CAREER: Understanding Spatial Heterogeneity in Biofilms Using Colloidal Engineering

职业：利用胶体工程了解生物膜的空间异质性

• 批准号: 1753352
• 负责人: Connie Chang
• 金额: $ 50万
• 依托单位: Montana State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1753352&HistoricalAwards=false
• 关键词: CAREER; Understanding; Spatial; Heterogeneity; Biofilms

Non-Technical Abstract:Many microorganisms, including bacteria and algae, form thin films on surfaces known as biofilms. Examples of these include dental plaque, the slime that forms on ship hulls, and wound-associated infections. Biofilms contain millions of microscopic cells, and the individual locations of these cells affect their ability to function. For example, the location of cells within a wound-associated biofilm affects their ability to resist antibiotics. To understand how cell location affects function, their locations must be controlled in three dimensions; however, because the microorganisms within a biofilm are microscopically small, they are difficult to manipulate at a single cell level. This CAREER award by the Biomaterials program in the Division of Materials Research to Montana State University, Bozeman, supports the investigation of biofilm structure and function by building biofilms from microscopic modules that can be assembled, disassembled, and analyzed in a controlled manner. This approach will enable researchers to synthetically construct multi-species biofilms in the laboratory to understand how bugs communicate, cooperate, or compete. The proposed work will benefit society as a whole by uncovering the physical mechanisms behind microbial virulence and antibiotic resistance. This understanding can lead to new methods that will aid in controlling these persistent, surface-associated microbial colonies. Impact in the state of Montana will be provided by a "Soft Materials Shop" (SMASH), which will be used for both outreach and teaching, and will strengthen the connection of sciences, engineering, architecture, and art on campus. This space will be the first in the state of Montana and will be accessible to the public. This proposal will help prepare Montana students for future careers in STEM using inquiry-based learning strategies and hands-on demos, educate the public about microfabrication and engineering, and provide opportunities for outreach that will engage public audiences from youth to adults. Technical AbstractBiofilms are ubiquitous in the natural world, are formed by nearly all microorganisms, including algae, bacteria, archaea, and can be both beneficial and harmful. The spatial locations of microbes in a biofilm impact biofilm function and physiology; however, methods for structuring biofilms in three dimensions are currently lacking. The ability to synthetically construct a heterogeneous, multi-species biofilm by placing microorganisms in discrete locations within a cultivated environment would allow researchers to bridge this gap and explore how bugs communicate, cooperate, or compete. To achieve this, colloidal engineering techniques will be applied to place microbes in spatially organized three-dimensional locations to understand how spatial heterogeneity influences structure-function relationships in biofilms. The aims of this proposal are to create monodisperse capsules of gels and shells to encapsulate microbes, to assemble capsules into three-dimensional structures, and finally to create modular synthetic consortia of bacteria to explore questions related to heterogeneity in biofilms. These constructs will be used to understand how biofilm behavior such as quorum sensing, matrix formation, and dormancy arises from heterogeneity in biofilms. This CAREER award by the Biomaterials program in the Division of Materials Research to Montana State University, Bozeman, will support the integration of soft matter science into classes, workshops, demonstrations, and tours within a dedicated space called "SMASH", the creation of K-8 after-school outreach modules, and an undergraduate/graduate course on "Soft Matter Engineering". This award will allow for creative, multidisciplinary collaboration, bringing together students in art, architecture, engineering, and science on campus to create, prototype, design, and build. The ideas, workshops, and demonstrations created in this space will be incorporated into current K-8 outreach efforts and extended to future outreach efforts to introduce soft matter science, a relatively young area of research, to students and the general public.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

