Single-cell elemental analysis of bacteria: quantitative analysis of polyphosphates in Mycobacterium tuberculosis.
Single-cell elemental analysis of bacteria: quantitative analysis of polyphosphates in Mycobacterium tuberculosis.
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DOI：
10.3389/fcimb.2012.00063
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发表时间：
2012
影响因子：
5.7
通讯作者：
Talaat AM
中科院分区：
文献类型：
--
作者：
Ward SK;Heintz JA;Albrecht RM;Talaat AM
每年有180多万人死于结核病的病原体结核分枝杆菌感染。结核分枝杆菌获取和分配微量营养物质（包括生物金属）的能力，已知在其细胞内存活和宿主内的毒力中发挥作用。检测结核分枝杆菌细胞内元素分布的技术以前仅限于批量检测方法或低分辨率分析。在这里，我们提出了一种在单细胞水平上以高（单个纳米）分辨率确定结核分枝杆菌内元素分布的方法，使用扫描透射电子显微镜（STEM）与能量色散x射线光谱学（EDS）相结合。结果显示，在所有菌株的分枝杆菌测试存在大聚磷酸盐颗粒。即使在饥饿的情况下，它们也会持续存在，并可能在结核分枝杆菌的元素稳态中发挥作用。与聚磷酸盐颗粒相关的是钙和镁等微量营养素。此外，我们将该技术扩展到分枝杆菌之外，表明STEM和EDS可以作为一种简单的筛选方法，在所有其他六种被测试的细菌类型中检测单细胞水平上浓缩元素的存在或缺失，而对细菌的处理最少。总的来说，我们认为这项技术代表了更好地理解细胞内环境成分（包括多磷酸盐和生物金属）在结核分枝杆菌发病机制中的作用的第一步，具有潜在的未来体内分析应用。
More than 1.8 million people die annually from infection with Mycobacterium tuberculosis, the causative agent of tuberculosis. The ability of M. tuberculosis to obtain and distribute micronutrients, including biometals, is known to play a role in its intracellular survival and virulence within a host. Techniques to detect elemental distributions within M. tuberculosis cells have previously been limited to bulk detection methods or low-resolution analyses. Here, we present a method for determining the elemental distribution within M. tuberculosis on a single-cell level, at high (individual nanometer) resolution, using scanning transmission electron microscopy (STEM) in concert with energy-dispersive X-ray spectroscopy (EDS). Results revealed the presence of large polyphosphate granules in all strains of Mycobacteria tested. These persisted even through starvation conditions, and might play a role connected to elemental homeostasis in M. tuberculosis. Associated with the polyphosphate granules were micronutrients such as calcium and magnesium. In addition, we expanded the technique beyond Mycobacteria to show that STEM and EDS could be used as a simple screen to detect the presence or absence of concentrated elements on a single-cell level within all six other bacterial types tested, with minimal processing to the bacteria. Overall, we believe that this technique represents a first step in developing a better understanding of the role that components of the intracellular milieu, including polyphosphates and biometals, play in the pathogenesis of M. tuberculosis, with potential future applications for in vivo analysis.
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影响因子：
2.2
作者：
Cosgriff, EC;Oxley, MP;Pennycook, SJ
通讯作者：
Pennycook, SJ
影响因子：
2.8
作者：
Meyer, Daryl A.;Oliver, Julie A.;Albrecht, Ralph M.
通讯作者：
Albrecht, Ralph M.
DOI：
10.1073/pnas.97.3.1252
发表时间：
2000-02-01
影响因子：
11.1
作者：
De Voss, JJ;Rutter, K;Barry, CE
通讯作者：
Barry, CE
影响因子：
3.2
作者：
Rodriguez, GM;Smith, I
通讯作者：
Smith, I
影响因子：
6.4
作者：
Rifat, Dalin;Bishai, William R.;Karakousis, Petros C.
通讯作者：
Karakousis, Petros C.