Moiré deflectometry-based position detection for optical tweezers.

Optical tweezers have proven to be indispensable tools for pico-Newton range force spectroscopy. A quadrant photodiode (QPD) positioned at the back focal plane of an optical tweezers' condenser is commonly used for locating the trapped object. In this Letter, for the first time, to the best of our knowledge, we introduce a moiré pattern-based detection method for optical tweezers. We show, both theoretically and experimentally, that this detection method could provide considerably better position sensitivity compared to the commonly used detection systems. For instance, position sensitivity for a trapped 2.17 µm polystyrene bead is shown to be 71% better than the commonly used QPD-based detection method. Our theoretical and experimental results are in good agreement.

Digital holography based submicron thermometry.

Here we introduce a phase-shifting digital holography-based method to determine the temperature profile around an irradiated (sub-)micron spherical bead. The method utilizes a Mach-Zehnder interferometer implemented into an open setup microscope. The results of irradiated gold spheres with diameter of 400 nm and also silver-coated micron-sized silica beads embedded in silicone oil are presented. We show that the applied method is able to accurately determine the surface temperature with accuracy of 1 °C. Our experimental results perfectly confirm the theoretical prediction of temperature profile around the irradiated bead.

Simultaneous three-dimensional tracking of individual signals from multi-trap optical tweezers using fast and accurate photodiode detection.

Multiple-beam optical traps facilitate advanced trapping geometries and exciting discoveries. However, the increased manipulation capabilities come at the price of more challenging position and force detection. Due to unrivaled bandwidth and resolution, photodiode based detection is preferred over camera based detection in most single/dual-beam optical traps assays. However, it has not been trivial to implement photodiode based detection for multiple-beam optical traps. Here, we present a simple and efficient method based on spatial filtering for parallel photodiode detection of multiple traps. The technique enables fast and accurate 3D force and distance detection of multiple objects simultaneously manipulated by multiple-beam optical tweezers.

Nanoscale phase behavior on flat and curved membranes.

The diverse physical properties of membranes play a critical role in many membrane associated biological processes. Proteins responsible for membrane transport can be affected by the lateral membrane order and lateral segregation of proteins is often controlled by the preference of certain membrane anchors for membrane phases having a physically ordered state. The dynamic properties of coexisting membrane phases are often studied by investigating their thermal behavior. Optical trapping of gold nanoparticles is a useful tool to generate local phase transitions in membranes. The high local temperatures surrounding an irradiated gold nanoparticle can be used to melt a part of a giant unilamellar lipid vesicle (GUV) which is then imaged using phase sensitive fluorophores embedded within the bilayer. By local melting of GUVs we reveal how a protein-free, one component lipid bilayer can mediate passive transport of fluorescent molecules by localized and transient pore formation. Also, we show how tubular membrane curvatures can be generated by optical pulling from the melted region on the GUV. This will allow us to measure the effect of membrane curvature on the phase transition temperature.

Assessment of Physicochemical Properties of Orange Juice Concentrate Formulated with Pectin, Xanthan, and CMC Hydrocolloids.

Orange concentrate (OC) is one of the main raw materials in the nonalcoholic beverage industry. Considering the difference in orange varieties, preserving its natural quality is essential to yield a product with favorable attributes and physical stability. Thus, the present study is aimed at assessing the effect of pectin, xanthan, and carboxymethyl cellulose (CMC) in a concentration range of 0-0.2% (w/v) along with mixing temperature on Brix, pH, acidity, density, turbidity, and viscosity of OC and at calculating the model equation for each attribute. The results showed that, except for CMC, the influence of concentration, type, and amount of hydrocolloid on pH changes was insignificant. Adding each hydrocolloid individually, in pairs, or threes reduced the density, and the measured density was lower at a mixing temperature of 4°C. Also, it was observed that mixing temperature was the only factor influencing turbidity, and the values were significantly lower at 80°C compared to 4°C. A significant interaction effect of xanthan concentration and mixing temperature on the Brix was observed. Adding hydrocolloids, except pectin, resulted in a significant (p < 0.05) increase in viscosity, and xanthan had the greatest effect on the viscosity. A suitable model was designed using pectin and xanthan, pectin and CMC, and all three gums, resulting in a final OC product with high stability and improved physical and chemical attributes. The optimized values for Brix, pH, acidity, density, turbidity, and OC viscosity were achieved using 0.08% pectin, 0.19% xanthan, and 0.08% CMC at 80°C mixing temperature.

