A Dual Rheometer-Microscope Instrument for the Quantitative Characterization of Bacterial Biofilms

用于细菌生物膜定量表征的双流变仪-显微镜仪器

• 批准号: RTI-2019-00860
• 负责人: Virgilio, Nick
• 金额: $ 10.93万
• 依托单位: École Polytechnique de Montréal
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=667829
• 关键词: Dual; Rheometer; Microscope; Instrument; Quantitative

It is currently estimated that 80% of the planet's bacteria live in communities inside biofilms - a medium comprised of exopolysaccharides, DNA, proteins and various small molecules. A biofilm is a mechanically robust material that surrounds and protects the bacteria from the external world. As a result, these bacteria are generally more resistant or tolerant to antibiotics and disinfectants. The development of biofilms on medical devices, such as catheter and implants, and in living tissues, now poses some important issues, since intensive and prolonged treatments with antibiotics still often lead to treatment failure, and higher morbidity and mortality rates - this is a worldwide issue that still needs the development of efficient solutions. Biofilms also have useful properties, for example in the development of treatment processes for water and wastewater. Biofilm flocs allow the suspension of bacteria in the medium, or their adherence to inert supports onto which the biomass develops. As a result, understanding and controlling the properties of biofilms is of critical importance in these areas of research. Very recently, both interfacial and bulk rheology have been introduced as techniques to monitor quantitatively the viscoelastic properties of bacterial biofilms, which are directly related to the stability of the film. The commercial rheometer that we request in this proposal will be equipped with interfacial geometries, along with other various bulk measurement geometries, that will allow time-resolved, quantitative measurements of biofilms' viscoelastic properties, with a fluorescence microscope allowing microstructure and composition analyses. Previous work has demonstrated that this technique is particularly sensitive to biofilm composition, which can be controlled by gene knockout and by the external environment - temperature, composition of the media, etc. Since time-resolved measurements will be possible, quantifying the effects of perturbations, such as the addition of antibiotics, on the viscoelastic, microstructural and composition properties of biofilms will be possible. Finally, the installation of the rheometer in a Biosafety level 2 laboratory will allow the study of pathological strains such as Salmonella - a unique configuration in Canada. This original multidisciplinary program is a first collaboration between seven co-applicants and 4 collaborators with complementary expertises, including rheology, bacterial microbiology, interfacial science, water/wastewater treatment processes, veterinary medecine, and pediatrics, from Polytechnique Montreal, Université de Montréal, INRS Institut Armand-Frappier, and Stanford University.

目前据估计，该地球的80％的细菌生活在生物膜内的社区中 - 培养基完成了外多糖，DNA，蛋白质和各种小分子。生物膜是一种机械强大的材料，可环境并保护细菌免受外部世界的影响。结果，这些细菌通常对抗生素和消毒剂更具耐药性或耐受性。在医疗设备（例如导管和直接的医疗设备以及生活时间）上的生物膜开发，现在提出了一些重要问题，因为对抗生素的密集和长时间治疗仍经常导致治疗失败，并且发病率和死亡率更高 - 这是全球问题，仍然需要高效解决方案的开发。生物膜也具有有用的特性，例如在水和废水的处理过程中。生物膜允许在培养基中悬浮细菌，或它们遵守生物量发展的惰性支持。结果，了解和控制生物膜的特性在这些研究领域至关重要。最近，已经引入了界面和散装流变，以定量监测细菌生物膜的粘弹性特性，这与膜的稳定性直接相关。我们在该提案中要求的商业流变仪将等效于界面几何形状，以及其他各种大量测量几何形状，这些几何形状将允许对生物膜的粘弹性特性进行时间分辨，定量测量，并具有荧光显微镜允许微观结构和组成分析的荧光显微镜。先前的工作表明，该技术对生物膜组成特别敏感，该技术可以通过基因敲除和外部环境控制 - 温度，媒体组成等。由于时间分辨的测量将是可能的，可以量化扰动的影响，例如添加抗生素，对粘弹性，微生物结构和组合特性的添加。最后，在生物安全2级实验室中安装流变仪将允许研究病理菌株，例如沙门氏菌 - 加拿大独特的构型。这个原始的多学科计划是七个共同申请者与4位具有完整专业知识的合作者之间的首次合作，包括流变学，细菌微生物学，互化科学，水/废水处理过程，兽医医学和儿科，来自蒙特利尔大学DeTrelal，Inrtrérefort，Inrs Institut and Intranford and Strappier，