Bio-imaging of the Bacillus anthracis spore coat

故意释放致病性细菌孢子被认为对平民健康构成重大威胁，构成重大经济风险，并对军事人员构成重大危害。解决这一威胁需要结合对炭疽芽孢杆菌孢子的弹性、传染性和超休眠的基础科学见解，再加上新兴的便携式传感器技术的力量，这将使检测生物危害并验证其净化的有效性远远超过今天的可能性。在目前的项目中，炭疽芽孢杆菌孢子外壳的结构将以前所未有的分辨率绘制，使用由PI和部门同事Eric Rees博士最近开发的椭球定位显微镜技术。用于鉴定炭疽杆菌污染存在的保守的近表面蛋白标记物将被表征，与去污相关的精细结构变化将被量化。从这些实验室研究中获得的见解将阐明去污的机制，以及潜在的孢子超休眠。这项工作的第二个方面将需要发展“喷雾和可视化”化学技术。点击化学指的是一套现代检测试剂，当探针分子遇到目标时，可以产生荧光染料。通过将这种方法与纳米体结合到项目第一部分中鉴定的保守的炭疽杆菌靶蛋白，将开发出一种“喷雾和可视化”方案，用于突出污染表面。这将首先通过对拭子的实验室分析来证明，但目的是在现场现场突出显示污染

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