Separation and detection of individual submicron particles by capillary electrophoresis with laser-light-scattering detection

Separation and detection of individual submicron polystyrene spheres using capillary electrophoresis with laser-light-scattering detection has been demonstrated. Electrophoretically separated particles were passed through a focused laser beam and light scattered from individual particles was collected at 90 degrees. Each diameter of polystyrene spheres injected (from 110 to 992 nm) resulted in the observation of a well-defined migration window containing multiple peaks, each arising from the light scattered by an individual particle. The migration time window for individual particles of a particular size corresponded well to the migration time of a peak from a population of particles of the same size detected using a UV absorbance detector. The electrophoretic mobility and scattered light intensity were determined for each particle detected. The average scattered light intensity for each particle size was consistent with Mie scattering theory. Particles as small as 110 nm in diameter were detected individually using this method, but particles with a diameter of 57 nm could not be individually detected. The number of single particle scattering events was counted and compared to the theoretical number of particles injected electrokinetically, and the detection efficiency determined ranged from 38 to 57% for polystyrene spheres of different sizes. The laser-light-scattering detection method was directly compared to laser-induced fluorescence detection using fluorescent polystyrene microspheres. The number of particles detected individually by each method was in agreement.     

Nanoparticle immunoagglutination Rayleigh scatter assay to complement microparticle immunoagglutination Mie scatter assay in a microfluidic device

In this work, particle immunoagglutination assays for pathogen detection, utilizing light scattering measurements at a fixed angle from incident light delivery, are explored in both Rayleigh and Mie scatter regimes through scatter intensity simulations and compared to experimental results. The average size of immunoagglutinated particles obtained from microscope images correspond to the particle size parameter from simulations. Mie scatter measurements yield a greater signal increase with increasing pathogen concentration than Rayleigh scatter measurements, but with a non-monotonic relationship that is not observed in the Rayleigh scatter regime. These two similar yet distinctly different sources of information could easily be integrated into a single device through fabrication of a simple microfluidic device containing two y-channels, each for performing the respective light scattering measurement. Escherichia coli was used as a representative target, and detected in a microfluidic device down to a concentration of 1 colony forming units (CFU) per mL.     

On the Dependence of the Light Scattering Intensity on the Averaged Size of Polydisperse Particles: Comments on the Paper by M.S. Dyuzheva et al. (Colloid J., 2002, vol. 64, no. 1, p. 39)

The second stage in the solution of the inverse problem of dynamic light scattering is analyzed; the dependence of the static scattering intensity on the average fraction size is used to calculate the particle size distribution at this stage. The results of the paper by M.S. Dyuzheva et al. (Colloid J., 2002, vol. 64, no. 1, p. 39) concerning this problem are shown to be erroneous. On the basis of calculations within the framework of the Mie theory, the possibility of a power approximation for the particle size dependence of the scattered intensity and the effect of the particle parameters on the power approximation exponents are discussed.     

The imaging ammeter

A method for measuring local current density, not requiring segmentation of the electrode or spatial scanning, is presented. The motion of colloidal particles in response to local current density, characterized by the intensity of the light they scatter, is the fundamental phenomenon of the technique. The scattering was produced and measured with the electrochemical total internal reflection microscope, a tool that places an electrochemical cell within a total internal reflection apparatus. The electrolysis of water and the oxidation of ferrocene monocarboxylic acid were used as test reactions. Light scattered by a probe particle produced an "image" of current density; scattered light was converted to local current density by a function derived herein. Numerical simulations supplemented experimental evidence that local current density controlled the probe particle's vertical motion. The spatial resolution of the method was approximately the length scale of the probe particle, in this case 5.7 μm. The resolution of current density was better than 100 nA cm(-2). The method might find use in high throughput screening of electrocatalysts.     

Differential Light Scattering Spectroscopy for Studying Biospecific Assembling of Gold Nanoparticles with Protein or Oligonucleotide Probes

A new method is proposed for studying biospecific interactions in systems of conjugates of colloidal gold nanoparticles. The method is based on measuring the differential spectra of light scattered at 90° within wavelength range 350–800 nm. Addition of complementary component to bioconjugate probe results in the nanoparticle aggregation that can be monitored by the light scattering or extinction spectra. To this end, we developed a special attachment to a Specord M-40 spectrophotometer and a corresponding measurement procedure called by us the differential light scattering spectroscopy. The method is compared with the common spectrophotometry as applied to colloidal gold conjugates to various polymers including proteins and oligonucleotides. Our experiments with the gold nanoparticles of different sizes showed a higher potential sensitivity of the suggested method as compared to spectrophotometry. It is expected that the differential light scattering spectroscopy can be used to develop sensitive analytical biospecific test for various biopolymers.     

