Burr David J, Drauschke Janina, Kanevche Katerina, Kümmel Steffen, Stryhanyuk Hryhoriy, Heberle Joachim, Perfumo Amedea, Elsaesser Andreas
Department of Physics, Experimental Biophysics and Space Sciences, Freie Universität Berlin, Arnimallee 14, 14195, Berlin, Germany.
Department of Physics, Experimental Molecular Biophysics, Freie Universität Berlin, Arnimallee 14, 14195, Berlin, Germany.
Small. 2024 Sep;20(36):e2400289. doi: 10.1002/smll.202400289. Epub 2024 May 6.
This study utilizes nanoscale Fourier transform infrared spectroscopy (nanoFTIR) to perform stable isotope probing (SIP) on individual bacteria cells cultured in the presence of C-labelled glucose. SIP-nanoFTIR simultaneously quantifies single-cell metabolism through infrared spectroscopy and acquires cellular morphological information via atomic force microscopy. The redshift of the amide I peak corresponds to the isotopic enrichment of newly synthesized proteins. These observations of single-cell translational activity are comparable to those of conventional methods, examining bulk cell numbers. Observing cells cultured under conditions of limited carbon, SIP- nanoFTIR is used to identify environmentally-induced changes in metabolic heterogeneity and cellular morphology. Individuals outcompeting their neighboring cells will likely play a disproportionately large role in shaping population dynamics during adverse conditions or environmental fluctuations. Additionally, SIP-nanoFTIR enables the spectroscopic differentiation of specific cellular growth phases. During cellular replication, subcellular isotope distribution becomes more homogenous, which is reflected in the spectroscopic features dependent on the extent of C-C mode coupling or to specific isotopic symmetries within protein secondary structures. As SIP-nanoFTIR captures single-cell metabolism, environmentally-induced cellular processes, and subcellular isotope localization, this technique offers widespread applications across a variety of disciplines including microbial ecology, biophysics, biopharmaceuticals, medicinal science, and cancer research.
本研究利用纳米级傅里叶变换红外光谱技术（nanoFTIR）对在含碳标记葡萄糖的环境中培养的单个细菌细胞进行稳定同位素探测（SIP）。SIP-nanoFTIR通过红外光谱同时定量单细胞代谢，并通过原子力显微镜获取细胞形态信息。酰胺I峰的红移对应于新合成蛋白质的同位素富集。这些单细胞翻译活性的观察结果与传统方法观察大量细胞数量的结果相当。在有限碳条件下培养细胞时，SIP-nanoFTIR用于识别环境诱导的代谢异质性和细胞形态变化。在不利条件或环境波动期间，在与邻近细胞竞争中胜出的个体可能在塑造种群动态方面发挥不成比例的重要作用。此外，SIP-nanoFTIR能够对特定细胞生长阶段进行光谱区分。在细胞复制过程中，亚细胞同位素分布变得更加均匀，这反映在依赖于C-C模式耦合程度或蛋白质二级结构内特定同位素对称性的光谱特征中。由于SIP-nanoFTIR能够捕捉单细胞代谢、环境诱导的细胞过程以及亚细胞同位素定位，该技术在微生物生态学、生物物理学、生物制药、医学和癌症研究等多个学科中具有广泛的应用。
