Label-free, rapid and quantitative phenotyping of stress response in E. coli via ramanome.
Label-free, rapid and quantitative phenotyping of stress response in E. coli via ramanome.
复制标题
通过拉马诺组对大肠杆菌应激反应进行无标记、快速和定量表型分析
DOI：
10.1038/srep34359
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发表时间：
2016-10-19
影响因子：
4.6
通讯作者：
Xu J
中科院分区：
文献类型：
--
作者：
Teng L;Wang X;Wang X;Gou H;Ren L;Wang T;Wang Y;Ji Y;Huang WE;Xu J
以单细胞分辨率快速分析应激反应，但以无标记，非破坏性和机制特异性的方式可以导致许多新的应用。我们提出了一种单细胞水平的生化指纹分析方法“拉曼组”（ramanome），它是在给定的时间和条件下，从一个等基因群体中随机选取一定数量的细胞，收集其单细胞拉曼光谱（SCRS），以快速、定量地检测和表征细胞群体的应激反应。大肠杆菌细胞的SCRS对乙醇的暴露时间（8个时间点）和剂量（6个剂量）均敏感，检测时间最早可达5min，对任一因素的识别率均在80%以上。此外，还对氨苄青霉素和卡那霉素抗生素、乙醇和正丁醇醇类以及Cu 2+和Cr 6+重金属等3类6种化合物的拉曼光谱进行了分析，得到了31条标记拉曼谱带，这些谱带可通过细胞毒性机制和细胞间异质性的变化来区分胁迫反应。此外，特异性，重现性和机制的基础ramanome进行了验证，通过跟踪应力诱导的代谢动力学和细胞之间的对比与无基因，传达抗应激。因此，ramanome能够以单细胞分辨率快速预测和基于机制的细胞毒性和应激反应程序筛选。
Rapid profiling of stress-response at single-cell resolution yet in a label-free, non-disruptive and mechanism-specific manner can lead to many new applications. We propose a single-cell-level biochemical fingerprinting approach named “ramanome”, which is the collection of Single-cell Raman Spectra (SCRS) from a number of cells randomly selected from an isogenic population at a given time and condition, to rapidly and quantitatively detect and characterize stress responses of cellular population. SCRS of Escherichia coli cells are sensitive to both exposure time (eight time points) and dosage (six doses) of ethanol, with detection time as early as 5 min and discrimination rate of either factor over 80%. Moreover, the ramanomes upon six chemical compounds from three categories, including antibiotics of ampicillin and kanamycin, alcohols of ethanol and n-butanol and heavy metals of Cu2+ and Cr6+, were analyzed and 31 marker Raman bands were revealed which distinguish stress-responses via cytotoxicity mechanism and variation of inter-cellular heterogeneity. Furthermore, specificity, reproducibility and mechanistic basis of ramanome were validated by tracking stress-induced dynamics of metabolites and by contrasting between cells with and without genes that convey stress resistance. Thus ramanome enables rapid prediction and mechanism-based screening of cytotoxicity and stress-response programs at single-cell resolution.