Angular-ratiometric plasmon-resonance based light scattering for bioaffinity sensing

We describe an exciting opportunity for affinity biosensing using a ratiometric approach to the angular-dependent light scattering from bioactivated and subsequently aggregated noble metal colloids. This new model sensing platform utilizes the changes in particle scattering from very small colloids, which scatter light according to traditional Rayleigh theory, as compared to the changes in scattering observed by much larger colloidal aggregates, formed due to a bioaffinity reaction. These larger aggregates no longer scatter incident light in a Cos(2) theta dependence, as is the case for Rayleigh scattering, but instead scatter light in an increased forward direction as compared to the incident geometry. By subsequently taking the ratio of the scattered intensity at two angles, namely 90 degrees and 140 degrees , relative to the incident light, we can follow the association of biotinylated bovine serum albumin-coated 20 nm gold colloids, cross-linked by additions of streptavidin. This new model system can be potentially applied to many other nanoparticle assays and has many advantages over traditional fluorescence sensing and indeed light-scattering approaches. For example, a single nanoparticle can have the equivalent scattered intensity as 10(5) fluorescing fluorescein molecules substantially increasing detection; the angular distribution of scattered light from noble metal colloids is substantially easier to predict as compared to fluorescence; the scattered light is not quenched by biospecies; the ratiometric measurements described here are not dependent on colloid concentration as are other scattering techniques; and finally, the noble metal colloids are not prone to photodestruction, as is the case with organic fluorophores.     

Small-angle scattering of polarized light. II. experiments with isotropic spheres

New experimental measurements are reported of small-angle polarized and depolarized light scattering from almost monodisperse isotropic, spherical particle, polystyrene latexes. The shape and intensity distribution of the scattering patterns is shown to compare closely with the calculated patterns based on the exact Mie scattering theory from a single sphere.     

The optical properties of nanodisperse silica in hydrothermal solutions

The optical properties of aqueous solutions of colloidal silica were studied by photon correlation spectroscopy and spectrophotometry. Photon correlation spectroscopy measurements were taken at a 633 nm wavelength of monochromatic laser radiation. Autocorrelation function and scattered light amplitude plots were constructed, and the mean radii and diffusion coefficients of particles were determined. Spectrophotometric measurements were taken over the electromagnetic radiation wavelength range 200–1000 nm. The positions of optical density maxima were determined. The influence of various factors, including concentration, the size of nanoparticles, temperature, and pH, on the optical properties of aqueous solutions of silica was estimated.     

Analysis of surface structure and hydrogen/deuterium exchange of colloidal silica suspension by contrast-variation small-angle neutron scattering

The microscopic surface structure and hydrogen/deuterium exchange effect were investigated by contrast-variation small-angle neutron scattering (CV-SANS) for three different-sized amorphous colloidal silica aqueous suspensions. The results show that the fraction of hydrogen/deuterium exchange per nanoparticle, phiH/D, strongly depends on the size of silica nanoparticles. This finding supports that the hydrogen/deuterium exchange occurs exclusively within a finite surface layer of silica nanoparticles, while the inner component remained unchanged. Detailed analyses of the scattering intensity functions led to the estimation of (1) phiH/D and (2) the thickness of the surface layer as functions of the particle radius. The surface layer thickness was found to increase from 18 to 35 A with decreasing the particle radius from 165 to 71.2 A. The surface area per unit weight of silica estimated with the CV-SANS results are comparable to those reported in the literature.     

Energy transfer and amplified spontaneous emission in temperature-controlled random scattering media

Hydroxypropyl cellulose (HPC) in aqueous solution forms nanoparticles and becomes highly scattering above its lower critical solution temperature (LCST approximately 41 degrees C). Enhancement of energy transfer (ET) and amplified spontaneous emission (ASE) have been observed in turbid HPC solutions containing Rhodamine 6G (RG) as an energy donor and Kiton Red 620 (KR) as an acceptor. A detailed analysis of self-absorption, absorption saturation, and multiple scattering effects has revealed the importance of photon diffusion in shutting down the intensity leakage. A 5-fold enhancement of ET in the turbid condition is estimated. Possible factors crucial for ET and ASE in random media are discussed, such as the donor-to-acceptor ASE energy pumping, the optical path elongation by multiple scattering, and the formation of "light pipes" in the near-field of the Mie scattering. The temperature-dependent colloidal formation is found to successfully control optical processes via multiple scattering with a sharp threshold and abrupt emergence of dense scatterers.     

