Pluhacek Tomas, Skriba Anton, Novak Jiri, Luptakova Dominika, Havlicek Vladimir
Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic.
Methods Mol Biol. 2019;1996:131-153. doi: 10.1007/978-1-4939-9488-5_12.
Siderophores represent important microbial virulence factors and infection biomarkers. Their monitoring in fermentation broths, bodily fluids, and tissues should be reproducible. Similar isolation, characterization, and quantitation studies can often have conflicting results, and without proper documentation of sample collection, data processing, and analysis methods, it is difficult to reexamine the data and reconcile these differences. In this Springer Nature Protocol, we present the procedure optimized for ferricrocin/triacetylfusarinine C extraction from biological material as well as for tissue fixation and cryosectioning for optical microscopy and for both elemental and molecular mass spectrometry imaging. Special attention is paid to siderophore data mining from conventional and product ion mass spectra, liquid chromatography, and mass spectrometry imaging datasets, performed here by our free software called CycloBranch.
铁载体是重要的微生物毒力因子和感染生物标志物。对其在发酵液、体液和组织中的监测应具有可重复性。类似的分离、表征和定量研究结果往往相互矛盾，而且如果没有对样本采集、数据处理和分析方法进行适当记录，就很难重新审视数据并协调这些差异。在本施普林格·自然实验方案中，我们展示了针对从生物材料中提取铁色素/三乙酰镰刀菌素C以及用于光学显微镜的组织固定和冷冻切片以及元素和分子质谱成像而优化的程序。特别关注从常规和产物离子质谱、液相色谱和质谱成像数据集中挖掘铁载体数据，此处通过我们名为CycloBranch的免费软件进行。
