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

项目摘要智力奖授予洛杉矶的加州大学，以开发高速荧光激活细胞分选（FACS）系统，以便在生物安全3级（BSL 3）设施内的生物安全柜中分选高度传染性和潜在危险的样品，从而首次实现活BSL 3亚细胞生物体的高纯度分选。该分选机采用紧密聚焦的脉冲激光束来产生爆炸性蒸汽气泡和目标物体附近的瞬时、局部、小体积流体扰动，以使它们在微通道中的轨迹偏转到收集或废物出口中。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以100，000个细胞/秒的速度实现90%的高纯度分选，以确保最高水平的生物安全。从109个细胞的群体中纯化108个细胞或亚细胞器只需要一个小时。该项目还旨在开发一种新型的无标记分选仪，利用介电电泳机制对含有各种不需要的亚细胞器和碎片的样品进行预纯化。Broad ImpactA BSL 3兼容的高通量FACS平台将为研究传染病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发展的专门知识也将纳入项目执行情况？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和医学院获得优秀的多学科培训环境。

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.