Experimental verification of an exact evanescent light scattering model for TIRM

Total internal reflection microscopy (TIRM) is a method for the precise measurement of interaction potentials between a spherical colloidal particle and a wall. The method is based on single-particle evanescent wave light scattering. The well-established model used to interpret TIRM data is based on an exponential relation between scattering intensity and particle wall distance. However, applying this model for a certain range of experimental parameters leads to significant distortions of the measured potentials. Using a TIRM setup based on a two-wavelength illumination technique, we were able to directly measure the intensity distance relation revealing deviations from an exponential decay. The intensity-distance relations could be compared to scattering simulations taking into account exact experimental parameters and multiple reflections between a particle and the wall. Converging simulation results were independently obtained by the T-matrix method and the discrete sources method (DSM) and show excellent agreement with experiments. Using the new scattering model for data evaluation, we could reconstruct the correct potential shape for distorted interaction potentials as we demonstrate. The comparison of simulations to experiment intrinsically yields a new method to determine absolute particle-wall distances, a highly desired quantity in TIRM experiments.     

Atomic force microscopy colloid-probe measurements with explicit measurement of particle-solid separation

We describe the use of evanescent wave scattering to measure the separation between the surface of a solid and a particle that is attached to an atomic force microscope (AFM) cantilever. Termed evanescent wave atomic force microscopy, our approach involves measuring the intensity of the light scattered from an evanescent field formed by the total internal reflection of a laser beam at a solid/fluid interface. In a conventional AFM "colloid probe" measurement, this separation must be inferred from an examination of the surface forces. Direct measurement of this separation with an evanescent wave atomic force microscope (EW-AFM) removes some ambiguity in the surface force measurement and, in addition, allows new types of measurements. For example, the force can be monitored at a constant separation. Our evanescent scattering apparatus is essentially identical to that used in total internal reflection microscopy (TIRM), except that we collect the light that scatters back into the incident medium, because the AFM partly obscures the forward scattered light (i.e., light scattered into the transmitted region). Compared to a conventional TIRM measurement, where the particle moves freely, attaching the particle to the cantilever in an EW-AFM gives much greater control of the particle position.     

Formation kinetics of nanoparticles as the basis for simulating the generation mechanism of Liesegang rings in gels

An approach has been proposed for determining the space-time parameters of the formation process of Liesegang rings. This approach relates the processes of the formation of nanoparticles and dense periodic precipitates from them. Dynamic light scattering is employed to reveal periodic variations in the intensity of light scattered by nanoparticles with different sizes, from which Liesegang rings are formed. A relationship between the periods of the variations in the intensity of light scattered by nanoparticles (microscopic process) and the times of the formation of Liesegang rings as a whole (macroscopic process) has been found.     

Cavity ring-down spectroscopy measurement of single aerosol particle extinction. II. Extinction of light by an aerosol particle in an optical cavity excited by a cw laser

The authors present an analytical derivation of the scattered power from a spherical, homogeneous, nonabsorbing particle in a plane standing wave. The scattered power changes significantly with the position of the particle with respect to the peaks and nodes of the standing wave, even for particles whose diameters are many times the wavelength of the light. The analysis is applicable to continuous-wave cavity ring-down spectroscopy on aerosol particles, and the structure of the standing wave is expected to affect both the measured ring-down time and the shape of the ring-down trace. The dependence of the extinction on the phase of the standing wave at the location of the particle is captured in a parameter zeta which connects the current treatment to standard Mie scattering theory. Methods for calculating zeta are presented.     

Monte Carlo simulation of X-ray fluorescence spectra: Part 4. Photon scattering at high X-ray energies

The photon scattering model of a Monte Carlo simulation code for synchrotron radiation X-ray fluorescence (SRXRF) spectrometers is evaluated at high X-ray energies (60–100 keV) by means of a series of validation experiments performed at Beamline BW5 of HASYLAB. Using monochromatic X-rays, Compton/Rayleigh multiple scattering experiments were performed on polypropylene, Al and Cu samples. Especially in the case of the first two matrices multiple Compton scattering occurs with high probability. This work demonstrates that the simulation model provides a reliable estimate of the spectral distribution of the multiply scattered linearly polarized photon beam as observed by an HPGe detector. Next to variations in sample composition and thickness, the ability of the code to simulate various detection geometries has also been verified. As an application of the code, the achievable detection limits of SRXRF for rare earth elements as obtained with white beam and monochromatic (80 keV) excitation are compared.     

