Vibrational spectroscopy—A powerful tool for the rapid identification of microbial cells at the single‐cell level

Harz, Michaela; Rösch, Petra; Popp, Jürgen

doi:10.1002/cyto.a.20682

Cited by 257 publications

(187 citation statements)

References 88 publications

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“…A number of studies have shown that Raman scattering-based methods allow for the assessment of the type of contamination, as well as the degree of contamination in such applications. [79][80][81][82][83][84] In early work, Choo-Smith et al 85 86 used Raman spectroscopy to quantify filtered waterborne pathogenic microbial contamination, and Pasteris et al 87 used Raman spectroscopy to identify sulfur inclusions in sulfur-oxidizing marine bacteria. Recently, Palchaudhuri et al 88 investigated the uptake and metabolism of the five-carbon sugar alcohol xylitol by gram-positive viridian group Streptococcus and two gram-negative Escherichia coli strains.…”

Section: Biological Applicationsmentioning

confidence: 99%

Methods and Applications of Raman Microspectroscopy to Single-Cell Analysis

2013

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Raman spectroscopy is a powerful biochemical analysis technique that allows for the dynamic characterization and imaging of living biological cells in the absence of fluorescent stains. In this review, we summarize some of the most recent developments in the noninvasive biochemical characterization of single cells by spontaneous Raman scattering. Different instrumentation strategies utilizing confocal detection optics, multispot, and line illumination have been developed to improve the speed and sensitivity of the analysis of single cells by Raman spectroscopy. To analyze and visualize the large data sets obtained during such experiments, sophisticated multivariate statistical analysis tools are necessary to reduce the data and extract components of interest. We highlight the most recent applications of single cell analysis by Raman spectroscopy and their biomedical implications that have enabled the noninvasive characterization of specific metabolic states of eukaryotic cells, the identification and characterization of stem cells, and the rapid identification of bacterial cells. We conclude the article with a brief look into the future of this rapidly evolving research area.

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Section: Biological Applicationsmentioning

confidence: 99%

Methods and Applications of Raman Microspectroscopy to Single-Cell Analysis

2013

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“…Bacteria were diluted to an OD of 0.1 in the particle spiked at ZnO concentrations of 5,10,20 and 50 mg/L. After 48 hours incubation, levels of total Zn and dissolved Zn were determined.…”

Section: Assessment Of Dissolutionmentioning

confidence: 99%

Metalloproteins and phytochelatin synthase may confer protection against zinc oxide nanoparticle induced toxicity in Caenorhabditis elegans

Polak

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Jurkschat

et al. 2014

Comparative Biochemistry and Physiology Part C: Toxicology &

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Spurgeon, David J.; Sturzenbaum, Stephen R.. 2014. Metalloproteins and phytochelatin synthase may confer protection against zinc oxide nanoparticle induced toxicity in Caenorhabditis elegansContact CEH NORA team at noraceh@ceh.ac.ukThe NERC and CEH trademarks and logos ('the Trademarks') are registered trademarks of NERC in the UK and other countries, and may not be used without the prior written consent of the Trademark owner. suggesting that the nominal exposure to pure ZnONPs represents in vivo, at best, a mixture exposure of ionic zinc and nanoparticles. Nevertheless, the analyzes provided evidence that the metallothioneins (mtl-1 and mtl-2), the pytochelatin synthase (pcs-1) and an apoptotic marker (cep-1) were transcriptionally activated. In addition, the DCFH-DA assay provided in vitro evidence of the oxidative potential of ZnONPs in the metal exposure sensitive triple mutant.Raman spectroscopy highlighted that the biomolecular phenotype changes significantly in the metal sensitive knockout worm upon ZnONP exposure, suggesting that these metalloproteins are instrumental in the protection against cytotoxic damage. Finally, ZnONP exposure was shown to decrease growth and development, reproductive capacity and lifespan, effects which were amplified in the triple knockout. By combining diverse toxicological strategies, we identified that individuals (genotypes) housing mutations in key metalloproteins are more susceptible to ZnONP exposure, which underlines the importance of fully functional metalloproteins to minimize ZnONP induced toxicity.

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“…A lthough some specificity could be found, particularly in the 9 -13 cm-1 and 21 -24 cm-1 ranges, better spectral resolution is required for more certain discrimination result. We chose to characterize the E.coli DNA, which constitutes 5 -15% of an entire cell weight [41]. Fig.…”

Section: Abs Orpti On S Pectra From Different Bi Ological Speciesmentioning

confidence: 99%

Sub-THz Vibrational Spectroscopy of Bacterial Cells and Molecular Components

Globus¹,

Dorofeeva²,

Sizov³

et al. 2012

AJBE

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In this work, sub-terahertz (THz) spectroscopy is applied to characterize lyophilized and in vitro cultured bacterial cells of non-pathogenic species of Escherichia coli (E. coli) and Bacillus subtilis (BG), spores of BG and DNA fro m E. coli. One of the goals of this research is to demonstrate that Fourier Transform (FT) spectroscopy in the frequency region of 10-25 cm -1 is sensitive enough to reveal characteristic spectral features fro m bio-cells and spores in different environment, to verify the d ifferences between species, and to show the response of spores to vacuum and response of cultured cells to heat. The experimental technique was significantly improved for sensitivity and reliab ility. Observed spectra taken with a spectral resolution of 0.25 cm -1 using FT spectrometer with a detector operating at 1.7 K are rich in well resolved features having spectral widths of ~0.5-1 cm -1 . The reproducibility of experimental results was verified and confirmed. Measured spectra from E. coli DNA and from the entire cell have many similarit ies, thus demonstrating that the cellu lar co mponents might contribute to the v ibrational spectrum of the cell. The results of this work confirm that observed spectroscopic features are caused by fundamental physical mechanism of interaction between THz rad iation and biological macro-mo lecules. Particu larly, the analysis of results indicates that the spectroscopic signatures of microorganis ms originate fro m the co mbination of low frequency vibrational modes or group of modes at close frequencies (vibrational bands) within mo lecular co mponents of bacterial cells/spores, with the significant contribution from the DNA. The significance of this study is justified by necessity for a fast and effective, label free and reagent free optical technology to protect against environmental and other biological threats, as well as for general medical research. The obtained results show that THz v ibrational spectroscopy promises to add quantitative genetic information to the characteristic signatures of biological objects, increasing the detection accuracy and selectivity when appropriate spectral resolution is used.

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Vibrational spectroscopy—A powerful tool for the rapid identification of microbial cells at the single‐cell level

Cited by 257 publications

References 88 publications

Methods and Applications of Raman Microspectroscopy to Single-Cell Analysis

Methods and Applications of Raman Microspectroscopy to Single-Cell Analysis

Metalloproteins and phytochelatin synthase may confer protection against zinc oxide nanoparticle induced toxicity in Caenorhabditis elegans

Sub-THz Vibrational Spectroscopy of Bacterial Cells and Molecular Components

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