Probing the dynamics of an optically trapped particle by phase sensitive back focal plane interferometry

The dynamics of an optically trapped particle are often determined by measuring intensity shifts of the back-scattered light from the particle using position sensitive detectors. We present a technique which measures the phase of the back-scattered light using balanced detection in an external Mach-Zender interferometer scheme where we separate out and beat the scattered light from the bead and that from the top surface of our trapping chamber. The technique has improved axial motion resolution over intensity-based detection, and can also be used to measure lateral motion of the trapped particle. In addition, we are able to track the Brownian motion of trapped 1 and 3 $\mu$m diameter beads from the phase jitter and show that, similar to intensity-based measurements, phase measurements can also be used to simultaneously determine displacements of the trapped bead as well as the spring constant of the trap. For lateral displacements, we have matched our experimental results with a simulation of the overall phase contour of the back-scattered light for lateral displacements by using plane wave decomposition in conjunction with Mie scattering theory. The position resolution is limited by path drifts of the interferometer which we have presently reduced to obtain a displacement resolution of around 2 nm for 1.1 $\mu$m diameter probes by locking the interferometer to a frequency stabilized diode laser.Comment: 10 pages, 7 figure

A multiplex oligonucleotide ligation-PCR as a complementary tool for subtyping of Salmonella Typhimurium

Subtyping below the serovar level is essential for surveillance and outbreak detection and investigation of Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) and its monophasic variant 1,4,[5],12:i:- (S. 1,4,[5],12:i:-), frequent causes of foodborne infections. In an attempt to overcome the intrinsic shortcomings of currently used subtyping techniques, a multiplex oligonucleotide ligation-PCR (MOL-PCR) assay was developed which combines different types of molecular markers in a high throughput microsphere suspension array. The 52 molecular markers include prophage genes, amplified fragment length polymorphism (AFLP) elements, Salmonella genomic island 1 (SGI1), allantoinase gene allB, MLVA locus STTR10, antibiotic resistance genes, single nucleotide polymorphisms (SNPs) and phase 2 flagellar gene fljB. The in vitro stability of these markers was confirmed in a serial passage experiment. The validation of the MOL-PCR assay for subtyping of S. Typhimurium and S. 1,4,[5],12:i:- on 519 isolates shows that the method is rapid, reproducible, flexible, accessible, easy to use and relatively inexpensive. Additionally, a 100 % typeability and a discriminatory power equivalent to that of phage typing were observed, and epidemiological concordance was assessed on isolates of 2 different outbreaks. Furthermore, a data analysis method is provided so that the MOL-PCR assay allows for objective, computerised data analysis and data interpretation of which the results can be easily exchanged between different laboratories in an international surveillance network

Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed

A multi-mycotoxin immunoassay—using the MultiAnalyte Profiling (xMAP) technology—is developed and evaluated. This technology combines a unique color-coded microsphere suspension array, with a dedicated flow cytometer. We aimed for the combined detection of aflatoxins, ochratoxin A, deoxynivalenol, fumonisins, zearalenone and T-2-toxin in an inhibition immunoassay format. Sets of six mycotoxin-protein conjugates and six specific monoclonal antibodies were selected, and we observed good sensitivities and no cross-interactions between the assays in buffer. However, detrimental effects of the feed extract on the sensitivities and in some cases on the slopes of the curves were observed and different sample materials showed different effects. Therefore, for quantitative analysis, this assay depends on calibration curves in blank matrix extracts or on the use of a suitable multi-mycotoxin cleanup. To test if the method was suitable for the qualitative detection at EU guidance levels, we fortified rapeseed meal, a feed ingredient, with the six mycotoxins, and all extracts showed inhibited responses in comparison with the non-fortified sample extract. Contaminated FAPAS reference feed samples assigned for a single mycotoxin showed strong inhibitions in the corresponding assays but also often in other assays of the multiplex. In most cases, the presence of these other mycotoxins was confirmed by instrumental analysis. The multiplex immunoassay can be easily extended with other mycotoxins of interest, but finding a suitable multi-mycotoxin cleanup will improve its applicability

Behavior of nanoparticle clouds around a magnetized microsphere under magnetic and flow fields

When a micron-sized magnetizable particle is introduced into a suspension of nanosized magnetic particles, the nanoparticles accumulate around the microparticle and form thick anisotropic clouds extended in the direction of the applied magnetic field. This phenomenon promotes colloidal stabilization of bimodal magnetic suspensions and allows efficient magnetic separation of nanoparticles used in bioanalysis and water purification. In the present work, size and shape of nanoparticle clouds under the simultaneous action of an external uniform magnetic field and the flow have been studied in details. In experiments, dilute suspension of iron oxide nanoclusters (of a mean diameter of 60 nm) was pushed through a thin slit channel with the nickel microspheres (of a mean diameter of 50$\mu$m) attached to the channel wall. The behavior of nanocluster clouds was observed in the steady state using an optical microscope. In the presence of strong enough flow, the size of the clouds monotonically decreases with increasing flow speed in both longitudinal and transverse magnetic fields. This is qualitatively explained by enhancement of hydrodynamic forces washing the nanoclusters away from the clouds. In the longitudinal field, the flow induces asymmetry of the front and the back clouds. To explain the flow and the field effects on the clouds, we have developed a simple model based on the balance of the stresses and particle fluxes on the cloud surface. This model, applied to the case of the magnetic field parallel to the flow, captures reasonably well the flow effect on the size and shape of the cloud and reveals that the only dimensionless parameter governing the cloud size is the ratio of hydrodynamic-to-magnetic forces - the Mason number. At strong magnetic interactions considered in the present work (dipolar coupling parameter $\alpha \geq 2$), the Brownian motion seems not to affect the cloud behavior

Nanostructuring GaN using microsphere lithography

The authors report on the fabrication and characterization of nanopillar arrays on GaN substrates using the technique of microsphere lithography. Self-assembled hexagonally packed silica microsphere arrays were formed on GaN wafers by spin coating and tilting. By precision control of process parameters, a monolayer can be formed over a wide region. The silica microspheres act as a hard mask for pattern transfer of the nanostructures. After dry etching, arrays of nanopillars were formed on the surface of the wafer. The ordered nanostructures can be clearly seen in the scanning electron microscopy images, while photoluminescence measurements revealed a twofold enhancement of light emission intensity. © 2008 American Vacuum Society.published_or_final_versio