Special Section on Vibrational Spectroscopy and Imaging

Single bacteria identification by Raman spectroscopy

[+] Author Affiliations
Samy Andrea Strola

CEA-LETI, Minatec Campus, 17, avenue des Martyrs, 38054 Grenoble Cedex 9, France

Jean-Charles Baritaux

CEA-LETI, Minatec Campus, 17, avenue des Martyrs, 38054 Grenoble Cedex 9, France

Emmanuelle Schultz

CEA-LETI, Minatec Campus, 17, avenue des Martyrs, 38054 Grenoble Cedex 9, France

Anne Catherine Simon

CEA, LIST, Gif-sur-Yvette, F-91191, France

Cédric Allier

CEA-LETI, Minatec Campus, 17, avenue des Martyrs, 38054 Grenoble Cedex 9, France

Isabelle Espagnon

CEA, LIST, Gif-sur-Yvette, F-91191, France

Dorothée Jary

CEA-LETI, Minatec Campus, 17, avenue des Martyrs, 38054 Grenoble Cedex 9, France

Jean-Marc Dinten

CEA-LETI, Minatec Campus, 17, avenue des Martyrs, 38054 Grenoble Cedex 9, France

J. Biomed. Opt. 19(11), 111610 (Jul 16, 2014). doi:10.1117/1.JBO.19.11.111610
History: Received February 20, 2014; Revised June 13, 2014; Accepted June 19, 2014
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Abstract.  We report on rapid identification of single bacteria using a low-cost, compact, Raman spectroscope. We demonstrate that a 60-s procedure is sufficient to acquire a comprehensive Raman spectrum in the range of 600 to 3300cm1. This time includes localization of small bacteria aggregates, alignment on a single individual, and spontaneous Raman scattering signal collection. Fast localization of small bacteria aggregates, typically composed of less than a dozen individuals, is achieved by lensfree imaging over a large field of view of 24mm2. The lensfree image also allows precise alignment of a single bacteria with the probing beam without the need for a standard microscope. Raman scattered light from a 34-mW continuous laser at 532 nm was fed to a customized spectrometer (prototype Tornado Spectral Systems). Owing to the high light throughput of this spectrometer, integration times as low as 10 s were found acceptable. We have recorded a total of 1200 spectra over seven bacterial species. Using this database and an optimized preprocessing, classification rates of 90% were obtained. The speed and sensitivity of our Raman spectrometer pave the way for high-throughput and nondestructive real-time bacteria identification assays. This compact and low-cost technology can benefit biomedical, clinical diagnostic, and environmental applications.

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© 2014 Society of Photo-Optical Instrumentation Engineers

Citation

Samy Andrea Strola ; Jean-Charles Baritaux ; Emmanuelle Schultz ; Anne Catherine Simon ; Cédric Allier, et al.
"Single bacteria identification by Raman spectroscopy", J. Biomed. Opt. 19(11), 111610 (Jul 16, 2014). ; http://dx.doi.org/10.1117/1.JBO.19.11.111610


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