Multiwavelength Resonance Raman Characterization of the Effect of Growth Phase and Culture Medium on Bacteria

Kunapareddy, Nagapratima; Grün, J.; Lunsford, R.; Nikitin, Sergei; Wang, Zheng; Gillis, David

doi:10.1366/14-07770

Cited by 9 publications

(24 citation statements)

References 23 publications

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“…In contrast, culture conditions of the human body vary from person to person. Researchers are aware of the discrepancy and have investigated the changes in Raman spectra due to factors such as temperature [29,30], growth phase [29][30][31][32][33][34][35][36][37][38], and growth media [29,30,33,38]. For the sake of this review, emphasis will be placed on the influence of growth media and phase on Raman spectra.…”

Section: Raman Spectroscopy For Bacterial Analysismentioning

confidence: 99%

“…The influence of growth media on Raman spectra of bacteria has been investigated by a number of researchers [29,30,33,39,40] with varied results. The majority of findings indicated differences in spectra due to growth media.…”

Section: Influence Of Growth Media On Raman Spectramentioning

confidence: 99%

“…The influence of growth time or growth phase (metabolic state) on Raman spectra of bacteria has been studied by a number of researchers [29][30][31][32][33][34][35][36][37][38] Other articles have collected and compared Raman spectra of bacteria from different growth times, but have not specified growth stage. Choo-Smith et al [34] collected and compared Raman spectra of bacterial colonies grown on agar plates for 6, 12, and 24 hours at various colony depths using a confocal Raman microscope.…”

Section: Influence Of Culture Time On Raman Spectramentioning

confidence: 99%

“…For example, Raman spectra from laser wavelengths in the ultraviolet region like 244 nm and 229 nm will primarily be rich in information about nucleic acids and aromatic amino acids, respectively [42]. Despite the spectral variations, Kunapareddy et al [33] found that bacteria could be distinguished regardless of growth phase or culture medium.…”

Section: Influence Of Culture Time On Raman Spectramentioning

confidence: 99%

“…Raman spectroscopy is sensitive to small changes in bacteria cells due to growth media [29,30,33,39,40,45]. For example, Mlynàrikovà et al [39] compared Raman spectra of bacteria and yeast cultured in various growth media to identify a media which lead to the least amount of spectral variance.…”

Section: Introductionmentioning

confidence: 99%

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The use of microfluidics and dielectrophoresis for separation, concentration, and identification of bacteria

et al. 2016

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Traditional bacterial analyses take one to two days under favorable conditions where the bulk of the time is spent waiting for bacteria to divide and grow until visual colonies can be observed for identification. In the case of bacteria with slow doubling times, this process can take weeks. This delay in analysis is unacceptable, especially in cases of life threatening diseases or emergencies. It is clear that in order to decrease the analysis time of the bacteria, the culturing and growth step must be circumvented. The goal of this research is to design, build, and test a device that could decrease the analysis time of bacteria using label-free methods of dielectrophoresis and Raman spectroscopy.Testing for device design was performed with clinical samples in mind, which consist of bacteria grown in a variety of environmental conditions (i.e. available food sources, growth stage, temperature, etc.) and accompanied by sample debris. Raman spectra of bacteria grown in varying media and metabolic stages were collected and analyzed. Results indicate that growth phase and media have an impact on Raman spectra iv and is distinguishable by linear discriminant analysis (LDA). Despite these spectral differences, it was found that LDA classification of closely related bacteria remains fairly high (90%) regardless of growth phase. Sample debris were also considered in device design and accommodated for by dielectrophoresis. Devices were built with the goal to isolate bacteria from a mixed sample and simultaneously acquire Raman spectra for identification.For this dissertation, a device was designed, built, and tested that incorporates dielectrophoresis for particle isolation and Raman spectroscopy for identification. The device was modeled in COMSOL to ensure that an appropriate electrical field gradient could be obtained to isolate bacteria from 5 µm diameter polystyrene spheres. The device was built and successfully trapped bacteria away from polystyrene spheres and Raman spectra of the bacteria were collected while trapped. These results indicate a clear potential for contactless dielectrophoresis-Raman devices to isolate and identify bacteria from sample debris, and thereby decrease the analysis time of bacteria. Typical bacterial analysis involves culturing and visualizing colonies on an array of agar plates. The growth patterns and colors among the array are used to identify the bacteria. For fast growing bacteria such as Escherichia coli, analysis will take one to two days. However, slow growing bacteria such as mycobacteria can take weeks to identify.In addition, there are some species of bacteria that are viable but nonculturable. This lengthy analysis time is unacceptable for life-threatening infections and emergency situations. It is clear that to decrease the analysis of the bacteria, the culturing and growth steps must be avoided. The goal of this research is to design, build, and test a device that could decrease the analysis time of bacteria.

