in clinics and resource-limited areas are highly sought but have not been realized. Many microfluidic FACS systems have been proposed over the past ten years, yet sorting live mammalian cells at high speeds with high sort purity and high cell viability remains a major challenge and the integrated system is not compact. Bulky optics such as PMT tubes for optical detection, and bulky external fluid pumping systems for fluid delivery are the two major barriers for FACS miniaturization. This proposal aims to develop a compact and high throughput FACS enabled by a novel ultrahigh sensitivity optoelectronic tweezers (OET) fabricated on a transparent sapphire substrate. This platform provides millions of light-actuated single cell trapping wells to provide a high throughput single cell sorting based on multicolor fluorescence signals. The massively parallel OET enabled optical manipulation gives O-FACS a throughput of 55,000 cells/sec on a smartphone size system, a throughput equivalent to current high-end benchtop FACS systems. Broader Impacts: The proposed compact, handheld, and high throughput O-FACS platform will provide paradigm-shifting impacts in cell sorting fields. It will have broad applications from fundamental biological research to clinical applications. Results and expertise developed during the course of this project will also be incorporated into the PIs? teaching activities at both the undergraduate and graduate levels. Minority graduate and undergraduate students will participate in these projects through independent research courses. Students involved in this project will be exposed to an excellent multidisciplinary training environment in the UCLA Engineering School. The PIs will also be involved with the outstanding outreach program (CEED) in UCLA to recruit underrepresented college students for short-term project through the Introduction to Engineering Disciplines (E 87) course.