A Single-Cell Micromanipulation Platform Relying on Optoelectronic Tweezers

我们已经进入了一个越来越依赖于操纵单个生物细胞的能力的研究时代。无论这些技术是应用于个体基因组的克隆扩增，卵母细胞的体外受精，还是仅仅用于研究支撑所有生物系统行为的细胞间异质性，单细胞微操作在现代生物分子研究中已经成为不可或缺的。******有许多技术正在改进，以支持“下一代”单细胞操作，包括光学镊子，电介质电泳和微流体。这一组的一个相对较新的成员，光电镊子（OET），以前没有在加拿大实施。在这里，Aaron Wheeler PI（多伦多大学化学系），Nazir Kherani co-PI（多伦多大学电气与计算机工程系）和Peter Zandstra合作者（UBC生物医学工程系）提议在加拿大建立一个独一无二的OET系统来操纵单细胞和小群细胞进行长期培养和分析。******一个由三名博士生和一名博士后组成的团队将共同建立和验证一个集成系统，该系统将精密光电镊子与长期细胞培养和微流体控制相结合。然后，该团队将把该系统应用到一长串需要对单细胞进行高精度操作的项目中，包括对内吞作用的系统评估，内吞作用是一种最近观察到的现象，在这种现象中，特定的细胞被触发侵入邻近细胞的细胞质。我们对内吞所知不多，但它被认为是某些类型肿瘤发生的关键阶段，因此我们有必要更多地了解它是如何发生的以及为什么发生的。这项工作将是高度跨学科的，并将产生显微镜，微流体，材料表征，下一代基因组测序，单细胞生物学等方面的市场技能和经验。******鉴于单细胞操作研究的日益普及（例如，2018年诺贝尔物理学奖授予了光学镊子的亚瑟·阿什金），这一提议非常紧迫。如果获得批准，这里提出的工作代表了一个独特的机会，使加拿大成为OET技术和单细胞微操作的新兴领导者，并培养下一代加拿大研究人员，发明家和就业创造者。*****

***We have entered an age of research that is increasingly dependent upon the capacity to manipulate individual biological cells. Whether these techniques are applied to clonal expansion of individual genomes, to in vitro fertilization of oocytes, or simply to study the cell-to-cell heterogeneity that underpins the behaviour of all biological systems, single-cell micromanipulation has become indispensable in modern biomolecular research. ******There are many techniques that are being refined to support the “next generation” of single-cell manipulation, including optical tweezers, dielectrophoresis, and microfluidics. A relatively new member of this group, optoelectronic tweezers (OET), has not been implemented in Canada before. Here, PI Aaron Wheeler (U of T Chemistry), co-PI Nazir Kherani (U of T Electrical and Computer Engineering), and collaborator Peter Zandstra (UBC Biomedical Engineering) propose to build a one-of-its-kind-in-Canada OET system to manipulate single cells and small groups of cells for long-term culture and analysis. ******A team of three Ph.D. students and a postdoc will work together to build and validate an integrated system coupling precision optoelectronic tweezers with long-term cell culture and microfluidic control. The team will then apply the system to a long list of projects requiring high-precision manipulation of single cells, including a systematic evaluation of entosis, a recently observed phenomenon in which particular cells are triggered to invade the cytoplasms of their neighbors. Not much is known about entosis, but it is believed to be a key stage in some types of tumorigenesis, making it imperative that we learn more about how and why it happens. This work will be highly interdisciplinary, and will result in marketable skills and experience with microscopy, microfluidics, materials characterization, next generation genome sequencing, single-cell biology, and beyond. ******There is great urgency for this proposal, given the surging popularity of single cell manipulation research (e.g., the 2018 Nobel Prize in Physics to Arthur Ashkin for optical tweezers). If granted, the work proposed here represents a unique opportunity to establish Canada as an emerging leader in OET technology and single-cell micromanipulation, and to train the next generation of Canadian researchers, inventors, and job creators.*****