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Label-free Raman characterization of bacteria calls for standardized procedures
García-Timermans, C.; Rubbens, P.; Kerckhof, F.-M.; Buysschaert, B.; Khalenkow, D.; Waegeman, W.; Skirtach, A.G.; Boon, N. (2018). Label-free Raman characterization of bacteria calls for standardized procedures. J. microbiol. methods 151: 69-75. https://dx.doi.org/10.1016/j.mimet.2018.05.027
In: Journal of Microbiological Methods. Elsevier: Amsterdam. ISSN 0167-7012; e-ISSN 1872-8359, more
Peer reviewed article  

Available in  Authors 

Author keywords
    Phenotypic identification; Label-free Raman spectroscopy; Single-cell analysis; Raman standardization

Authors  Top 
  • García-Timermans, C., more
  • Rubbens, P., more
  • Kerckhof, F.-M., more
  • Buysschaert, B.
  • Khalenkow, D.
  • Waegeman, W., more
  • Skirtach, A.G.
  • Boon, N., more

Abstract
    Raman spectroscopy has gained relevance in single-cell microbiology for its ability to detect bacterial (sub)populations in a non-destructive and label-free way. However, the Raman spectrum of a bacterium can be heavily affected by abiotic factors, which may influence the interpretation of experimental results. Additionally, there is no publicly available standard for the annotation of metadata describing sample preparation and acquisition of Raman spectra. This article explores the importance of sample manipulations when measuring bacterial subpopulations using Raman spectroscopy. Based on the results of this study and previous findings in literature we propose a Raman metadata standard that incorporates the minimum information that is required to be reported in order to correctly interpret data from Raman spectroscopy experiments. Its aim is twofold: 1) mitigate technical noise due to sample preparation and manipulation and 2) improve reproducibility in Raman spectroscopy experiments studying microbial communities.

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