MRI: Development of a Microfluidic Instrument for High-throughput Production of Asymmetric Vesicles to Support Membrane Biology Research

授予位于宾厄姆顿的纽约州立大学开发用于构建具有不对称脂质分布的可定制囊泡的高通量微流体仪器。这种囊泡上级现有的脂质体，因为它们可以被定制为精确复制天然膜，因此更具生理相关性。该仪器提供的创新将对基础生物膜和膜生物学研究产生广泛影响。特别是，生理相关的囊泡将使研究是不可能的与目前的脂质体技术。该项目的跨学科性质为各级学生提供了极好的培训机会。生物学家和工程师往往不具备其他学科中最基本的技能，限制了承担重要研究问题的能力。这一差距将通过为学生提供发展跨学科技能的机会来解决，同时为该项目的目标做出贡献。各级学生将与他们的跨学科的同事在实验的规划和执行整合，并接受由他们的研究同行使用的实验室技术的培训。为了支持更广泛社区的跨学科技能发展，将创建一个网站，提供与微流体，生物膜和一般实验室实践相关的协议的指导。本网站将针对没有相关学科背景的个人。它还将用于传播新文书。该项目将协助在纽约州立大学宾厄姆顿分校建立一个生物膜微流体计划，该计划将在新型微流体工具的帮助下解决生物膜社区的基本问题。该仪器将产生生理相关的合成囊泡，以实现基础膜生物学研究。该仪器的能力将通过使用合成囊泡来研究外膜囊泡在生物膜形成中的作用来证明。细菌生物膜在自然界中普遍存在，对人类健康和工业有很大影响。必须了解有助于生物膜结构发展的过程，以便可以利用它们来控制生物膜生长。使用传统的脂质体无法实现这一目标，因为它们不能充分模拟天然囊泡。因此，它们不能用于研究脂质组成和结构的细微差异很重要的复杂现象。新兴技术不能以高通量合成具有受控尺寸的均匀、单层、不对称囊泡。该仪器产生具有所有这些特征的合成囊泡的能力是膜生物学和生物膜研究中的范式转变进步。从长远来看，使用该仪器构建的囊泡有可能用于疫苗开发和作为抗菌剂的递送载体。作为仪器开发的一部分，与微尺度多相流体流动相关的重要基本问题也将得到解决。这些包括：流动聚焦和界面稳定性，脂质自组装的时间尺度，和乳液运动学。

An award is made to the State University of New York at Binghamton to develop a high throughput microfluidic instrument for constructing customizable vesicles with asymmetric lipid distributions. Such vesicles are superior to existing liposomes because they can be tailored to exactly replicate natural membranes and are therefore more physiologically relevant. The innovations provided by the instrument will have a broad impact on fundamental biofilm and membrane biology research. In particular, the physiologically relevant vesicles will enable studies that are impossible with current liposome technology. The interdisciplinary nature of this project provides an excellent training opportunity for students at all levels. Biologists and engineers often do not possess even the most fundamental skills in the others' discipline, limiting the ability to take on important research questions. This gap will be addressed by providing students with the opportunity to develop their interdisciplinary skills while contributing to the objectives of this project. Students at all levels will integrate with their cross disciplinary colleagues in the planning and execution of experiments and receive training in the laboratory techniques used by their research counterparts. To support interdisciplinary skills development in the wider community, a website will be created that provides instruction on protocols relevant to microfluidics, biofilms, and general laboratory practice. This website will be targeted to individuals with no background in the relevant discipline. It will also be used to disseminate the new instrument. This project will assist in establishing the formation of a Biofilm Microfluidics Initiative at SUNY Binghamton, which will tackle fundamental questions in the biofilms community with the aid of novel microfluidic tools.The instrument will produce physiologically relevant synthetic vesicles to enable fundamental membrane biology research. The capabilities of the instrument will be demonstrated by using the synthetic vesicles to study the role of outer membrane vesicles in biofilm formation. Bacterial biofilms are ubiquitous in nature and have a large impact on human health and industry. It is imperative to understand the processes that contribute to the development of biofilm structure so that they can be exploited to control biofilm growth. This objective would not be achievable using traditional liposomes because they do not adequately mimic natural vesicles. Therefore they cannot be used to study complex phenomena where subtle differences in lipid composition and architecture are important. Emerging technologies are unable to synthesize uniform, unilamellar, asymmetric vesicles with controlled size at high throughput. The ability of the instrument to produce synthetic vesicles possessing all of these features is a paradigm shifting advancement in membrane biology and biofilm research. Long term, the vesicles built using the instrument have the potential to be used in vaccine development and as delivery vehicles for antimicrobial agents. As part of the instrument development, important fundamental issues relevant to microscale multiphase fluid flows will also be addressed. These include: flow focusing and interfacial stability, timescales for lipid self assembly, and emulsion kinematics.