Flow cytometric viability assessment and transmission electron microscopic morphological study of bacteria in glycerol.

Human cadaveric skin allografts are used in the treatment of burns and can be preserved in glycerol at high concentrations. Previously, glycerol has been attributed some antimicrobial effect. In an experimental set-up, we aimed at investigating this effect of prolonged incubation of bacteria in 85% glycerol. Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus subtilis were incubated in 85% glycerol. The influence of duration of incubation and temperature on ultrastructure and viability were investigated. Unstressed cultures served as controls. Survival was studied after 24-36 h and 10 days incubation in 85% glycerol at 4 degrees C and 36 degrees C with transmission electron microscopy (TEM) and flow cytometry using viability stains indicating membrane damage (SYTO9, propidium iodide) or esterase activity (carboxyfluorescein diacetate). TEM clearly demonstrated variability in morphological changes of bacteria suggesting different mechanisms of damage. Viability stains supported these findings with faster declining viable cell populations in 85% glycerol at 36 degrees C compared with 4 degrees C. Both methods demonstrated that Gram-negative species were more susceptible than Gram-positive species. In conclusion, 85% glycerol may have some additional antimicrobial effect. Temperature is an important factor herein and Gram-negatives are most susceptible. The latter finding probably reflects the difference in cell wall composition between Gram-positive and Gram-negative bacteria.     

New Approach to Determine the Phase Transition Temperature,Cloud Point,of Thermoresponsive Polymers

In this study, a novel method to determine the cloud point temperature variation in aqueous solutions of thermoresponsive homo- and copolymers was developed. Poly(N-vinylcaprolactam) (PVCL) and triblock copolymers of poly(t-butyl acrylate-co-acrylic acid)-b-poly(N-vinylcaprolactam)-b-(t-butyl acrylate-co-acrylic acid) (P[(tBA-co-AA)-b-PVCL-b-P(tBA-co-AA)] were synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization and used as models. The incorporation of AA units (hydrophilic segments) into the polymeric chain of PVCL influenced the phase transition, increasing the cloud point temperature of the final copolymer. The cloud point temperatures of the PVCL and the triblock copolymer P(tBA-co-AA)-b-PVCL-b-P(tBA-co-AA) were determined by measuring the transmittance of aqueous solutions of the polymers in a Turbiscan Lab instrument in the range of 29 to 40 C. This is the first study in which Turbiscan Lab is used to determine the cloud point temperature.     

Speckle decorrelation influence on measurements quality in vortex metrology

We study speckle decorrelation effects in connection with conventional vortex metrology techniques. Our proposal is based on processing speckled images recorded by using two different experimental set-ups. In both schemes two laterally displaced patterns are generated: one scheme allows for obtaining undecorrelated speckle distributions and the other for decorrelated ones. Vortex networks associated with speckle patterns are analyzed by employing the usual tools developed for vortex metrology. For each recorded image, a 2D pseudo-phase map is generated on the basis of the Reisz transform. Then the vortices are located, and parameterized in terms of their topological charge, eccentricity, vorticity and angles between the zero crossing lines from the real and the imaginary parts of the analytical signal. After tracking the homologous vortices onto the maps, the histograms corresponding to the coordinate displacements are analyzed. We show that histograms interpretation is prone to failure due to its high sensitivity to decorrelation. Experimental evidences are presented to support the restrictions imposed by decorrelation of actual speckles due to uniform in-plane displacements.     

Analysis of reflection speckle holograms in a BSO crystal

A theoretical and experimental study of speckle registering in photorefractive BSO crystals through reflection hologram geometry is implemented. The three-dimensional speckle nature is considered in the diffraction efficiency calculations. The interference process produces index-of-refraction gratings in the speckle volume via the photorefractive effect. It is demonstrated that the coupled-wave theory for reflection geometry allows explaining the diffraction efficiency behavior when the interaction length is properly taken into account. It means that the speckle depth is related with the interaction length. The speckle depth can be controlled by the imaging system pupil aperture diameter. Under this condition, the influence of the speckle depth on the diffraction efficiency is analyzed. An agreement between the numerical model based on the coupled-wave theory and the experimental results is obtained.     

Multiplexing encryption-decryption via lateral shifting of a random phase mask

We present an holographic memory optical arrangement based on the successive shifting of a random pure-phase mask to achieve encrypted images multiplexing. The input images are encrypted to a stationary white noise using the usual double random encoding in the Fresnel domain. The encrypted information is imaged in a photorefractive crystal where also a reference beam impinges. In the holographic memory, a BSO crystal is used to provide both a recording medium and a phase conjugate mirror. The combination of these two features supplies at the same time the necessary exact cancellation of the random pure-phase mask as well as allows a real-time decryption process. Successive images are encrypted and position-encoded by speckle patterns arising from the random pure-phase mask in-plane shifting between exposures. We include experimental results to corroborate the multiplexing capability and the read-out fidelity of the proposed arrangement.     

Dynamic behaviour of speckle cluster formation

In this work, we analyse the speckle cluster structure generated when a coherently illuminated diffuser is imaged by introducing a multiple aperture pupil mask in front of the lens plane. We demonstrate that the speckle cluster originates from the complex speckle modulation generated by multiple interferences among the wavefront passing through each aperture. The auto-correlation function of the intensity distribution when using a multiple aperture pupil arrangement is calculated. Besides, we demonstrate that the autocorrelation function and the intensity corresponding to a single scattering element of the input are coincident. This result allows interpretation of the dynamics behaviour of the speckle cluster formation by considering the result obtained by a single scattering element. Then, we determine the pupil mask geometrical parameters that control the cluster behaviour and therefore the condition for obtaining a highly repetitive cluster structure that we define as a 'regular cluster'. The theoretical simulations based on the random walk model are in agreement with the experimental results supporting the validity of our approach.     

Code retrieval via undercover multiplexing

The purpose of this research is to develop an undercover multiplexing technique to give additional protection for optical information encryption. We employ the double random phase mask as our basic optical encryption system. The holographic storage medium of choice is a photorefractive crystal. To achieve the multiplexing we use the aperture size of the pupil in the optical system, as it governs the speckle size. We introduce such variation in order to produce a decorrelation between two consecutively stored speckle patterns. Each stored speckle pattern is associated to an input encrypted image, thus producing a multiplexing of the encrypted information. We implement this operation without altering the setup architecture and the random phase masks. This multiplexing is our undercover operation to encipher a true code behind a fake code. Under this approach, the user can only recover the bulk information stored in the volume hologram. However, he cannot recover the true code without the additional information on the pupil size key, even if accessed in position of the original decoding mask.     

Sequences of infinite bifurcations and turbulence in a five-mode truncation of the Navier-Stokes equations

Two infinite sequences of orbits leading to turbulence in a five-mode truncation of the Navier-Stokes equations for a 2-dimensional incompressible fluid on a torus are studied in detail. Their compatibility with Feigenbaum's theory of universality in certain infinite sequences of bifurcations is verified and some considerations on their asymptotic behavior are inferred. An analysis of the Poincaré map is performed, showing how the turbulent behavior is approached gradually when, with increasing Reynolds number, no stable fixed point or periodic orbit is present and all the unstable ones become more and more unstable, in close analogy with the Lorenz model.