Rapid identification of single microbes by various Raman spectroscopic techniques

Petra Rösch; Michaela Harz; Michael Schmitt; Klaus-Dieter Peschke; Olaf Ronneberger; Hans Burkhardt; Hans-Walter Motzkus; Markus Lankers; Stefan Hofer; Hans Thiele; Jürgen Popp

doi:10.1117/12.645632

27 February 2006 Rapid identification of single microbes by various Raman spectroscopic techniques

Petra Rösch, Michaela Harz, Michael Schmitt, Klaus-Dieter Peschke, Olaf Ronneberger, Hans Burkhardt, Hans-Walter Motzkus, Markus Lankers, Stefan Hofer, Hans Thiele, Jürgen Popp

Author Affiliations +

Proceedings Volume 6093, Biomedical Vibrational Spectroscopy III: Advances in Research and Industry; 60930D (2006) https://doi.org/10.1117/12.645632
Event: SPIE BiOS, 2006, San Jose, California, United States

Abstract

A fast and unambiguous identification of microorganisms is necessary not only for medical purposes but also in technical processes such as the production of pharmaceuticals. Conventional microbiological identification methods are based on the morphology and the ability of microbes to grow under different conditions on various cultivation media depending on their biochemical properties. These methods require pure cultures which need cultivation of at least 6 h but normally much longer. Recently also additional methods to identify bacteria are established e.g. mass spectroscopy, polymerase chain reaction (PCR), flow cytometry or fluorescence spectroscopy. Alternative approaches for the identification of microorganisms are vibrational spectroscopic techniques. With Raman spectroscopy a spectroscopic fingerprint of the microorganisms can be achieved. Using UV-resonance Raman spectroscopy (UVRR) macromolecules like DNA/RNA and proteins are resonantly enhanced. With an excitation wavelength of e.g. 244 nm it is possible to determine the ratio of guanine/cytosine to all DNA bases which allows a genotypic identification of microorganisms. The application of UVRR requires a large amount of microorganisms (> 10⁶ cells) e.g. at least a micro colony. For the analysis of single cells micro-Raman spectroscopy with an excitation wavelength of 532 nm can be used. Here, the obtained information is from all type of molecules inside the cells which lead to a chemotaxonomic identification. In this contribution we show how wavelength dependent Raman spectroscopy yields significant molecular information applicable for the identification of microorganisms on a single cell level.

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Petra Rösch, Michaela Harz, Michael Schmitt, Klaus-Dieter Peschke, Olaf Ronneberger, Hans Burkhardt, Hans-Walter Motzkus, Markus Lankers, Stefan Hofer, Hans Thiele, and Jürgen Popp "Rapid identification of single microbes by various Raman spectroscopic techniques", Proc. SPIE 6093, Biomedical Vibrational Spectroscopy III: Advances in Research and Industry, 60930D (27 February 2006); https://doi.org/10.1117/12.645632

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