Differentiation of bacterial spores via 2D-IR spectroscopy

Barbara Procacci, Samantha H. Rutherford, Gregory M. Greetham, Michael Towrie, Anthony W. Parker, Camilla V. Robinson, Christopher R. Howle, Neil T. Hunt

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)


Ultrafast 2D-IR spectroscopy is a powerful tool for understanding the spectroscopy and dynamics of biological molecules in the solution phase. A number of recent studies have begun to explore the utility of the information-rich 2D-IR spectra for analytical applications. Here, we report the application of ultrafast 2D-IR spectroscopy for the detection and classification of bacterial spores. 2D-IR spectra of Bacillus atrophaeus and Bacillus thuringiensis spores as dry films on CaF2 windows were obtained. The sporulated nature of the bacteria was confirmed using 2D-IR diagonal and off-diagonal peaks arising from the calcium dipicolinate CaDP·3H2O biomarker for sporulation. Distinctive peaks, in the protein amide I region of the spectrum were used to differentiate the two types of spore. The identified marker modes demonstrate the potential for the use of 2D-IR methods as a direct means of spore classification. We discuss these new results in perspective with the current state of analytical 2D-IR measurements, showing that the potential exists to apply 2D-IR spectroscopy to detect the spores on surfaces and in suspensions as well as in dry films. The results demonstrate how applying 2D-IR screening methodologies to spores would enable the creation of a library of spectra for classification purposes.

Original languageEnglish
Article number119319
Number of pages8
JournalSpectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy
Early online date14 Dec 2020
Publication statusPublished - 15 Mar 2021


  • 2D-IR analysis
  • 2D-IR spectroscopy
  • Bacterial spores


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