Quantitative determination of optical trapping strength and viscoelastic moduli inside living cells.

With the success of in vitro single-molecule force measurements obtained in recent years, the next step is to perform quantitative force measurements inside a living cell. Optical traps have proven excellent tools for manipulation, also in vivo, where they can be essentially non-invasive under correct wavelength and exposure conditions. It is a pre-requisite for in vivo quantitative force measurements that a precise and reliable force calibration of the tweezers is performed. There are well-established calibration protocols in purely viscous environments; however, as the cellular cytoplasm is viscoelastic, it would be incorrect to use a calibration procedure relying on a viscous environment. Here we demonstrate a method to perform a correct force calibration inside a living cell. This method (theoretically proposed in Fischer and Berg-Sørensen (2007 J. Opt. A: Pure Appl. Opt. 9 S239)) takes into account the viscoelastic properties of the cytoplasm and relies on a combination of active and passive recordings of the motion of the cytoplasmic object of interest. The calibration procedure allows us to extract absolute values for the viscoelastic moduli of the living cell cytoplasm as well as the force constant describing the optical trap, thus paving the way for quantitative force measurements inside the living cell. Here, we determine both the spring constant of the optical trap and the elastic contribution from the cytoplasm, influencing the motion of naturally occurring tracer particles. The viscoelastic moduli that we find are of the same order of magnitude as moduli found in other cell types by alternative methods.

Axial potential mapping of optical tweezers for biopolymer stretching: the bead size matters.

Optical tweezers (OT) are widely used for pico (and femto)-Newton range force measurements. The appropriate choice of the bead size is not well understood for biopolymer stretching applications of OT. We have shown, both by theory and experiment, that wrong choice of the bead size could cause errors as large as 295% in the measured force. We provide a simple map for correct choice of the bead size and the direction of pulling for such applications. There is a good agreement between our theoretical and experimental results.

Polarization-induced stiffness asymmetry of optical tweezers.

A tightly focused, linearly polarized laser beam, so-called optical tweezers, is proven to be a useful micromanipulation tool. It is known that there is a stiffness asymmetry in the direction perpendicular to the optical axis inherited from the polarization state of the laser. In this Letter, we report our experimental results of stiffness asymmetry for different bead sizes measured at the optimal trapping condition. We also provide the results of our generalized Lorenz-Mie based calculations, which are in good agreement with our experimental results. We also compare our results with previous reports.

Survey of the salt (NaCl) Contents of Traditional Breads in Tehran, 2016‒2018: Implication for Public Health.

Background: Considering the importance of non-communicable diseases (NCDs) in Iran, the aim of this study is to identify the trend in the salt (NaCl) levels of various types of traditional bread (sangak, barbari, taftoon, and lavash breads) in Tehran in 2016 and 2018 and its implication for public health. Methods: A total of 777 samples of various traditional breads were randomly collected from various districts located in Tehran in 2018. The salt content (expressed as g/100 g dry weight) in these breads were determined according to Volhard method. Results from this study were compared with those reported in 2016. Results: The present study indicated that the mean salt content in traditional breads in 2018 was significantly higher than that reported in 2016. Salt content in traditional breads collected in 2018 ranged from 0.03 to 6.52/100 g dry weight, with mean value of 1.43 g/100 g dry weight. When comparing with the permitted limit set by Institute of Standards and Industrial Research of Iran (ISIRI), there was an increase in the percentage of samples complying with the permitted limit; 50.8% (2016) vs 54.1% (2018). Conclusions: The daily salt intake increased from 1.56 g per person in 2016 to 2.31 g per person in 2018. Considering the high bread per capita consumption in Iran, it seems that half of the daily recommended salt intake could be reached exclusively through breads. Hence, the main strategies for salt intake reduction from bread could be achieved through evaluation of salt reduction programs and development of technological factors in bread baking.

