Sub-THz Vibrational Spectroscopy of Bacterial Cells and Molecular Components

T. Globus, T. Dorofeeva, I. Sizov, B. Gelmont, M. Lvovska, T. Khromova, O. Chertihin, Yulia A Koryakina
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引用次数: 36

Abstract

In this work, sub-terahertz (THz) spectroscopy is applied to characterize lyophilized and in vitro cultured bacterial cells of non-pathogenic species of Escherichia coli (E. coli) and Bacillus subtilis (BG), spores of BG and DNA fro m E. coli. One of the goals of this research is to demonstrate that Fourier Transform (FT) spectroscopy in the frequency region of 10- 25 cm -1 is sensitive enough to reveal characteristic spectral features fro m bio-cells and spores in different environment, to verify the d ifferences between species, and to show the response of spores to vacuum and response of cultured cells to heat. The experimental technique was significantly improved for sensitivity and reliab ility. Observed spectra taken with a spectral resolution of 0.25 cm -1 using FT spectrometer with a detector operating at 1.7 K are rich in well resolved features having spectral widths of ~0.5-1 cm -1 . The reproducibility of experimental results was verified and confirmed. Measured spectra from E. coli DNA and from the entire cell have many similarit ies, thus demonstrating that the cellu lar co mponents might contribute to the v ibrational spectrum of the cell. The results of this work confirm that observed spectroscopic features are caused by fundamental physical mechanism of interaction between THz rad iation and biological macro-mo lecules. Particu larly, the analysis of results indicates that the spectroscopic signatures of microorganis ms originate fro m the co mbination of low frequency vibrational modes or group of modes at close frequencies (vibrational bands) within mo lecular co mponents of bacterial cells/spores, with the significant contribution from the DNA. The significance of this study is justified by necessity for a fast and effective, label free and reagent free optical technology to protect against environmental and other biological threats, as well as for general medical research. The obtained results show that THz v ibrational spectroscopy promises to add quantitative genetic information to the characteristic signatures of biological objects, increasing the detection accuracy and selectivity when appropriate spectral resolution is used.
细菌细胞和分子成分的亚太赫兹振动光谱
在这项工作中,亚太赫兹(THz)光谱应用于冻干和体外培养的非致病性大肠杆菌(E. coli)和枯草芽孢杆菌(BG), BG孢子和大肠杆菌的DNA。本研究的目的之一是证明傅里叶变换(FT)光谱在10- 25 cm -1的频率区域足够灵敏,可以揭示不同环境下生物细胞和孢子的特征光谱特征,验证物种之间的差异,并显示孢子对真空的响应和培养细胞对热的响应。实验技术的灵敏度和可靠性有了明显提高。使用光谱分辨率为0.25 cm -1的FT光谱仪和工作在1.7 K的探测器所观察到的光谱具有丰富的光谱宽度为~0.5-1 cm -1的良好分辨率特征。对实验结果的重复性进行了验证和确认。大肠杆菌DNA的测量光谱与整个细胞的测量光谱有许多相似之处,从而表明细胞组分可能对细胞的振动光谱有贡献。本研究结果证实了观测到的光谱特征是由太赫兹辐射与生物大分子相互作用的基本物理机制引起的。特别是,分析结果表明,微生物的光谱特征源于细菌细胞/孢子的非分子组分中的低频振动模式或近频率(振动带)模式的组合,其中DNA的贡献很大。这项研究的意义是合理的,因为需要一种快速有效、无标签和无试剂的光学技术,以防止环境和其他生物威胁,以及一般医学研究。结果表明,太赫兹振动光谱有望为生物物体的特征特征增加定量遗传信息,在适当的光谱分辨率下提高检测精度和选择性。
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