Cell-selective intracellular targeting is a key element of more specific and safe enzyme, toxin, and gene therapies. Endothelium poorly internalizes certain candidate carriers for vascular immunotargeting, such as antibodies to platelet endothelial cell adhesion molecule 1 (PECAM-1). Conjugation of poorly internalizable antibodies with streptavidin (SA) facilitates the intracellular uptake. Although both small and large (100-nm versus 1000-nm diameter) anti-PECAM/SA-beta galactosidase (SA-beta-gal) conjugates bound selectively to PECAM-expressing cells, only small conjugates showed intracellular accumulation of active P-gal. To study whether size of the conjugates controls the uptake, a series of anti-PECAM/SA and anti-PECAM/bead conjugates ranging from 80 nm to 5 mum in diameter were produced. Human umbilical vein endothelial cells and PECAM-transfected mesothelioma cells internalized 80- to 350-nm anti-PECAM conjugates, but not conjugates larger than 500 nm. Further, size controls intracellular targeting of active therapeutic cargoes in vitro and in vivo. Small anti-PECAM/DNA conjugates transfected target cells in culture 5-fold more effectively than their large counterpart (350- versus 4200-nm diameter). To evaluate the practical significance of the size-controlled subcellular addressing, we coupled glucose oxidase (GOX) to anti-PECAM and antithrombomodulin. Both types of conjugates had equally high pulmonary uptake after intravenous injection in mice, yet only small (200- to 250-nm), not large (600- to 700-nm), GOX conjugates caused profound oxidative vascular injury in the lungs, presumably owing to intracellular generation of H2O2. Thus, engineering of affinity carriers of specific size permits intracellular delivery of active cargoes to endothelium in vitro and in vivo, a paradigm useful for the targeting of drugs, genes, and toxins. (C) 2002 by The American Society of Hematology.
We use confocal microscopy to study particle motion in colloidal systems. Near the glass transition, motion is inhibited, as particles spend time trapped in transient "cages" formed by neighboring particles. We measure the cage sizes and lifetimes, which, respectively, shrink and grow as the glass transition approaches. Cage rearrangements are more prevalent in regions with lower concentrations and higher disorder. Neighboring rearranging particles typically move in parallel directions, although a nontrivial fraction moves in antiparallel directions, usually from particle pairs with initial separations corresponding to local maxima and minima of the pair correlation function g(r), respectively.
The dynamics of a glass-forming material slow greatly near the glass transition, and molecular motion becomes inhibited. We use confocal microscopy to investigate the motion of colloidal particles near the colloidal glass transition. As the concentration in a dense colloidal suspension is increased, particles become confined in transient cages formed by their neighbors. This prevents them from diffusing freely throughout the sample. We quantify the properties of these cages by measuring temporal anticorrelations of the particles' displacements. The local cage properties are related to the subdiffusive rise of the mean square displacement: over a broad range of time scales, the mean square displacement grows slower than linearly in time. (C) 2002 Elsevier Science B.V. All rights reserved.
We have combined the. drug release and delivery potential of nanoparticle (NP) systems with the ease of flow, processing, and aerosolization potential of large porous particle (LPP) systems by spray drying solutions of polymeric and nonpolymeric NPs into extremely thin-walled macroscale structures. These hybrid LPPs exhibit much better flow and aerosolization properties than the NPs; yet, unlike the LPPs, which dissolve in physiological conditions to produce molecular constituents, the hybrid LPPs dissolve to produce NPs, with the drug release and delivery advantages associated with NP delivery systems. Formation of the large porous NP (LPNP) aggregates occurs via a spray-drying process that ensures the drying time of the sprayed droplet is sufficiently shorter than the characteristic time for redistribution of NPs by diffusion within the drying droplet, implying a local Peclet number much greater than unity. Additional control over LPNPs physical characteristics is achieved by adding other components to the spray-dried solutions, including sugars, lipids, polymers, and proteins. The ability to produce LPNPs appears to be largely independent of molecular component type as well as the size or chemical nature of the NPs.
