Analytical and Bioanalytical Chemistry

, Volume 405, Issue 5, pp 1577–1591 | Cite as

Characterization and analysis of mycobacteria and Gram-negative bacteria and co-culture mixtures by Raman microspectroscopy, FTIR, and atomic force microscopy

  • Mingjie Tang
  • Gerald D. McEwen
  • Yangzhe Wu
  • Charles D. Miller
  • Anhong Zhou
Original Paper


The molecular composition of mycobacteria and Gram-negative bacteria cell walls is structurally different. In this work, Raman microspectroscopy was applied to discriminate mycobacteria and Gram-negative bacteria by assessing specific characteristic spectral features. Analysis of Raman spectra indicated that mycobacteria and Gram-negative bacteria exhibit different spectral patterns under our experimental conditions due to their different biochemical components. Fourier transform infrared (FTIR) spectroscopy, as a supplementary vibrational spectroscopy, was also applied to analyze the biochemical composition of the representative bacterial strains. As for co-cultured bacterial mixtures, the distribution of individual cell types was obtained by quantitative analysis of Raman and FTIR spectral images and the spectral contribution from each cell type was distinguished by direct classical least squares analysis. Coupled atomic force microscopy (AFM) and Raman microspectroscopy realized simultaneous measurements of topography and spectral images for the same sampled surface. This work demonstrated the feasibility of utilizing a combined Raman microspectroscopy, FTIR, and AFM techniques to effectively characterize spectroscopic fingerprints from bacterial Gram types and mixtures.


AFM deflection images, Raman spectra, SEM images, and FTIR of Mycobacterium sp. KMS


Mycobacteria Gram-negative bacteria Raman microspectroscopy Fourier transform infrared (FTIR) spectroscopy Atomic force microscopy (AFM) 



This work is partially supported by Huntsman Environmental Research Center and Utah Water Research Laboratory, Logan, UT, USA. We also thank Mr. Juan Ciorciari from Thermo Fisher Scientific for helping in the FTIR measurement of bacterial cells. We also thank Dr. Fen-Ann Shen from the Center for Surface Analysis and Applications, USU for the SEM imaging.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Mingjie Tang
    • 1
  • Gerald D. McEwen
    • 1
  • Yangzhe Wu
    • 1
  • Charles D. Miller
    • 1
  • Anhong Zhou
    • 1
  1. 1.Department of Biological EngineeringUtah State UniversityLoganUSA

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