Bio-imaging of the Bacillus anthracis spore coat

故意释放致病细菌孢子被认为是对平民健康的重大威胁、重大经济风险和对军事人员的重大危害。应对这一威胁将需要对炭疽芽胞的韧性、传染性和超级休眠的基本科学洞察力的结合，以及新兴便携式传感器技术的力量，该技术将使检测生物危害和验证其去污的有效性远远超过今天的可能。在目前的项目中，炭疽芽胞被的结构将以前所未有的分辨率使用由PI和部门同事Eric Rees博士最近开发的椭球体定位显微镜技术来绘制。将确定用于识别炭疽杆菌污染存在的保守的近表面蛋白质标记的特征，并将量化与净化相关的细微结构变化。这些实验室研究的洞察力将阐明清除污染的机制，以及可能的孢子超休眠。这项工作的第二个方面将涉及“喷雾和可视化”化学技术的发展。点击化学是指一套现代检测试剂，当探针分子与其目标相遇时，可能会产生荧光染料。通过将这种方法与该项目第一部分中确定的与保守的炭疽杆菌目标蛋白相结合的纳米体结合，将开发出一种用于突出受污染表面的“喷雾和可视化”方案。这将首先通过实验室对拭子的分析来证明，但目的是在现场突出污染情况。

The deliberate release of pathogenic bacterial spores is recognised as presenting a major threat to civilian health, a major economic risk, and a significant hazard to military personnel. Addressing this threat will require a combination of fundamental scientific insight to the resilience, infectivity, and super-dormancy of Bacillus anthracis spores, coupled with the power of emerging portable sensor technology that will make it possible to detect the biohazard and to validate its decontamination with far greater effectiveness than is possible today. In the current project, the structure of the B. anthracis spore coat will be mapped with unprecedented resolution using the ellipsoid localisation microscopy technique developed recently by the PI and departmental colleague Dr Eric Rees. Conserved near-surface protein markers for identifying the presence of B. anthracis contamination will be characterised, and fine structural changes associated with decontamination will be quantified. Insight from these laboratory studies will elucidate mechanisms of decontamination, and potentially super-dormancy of spores. The second aspect of the work will entail the development of 'spray and visualise' chemical technology. Click-chemistry refers to a suite of modern detection reagents that can involve the creation of a fluorescent dye when a probe molecule encounters its target. By combining this method with nanobody-binding to conserved B. anthracis target proteins identified in the first part of the project, a 'spray and visualise' protocol for highlighting contaminated surfaces will be developed. This will first be demonstrated with laboratory analysis of swabs, but with a view to in-situ highlighting of contamination in the field