Review of methods to probe single cell metabolism and bioenergetics.

Review of methods to probe single cell metabolism and bioenergetics.
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DOI:
10.1016/j.ymben.2014.09.007
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发表时间:
2015-01
影响因子:
8.4
通讯作者:
Stephanopoulos, Gregory
Stephanopoulos, Gregory
中科院分区:
工程技术1区
文献类型:
--
作者:
Vasdekis, Andreas E.;Stephanopoulos, Gregory

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单细胞研究带来了意想不到的发现,例如在感染、代谢和治疗中存在生物噪声和表型转换。在这里,我们回顾的方法,使这样的单细胞研究特定的代谢和生物能量学。首先,我们讨论了如何从细胞悬液中分离和培养个体,包括永久性和可逆性方法。我们还强调了微生物学在代谢工程方面的具体进展。探测单细胞生理和代谢的方法随后进行审查。其中的主要重点是动态和高含量的分析策略的基础上,无标记和荧光显微光谱和显微镜。非动态的方法,如质谱和核磁共振,也简要讨论。
Single cell investigations have enabled unexpected discoveries, such as the existence of biological noise and phenotypic switching in infection, metabolism and treatment. Herein, we review methods that enable such single cell investigations specific to metabolism and bioenergetics. Firstly, we discuss how to isolate and immobilize individuals from a cell suspension, including both permanent and reversible approaches. We also highlight specific advances in microbiology for its implications in metabolic engineering. Methods for probing single cell physiology and metabolism are subsequently reviewed. The primary focus therein is on dynamic and high-content profiling strategies based on label-free and fluorescence microspectroscopy and microscopy. Non-dynamic approaches, such as mass spectrometry and nuclear magnetic resonance, are also briefly discussed.
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