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High-Wavenumber Raman Spectroscopy for the Identification and Biochemical Characterization of Microbial Species
Alec B Walter1,2, Ezekiel Haugen1,2, Anna S Rourke-Funderburg1,2
1Vanderbilt Biophotonics Center, Vanderbilt University, Nashville, Tennessee 37240, United States.
Analytical Chemistry
|December 17, 2025
Summary
High-wavenumber Raman spectroscopy effectively identifies and characterizes microbes by analyzing their biochemical makeup. This method overcomes autofluorescence issues, enabling accurate microbial discrimination and analysis.
Area of Science:
- Microbiology
- Spectroscopy
- Biochemistry
Background:
- Raman spectroscopy is vital for microbial research and infectious disease diagnostics.
- Traditional fingerprint region analysis is hindered by high autofluorescence backgrounds.
- Investigating alternative spectral regions is crucial for improving signal quality.
Purpose of the Study:
- To evaluate the high-wavenumber Raman region (2800-3800 cm-1) for microbial identification and biochemical characterization.
- To develop and validate a classification model for microbial discrimination using high-wavenumber spectra.
- To determine the biochemical composition of microbial species non-destructively.
Main Methods:
- Collected high-wavenumber Raman spectra from 14 microbial species.
- Trained and validated a multitiered classification model for microbial identification.
- Employed spectral unmixing to quantify Raman contributions from cellular components.
- Converted spectral fractions to relative dry mass for biochemical characterization.
Main Results:
- Achieved high accuracy in identifying cell wall type (100%), genus (98.9%), and species (97.4%).
- Identified relative contributions of proteins, carbohydrates, nucleic acids, lipids, and cell wall components.
- Biochemical characterization using Raman dry mass closely matched existing data for E. coli.
Conclusions:
- High-wavenumber Raman spectroscopy offers a low-background, high-performance alternative for microbial analysis.
- This technique enables both accurate microbial discrimination and non-destructive biochemical profiling.
- The findings support the utility of high-wavenumber Raman spectroscopy in microbiology and infectious disease research.
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