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首页> 外文期刊>FEMS Microbiology Reviews >Detecting metabolic activities in single cells, with emphasis on nanoSIMS
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Detecting metabolic activities in single cells, with emphasis on nanoSIMS

机译:检测单细胞的代谢活性,重点是nanoSIMS

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Investigating the contribution of microbial populations to biochemical processes of global significance is challenging as there are few approaches that can detect microbial metabolic activities on single-cell level. Given the widespread distribution and importance of microorganisms in elemental transformations, improved methods for measuring microbial activities in naturally occurring microbial communities is essential. In this article, microautoradiography (MAR), Raman microspectroscopy, and Secondary Ion Mass Spectrometry (SIMS) and their combination with isotope labeling and molecular genetic methods for cell identification (i.e. FISH and related methods) are reviewed. We focus our review on the application of MAR-FISH, Raman-FISH, and FISH-SIMS to environmental samples, with a more detailed description of the use of nanoSIMS-based methodologies to identify, quantify, and visualize the incorporation of labeled substrates of single microorganisms in complex microbial communities. We highlight examples from the marine habitat. In addition, relevant technical aspects as well as important considerations concerning sample preparation and handling are presented. We conclude with a perspective on the usefulness of such tools to study the role of microorganisms in biogeochemical cycling from micron to global scales.
机译:调查微生物种群对具有全球意义的生化过程的贡献是具有挑战性的,因为很少有方法可以在单细胞水平上检测微生物的代谢活性。鉴于微生物在元素转化中的广泛分布和重要性,改进用于测量天然存在的微生物群落中微生物活性的方法至关重要。本文对微放射自显影(MAR),拉曼光谱和二次离子质谱(SIMS)及其与同位素标记和分子遗传学方法(用于FISH和相关方法)的结合进行了综述。我们将重点放在MAR-FISH,Raman-FISH和FISH-SIMS在环境样品中的应用上,并对基于nanoSIMS的方法用于识别,定量和可视化标记的底物掺入的更详细描述。复杂微生物群落中的单一微生物。我们重点介绍海洋栖息地的例子。此外,还介绍了相关的技术方面以及有关样品制备和处理的重要注意事项。我们以这种工具的有用性来得出结论,以研究微生物在从微米到全球规模的生物地球化学循环中的作用。

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