A new experimental platform to study biofilms: Microfluidic-DHM

描述（由申请人提供）：我们建议开发一个新的生物膜研究实验平台。生物膜由生活在表面上的细菌联合体组成。它们对宿主防御的固有耐受性和对抗生素的耐药性日益增强，在许多临床应用中引起越来越多的关注，包括传染病的发病机制和假体植入物和生物材料的临床感染。迄今为止，小的空间尺度、异质性和时间依赖性已经使生物膜过程在一般机制原则方面的合理化失败，迫使治疗主要诉诸于有限成功的经验策略。目前的实验技术仅提供了控制生物膜微环境的粗糙手段，并且在单细胞水平上以适当的时空分辨率量化微尺度过程方面受到严重限制。我们的方法是整合两种最先进的实验技术，微流体和数字全息显微镜，为生物膜研究创造一个强大的新平台。我们称之为5Fluidic-DHM。5 . fluidic - dhm将利用微流体的多功能性来精确控制微环境条件，包括几何、化学和流体动力学参数，再加上数字全息技术在单细胞水平和高时间分辨率上捕捉三维动力学的能力。我们这个R21项目的目标是开发、验证和优化一个5Fluidic-DHM平台。我们将通过在两个重要的生物膜过程中进行测试来展示这种方法的优势：细胞附着于表面和通过生物膜水通道流动。该项目与人类健康直接相关，因为它将通过使用前所未有的准确性、分辨率和灵活性的仪器推进生物膜实验的最新技术，提高我们治疗生物膜源性感染的能力。这将最终导致在广泛的临床应用中增强治疗策略。

DESCRIPTION (provided by applicant): We propose to develop a new experimental platform for the study of biofilms. Biofilms consist of bacterial consortia living on surfaces. Their inherent tolerance to host defenses and increasing resistance to antibiotics cause growing concern in many clinical applications, including pathogenesis of infectious diseases and clinical infection of prosthetic implants and biomaterials. To date, the small spatial scales, heterogeneity and time-dependence have defeated the rationalization of biofilm processes in terms of general mechanistic principles, forcing therapeutics to resort primarily to empirical strategies of limited success. Current experimental techniques provide only crude means of controlling a biofilm's microenvironment and are severely limited in quantifying microscale processes at the single-cell level with appropriate spatiotemporal resolution. Our approach is to integrate two state-of-the-art experimental techniques, microfluidics and digital holographic microscopy, to create a powerful new platform for biofilm studies. We call this 5Fluidic-DHM. 5Fluidic-DHM will exploit the versatility of microfluidics in accurately manipulating microenvironmental conditions, including geometrical, chemical and fluid dynamical parameters, coupled with the ability of digital holography to capture three-dimensional dynamics at single-cell level and high temporal resolution. Our goal for this R21 project is to develop, validate and optimize a 5Fluidic-DHM platform. We will showcase the advantages of this approach by testing it on two important biofilm processes: cell attachment to surfaces and flow through biofilm water channels. This project is directly relevant to human health, as it will improve our ability to treat biofilm- originated infection by advancing the state-of-the-art in biofilm experimentation with an instrument of unprecedented accuracy, resolution and flexibility. This will ultimately lead to enhanced therapeutic strategies in a wide range of clinical applications.