Impact of polymerization and storage on the degree of conversion and mechanical properties of veneering resin composites.

To investigate the degree of conversion (DC), Martens hardness (HM), elastic indentation modulus (EIT), and flexural strength (FS) of veneering resin composites (SR Nexco Paste (NP), Ceramage Incisal (CI), Gradia Plus (GP); n=60/group) cured with different polymerization devices (bre.Lux Power Unit, Labolight DUO, Otoflash G171, LC-3DPrint Box, PCU LED; n=12/subgroup) after storage. Otoflash G171 and Labolight DUO showed increased DC/HM/EIT. CI presented the lowest DC and highest HM/EIT. NP showed the highest DC and lowest HM/EIT. Within Otoflash G171, Laboligth DUO and PCU LED, highest FS was observed for CI. Storage did not affect DC/HM/EIT for specimens cured with Otoflash G171 or Labolight DUO. With storage not showing an influence on the tested parameters for polymerization devices that otherwise presented superior results, increased storage time cannot be recommended. For the tested resin composites, this study observed a high/low degree of conversion to coincide with respectively low/high amounts of fillers/mechanical properties.

Raman microspectroscopy, surface-enhanced Raman scattering microspectroscopy, and stable-isotope Raman microspectroscopy for biofilm characterization.

Biofilms represent the predominant form of microbial life on our planet. These aggregates of microorganisms, which are embedded in a matrix formed by extracellular polymeric substances, may colonize nearly all interfaces. Detailed knowledge of microorganisms enclosed in biofilms as well as of the chemical composition, structure, and functions of the complex biofilm matrix and their changes at different stages of the biofilm formation and under various physical and chemical conditions is relevant in different fields. Important research topics include the development and improvement of antibiotics and medical devices and the optimization of biocides, antifouling strategies, and biological wastewater treatment. Raman microspectroscopy is a capable and nondestructive tool that can provide detailed two-dimensional and three-dimensional chemical information about biofilm constituents with the spatial resolution of an optical microscope and without interference from water. However, the sensitivity of Raman microspectroscopy is rather limited, which hampers the applicability of Raman microspectroscopy especially at low biomass concentrations. Fortunately, the resonance Raman effect as well as surface-enhanced Raman scattering can help to overcome this drawback. Furthermore, the combination of Raman microspectroscopy with other microscopic techniques, mass spectrometry techniques, or particularly with stable-isotope techniques can provide comprehensive information on monospecies and multispecies biofilms. Here, an overview of different Raman microspectroscopic techniques, including resonance Raman microspectroscopy and surface-enhanced Raman scattering microspectroscopy, for in situ detection, visualization, identification, and chemical characterization of biofilms is given, and the main feasibilities and limitations of these techniques in biofilm research are presented. Future possibilities of and challenges for Raman microspectroscopy alone and in combination with other analytical techniques for characterization of complex biofilm matrices are discussed in a critical review. Graphical Abstract Applicability of Raman microspectroscopy for biofilm analysis.

The origin of the band at around 730 cm(-1) in the SERS spectra of bacteria: a stable isotope approach.

Raman microspectroscopy is an emerging tool to analyze the molecular and isotopic composition of single microbial cells. It can be used to achieve an in situ understanding of metabolic processes. Due to the low sensitivity of the Raman effect, surface-enhanced Raman scattering (SERS) is utilized to enhance the Raman signal. The SERS spectra of bacteria are usually characterized by a pronounced band at around 730 cm(-1), which is assigned to glycosidic ring vibrations or to adenine or even to CH2 deformation in different studies. In order to clarify the origin of this band, we employed a stable isotope approach and performed a SERS analysis of Escherichia coli bacteria using in situ prepared Ag nanoparticles. The cells were grown on unlabeled ((12)C, (14)N) and labeled ((13)C, (15)N) carbon and nitrogen sources in different combinations. The SERS band of the stable isotope labeled microorganisms showed a characteristic red-shift in the SERS spectra, which solely depends on the isotopic composition. It was therefore possible to confidently assign this band to adenine-related compounds. Furthermore, by utilizing the fingerprint area of single-cell SERS spectra as the input for the principal component analysis, one can clearly differentiate between E. coli bacteria incorporating different stable isotopes.

Exploring the Potential of Stable Isotope (Resonance) Raman Microspectroscopy and Surface-Enhanced Raman Scattering for the Analysis of Microorganisms at Single Cell Level.

Raman microspectroscopy is a prime tool to characterize the molecular and isotopic composition of microbial cells. However, low sensitivity and long acquisition times limit a broad applicability of the method in environmental analysis. In this study, we explore the potential, the applicability, and the limitations of stable isotope Raman microspectroscopy (SIRM), resonance SIRM, and SIRM in combination with surface-enhanced Raman scattering (SERS) for the characterization of single bacterial cells. The latter two techniques have the potential to significantly increase sensitivity and decrease measurement times in SIRM, but to date, there are no (SERS-SIRM) or only a limited number (resonance SIRM) of studies in environmental microbiology. The analyzed microorganisms were grown with substrates fully labeled with the stable isotopes (13)C or (2)H and compounds with natural abundance of atomic isotopes ((12)C 98.89% or (1)H 99.9844%, designated as (12)C or (1)H, respectively). Raman bands of bacterial cell compounds in stable isotope-labeled microorganisms exhibited a characteristic red-shift in the spectra. In particular, the sharp phenylalanine band was found to be an applicable marker band for SIRM analysis of the Deltaproteobacterium strain N47 growing anaerobically on (13)C-naphthalene. The study of G. metallireducens grown with (13)C- and (2)H-acetate showed that the information on the chromophore cytochrome c obtained by resonance SIRM at 532 nm excitation wavelength can be successfully complemented by whole-organism fingerprints of bacteria cells achieved by regular SIRM after photobleaching. Furthermore, we present here for the first time the reproducible SERS analysis of microbial cells labeled with stable isotopes. Escherichia coli strain DSM 1116 cultivated with (12)C- or (13)C-glucose was used as a model organism. Silver nanoparticles synthesized in situ were applied as SERS media. We observed a reproducible red-shift of an adenine-related marker band from 733 to 720 cm(-1) in SERS spectra for (13)C-labeled cells. Additionally, Raman measurements of (12)C/(13)C-glucose and -phenylalanine mixtures were performed to elucidate the feasibility of SIRM for nondestructive quantitative and spatially resolved analysis. The performed analysis of isotopically labeled microbial cells with SERS-SIRM and resonance SIRM paves the way toward novel approaches to apply Raman microspectroscopy in environmental process studies.