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Biology of Microbial Communities - Interview
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Quantitative SIMS Imaging of Agar-Based Microbial Communities.

Sage J B Dunham1, Joanna F Ellis1, Nameera F Baig2

  • 1Department of Chemistry and Beckman Institute for Advanced Science and Technology , University of Illinois at Urbana-Champaign , Urbana , Illinois 61801 , United States.

Analytical Chemistry
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Summary

This study introduces a new quantitative method for secondary ion mass spectrometry (SIMS) imaging of small molecules in microbial communities. This technique allows for accurate measurement of molecular abundance, overcoming previous limitations in biomolecular analysis.

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Area of Science:

  • Analytical Chemistry
  • Microbiology
  • Surface Science

Background:

  • Secondary ion mass spectrometry (SIMS) is widely used for surface analysis.
  • Biomolecular SIMS imaging has been primarily qualitative, limiting quantitative comparisons.
  • Accurate quantification of small molecules in biological samples remains a challenge.

Purpose of the Study:

  • To develop a quantitative secondary ion mass spectrometry (SIMS) imaging method for small molecules in microbial communities.
  • To enable accurate comparison of relative molecular abundance in biofilms.
  • To validate the quantitative imaging methodology using specific signaling molecules in Pseudomonas aeruginosa.

Main Methods:

  • Microbes were cultivated on agar, dehydrated, and imaged using SIMS.
  • A quantitative imaging methodology involving normalization, interference correction, and noise filtering was applied.
  • Optical microscopy confirmed minimal morphological changes during dehydration.

Main Results:

  • The developed method provides two-dimensional surface density images for analytes.
  • Quantitative imaging revealed significant differences in relative surface densities compared to direct intensity comparisons.
  • The methodology was validated by accurately quantifying alkyl-quinolone and alkyl-quinoline N-oxide signaling molecules.

Conclusions:

  • The new SIMS imaging method enables quantitative analysis of small molecules in microbial biofilms.
  • This approach overcomes limitations of qualitative SIMS imaging, allowing for precise relative abundance comparisons.
  • The validated methodology can be applied to quantitatively compare multiple ions with available standards.