The effect of immersion oil in optical tweezers.

Optimized optical tweezers are of great importance for biological micromanipulation. In this paper, we present a detailed electromagnetic-based calculation of the spatial intensity distribution for a laser beam focused through a high numerical aperture objective when there are several discontinuities in the optical pathway of the system. For a common case of 3 interfaces we have shown that 0.01 increase in the refractive index of the immersion medium would shift the optimal trapping depth by 3-4 µm (0.2-0.6 µm) for aqueous (air) medium. For the first time, We have shown that the alteration of the refractive index of the immersion medium can be also used in aerosol trapping provided that larger increase in the refractive index is considered.

Potential mapping of optical tweezers.

Optical tweezers are very often used for measurement of piconewton range forces. Depending on the displacement of the trapped bead, the trap may become stiffer which causes considerable underestimation of the measured force. We have shown, both by theory and experiment, that such a stiffening occurs for beads larger than 0.5 µm in radius. For the first time, we have shown that the displacement at which the stiffening starts is size dependent and that the stiffening starts at higher forces for larger beads. We have shown that for the applications, which simultaneous force measurement and position sensing are on demand (such as biopolymer stretching), mid-range sized (∼1.5 µm in radius) beads could be the best choice.

Role of condenser iris in optical tweezer detection system.

Optical tweezers have proven to be very useful in various scientific fields, from biology to nanotechnology. In this Letter we show, both by theory and experiment, that the interference intensity pattern at the back focal plane of the condenser consists of two distinguishable areas with anticorrelated intensity changes when the bead is moved in the axial direction. We show that the space angle defining the border of two areas linearly depends on the NA of the objective. We also propose a new octant photodiode, which could significantly improve the axial resolution compared to the commonly used quadrant photodiode technique.

Optimized optical trapping of gold nanoparticles.

Metallic nanoparticles are of significant interest due to their particular optical and biological applications. Gold nanoparticles are proven to be excellent candidate for in vivo micro-manipulation using Optical Tweezers. This manuscript reports on stable 3-D trapping of 9.5-254nm gold nanospheres using substantially decreased laser power. The lower limit is approximately 2 times smaller than previous record. 5.4nm gold nanospheres were trapped for only 2-3 seconds. For the first time, our experimental data verify the volume corrected Rayleigh model for particles smaller than 100nm in diameter. Measuring the maximum applicable force for gold nanoparticles, we have shown that a few tens of milli-Watts of laser power can produce pico-Newton range forces.

Phase contrast optical tweezers.

In this paper, for the first time, we report on systematic theoretical and experimental investigation of Phase Contrast Optical Tweezers (PCOT) which could be an indispensable tool for micromanipulation of the transparent micro and nano objects such as biological tissues and vesicles. The quadrant photodiode detection scheme and the power-spectrum calibration method is shown to be valid for this case. We have shown that the phase objective with new designed phase plates can provide nearly aberration-free condition at a desired depth. This could be a valuable advantage for simultaneous in-depth micro-manipulations and visualization of the sample.

Effect of alumina nanoparticles on hot strength and deformation behaviour of AI-5vol% Al2O3 nanocomposite: experimental study and modelling.

Hot deformation behaviour of as-extruded Al-5vol% Al2O3 nanocomposite was investigated at temperatures range 350 to 450 degrees C and initial strain rates of 5.5 x 10(-4) to 10(-1) s(-1) and compared with those of monolithic (unreinforced) aluminium. Both extruded materials exhibited work-softening during hot deformation. The results showed that with the addition of 5 vol% alumina nanoparticles with an average particle size of 35 nm, a significant increase in compressive strength of aluminium was obtained. For instance, at 350 degrees C an abrupt rise of approximately 340% in hot strength of the nanocomposite relative to monolithic aluminium was achieved. TEM investigation of microstructure of the nanocomposite after hot deformation showed formation of equiaxed grains from elongated ones indicating the occurrence of dynamic recrystallization. Considering experimental data, deformation behaviour of Al-5vol% Al2O3 nanocomposite and monolithic Al was modelled via trained artificial neural network (ANN). The results showed that ANN can predict complex flow behaviour of the nanocomposite as well as the monolithic aluminium.

