Imaging microorganisms and pollutants at the microscale

微生物和污染物的微观成像

• 批准号: NE/D00683X/1
• 负责人: David Werner
• 金额: $ 3.34万
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=NE%2FD00683X%2F1
• 关键词: Imaging; microorganisms; pollutants; microscale

Fluorescence microscopy is a powerful tool for studying microorganisms in their natural environment without cultivation. The nucleic acids inside microbial cells can be stained with fluorescing dyes which specifically bind to targeted organisms. This technique can stain pollutant degrading microorganisms in natural samples for their identification, localisation and enumeration. Microbial degradation can break down and detoxify polycyclic aromatic hydrocarbons (PAHs). PAHs are carcinogenic pollutants present in organic liquids like crude oil or tar and also in numerous soils or sediments in the United Kingdom impacted by industrial activity. An interesting property of PAHs is their autofluorescence under ultraviolet light which means that they can be visualised with fluorescence microscopy. Fluorescence microscopy may thus be suitable for the simultaneous microscale imaging of PAH-rich patches in soils and sediments and their associated microbial communities. To this end criteria will be derived for the identification of PAH-rich patches in soil or sediment by fluorescence microscopy. Once the unambiguous distinction between the fluorescence of PAH-rich patches and other natural soil or sediment constituents has been achieved, a suitable methodology will be derived to stain and image the associated microorganisms using fluorescence in situ hybridisation (FISH), conventional epifluorescence microscopy and confocal laser scanning microscopy. The derived methodology could be used to study in detail and at the microscale the co-localisation of microorganisms and pollutants such as PAHs in the natural environment. A microscale investigation may shed light on the exact mechanisms controlling the microbial breakdown and attenuation of PAHs in polluted soils and sediments. Such insight is of fundamental importance for the design of bioremediation technologies and for the risk assessment for impacted ecosystems.

荧光显微镜是在没有培养的情况下在自然环境中研究微生物的强大工具。微生物细胞内部的核酸可以用荧光染料染色，这些染料特异性结合到靶向生物。该技术可以在天然样品中降解微生物的污染物，以降解其识别，定位和枚举。微生物降解可以分解并解毒多环芳烃（PAHS）。 PAH是在有机液体中存在的致癌污染物，例如原油或焦油，也存在于英国的许多土壤或沉积物中，受工业活动影响。 PAH的一个有趣的特性是它们在紫外线下的自动荧光，这意味着它们可以用荧光显微镜可视化。因此，荧光显微镜可以适用于土壤和沉积物及其相关微生物群落中富含PAH的斑块的同时微观成像。为此，将得出通过荧光显微镜在土壤或沉积物中鉴定富含PAH的斑块的标准。一旦实现了富含PAH的斑块的荧光与其他天然污垢或沉积物成分之间的明确区分，将得出一种合适的方法，以使用荧光原位杂交（FISH），常规的表现显微镜显微镜和共聚焦激光扫描来染色和图像相关的微生物。派生的方法可用于详细研究和微观研究，微生物和污染物（例如自然环境中的PAH）的共定位。微观研究可以阐明控制污染土壤和沉积物中PAH的微生物分解和衰减的确切机制。这种洞察力对于生物修复技术的设计和受影响的生态系统的风险评估至关重要。