Mechanical Characterization of the Erythrocyte Membrane Using a Capacitor-Based Technique.

Pathological processes often change the mechanical properties of cells. Increased rigidity could be a marker of cellular malfunction. Erythrocytes are a type of cell that deforms to squeeze through tiny capillaries; changes in their rigidity can dramatically affect their functionality. Furthermore, differences in the homeostatic elasticity of the cell can be used as a tool for diagnosis and even for choosing the adequate treatment for some illnesses. More accurate types of equipment needed to study biomechanical phenomena at the single-cell level are very costly and thus out of reach for many laboratories around the world. This study presents a simple and low-cost technique to study the rigidity of red blood cells (RBCs) through the application of electric fields in a hand-made microfluidic chamber that uses a capacitor principle. As RBCs are deformed with the application of voltage, cells are observed under a light microscope. From mechanical force vs. deformation data, the elastic constant of the cells is determined. The results obtained with the capacitor-based method were compared with those obtained using optical tweezers, finding good agreement. In addition, P. falciparum-infected erythrocytes were tested with the electric field applicator. Our technique provides a simple means of testing the mechanical properties of individual cells.

Trapping and manipulation of bubbles with holographic optical tweezers.

A methodology to manipulate bubbles and measure adhesion forces is presented and validated. Holographic optical tweezers are employed to establish a circular array of high intensity points to effectively trap a gas bubble within a liquid medium. This approach includes an efficient calibration protocol based on a theoretical framework for the calculation of optical forces using a ray tracing algorithm, which allows enhancing the versatility of optical manipulation to micro-objects with a lower refractive index than the surrounding medium. As an initial application, the adhesion force between two stable bubbles at different sizes is measured, finding a minimum when they have the same diameter.

Activation energy, spatial confinement, and mean first passage and escape times of a tracer in a wormlike micellar fluid: an effective potential approach.

Wormlike micelles are long semiflexible cylindrical polymer structures formed by amphiphiles. In solution, these linear micelles percolate in multiconected entangled networks, where cross-links can break and recombine dynamically. Technological applications of wormlike micellar fluids include tunable encapsulation/delivery of molecules or colloids in biomedicine, oil industry, and/or cleaning processes. In this work, we propose that the experimental activation energy, the spatial confinement, and the mean first passage and escape times of a spherical tracer immersed in wormlike micellar network, in which caging effects are observed, can be estimated from economic Brownian dynamics simulations of a single particle interacting with an effective one-dimensional cosine-like potential of amplitudeU0and periodicityL. The proposed one-fitting parameter method has been used to characterize the long-time dynamics of wormlike micellar solutions formed by the self-assembly of a mixture of zwitterionic and anionic surfactants at several temperatures and different concentrations of surfactant and brine. The amplitudeU0has displayed a good agreement regarding the corresponding experimental activation energy at different temperatures. The periodicityLhas shown to be an upper bound of the mesh sizeξand of the same order of magnitude regarding the entanglement lengthle, obtained from rheology and microrheology experiments. The escape time of the tracer in the effective potentialτescapeand the timet*, at which a change of curvature in the mean square displacement occurs, are upper and lower limits, respectively, of the experimental relaxation time. Our method is simple and fast, and we foresee that it should be applicable to model the long-time behaviour of tracers in other polymer systems, in which caging effects are present.

Fluorescent organic particle doped polymer-based gel dosimeter for neutron detection.

The purpose of this work is to develop a material capable of detecting neutrons produced by photodisintegration in a linear accelerator for its medical use. In this study, we have developed a gel-like material doped with fluorescent organic particles. PPO at 1 wt% is used as primary dopant and POPOP as secondary one at 0.03 wt%. A set of four samples is produced, with boric acid concentrations of 0, 400, 800 and 1200 ppm. The viscoelastic properties of the material are characterized with rheological measurements, finding a gel-like behavior, i.e., a material that can keep its original shape if no stresses are applied, but can also be deformed by applying a moderate shear rate. Furthermore, the material was irradiated with gamma, electron, and neutron emission sources from 137Cs, 22Na, 60Co, 210Po, 90Sr and 241AmBe, and its response was measured in two different experimental settings, in two different institutions, for comparative purposes. From these measurements, one can clearly establish that the new material detects neutrons, electrons, and gammas within the MeV regions and below. Thus, our findings show that the developed material and its properties make it a promising technology for its use in a neutron detector.

Two-dimensional Brownian motion of anisotropic dimers.

We study the two-dimensional motion of colloidal dimers by single-particle tracking and compare the experimental observations obtained by bright-field microscopy to theoretical predictions for anisotropic diffusion. The comparison is based on the mean-square displacements in the laboratory and particle frame as well as generalizations of the self-intermediate scattering functions, which provide insights into the rotational dynamics of the dimer. The diffusional anisotropy leads to a measurable translational-rotational coupling that becomes most prominent by aligning the coordinate system with the initial orientation of the particles. In particular, we find a splitting of the time-dependent diffusion coefficients parallel and perpendicular to the long axis of the dimer which decays over the orientational relaxation time. Deviations of the self-intermediate scattering functions from pure exponential relaxation are small but can be resolved experimentally. The theoretical predictions and experimental results agree quantitatively.

