Microfluidic Platforms for Biological Investigations

微制片上实验室(LOC)器件正被广泛应用于细胞、组织、器官和整个生物系统的生物学研究，例如蠕虫(线虫)和果蝇(果蝇)。由于其诸多优点，全生物系统为研究感知和响应各种刺激信号的神经行为机制提供了经济有效和相关的手段。然而，它们的小尺寸(微米到毫米)给它们在细胞和行为分析中的操纵带来了挑战。因此，目前的人工工作不仅缓慢和费力，而且有时在研究整个生物系统时也不准确和不一致。这项研究计划建议开发创新的LOC技术，以改进生物有机体的分析方法，以回答基本和复杂的神经行为问题。 这项研究计划的总体目标是开发各种LOC技术(通过进行两个创新的研究主题)，以在神经元和行为水平上准确操纵和测量生物标本对相关刺激(例如，化学和电)的反应。这些LOC将使以自动化、高通量、定量和可重复的方式筛选果蝇和线虫上的化学物质成为可能。 在研究主题1中，我们将开发LOC，使不同化学品对果蝇幼虫的毒性效应能够自动测量。为了能够做到这一点，我们将发展对表面属性对刺激成年果蝇产卵空间偏好的影响的基本理解。我们将利用获得的知识和专业知识开发分类和分隔模块，将鸡蛋并列放入微室，在那里它们将可控地暴露在化学品中。研究流动特性对卵子存活和发育成幼虫的影响具有重要意义。然后将研究刺激和参数研究幼虫运动和神经元活动的技术。 在研究主题2中，我们将开发独特的操作(例如固定和定位)和外科(例如解剖)微工具，并将它们集成到芯片中，使即使是非专业研究人员也能够以及时自动化的方式访问小型生物物质(例如蠕虫和果蝇)的内部环境。为了能够设计和开发上述工具，我们需要从根本上研究这些生物体外支架的机械性能，以及使用各种力(例如介电和声学)来操纵它们。待开发的知识和技术可直接应用于许多研究，如器官/卵子提取和/或生物标本中神经肌肉活动的体内记录。 六名研究生和五名本科生将在这个研究项目中接受培训，并将获得技术、科学、跨学科和软技能。所开发的知识和技术可应用于药物发现、毒理学、神经科学和系统生物学。通过自动化，与现有技术相比，可以以更容易的方式从这些样本中以更高的吞吐量获得定量数据。从长远来看，这项研究将加快和降低毒性测试、临床前药物发现、神经元图谱和生物过程基础研究的成本，从而使加拿大受益。

Microfabricated Lab-On-a-Chip (LOC) devices are being applied to a wide range of biological studies on cells, tissues, organs and whole biological systems, e.g. the worm (C. elegans) and the fruit fly (Drosophila). Due to their many advantages, whole biological systems offer cost-effective and relevant means to study the neurobehavioral mechanisms of sensing and responding to various stimulating signals. However, their small sizes (micrometers to millimeters) impose challenges for their manipulation in cellular and behavioral assays. Hence, current manual efforts are not only slow and laborious, but also sometimes inaccurate and inconsistent in studying whole biological systems. This research program proposes developing innovative LOC techniques that will improve the ways bio-organisms are assayed to answer fundamental and complex neurobehavioral questions. The overall goal of this research program is to develop various LOC techniques (by conducting two innovative research themes) to accurately manipulate and measure the response of biological specimens to relevant stimuli (e.g. chemical and electrical) at neuronal and behavioural levels. These LOCs will make it possible to screen chemicals on Drosophila and C. elegans in automated, high-throughput, quantitative and repeatable manners. In research theme 1, we will develop LOCs that enable automated measurements of the toxic effects of different chemicals on Drosophila larvae. To be able to accomplish this, we will develop fundamental understanding of the effects of surface properties on stimulating spatial preference of egg-laying in adult Drosophila. We will use the knowledge and expertise acquired to develop sorting and compartmentalization modules to parallelize eggs into microchambers where they will be exposed controllably to chemicals. It is important to investigate the effects of flow characteristics on the survival and development of eggs into larvae. Techniques to stimulate and parametrically study the movement and neuronal activities of larvae will then be investigated. In research theme 2, we will develop unique manipulation (e.g. immobilization and orientation) and surgical (e.g. dissection) micro-tools and integrate them inside chips that can enable even non-expert researchers access the internal environment of small biological substances (e.g. worm and fruit fly) in a timely automated manner. We need to fundamentally investigate the mechanical properties of the outer body shelves of these organisms and the use of various forces (e.g. dielectrophoretic and acoustic) to manipulate them in order to be able to design and develop the abovementioned tools. Knowledge and techniques to be developed are directly applicable to many studies such as organ/egg extraction and/or inner-body recording of neuromuscular activities in bio-specimens. Six graduate and five undergraduate students will be trained in this research program and will gain technical, scientific, interdisciplinary and soft skills. The developed knowledge and techniques can be applied in drug discovery, toxicology, neurosciences and systems biology. By automation, quantitative data from these specimens in higher throughputs can be obtained in easy manners as compared to existing techniques. In the long term, the research will benefit Canada by accelerating and lowering the cost of toxicity tests, preclinical drug discovery, neuronal mapping and fundamental investigation of bio-processes.