Confocal microscopy of thick specimens.

Confocal microscopy is an excellent tool to gain structural information from deep within a biological sample. The depth from which information can be extracted as well as the resolution of the detection system are limited by spherical aberrations in the laser pathway. These spherical aberrations of the visible light can be efficiently canceled by optimizing the refractive index of the immersion media. Another way of cancelling spherical aberrations is by changing tube length, or alternatively, by changing the objective from infinite correction to finite correction, or vice versa, depending on which microscope is used. A combination of these two methods allows for confocal imaging at continuous depths. Presently, confocal microscopes typically operate at a maximum depth of 40 microm in the sample, but with the methods presented here, we show that information can easily be gained from depths up to 100 microm. Additionally, the precision of localization of a single fluorophore in the axial direction, limited by spherical aberrations, can be significantly improved, even if the fluorophore is located deep within the sample. In principle, this method can improve the efficiency of any kind of microscopy based on visible light.

Proper measurement of pure dielectrophoresis force acting on a RBC using optical tweezers.

The force experienced by a neutral dielectric object in the presence of a spatially non-uniform electric field is referred to as dielectrophoresis (DEP). The proper quantification of DEP force in the single-cell level could be of great importance for the design of high-efficiency micro-fluidic systems for the separation of biological cells. In this report we show how optical tweezers can be properly utilized for proper quantification of DEP force experienced by a human RBC. By tuning the temporal frequency of the applied electric field and also performing control experiments and comparing our experimental results with that of theoretically calculated, we show that the measured force is a pure DEP force. Our results show that in the frequency range of 0.1-3 M H z the DEP force acting on RBC is frequency independent.

Non-harmonic potential of a single beam optical trap.

Since the invention of optical traps based on a single laser beam, the potential experienced by a trapped specimen has been assumed harmonic, in the central part of the trap. It has remained unknown to what extent the harmonic region persists and what occurs beyond. By employing a new method, we have forced the trapped object to extreme positions, significantly further than previously achieved in a single laser beam, and thus experimentally explore an extended trapping potential. The potential stiffens considerably as the bead moves to extreme positions and therein is not well described by simple Uhlenbeck theories.

Stochastic analysis of time series for the spatial positions of particles trapped in optical tweezers.

We propose a nonlinear method for the analysis of the time series for the spatial position of a bead trapped in optical tweezers, which enables us to reconstruct its dynamical equation of motion. The main advantage of the method is that all the functions and parameters of the dynamics are determined directly (non-parametrically) from the measured series. It also allows us to determine, for the first time to our knowledge, the spatial-dependence of the diffusion coefficient of a bead in an optical trap, and to demonstrate that it is not in general constant. This is in contrast with the main assumption of the popularly-used power spectrum calibration method. The proposed method is validated via synthetic time series for the bead position with spatially-varying diffusion coefficients. Our detailed analysis of the measured time series reveals that the power spectrum analysis overestimates considerably the force constant.

Dynamics of membrane nanotubes coated with I-BAR.

Membrane deformation is a necessary step in a number of cellular processes such as filopodia and invadopodia formation and has been shown to involve membrane shaping proteins containing membrane binding domains from the IRSp53-MIM protein family. In reconstituted membranes the membrane shaping domains can efficiently deform negatively charged membranes into tubules without any other proteins present. Here, we show that the IM domain (also called I-BAR domain) from the protein ABBA, forms semi-flexible nanotubes protruding into Giant Unilamellar lipid Vesicles (GUVs). By simultaneous quantification of tube intensity and tubular shape we find both the diameter and stiffness of the nanotubes. I-BAR decorated tubes were quantified to have a diameter of ~50 nm and exhibit no stiffening relative to protein free tubes of the same diameter. At high protein density the tubes are immobile whereas at lower density the tubes diffuse freely on the surface of the GUV. Bleaching experiments of the fluorescently tagged I-BAR confirmed that the mobility of the tubes correlates with the mobility of the I-BAR on the GUV membrane. Finally, at low density of I-BAR the protein upconcentrates within tubes protruding into the GUVs. This implies that I-BAR exhibits strong preference for negatively curved membranes.