Optimized preparation of pDNA/poly(ethylene imine) polyplexes using a microfluidic system

Poly(ethylene imine) (PEI) is an established non-viral vector system for the delivery of various nucleic acids in gene therapy applications. Polyelectrolyte complexes between both compounds, so called polyplexes, are formed by electrostatic interactions of oppositely charged macromolecules and are thought to facilitate uptake into cells. Such complexes form spontaneously and on lab scale they are usually prepared by mixing solutions through pipetting. Hence, an optimized preparation procedure allowing the scale-up of well-defined polyplexes would be of general interest. We developed a new method for microfluidic polyplex preparation on a chip. The mixing behaviour within the microfluidic channels was evaluated. Polyplexes with PEI and plasmid DNA were prepared using this method, in comparison to the standard pipetting procedure. Sizes and polydispersity indices of these complexes were examined. The influence of various parameters on the polyplex characteristics and the suitability of this production procedure for other PEI-based complexes were also evaluated. It was shown that polyplexes could easily be prepared by microfluidics. The ratio of PEI to DNA was most important for the formation of small polyplexes, whereas other parameters had minor influence. The size of polyplexes prepared with this new method was observed to be relatively constant between 140 nm and 160 nm over a wide range of complex concentrations. In comparison, the size of polyplexes prepared by pipetting (approximately 90 nm to 160 nm) varied considerably. The versatility of this system was demonstrated with different (targeted) PEI-based vectors for the formation of complexes with pDNA and siRNA. In conclusion, polyplex preparation using microfluidics could be a promising alternative to the standard pipetting method due to its suitability for preparation of well-defined complexes with different compositions over a wide range of concentrations.     

School students' reasoning about conjunction and conditional events

The objective of this study was to provide baseline data in an area of the mathematics curriculum that is beginning to receive greater attention than previously. Four survey items were completed by 2615 students in grades 5 to 11. Two survey items asked for estimates of probability or frequency for everyday events (A), (B), and their conjunction (A and B). Two survey items asked for estimates of probability or frequency for conditional events, (X|Y) and (Y|X). Cross-sectional and longitudinal analyses revealed improvement with grade in expressing probability numerically and in distinguishing conditional events, but no change in incidence of conjunction errors. The relationships of responses to conjunction items with those to conditional items, and of both with responses to other items of basic chance measurement were considered. Implications were related to interpretation of the results in terms of previous research and suggestions for educators.     

Combined surface-focused acoustic microscopy in transmission and scanning ultrasonic holography

Employment of ultrasound techniques in nondestructive testing may require identification of the acoustic modes contributing to imaging. Such identification can be achieved, with some restrictions, by time-of-flight analysis. Another approach is acoustic holography that reveals the propagation properties of any selected mode. In anisotropic media, the propagation features are distinct and allow for a reliable classification of the selected mode. Both techniques were applied for classification of bonded, disbonded, and weakly bonded areas in directly bonded semiconductor wafers.     

Construction of environment states in quantum-chemical density-matrix renormalization group calculations

The application of the quantum-chemical density-matrix renormalization group (DMRG) algorithm is cumbersome for complex electronic structures with many active orbitals. The high computational cost is mainly due to the poor convergence of standard DMRG calculations. A factor which affects the convergence behavior of the calculations is the choice of the start-up procedure. In this start-up step matrix representations of operators have to be calculated in a guessed many-electron basis of the DMRG environment block. Different possibilities for the construction of these basis states exist, and we first compare four procedures to approximate the environment states using Slater determinants explicitly. These start-up procedures are applied to DMRG calculations on a sophisticated test system: the chromium dimer. It is found that the converged energies and the rate of convergence depend significantly on the choice of the start-up procedure. However, since already the most simple start-up procedure, which uses only the Hartree-Fock determinant, is comparatively good, Slater determinants, in general, appear not to be a good choice as approximate environment basis states for convergence acceleration. Based on extensive test calculations it is demonstrated that the computational cost can be significantly reduced if the number of total states m is successively increased. This is done in such a way that the environment states are built up stepwise from system states of previous truncated DMRG sweeps for slowly increasing m values.     

Sequential penning ionization: harvesting energy with ions

We report the observation of the ejection of electrons caused by collisions of excited atoms with ions, rather than neutrals, leading to the production of doubly charged ions. Doping superfluid He droplets with methyl iodide and exposing them to electrons enhances the formation of doubly charged iodine atoms at the threshold for the production of two metastable He atoms. These observations point toward a novel ionization process where doubly charged ions are produced by sequential Penning ionization. In some cases, depending on the neutral target, the process also leads to a subsequent Coulomb explosion of the dopant.     

