Portable Raman Spectroscopy for

Identifying Bacterial Growth Media

As published on spectroscopyonline.com on August 15 th, 2013
http://www.spectroscopyonline.com/spectroscopy/Feature/Portable-Raman-Spectroscopy-for-IdentifyingBacter/ArticleStandard/Article/detail/820547?contextCategoryId=36789

Handheld Raman spectrometers have been used to identify a variety of chemical materials, including
toxic industrial chemicals, chemical warfare agents, explosives, narcotics, pharmaceutical compounds,
and plastics. Researcher Mathew Lyman of the Chemical, Biological, Radiological Nuclear and
Explosives Division at Oklahoma State University University Multispectral Laboratories (OSU-UML)
spoke to Spectroscopy about his and his colleague Jessica Randalls use of a handheld Raman
spectrometer for identifying bacterial growth media.
Given that Raman spectroscopy has historically been problematic for the identification of
biological samples, why did you decide to investigate its use for applications of this type?
There are three reasons why this project was initiated. First, Jessica and I are microbiologists by
training, and we are always impressed with the analytical instrumentation available to the chemists in
our division: Fourier transform infrared (FT-IR) spectroscopy, Raman spectroscopy, inductively coupled
plasma mass spectrometry (ICPMS), gas chromatographymass spectrometry (GCMS), liquid
chromatographymass spectrometry (LCMS), and X-ray diffraction (XRD). An obvious question arises:
How can we apply some of this instrumentation to study forensic biology?
Second, Oklahoma State University University Multispectral Laboratories (OSU-UML) provides
realistic training scenarios for Weapons of Mass Destruction Civil Support Teams (WMD-CSTs)
throughout the region. While observing CST training evolutions, we noticed that handheld Raman
instruments were extremely useful for identifying anything chem for example, toxic industrial
chemicals, toxic industrial materials, solvents, explosives, and chemical precursors. However, anything
bio was impossible to analyze by Raman spectroscopy; other techniques had to be employed, such
as lateral flow assays, polymerase chain reaction, or chemiluminescent enzyme linked immunosorbent
assays (ELISA) (common analytical techniques for microbial agents). It takes significantly more time to
obtain results from these assays compared to Raman spectroscopy; in addition, these assays are
limited by the antibodies or primer sets that are on hand. If the background fluorescence from
biological samples could be mitigated, Raman spectroscopy would be a clear choice!
We also read several papers by the Ziegler lab in the Chemistry Department at Boston University. They
have used surface-enhanced Raman spectroscopy (SERS) to differentiate between multiple types of
19 Shea Way, Newark, DE 19713 Tel: (302) 368-7824 Fax: (302) 368-7830 Web: www.bwtek.com

bacteria, and theres potential for using this technique as a clinical diagnostic tool. It is interesting
work, and it helped us frame the question that led to our paper in Spectroscopy (1).
What were the results of your study of using a handheld Raman spectrometer to identify
bacterial media?
We found that a handheld Raman spectrometer, with a fluorescence background subtraction
algorithm, was able to distinguish between (and correctly identify) several commonly used bacterial
growth media. We also observed that three different types of bacteria culture bacteria propagated
in growth medium had unique Raman signatures. Although this was a small study, it encouraged
us to expand our testing to other types of bacteria and growth media.
Considering the results youve seen, do you think a handheld Raman spectrometer can be a
useful tool for the military and first responders for the identification of biological materials?
Eventually handheld Raman will be useful for that purpose. Most of the handheld Raman
spectrometers currently utilized by the military do not subtract background fluorescence, thereby
limiting their capability to identify biological samples. This should gradually change as newer
instruments and subtraction algorithms become available. The development of a handheld SERS
instrument for the military is also a possibility. Of course, additional reference libraries will need to be
constructed once these technologies arise.
What are your next steps in this work?
Since completing our study with bacterial growth media, we plan to test a limited number of purified
bacteria and spores using our handheld Raman spectrometer. Ultimately we hope to build a spectral
library from purified bacteria and bacterial cultures for database matching (for identifying unknown
biological cultures). It would be rewarding to attend a WMD-CST training in five years and see that
they are having as much success with Raman spectroscopy on biological agents as they do on
chemical agents.

References
(1) J.A. Randall and M.G. Lyman, Raman Technology for Todays Spectroscopists, supplement to
Spectroscopy, pp. 820 (June 2013).
For further reading on Lyman and Randalls work, please visit www.spectroscopyonline.com/Lyman

19 Shea Way, Newark, DE 19713 Tel: (302) 368-7824 Fax: (302) 368-7830 Web: www.bwtek.com