A few-layer graphene/chlorin e6 hybrid nanomaterial and its application in photodynamic therapy against Candida albicans.

The global emergence of multidrug resistance of fungal infections and the decline in the discovery of new antibiotics are increasingly prevalent causes of hospital-acquired infections, among other major challenges in the global health care sector. There is an urgent need to develop noninvasive, nontoxic, and new antinosocomial approaches that work more effectively and faster than current antibiotics. In this work, we report on a biocompatible hybrid nanomaterial composed of few-layer graphene and chlorin e6 (FLG-Ce6) for the photodynamic treatment (PDT) of Candida albicans. We show that the FLG-Ce6 hybrid nanomaterial displays enhanced reactive oxygen species (ROS) generation compared with Ce6. The enhancement is up to 5-fold when irradiated for 15 min at 632 nm with a red light-emitting diode (LED). The viability of C. albicans in the presence of FLG-Ce6 was measured 48 h after photoactivation. An antifungal effect was observed only when the culture/FLG-Ce6 hybrid was exposed to the light source. C. albicans is rendered completely unviable after exposure to ROS generated by the excited FLG-Ce6 hybrid nanomaterial. An increased PDT effect was observed with the FLG-Ce6 hybrid nanomaterial by a significant reduction in the viability of C. albicans, by up to 95%. This is a marked improvement compared to Ce6 without FLG, which reduces the viability of C. albicans to only 10%. The antifungal action of the hybrid nanomaterial can be activated by a synergistic mechanism of energy transfer of the absorbed light from Ce6 to FLG. The novel FLG-Ce6 hybrid nanomaterial in combination with the red LED light irradiation can be used in the development of a wide range of antinosocomial devices and coatings.

Retrieval of Absorption or Scattering Coefficient Spectrum (RASCS) Program: A Tool to Monitor Optical Properties in Real Time.

BACKGROUND AND OBJECTIVES: Optical properties characterize light propagation in turbid media, such as tissue. Recovery of optical properties is of great importance in a wide variety of biomedical applications, including both therapeutic treatments and diagnosis. Most of the available methodologies are well established, however, these are not optimized for real-time measurements. STUDY DESIGN/MATERIALS AND METHODS: Optical properties are recovered using the Inverse Adding Doubling program from reflectance measurements measured with an integrating sphere and light in the visible range. A user-friendly interface was programmed in Visual Studio and the libraries of a particular spectrophotometer were used. To achieve real-time measurements, a parallel computing routine was implemented, splitting the whole spectra in threads to be computed independently. Several tests using living tissue and inorganic materials were carried out to validate the proposed algorithm. RESULTS: Recovery of absorption/scattering coefficient spectrum in the visible range with high precision in a couple of seconds was achieved, demonstrating its capabilities for real-time monitoring in biomedical applications. The absorption coefficient spectrum shows the expected characteristics according to the different melanin and blood concentration of various volunteers, also showing the expected changes during a thermoregulation process. CONCLUSIONS: A real-time monitoring of optical properties algorithm was developed, including parallel computing and a user-friendly interface. The proposed algorithm would be of help in biomedical applications, where real-time monitoring optical properties is required. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

Finding trap stiffness of optical tweezers using digital filters.

Obtaining trap stiffness and calibration of the position detection system is the basis of a force measurement using optical tweezers. Both calibration quantities can be calculated using several experimental methods available in the literature. In most cases, stiffness determination and detection system calibration are performed separately, often requiring procedures in very different conditions, and thus confidence of calibration methods is not assured due to possible changes in the environment. In this work, a new method to simultaneously obtain both the detection system calibration and trap stiffness is presented. The method is based on the calculation of the power spectral density of positions through digital filters to obtain the harmonic contributions of the position signal. This method has the advantage of calculating both trap stiffness and photodetector calibration factor from the same dataset in situ. It also provides a direct method to avoid unwanted frequencies that could greatly affect calibration procedure, such as electric noise, for example.

Short-time dynamics of monomers and dimers in quasi-two-dimensional colloidal mixtures.

We report on the short-time dynamics in colloidal mixtures made up of monomers and dimers highly confined between two glass plates. At low concentrations, the experimental measurements of colloidal motion agree well with the solution of the Navier-Stokes equation at low Reynolds numbers; the latter takes into account the increase in the drag force on a colloidal particle due to wall-particle hydrodynamic forces. More importantly, we find that the ratio of the short-time diffusion coefficient of the monomer and that of the center of mass of the dimmer is almost independent of both the dimer molar fraction, x_{d}, and the total packing fraction, ϕ, up to ϕ≈0.5. At higher concentrations, this ratio displays a small but systematic increase. A similar physical scenario is observed for the ratio between the parallel and the perpendicular components of the short-time diffusion coefficients of the dimer. This dynamical behavior is corroborated by means of molecular dynamics computer simulations that include explicitly the particle-particle hydrodynamic forces induced by the solvent. Our results suggest that the effects of colloid-colloid hydrodynamic interactions on the short-time diffusion coefficients are almost identical and factorable in both species.

