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A novel method for single bacteria identification by Raman spectroscopy

机译：拉曼光谱法鉴定单一细菌的新方法

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In this paper we present results on single bacteria rapid identification obtained with a low-cost and compact Raman spectrometer. At present, we demonstrate that a 1 minute procedure, including the localization of single bacterium, is sufficient to acquire comprehensive Raman spectrum in the range of 600 to 3300 cm~(-1). Localization and detection of single bacteria is performed by means of lensfree imaging over a large field of view of 24 mm~2. An excitation source of 532 nm and 30 mW illuminates single bacteria to collect Raman signal into a Tornado Spectral Systems prototype spectrometer (HTVS technology). The acquisition time to record a single bacterium spectrum is as low as 10 s owing to the high light throughput of this spectrometer. The spectra processing features different steps for cosmic spikes removal, background subtraction, and gain normalization to correct the residual inducted fluorescence and substrate fluctuations. This allows obtaining a fine chemical fingerprint analysis. We have recorded a total of 1200 spectra over 7 bacterial species (E. coli, Bacillus species, S. epidermis, M. luteus, S. marcescens). The analysis of this database results in a high classification score of almost 90 %. Hence we can conclude that our setup enables automatic recognition of bacteria species among 7 different species. The speed and the sensitivity (＜30 minutes for localization and spectra collection of 30 single bacteria) of our Raman spectrometer pave the way for high-throughput and non-destructive real-time bacteria identification assays. This compact and low-cost technology can benefit biomedical, clinical diagnostic and environmental applications.
机译：在本文中，我们介绍了使用低成本紧凑型拉曼光谱仪对单个细菌进行快速鉴定的结果。目前，我们证明了包括单个细菌定位在内的1分钟的程序足以获取600至3300 cm〜（-1）范围内的综合拉曼光谱。单个细菌的定位和检测通过无透镜成像在24 mm〜2的大视野内进行。 532 nm和30 mW的激发源照射单个细菌，以将拉曼信号收集到龙卷风光谱系统原型光谱仪（HTVS技术）中。由于该光谱仪的高光通量，记录单个细菌光谱的采集时间可低至10 s。光谱处理采用不同的步骤来消除宇宙尖峰，消除背景并进行增益归一化，以校正残留的感应荧光和底物波动。这允许获得精细的化学指纹分析。我们已经记录了7个细菌物种（大肠杆菌，芽孢杆菌物种，表皮葡萄球菌，黄体分枝杆菌，marcescens）的1200张光谱。该数据库的分析导致近90％的高分类得分。因此，我们可以得出结论，我们的设置可以自动识别7种不同细菌中的细菌。我们的拉曼光谱仪的速度和灵敏度（对30种单一细菌进行定位和收集光谱，均小于30分钟）为高通量和无损实时细菌鉴定检测铺平了道路。这种紧凑而低成本的技术可以使生物医学，临床诊断和环境应用受益。

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