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

Pi：Chiou，Pei-Yu研究所：加州大学洛杉矶分校题目：蓝宝石上用于紧凑型高通量荧光激活细胞分类器的光电镊子智能优点：高通量单细胞分析是定量生物学的未来趋势。荧光激活细胞分类器(FACS)是用于高速多参数单细胞分析的最强大和最金标准的工具。从免疫学、细胞生物学、分子生物学的基础研究到艾滋病和癌症检测分析等临床应用，它有着广泛的应用。然而，目前的FACS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。高性能和低成本的流式细胞仪在临床和资源有限的领域得到了极大的应用，但尚未实现。在过去的十年里，已经提出了许多微流控FACS系统，但高速分选具有高分类纯度和高细胞活力的活哺乳动物细胞仍然是一个主要的挑战，并且集成系统并不紧凑。体积庞大的光学元件，如用于光学检测的PMT管，以及用于流体输送的笨重的外部流体泵系统，是FACS小型化的两大障碍。该方案旨在开发一种基于透明蓝宝石衬底上的新型超高灵敏度光电镊子(OET)来实现紧凑和高通量的流式细胞仪。该平台提供数百万个光驱动的单细胞捕获井，以提供基于多色荧光信号的高通量单细胞分选。大规模并行的OET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：拟议的紧凑型、手持式和高吞吐量O-FACS平台将在细胞分选领域带来范式转换的影响。从基础生物学研究到临床应用，都将有广阔的应用前景。在这个项目过程中形成的结果和专业知识也将被纳入绩效指标？本科生和研究生的教学活动。少数民族研究生和本科生将通过自主研究课程参与这些项目。参与该项目的学生将接触到加州大学洛杉矶分校工程学院良好的多学科培训环境。PIS还将参与加州大学洛杉矶分校(UCLA)的杰出外展计划(CEED)，通过工程学科导论(E87)课程招募代表人数不足的大学生参加短期项目。

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.