Absorption effects in diffusing wave spectroscopy.

The effect of absorption in diffusing wave spectroscopy (DWS) was studied using an absorption-dependent diffusive equation for describing the light propagation within a turbid liquid where dielectric microspheres have been embedded. Here, we propose an expression for the time-averaged light intensity autocorrelation function that correctly describes the time fluctuations for the scattered light, in the regime where the diffusion approximation accurately describes the light propagation. This correction was suspected previously, but it was not formally derived from a light diffusive equation. As in the case of no absorption, we obtained that time fluctuations of the scattered light can be related to the mean square displacement of the embedded particles. However, if a correction for absorption is not taken into account, the colloidal dynamics can be misinterpreted. Experimental results show that this new formulation correctly describes the time fluctuations of scattered light. This new procedure extends the applicability of DWS, and it opens the possibility of doing microrheology with this optical method in systems where absorption cannot be avoided.

Mean-square displacement of particles in slightly interconnected polymer networks.

Structural and viscoelastic properties of slightly interconnected polymer networks immersed in a solvent have been studied in two cases: when the polymer network is building up and when the polymer network is shrinking stepwise in a controlled way. To accomplish this goal, the mean square displacement (MSD) of embedded microspheres in the polymer network was measured as a function of time, with diffusive wave spectroscopy. Particle motion was analyzed in terms of a model, based on a Fokker-Planck type equation, developed for describing particles in Brownian motion within a network that constrain their movement. The model reproduces well the experimental features observed in the MSD vs t curves. The variation of the parameters describing the structure of the network can be understood as the polymerization comes about, and also after the successive volume contractions. In addition, from the MSD curves, the complex shear moduli were obtained in a wide range of frequencies when the network is building up, and at the different shrinking states of the network. Our microrheological results give an insight about the dynamics of embedded particles in slightly interconnected networks, which were also compared with similar results for polymers without interconnections and polymer gels.

Microrheology and characteristic lengths in wormlike micelles made of a zwitterionic surfactant and SDS in brine.

We study the Brownian motion of probe particles embedded in a wormlike micellar fluid made of a zwitterionic surfactant N-tetradecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (TDPS), sodium dodecyl sulfate (SDS), and salty water to get structural and dynamical information of the micellar network. The motion of the probe particles was tracked with diffusing wave spectroscopy, and the mean square displacement as a function of time for the particles was obtained. This allowed us to obtain the long-time diffusion coefficient for microspheres moving in the micellar network and the cage size where each particle is harmonically bound at short times in that network. The bulk mechanical susceptibility of the fluid determines the response of the probe particles excited by the thermal stochastic forces. As a consequence, the mean square displacement curves allowed us to calculate the elastic (storage) and the viscous (loss) moduli as a function of the frequency. From these curves, spanning a wide frequency range, we estimated the characteristic lengths as the mesh size, the entanglement length, the persistence length, and the contour length for micellar solutions of different zwitterionic surfactant concentration, surfactant ratio ([SDS]/[TDPS]), salt concentration, and temperature. Mesh size, entanglement length, and persistence length are almost insensitive to the change of these variables. In contrast, the contour length changes in an important way. The contour length becomes shorter as the temperature increases, and it presents a peak at a surfactant ratio of ∼0.50-0.55. When salt is added to the solution, the contour length presents a peak at a salt concentration of ∼0.225 M, and in some solutions, this length can reach values of ∼12 µm. Scission energies help us to understand why the contour length first increases and then decreases when salt is added.

A rheological study in the dilute regime of the worm-micelle fluid made of zwitterionic surfactant (TDPS), anionic surfactant (SDS), and brine.

We present a rheological study for a system made of zwitterionic surfactant N-tetradecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (TDPS), sodium dodecyl sulfate (SDS), and water (0.5M NaCl). We found that this system forms wormlike micelles. This study is focused in the dilute regime below the overlap concentration, where micelles are not entangled. The overlap concentration was determined using dynamic light scattering. The behavior of the apparent viscosity and the shear stress, both as a function of the shear rate, was determined for different zwitterionic surfactant concentrations, temperatures, and two surfactant ratios (R=[SDS]/[TDPS]). The shear-thickening transition and its temperature dependence was also studied. Finally, we were able to observe the shear-induced structures by using the scattered light from a sheet of light perpendicular to the flow that is installed in the gap of a transparent Couette cell filled with the micellar fluid.