非技术摘要：许多微生物，包括细菌和藻类，在表面形成薄膜，称为生物膜。这些疾病的例子包括牙菌斑、船体上形成的粘液以及与伤口相关的感染。生物膜包含数百万个微观细胞，这些细胞的单个位置影响它们的功能。例如，细胞在伤口相关生物膜内的位置影响它们抵抗抗生素的能力。为了了解细胞位置如何影响功能，必须在三维空间中控制它们的位置;然而，由于生物膜内的微生物在显微镜下很小，因此很难在单细胞水平上进行操作。蒙大拿州立大学博兹曼分校材料研究部生物材料项目的这一职业奖支持通过从微观模块构建生物膜来研究生物膜的结构和功能，这些模块可以以受控的方式组装、拆卸和分析。这种方法将使研究人员能够在实验室中合成构建多物种生物膜，以了解昆虫如何交流，合作或竞争。这项拟议的工作将通过揭示微生物毒力和抗生素耐药性背后的物理机制而使整个社会受益。这种理解可以导致新的方法，这将有助于控制这些持久的，表面相关的微生物菌落。蒙大拿州的影响将由“软材料商店”（SMASH）提供，该商店将用于推广和教学，并将加强校园内科学，工程，建筑和艺术的联系。 这个空间将是蒙大拿州的第一个，将向公众开放。该提案将帮助蒙大拿州的学生为未来的STEM职业做好准备，使用基于探究的学习策略和动手演示，教育公众有关微细加工和工程，并提供机会，吸引从青年到成年人的公众观众。生物膜在自然界中普遍存在，由几乎所有的微生物形成，包括藻类、细菌、古生菌，并且可以是有益的和有害的。生物膜中微生物的空间位置影响生物膜功能和生理学;然而，目前缺乏用于在三维中构建生物膜的方法。通过将微生物放置在培养环境中的离散位置来合成构建异质，多物种生物膜的能力将使研究人员能够弥合这一差距，并探索微生物如何交流，合作或竞争。为了实现这一目标，胶体工程技术将被应用于将微生物放置在空间组织的三维位置，以了解空间异质性如何影响生物膜中的结构-功能关系。该提案的目的是创建凝胶和壳的单分散胶囊来封装微生物，将胶囊组装成三维结构，并最终创建细菌的模块化合成财团来探索与生物膜异质性相关的问题。这些构建体将被用来了解生物膜的行为，如群体感应，基质形成和休眠是如何从生物膜的异质性产生的。蒙大拿州立大学材料研究部的生物材料计划向博兹曼颁发的职业奖将支持将软物质科学整合到称为“SMASH”的专用空间内的课程，研讨会，演示和图尔斯，创建K-8课后外展模块，以及关于“软物质工程”的本科生/研究生课程。该奖项将允许创造性的，多学科的合作，汇集在校园艺术，建筑，工程和科学的学生创造，原型，设计和建造。在这个空间中创建的想法，研讨会和演示将被纳入目前的K-8外展工作，并扩展到未来的外展工作，向学生和公众介绍软物质科学，一个相对年轻的研究领域。这个奖项反映了NSF的法定使命，并被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

项目成果

Screening of Additive Formulations Enables Off-Chip Drop Reverse Transcription Quantitative Polymerase Chain Reaction of Single Influenza A Virus Genomes

• DOI: 10.1021/acs.analchem.0c03455
• 发表时间: 2021-02-26
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Loveday, Emma Kate;Zath, Geoffrey K.;Chang, Connie B.
• 通讯作者: Chang, Connie B.

DropSOAC: Stabilizing Microfluidic Drops for Time-Lapse Quantification of Single-Cell Bacterial Physiology

• DOI: 10.3389/fmicb.2019.02112
• 发表时间: 2019-09-24
• 期刊: FRONTIERS IN MICROBIOLOGY
• 影响因子: 5.2
• 作者: Pratt, Shawna L.;Zath, Geoffrey K.;Chang, Connie B.
• 通讯作者: Chang, Connie B.

Measuring colloid–surface interaction forces in parallel using fluorescence centrifuge force microscopy

使用荧光离心力显微镜并行测量胶体与表面相互作用力

• DOI: 10.1039/d1sm00461a
• 发表时间: 2021
• 期刊: Soft Matter
• 影响因子: 3.4
• 作者: LeFevre, Thomas B.;Bikos, Dimitri A.;Chang, Connie B.;Wilking, James N.
• 通讯作者: Wilking, James N.