Calibration of Polarization-Sensitive and Dual-Angle Laser Light Scattering Methods Using Standard Latex Particles

A polarization-sensitive laser light scattering (PSLLS) method and a dual-angle laser light scattering (DALLS) method have been studied for in situ measurement of submicrometer hydrosol and aerosol particles. By using standard monodisperse polystyrene latex particles suspended in water and air as test particles, calibration of systems built based on the above methods have been performed. The effects of light scattered by agglomerated aerosol particles (multiplets) were corrected by considering the fraction of multiplets as determined with an aerosol measurement technique using a differential mobility analyzer. The change in the measured intensities of scattered light with particle diameter was then determined by calculations based on Mie theory. It was shown that the PSLLS system can determine particle diameters as small as approximately 60 nm for the test hydrosol particles and approximately 100 nm for test aerosol particles, respectively. The DALLS system can determine smaller diameters than the PSLLS system for test particles with no light absorption. The change in scattered light intensities with particle diameter was also investigated by theoretical calculations with various refractive indexes and scattering angles. The PSLLS and DALLS systems promise to become routine measurement tools for absorbing and nonabsorbing particles, respectively. Copyright 2001 Academic Press.     

双组分偶氮聚合物胶体球制备及其光致形变行为研究

研究了2种环氧树脂类含4-氨基-4′-硝基偶氮苯和4-氨基-4′-羧基偶氮苯生色团聚合物(BP-AZ-NT、BP-AZ-CA)双组分胶体球的制备和光致形变行为.通过在上述聚合物的四氢呋喃溶液中逐步加水诱导自组装的方法,得到了BP-AZ-CA/BP-AZ-NT双组分胶体球.在上述两种聚合物自组装形成胶体球过程中,较为疏水的BP-AZ-NT分子先发生聚集,而较亲水的BP-AZ-CA则在形成的胶体颗粒表面发生进一步聚集.在胶体球形成过程中,体系的临界水含量(CWC)主要由BP-AZ-NT发生聚集时的水含量决定,双组分胶体球的外层则含较多的BP-AZ-CA分子.比较单组分胶体球与双组分胶体球在线偏振Ar+激光(488nm,100mW/cm2)照射下的形变行为,进一步证实了通过上述方法可以制备BP-AZ-CA和BP-AZ-NT双组份的胶体球;胶体球形变时的初始拉伸速率由胶体球的外层聚合物分子的性质所决定。     

Angle‐dependent light scattering by highly uniform colloidal rod‐shaped microparticles: Experiment and simulation

While extensive theoretical work has been devoted to analyzing scattering behavior for nonspherical particles, few experimental studies of the light‐scattering properties of such particles are available, largely because of the difficulty of synthesizing such particles with uniform geometries. Here we report the synthesis of highly uniform, volume‐equivalent rod‐shaped colloidal particles prepared from their commercial spherical counterparts, on which we performed light scattering experiments as a function of scattering angle for micro rods with varying aspect ratio and volume. These results were compared to values calculated using the T‐Matrix method. Good agreement with theoretical predictions was found for the experimentally measured scattering cross sections and the angular dependence of the scattering intensity. An increase in the forward scattering intensity is observed and predicted for particles with larger aspect ratios relative to their volume equivalent spheres, with only minor differences observed at both mid‐range and backscattering angles. Furthermore, the light scattering results for the rod‐shaped particles did not show the scattering fringes seen in scattering by the spheres, indicating that as three‐dimensional symmetry is broken, the associated Lorenz–Mie resonances are strongly attenuated. This observation also was predicted by theory. © 2016 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2016 , 54, 1889–1895     

Light scattering from coloured systems: an example of polyaniline dispersions

Static light scattering of highly diluted dispersions in 0.005 M H₂SO₄ was used to determine particle parameters of polyaniline dispersions stabilized with colloidal silica. The refractive index of polyaniline n = 1.8 and the refractive index increment (dn/dc) = 0.22 cm³g⁻¹ at λ = 532 nm were determined. The light scattering data are affected by the absorption of the green polyaniline and by a small amount of aggregates. The absorption has a negligible effect on the results. The influence of the aggregates was corrected by using the intensity‐weighed size distributions determined by dynamic light scattering at different angles and by the two‐component separation in static light scattering. Both procedures yield the same result.     

Light scattering from tilted two-dimensional spherulites

The theory of small-angle light scattering was developed for oblique incidence of the light beam on the surface of a two-dimensional spherulite. Results of the theory were compared with previously reported results of light scattering from two-dimensional and three-dimensional spherulites for normal incidence, and with some experimental patterns. The comparisons suggest that the scattering intensity distributions of two-dimensional spherulites deviate from those of three-dimensional spherulites when the sample surface is tilted with respect to the propagation direction of the incident beam, although they are almost identical when the sample surface is normal to the incident beam. Observation of the change of scattered intensity distributions upon tilting the samples thus provides a method of distinguishing between two-dimensional and three-dimensional spherulites. Moreover, this observation makes it possible to determine the degree of planar orientation of the optic axes of optically anisotropic scattering elements within two-dimensional spherulites. The calculations were carried out for special cases of two-dimensional spherulites with the optic axis orientation confined to the two-dimensional plane and randomly or helicoidally rotated around the spherulite radii.