Velocity fluctuations in sedimentation are studied to investigate the origin of a hypothesized universal scale [P. N. Segre, E. Herbolzheimer, and P. M. Chaikin, Phys. Rev. Lett. 79, 2574 (1997)]. Our experiments show that fluctuations decay continuously in time for sufficiently thick cells, never reaching steady state. Simulations and scaling arguments suggest that the decay arises from increasing vertical stratification of particle concentration due to spreading of the sediment front. The results suggest that the velocity fluctuations in sedimentation depend sensitively on cell geometry.
The rheological properties of cholesteric liquid crystals containing networks of defects are investigated. A network of linear defects of the "oily-streak" type is stabilized when colloidal particles are dispersed into the cholesteric liquid crystals. This network converts the rheological response of a presheared cholesteric liquid crystal from fluidlike to solidlike and leads to the formation of a "defect-mediated" solid. The frequency-dependent complex shear modulus G(*)(omega) is measured, for samples with and without inclusions, in both the linear and nonlinear viscoelastic regimes. The linear elastic response mediated by the defect network is discussed in terms of a model analogous to the theories of rubber elasticity. All our data for G(*)(omega) are fitted to a simplified theoretical form, and the values and variations of the fitting parameters, in the various regimes investigated, are discussed in terms of the properties of defect structure present in the samples. Similar rheological properties are expected to arise from particle-stabilized oily-streak defect networks in layered systems such as smectic-A and lyotropic L(alpha) phases.
Optical imaging of ex vivo tissue models to study heart fibrillation is normally performed using voltage-sensitive dyes. Upon stimulation by an electrode, time-dependent fluorescence or absorption signals are recorded, often in trans-illumination geometry. In order to provide quantification of the origins of these signals inside the tissue, the locally varying optical properties of the tissue have to be known and their change due to the presence of the dyes. To provide experimental input for further modeling efforts, we have performed depth dependent measurements with a fiber optic laser source inside the tissue, recording light profiles on the tissue surface, mainly in transmission geometry. From these measurements, optical properties have been extracted and the obtained profiles have been used as input into a preliminary image reconstruction scheme, together with Monte Carlo simulations. Experiments at different locations in the same sample show the variation of optical properties. Additionally, effects from the presence of heterogeneities on the signal have been investigated.
Times Cited: 0 Conference on Optical Biopsy IV Jan 21-23, 2002 San jose, ca Spie
Nanometre- and micrometre-sized charged particles at aqueous interfaces are typically stabilized by a repulsive Coulomb interaction. If one of the phases forming the interface is a nonpolar substance ( such as air or oil) that cannot sustain a charge, the particles will exhibit long-ranged dipolar repulsion(1); if the interface area is confined, mutual repulsion between the particles can induce ordering(2) and even crystallization(3,4). However, particle ordering has also been observed in the absence of area confinement(5), suggesting that like-charged particles at interfaces can also experience attractive interactions(6). Interface deformations are known to cause capillary forces that attract neighbouring particles to each other, but a satisfying explanation for the origin of such distortions remains outstanding(7,8). Here we present quantitative measurements of attractive interactions between colloidal particles at an oil - water interface and show that the attraction can be explained by capillary forces that arise from a distortion of the interface shape that is due to electrostatic stresses caused by the particles' dipolar field. This explanation, which is consistent with all reports on interfacial particle ordering so far, also suggests that the attractive interactions might be controllable: by tuning the polarity of one of the interfacial fluids, it should be possible to adjust the electrostaticstresses of the system and hence the interparticle attractions.
We have incorporated nematic liquid crystal into periodic, polymer host structures templated from self-assembled colloids. Using these composite materials, we demonstrate the first electrically switchable three-dimensional Bragg diffraction. The switchable beam deflection is potentially useful for non-mechanical beam steering and optical beam splitting devices. We compare the electro-optic response of our templated liquid-crystal/polymer composites with conventional polymer-dispersed liquid crystals (PDLCs). Our data reveal a qualitatively different and faster response for liquid crystal distributed within a connected cavity network, as compared to isolated liquid-crystal droplets within a polymer matrix.
We report optical switching studies on nematic liquid crystal incorporated into structures based on self-assembled colloids. We compare the electro-optic responses of liquid crystal imbibed into colloid-templated polymers, liquid crystal imbibed in the interstitial space of colloid crystals, and conventional polymer-dispersed liquid crystals. We characterize the Bragg diffraction of our templated liquid-crystal/polymer composites as a function of electric field and measure switching times. The response of liquid crystal in connected networks differs qualitatively from that of liquid crystal in isolated cavities.
