Collaborative Proposal: Instrumental Development of Microfluidics-Based Fluorescence Activated Cell Sorting Device for Research and Education

该奖项用于开发基于微流体的荧光激活细胞分选装置（M-FACS）。M-FACS将微流体的单液滴控制与FACS的细胞筛选和分选相结合。如果成功，M-FACS将允许人们封装单个细胞，向每个细胞添加额外的化合物，孵育这些细胞，然后对结果进行排序，所有这些都以每秒1000-10,000个细胞的速度进行，从而获得细胞种群分布的良好统计数据。M-FACS将能够研究细胞排出或呈现在其表面的化学物质。同样，它也可以在筛选大型文库的试验中使用单个细胞。这将显著提高FACS的效用。将单个细胞封装在滴液中的一种方法是使用微流体技术。单个细胞可以以滴状悬浮在惰性载体液中。此外，完全控制这些液滴也是可行的：它们可以与其他液滴组合，可以将它们分成更小的液滴，可以用光学探针对它们进行询问，可以用电场对它们进行分类。这项新技术将得到必要的理论工作的支持，以充分理解即将出现的新可能性。M-FACS克服了FACS的一个重要限制：使用FACS，细胞必须在封装后立即进行分类。M-FACS将从质量上改变FACS的适用性。它将允许使用细胞作为分析的一部分，将它们与化合物库进行测试，并根据结果对细胞进行分类。它将允许细胞生长作为分选的标准。这将使无数的新问题得以研究。台式荧光活化细胞分选机具有极大的扩展能力，将为更广泛的科学界带来巨大的利益。这台机器将由学生和博士后进行实验和理论研究。他们将接受工程学与生物学相结合的领域的培训，以创造出能够推动新科学的新设备。这项工作还将在一个讲习班上传播，说明已完成的装置。

This award is for the development of a microfluidic-based fluorescent-activated cell sorting device (M-FACS). The M-FACS will combine the single droplet control of microfluidics with the cell screening and sorting of a FACS. If successful, the M-FACS will allow one to encapsulate individual cells, add additional compounds to each cell, incubate these and then sort the results, all at 1000-10,000 cells per second, allowing good statistics to be obtained on a distribution of a population of cells. M-FACS will enable the study of chemicals excreted by cells or presented on their surface. It will likewise enable using individual cells in an assay that screens a large library. This would significantly increase the utility of FACS. One way to encapsulate individual cells within drops is through the use of microfluidic techniques. Individual cells can be suspended in drops in an inert carrier fluid. Moreover, it is also feasible to completely control these drops: they can be combined with other drops, they can be divided into smaller drops, they can be interrogated with optical probes, and they can be sorted with electric fields. This new technology will be supported with essential theoretical work to fully understand the new possibilities that will be available. The M-FACS overcomes one significant limitation of FACS: with FACS, the cells must be sorted as soon as they are encapsulated. The M-FACS will qualitatively change the applicability of FACS. It will allow using cells as part of an assay, testing them against a library of compounds, and sorting the cells based on the results. It will allow cell growth to be a criterion of the sorting. This will allow myriad new problems to be investigated. A desk-top Fluorescence-Activated Cell Sorting machine with greatly extended capabilities will be of great benefit to the broader scientific community. The machine will be developed by students and post docs, both experimental and theoretical. They will receive training in a field that merges engineering with biology to create novel devices that enable new science. The work will also be disseminated at a workshop that will describe the completed device.