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Section: Raman Spectroscopy For Bacterial Analysismentioning

confidence: 99%

Section: Influence Of Growth Media On Raman Spectramentioning

confidence: 99%

Section: Influence Of Culture Time On Raman Spectramentioning

confidence: 99%

Section: Influence Of Culture Time On Raman Spectramentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Markerfreie molekulare Bildgebung biologischer Zellen und Gewebe durch lineare und nichtlineare Raman‐spektroskopische Ansätze

et al. 2017

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Raman‐Spektroskopie ist eine aufstrebende Technik in der Analyse und Bildgebung von Biomaterialien, da sie in der Lage ist, einen einzigartigen spektroskopischen Fingerabdruck zu erzeugen. Die Darstellung von Zellen und Geweben durch Raman‐Mikrospektroskopie stellt einen zerstörungsfreien und markerfreien Ansatz dar. Dabei tragen alle Komponenten zu den Raman‐Signalen bei, die komplexe spektrale Signaturen liefern. Verfahren wie Resonanz‐Raman‐Streuung und oberflächenverstärkte Raman‐Streuung verstärken die Signale und ergeben einfachere Spektren. Raman‐aktive Markierungen können eingeführt werden, um die Spezifität und die Multimodalität zu erhöhen. Nichtlineare kohärente Raman‐Streuung bietet höhere Empfindlichkeiten, die eine schnelle Abbildung größerer Probenflächen ermöglichen. Schließlich eröffnen faserbasierte Bildgebungsverfahren den Weg zu In‐vivo‐Anwendungen der Raman‐Spektroskopie. Dieser Aufsatz fasst die Grundlagen der medizinischen Raman‐Bildgebung und ihre Fortschritte seit 2012 zusammen.

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Label‐Free Molecular Imaging of Biological Cells and Tissues by Linear and Nonlinear Raman Spectroscopic Approaches

et al. 2017

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Raman spectroscopy is an emerging technique in bioanalysis and imaging of biomaterials owing to its unique capability of generating spectroscopic fingerprints. Imaging cells and tissues by Raman microspectroscopy represents a nondestructive and label-free approach. All components of cells or tissues contribute to the Raman signals, giving rise to complex spectral signatures. Resonance Raman scattering and surface-enhanced Raman scattering can be used to enhance the signals and reduce the spectral complexity. Raman-active labels can be introduced to increase specificity and multimodality. In addition, nonlinear coherent Raman scattering methods offer higher sensitivities, which enable the rapid imaging of larger sampling areas. Finally, fiber-based imaging techniques pave the way towards in vivo applications of Raman spectroscopy. This Review summarizes the basic principles behind medical Raman imaging and its progress since 2012.

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Multiwavelength Resonance Raman Characterization of the Effect of Growth Phase and Culture Medium on Bacteria

Cited by 9 publications

References 23 publications

The use of microfluidics and dielectrophoresis for separation, concentration, and identification of bacteria

The use of microfluidics and dielectrophoresis for separation, concentration, and identification of bacteria

Markerfreie molekulare Bildgebung biologischer Zellen und Gewebe durch lineare und nichtlineare Raman‐spektroskopische Ansätze

Label‐Free Molecular Imaging of Biological Cells and Tissues by Linear and Nonlinear Raman Spectroscopic Approaches

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