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Analytical and Bioanalytical Chemistry

, Volume 407, Issue 29, pp 8919–8923 | Cite as

Raman spectroscopic monitoring of the growth of pigmented and non-pigmented mycobacteria

  • Stephan Stöckel
  • Andrei Sebastian Stanca
  • Jonathan Helbig
  • Petra Rösch
  • Jürgen Popp
Note

Abstract

Raman microspectroscopy has increased in popularity in the field of microbiology because it allows a spectral fingerprinting of bacterial pathogens at an unrivaled speed, which is important for the early treatment of infectious diseases such as tuberculosis. An indispensable prerequisite for the success of this method is a profound knowledge, how the spectral profiles depend on the age of the bacteria. We therefore followed the growth of two rapidly growing Mycobacterium tuberculosis relatives, the pigmented Mycobacteriumaurum, and the non-pigmented Mycobacteriumsmegmatis, by means of Raman microspectroscopy. Both species showed remarkable temporal changes in the single-bacteria Raman spectra: In the signatures of M. aurum, pigment-associated Raman signals could be detected not until 72 h of growth and also remained highly variable thereafter. The Raman spectra of M. smegmatis exhibited lipid signals presumably arising from mycolic acids, which are a hallmark feature of mycobacteria, but only after the bacteria reached the late stationary growth phase (>48 h). A principal component analysis thus classified the Raman spectra according to the cultivation age. In summary, these findings have to be reckoned with in future studies dealing with the identification of mycobacteria via Raman microspectroscopy.

Graphical abstract

Changes in the chemical composition of bacterial cells over growth time may influence the results of Raman spectroscopic studies of bacteria

Keywords

Raman spectroscopy Microscopy Mycobacteria Growth pigments Analytical methods 

Notes

Acknowledgment

The funding of the research projects Fast-TB (2013FE9057) and BioInter (13022-715) by the Free State of Thuringia and the European Union (EFRE) as well as the Deutsche Forschungsgemeinschaft (DFG) for the Collaborative Research Center ChemBioSys (SFB 1127) is highly acknowledged. The authors also thank Sophie Friedrich for technical assistance.

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Stephan Stöckel
    • 1
    • 2
  • Andrei Sebastian Stanca
    • 1
  • Jonathan Helbig
    • 1
  • Petra Rösch
    • 1
    • 2
  • Jürgen Popp
    • 1
    • 2
    • 3
  1. 1.Institute of Physical Chemistry and Abbe School of PhotonicsFriedrich Schiller University JenaJenaGermany
  2. 2.InfectoGnostics Forschungscampus JenaJenaGermany
  3. 3.Leibniz-Institute of Photonic TechnologyJenaGermany

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