Noisy optical pulses enhance the temporal resolution of pump-probe spectroscopy

Time-resolved measurements of quantum dynamics are based on the availability of controlled events that are shorter than the typical evolution time scale of the processes to be observed. Here we introduce the concept of noise-enhanced pump-probe spectroscopy, allowing the measurement of dynamics significantly shorter than the average pulse duration by exploiting randomly varying, partially coherent light fields consisting of bunched colored noise. These fields are shown to be superior by more than a factor of 10 to frequency-stabilized fields, with important implications for time-resolved experiments at x-ray free-electron lasers and, in general, for measurements at the frontiers of temporal resolution (e.g., attosecond spectroscopy). As an example application, the concept is used to explain the recent experimental observation of vibrational wave-packet motion in D(2)(+) on time scales shorter than the average pulse duration.     

Ordered structures of two sulfur containing donor molecules on the Au(111) surface

We report experiments by low energy electron diffraction (LEED) and scanning tunneling microscopy (STM) on ordered structures of two sulfur-containing π-conjugated molecules on the Au(111)-surface, namely tetrabenzothianthrene (TBTA) and tetrathiotetracene (TTT). These molecules are candidates for donors in charge transfer salts. On Au(111) both molecules form long-range ordered structures that are commensurate to the top-most surface layer. For TBTA, the reconstruction of the Au(111) surface is maintained, whereas it is lifted by TTT. Both molecules lie flat on the surface. For TBTA, the structure indicates that the molecule is planarized upon adsorption.     

Continuous-wave nonclassical light with gigahertz squeezing bandwidth

Squeezed states can be employed for entanglement-based continuous-variable quantum key distribution, where the secure key rate is proportional to the bandwidth of the squeezing. We produced a nonclassical cw laser field at the telecommunication wavelength of 1550 nm, which showed squeezing over a bandwidth of more than 2 GHz. The experimental setup used parametric downconversion via a periodically poled potassium titanyl phosphate crystal. We did not use any resonant enhancement for the fundamental wavelength, which should in principle allow a production of squeezed light over the full phase-matching bandwidth of several nanometers. We measured the squeezing to be up to 0.3 dB below the vacuum noise from 50 MHz to 2 GHz limited by the measuring bandwidth of the homodyne detector. The squeezing strength was possibly limited by thermal lensing inside the nonlinear crystal.     

Absolute, spatially resolved, in situ CO profiles in atmospheric laminar counter-flow diffusion flames using 2.3?μm TDLAS

We developed a new, spatially traversing, direct tunable diode laser absorption spectrometer (TDLAS) for quantitative, calibration-free, and spatially resolved in situ measurements of CO profiles in atmospheric, laminar, non-premixed CH₄/air model flames stabilized at a Tsuji counter-flow burner. The spectrometer employed a carefully characterized, room temperature distributed feedback diode laser to detect the R20 line of CO near 2,313?nm (4,324.4?cm^?1), which allows to minimize spectral CH₄ interference and detect CO even in very fuel-rich zones of the flame. The burner head was traversed through the 0.5?mm diameter laser beam in order to derive spatially resolved CO profiles in the only 60-mm wide CH₄/air flame. Our multiple Voigt line Levenberg?CMarquardt fitting algorithm and the use of highly efficient optical disturbance correction algorithms for treating transmission and background emission fluctuations as well as careful fringe interference suppression permitted to achieve a fractional optical resolution of up to 2.4?×?10^?4 OD (1??) in the flame (T up to 1,965?K). Highly accurate, spatially resolved, absolute gas temperature profiles, needed to compute mole fraction and correct for spectroscopic temperature dependencies, were determined with a spatial resolution of 65???m using ro-vibrational N₂-CARS (Coherent anti-Stokes Raman spectroscopy). With this setup we achieved temperature-dependent CO detection limits at the R20 line of 250?C2,000?ppmv at peak CO concentrations of up to 4?vol.%. This permitted local CO detection with signal to noise ratios of more than 77. The CO TDLAS spectrometer was then used to determine absolute, spatially resolved in situ CO concentrations in the Tsuji flame, investigate the strain dependence of the CO Profiles and favorably compare the results to a new flame-chemistry model.