<i>Bacillus</i> Spore Classification via Surface-Enhanced Raman Spectroscopy and Principal Component Analysis

Guicheteau, Jason A.; Argue, Leanne; Emge, Darren K.; Hyre, Aaron M.; Jacobson, Michele L.; Christesen, Steven D.

doi:10.1366/000370208783759623

Cited by 75 publications

(63 citation statements)

References 31 publications

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“…Guicheteau and his coworkers analyzed pathogens directly using a metal NPs solution, i.e. gold and silver (Guicheteau et al, 2008). Huang et al used a heatinduced SERS sensing method to directly dry a glutathione-AgNPs mixture, and found a detection limit of ca.…”

Section: Introductionmentioning

confidence: 99%

AuNPs@mesoSiO2 composites for SERS detection of DTNB molecule

Lin

Chang

2014

Biosensors and Bioelectronics

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Section: Introductionmentioning

confidence: 99%

AuNPs@mesoSiO2 composites for SERS detection of DTNB molecule

Lin

Chang

2014

Biosensors and Bioelectronics

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“…Several Raman-based methods dealing with bacterial spores have already verified their feasibility in this respect, relying on normal Raman, [9,32 -36] UV-resonance Raman, [37,38] surface-enhanced Raman (SERS) [39,40] and nonlinear Raman spectroscopies [41,42] as well as Raman imaging. [43 -46] In some of these publications, the possibility to perform the detection and identification at the single organism level via a micro-Raman setup was exploited, making cultivation steps prior to the actual measurements redundant.…”

Section: Introductionmentioning

confidence: 99%

Effect of supplementary manganese on the sporulation of Bacillus endospores analysed by Raman spectroscopy

Stöckel

Meisel

Böhme

et al. 2009

J Raman Spectroscopy

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It is a common practice in microbiology to induce and accelerate sporulation of spore-forming bacteria by adding small amounts of divalent manganese to the cultivation medium. By micro-Raman spectroscopy the effect of supplementary divalent manganese during the growth and sporulation of Bacillus spp. bacteria was studied. The spectral alterations in the Raman spectra of single endospores due to this cultivation parameter comprised slight alterations of the bands attributed to intracellular, abundantly present calcium dipicolinate (CaDPA). Those signals suffered a loss of intensity or partial band broadening because of the appearance of new weak signals next to them. Exclusively in Raman spectra of single B. sphaericus endospores, the band at 1485 cm −1 vanished. The theoretical spectra of CaDPA and manganese dipicolinate (MnDPA) were calculated and compared with the experimental spectra to prove the hypothesis that, while the overall intracellular DPA content decreased, an intracellular assembly of MnDPA in the endospores might also occur. Band shifts of the COO − vibrations in the salt's spectra as well as in the endospore's spectra, and the decrease of the two CaDPA bands, confirmed this proposal. The appearance of the 1030 cm −1 band in all Bacillus spectra as well as the disappearance of the 1485 cm −1 band in the B. sphaericus spectra still needs to be clarified. With the help of two multivariate chemometric methods, these spectral alterations allowed discrimination between single endospores of different Bacillus strains cultivated on normal nutrient agar (NA) and those grown on NA with MnSO 4 · xH 2 O addition. With these investigations, a possible strategy is shown to trace back the cultivation environment of matured single endospores. Utilizing the joint concept of micro-Raman spectroscopy and chemometric analysis, the differentiation between natively grown endospores and those cultivated in a laboratory with the help of manganous salts as a common sporulation accelerator seems accomplishable.

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“…Along with Raman spectroscopy, other methods generate complex spectra such as Fourier transform infrared spectroscopy (FT-IR) [34,98,99] and surface-enhanced Raman spectroscopy [61,71,94,100]. Due to the complex nature of the spectra, many researchers ease data processing by truncating data using principal component analysis (PCA) prior to classifying bacteria [34,92,[101][102][103][104][105][106].…”

Section: Introductionmentioning

confidence: 99%

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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Bacillus Spore Classification via Surface-Enhanced Raman Spectroscopy and Principal Component Analysis

Cited by 75 publications

References 31 publications

AuNPs@mesoSiO2 composites for SERS detection of DTNB molecule

AuNPs@mesoSiO2 composites for SERS detection of DTNB molecule

Effect of supplementary manganese on the sporulation of Bacillus endospores analysed by Raman spectroscopy

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

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