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Influence of Sterilization and Preservation Procedures on the Integrity of Serum Protein-Coated Magnetic Nanoparticles
2017, Dutz, Silvio, Wojahn, Stephanie, Gräfe, Christine, Weidner, Andreas, Clement, Joachim H.
Protein-coated magnetic nanoparticles are promising candidates for various medical applications. Prior to their application into a biological system, one has to guarantee that the particle dispersions are free from pathogens or any other microbiologic contamination. Furthermore, to find entrance into clinical routine, the nanoparticle dispersions have to be storable for several months. In this study, we tested several procedures for sterilization and preservation of nanoparticle containing liquids on their influence on the integrity of the protein coating on the surface of these particles. For this, samples were treated by freezing, autoclaving, lyophilization, and ultraviolet (UV) irradiation, and characterized by means of dynamic light scattering, determination of surface potential, and gel electrophoresis afterwards. We found that the UV sterilization followed by lyophilization under the addition of polyethylene glycol are the most promising procedures for the preparation of sterilized long-term durable protein-coated magnetic nanoparticles. Ongoing work is focused on the optimization of used protocols for UV sterilization and lyophilization for further improvement of the storage time.
Evaluation of shifted excitation Raman difference spectroscopy and comparison to computational background correction methods applied to biochemical Raman spectra
2017, Cordero, Eliana, Korinth, Florian, Stiebing, Clara, Krafft, Christoph, Schie, Iwan W., Popp, Jürgen
Raman spectroscopy provides label-free biochemical information from tissue samples without complicated sample preparation. The clinical capability of Raman spectroscopy has been demonstrated in a wide range of in vitro and in vivo applications. However, a challenge for in vivo applications is the simultaneous excitation of auto-fluorescence in the majority of tissues of interest, such as liver, bladder, brain, and others. Raman bands are then superimposed on a fluorescence background, which can be several orders of magnitude larger than the Raman signal. To eliminate the disturbing fluorescence background, several approaches are available. Among instrumentational methods shifted excitation Raman difference spectroscopy (SERDS) has been widely applied and studied. Similarly, computational techniques, for instance extended multiplicative scatter correction (EMSC), have also been employed to remove undesired background contributions. Here, we present a theoretical and experimental evaluation and comparison of fluorescence background removal approaches for Raman spectra based on SERDS and EMSC.
An optically pumped magnetometer working in the light-shift dispersed Mz mode
2017, Schultze, Volkmar, Schillig, Bastian, IJsselsteijn, Rob, Scholtes, Theo, Woetzel, Stefan, Stolz, Ronny
We present an optically pumped magnetometer working in a new operational mode—the light-shift dispersed Mz (LSD-Mz) mode. It is realized combining various features; (1) high power off-resonant optical pumping; (2) Mz configuration, where pumping light and magnetic field of interest are oriented parallel to each other; (3) use of small alkali metal vapor cells of identical properties in integrated array structures, where two such cells are pumped by circularly polarized light of opposite helicity; and (4) subtraction of the Mz signals of these two cells. The LSD-Mz magnetometer’s performance depends on the inherent and very complex interplay of input parameters. In order to find the configuration of optimal magnetometer resolution, a sensitivity analysis of the input parameters by means of Latin Hypercube Sampling was carried out. The resulting datasets of the multi-dimensional parameter space exploration were assessed by a subsequent physically reasonable interpretation. Finally, the best shot-noise limited magnetic field resolution was determined within that parameter space. As the result, using two 50 mm3 integrated vapor cells a magnetic field resolution below 10 fT/√Hz at Earth’s magnetic field strength is possible
A new strategy for silver deposition on Au nanoparticles with the use of peroxidase-mimicking DNAzyme monitored by Localized Surface Plasmon Resonance technique
2017, Kosman, Joanna, Jatschka, Jacqueline, Csáki, Andrea, Fritzsche, Wolfgang, Juskowiak, Bernard, Stranik, Ondrej
Peroxidase-mimicking DNAzyme was applied as a catalyst of silver deposition on gold nanoparticles. This DNAzyme is formed when hemin binds to the G-quadruplex-forming DNA sequence. Such a system is able to catalyze a redox reaction with a one- or two-electron transfer. The process of silver deposition was monitored via a localized surface plasmon resonance technique (LSPR), which allows one to record scattering spectrum of a single nanoparticle. Our study showed that DNAzyme is able to catalyze silver deposition. The AFM experiments proved that DNAzyme induced the deposition of silver shells of approximately 20 nm thickness on Au nanoparticles (AuNPs). Such an effect is not observed when hemin is absent in the system. However, we noticed non-specific binding of hemin to the capture oligonucleotides on a gold NP probe that also induced some silver deposition, even though the capture probe was unable to form G-quadruplex. Analysis of SEM images indicated that the surface morphology of the silver layer deposited by DNAzyme is different from that obtained for hemin alone. The proposed strategy of silver layer synthesis on gold nanoparticles catalyzed by DNAzyme is an innovative approach and can be applied in bioanalysis (LSPR, electrochemistry) as well as in material sciences.
Detection of Pseudomonas aeruginosa Metabolite Pyocyanin in Water and Saliva by Employing the SERS Technique
2017, Zukovskaja, Olga, Jahn, Izabella-Jolan, Weber, Karina, Cialla-May, Dana, Popp, Jürgen
Pyocyanin (PYO) is a metabolite specific for Pseudomonas aeruginosa. In the case of immunocompromised patients, it is currently considered a biomarker for life-threating Pseudomonas infections. In the frame of this study it is shown, that PYO can be detected in aqueous solution by employing surface-enhanced Raman spectroscopy (SERS) combined with a microfluidic platform. The achieved limit of detection is 0.5 μM. This is ~2 orders of magnitude below the concentration of PYO found in clinical samples. Furthermore, as proof of principle, the SERS detection of PYO in the saliva of three volunteers was also investigated. This body fluid can be collected in a non-invasive manner and is highly chemically complex, making the detection of the target molecule challenging. Nevertheless, PYO was successfully detected in two saliva samples down to 10 μM and in one sample at a concentration of 25 μM. This indicates that the molecules present in saliva do not inhibit the efficient adsorption of PYO on the surface of the employed SERS active substrates.