We present an approach to fabricate solid capsules with precise control of size, permeability, mechanical strength, and compatibility. The capsules are fabricated by the self-assembly of colloidal particles onto the interface of emulsion droplets. After the particles are locked together to form elastic shells, the emulsion droplets are transferred to a fresh continuous-phase fluid that is the same as that inside the droplets. The resultant structures, which we call "colloidosomes", are hollow, elastic shells whose permeability and elasticity can be precisely controlled. The generality and robustness of these structures and their potential for cellular immunoisolation are demonstrated by the use of a variety of solvents, particles, and contents.
We present novel measurements of the structure of colloidal gels. Using confocal microscopy, we obtain the precise three-dimensional positions of a large number of particles. We develop quantitative descriptions of the topology of the gel, including the number of bonds per particle, the chemical or bond fractal dimension, the number of flexible pivot points and other topological parameters that describe the chainlike structure. We investigate the dependence of these parameters on the particle volume fraction and the strength of the attraction that holds the particles together. While all samples have approximately the same fractal and chemical dimensions, we find that gels formed with stronger attraction or larger volume fraction have fewer bonds per particle, more filamentous chains and a greater number of flexible pivot points. Finally, we discuss the topological results in the context of the gel's elasticity. Measurements of the elastic constants of individual chainlike segments are explained with a simple model. The distribution of elastic constants, however, has a general form that is not understood.
We have developed a technique in ultrasonic correlation spectroscopy called diffusing acoustic wave spectroscopy (DAWS). In this technique, the motion of the scatterers (e.g., particles or inclusions) is determined from the temporal fluctuations of multiply scattered sound. In DAWS, the propagation of multiply scattered sound is described using the diffusion approximation, which allows the autocorrelation function of the temporal field fluctuations to be related to the dynamics of the multiply scattering medium. The expressions relating the temporal field autocorrelation function to the motion of the scatterers are derived, focusing on the types of correlated motions that are most likely to be encountered in acoustic measurements. The power of this technique is illustrated with ultrasonic data on fluidized suspensions of particles, where DAWS provides a sensitive measure of the local relative velocity and strain rate of the suspended particles over a wide range of time and length scales. In addition, when combined with the measurements of the rms velocity of the particles using dynamic sound scattering, we show that DAWS can be used to determine the spatial extent of the correlations in the particle velocities, thus indirectly measuring the particle velocity correlation function. Potential applications of diffusing acoustic wave spectroscopy are quite far reaching, ranging from the ultrasonic nondestructive evaluation of the dynamics of inhomogeneous materials to geophysical studies of mesoscopic phenomena in seismology.
Experiments investigating the local viscoelastic properties of a simple uncross-linked flexible polymer are performed on polyethylene oxide solutions in the semidilute regime using polystyrene beads of varying sizes and surface chemistry as probes. We measure the thermal motions of the beads to obtain the elastic and viscous moduli of our sample. Two different dynamic light scattering techniques, diffusing wave spectroscopy and quasielastic light scattering (QELS), are used to determine the dynamics of the probe particles. Diffusing wave spectroscopy probes the short time dynamics of the scatterers while QELS or single scattering measures the dynamics at larger times. This results in a larger frequency overlap of the data obtained from the microrheological techniques with the data obtained from the conventional bulk measurements. The moduli are estimated using a modified algebraic form of the generalized Stokes-Einstein equation. Comparison of microrheology with,bulk measurements shows excellent similarity confirming the applicability of this method for simple, uncross-linked polymeric systems.
Using particle tracking routines the location of single point light sources can be determined with an accuracy of a few nanometers. By using quantum dots (QDs) emitting at different wavelengths, the measurement of the distance between these point light sources, which are closer than the optical resolution of 200 nm, was achieved. The nanocrystals have major advantages over conventional chromophores in higher quantum yield, more photostability, and the possibility of different emission wavelengths by an excitation with a single wavelength. The colocalization of two single QDs at the least distance of 40 nm can be measured with a standard deviation of 5 nm and a time resolution of 117 ms using one excitation wavelength.