IDBR: Development of a High Speed FACS for Sorting BSL3 Infectious Agents

美国加州大学洛杉矶分校（University of California at Los Angeles）因开发一种高速荧光激活细胞分选（FACS）系统而获得梅里坦奖，该系统用于在生物安全级别3 （BSL3）设施的生物安全柜中分选高传染性和潜在危险的样品，首次实现对活的BSL3亚细胞生物的高纯度分选。该分拣机采用紧密聚焦的脉冲激光束，在目标物体附近产生爆炸性蒸汽泡和瞬态、局部、小体积流体扰动，使目标物体在微通道中的轨迹偏转，进入收集或废物出口。它的目标是在生物安全柜内的全封闭一次性微流控芯片上实现90%的高纯度分选，分选速度为100,000个细胞/秒，以确保最高水平的生物安全。从109个细胞中纯化108个细胞或亚细胞细胞器只需要一个小时。该项目还旨在开发一种新的、无标签的分选器，利用介电电泳机制来预纯化含有各种不需要的亚细胞细胞器和碎片的样品。BSL 3兼容的高通量FACS平台将为传染病研究提供范式转换的影响。它将有广泛的应用，从基础生物学研究到临床应用。在这个项目过程中产生的成果和专业知识也将被纳入pi。本科和研究生阶段的教学活动。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和医学院获得良好的多学科培训环境。

Project AbstractIntellectual MeritAn award is made to the University of California at Los Angeles to develop a high speed fluorescence activated cell sorting (FACS) system in order to sort highly infectious and potentially dangerous samples in a biosafety cabinet within a Biosafety Level 3 (BSL3) facility to enable for the first time high purity sorting of live BSL3 subcellular organisms. The sorter employs a tightly focused pulsed laser beam to generate an explosive vapor bubble and transient, local, small volume fluid perturbation near target objects to deflect their trajectories in a microchannel into collection or waste outlets. It aims to achieve 90% high purity sort at 100,000 cells/s on a fully enclosed, disposable, microfluidic chip inside a biosafety cabinet to ensure the highest level of biosafety. It requires only one hour to purify 108 cells or subcellular organelles from a population of 109 cells. This project also aims to develop a novel, label free sorter utilizing dielectrophoresis mechanism to pre-purify samples containing various unwanted subcellular organelles and debris.Broad ImpactA BSL 3 compatible high throughput FACS platform will provide paradigm-shifting impacts in studying infectious disease. 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 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 UCLA Engineering School and Medical School.