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Monitoring the temperature distribution within a local environment at the micro and nanoscale is vital as many processes are solely thermal. Various thermometric techniques have been explored in the community, and out of these, fluorescent nano/micro particle-based mechanisms are accepted widely (fluorescence intensity ratio (FIR) techniques, where the ratio of populations in two consecutive energy levels is compared with Boltzmann distribution). We describe a new technique to account for the temperature rise near an illuminated upconverting particle (UCP) using wavefront imaging, which is more sensitive than the conventional thermometric techniques on the microscale. We rely on a thermo-optical phase microscopic technique by reconstructing the wavefront of emission from an upconverting particle using a Shack-Hartmann wavefront sensor. The wavefront maps the local phase distribution, which is an indicator of the surroundings' optical parameters, particularly the suspended medium's temperature-induced refractive index in the presence of convection currents. We describe how these extracted phase values can provide information about the optical heating due to the particle and hence its local environment along the direction of the emission. Our findings demonstrate the detection of a minimum temperature rise of 0.23 K, while the FIR methods indicate a minimum of 0.3 K rise. This technique is used to study the temperature increase in the backscattered direction for an upconverting particle illuminated on pump resonance. We also estimate the Soret coefficient for an upconverting particle optically trapped on pump resonance and experiencing anisotropic heating across the body.
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A rigid body can have six degrees of freedom, of which three are with rotational origin. In the nomenclature of the airlines, the in-plane degree of rotational freedom can be called yaw while the first out-of-plane degree of freedom can be called pitch with the second one being called roll. Among these, only the yaw sense has been studied extensively in the optical tweezers literature, while the pitch rotation is starting to be explored. In this paper, we show a way to detect the pitch rotation in a hexagonal-shaped particle using photonic force microscopy using the forward scattered light under crossed polarizers and making it incident on a split photodiode. In this way, the pitch angle can be detected at high resolution and bandwidth. We apply this technique to detect continuous pitch rotation and also exhibit a power spectral density for an anisotropic particle optically trapped in a linearly polarized light and exhibiting Brownian motion.
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In recent years, there has been a growing interest in studying the trajectories of microparticles inside living cells. Among other things, such studies are useful in understanding the spatio-temporal properties of a cell. In this work, we study the stochastic trajectories of a passive microparticle inside a cell using experiments and theory. Our theory is based on modeling the microparticle inside a cell as an active particle in a viscoelastic medium. The activity is included in our model from an additional stochastic term with non-zero persistence in the Langevin equation describing the dynamics of the microparticle. Using this model, we are able to predict the power spectral density (PSD) measured in the experiment and compute active forces. This caters to the situation where a tracer particle is optically confined and then yields a PSD for positional fluctuations. The low frequency part of the PSD yields information about the active forces that the particle feels. The fit to the model extracts such active force. Thus, we can conclude that trapping the particle does not affect the values of the forces extracted from the active fits if accounted for appropriately by proper theoretical models. In addition, the fit also provides system properties and optical tweezers trap stiffness.
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In recent years, there has been a growing interest in controlling the motion of microparticles inside and outside a focused laser beam. A hydro-thermophoretic trap was recently reported [Nalupurackal et al., Soft Matter 18, 6825 (2022)], which can trap and manipulate microparticles and living cells outside a laser beam. Briefly, a hydro-thermophoretic trap works by the competition between thermoplasmonic flows due to laser heating of a substrate and thermophoresis away from the hotspot of the laser. Here, we extend that work to demonstrate the controlled roll rotation of a microparticle in a hydro-thermophoretic trap using experiments and theory. We experimentally measure the roll angular velocity of the trapped particle. We predict this roll rotation from theoretical computation of the fluid flow. The expression for the angular velocity fits the experimental data. Our method has potential applications in microrheology by employing a different mode of rotation.
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Hematite particles, which exhibit a high magnetic moment, are used to apply large forces on physical and biological systems under magnetic fields to investigate various phenomena, such as those of rheology and micromanipulation. However, the magnetic confinement of these particles requires complicated field configurations. On the other hand, laser-assisted optical confinement of single hematite particles results in thermophoresis and subsequent ejection of the particle from the laser spot. Herein, we explore an alternative strategy to induce the self-assembly of hematite. In this strategy, with indirect influence from an optically confined and heated upconverting particle (UCP) at an air-water interface, there is the generation of convection currents that facilitate assembly. We also show that the assembly remains at the interface even after removal of the laser light. The hematite particle assemblies can then be moved using magnetic fields and employed to perform interfacial rheology.
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Conventionally, only the normal cell membrane fluctuations have been studied and used to ascertain membrane properties like the bending rigidity. A new concept, the membrane local slope fluctuations was introduced recently (Vaippullyet al2020Soft Matter167606), which can be modelled as a gradient of the normal fluctuations. It has been found that the power spectral density (PSD) of slope fluctuations behave as (frequency)-1while the normal fluctuations yields (frequency)-5/3even on the apical cell membrane in the high frequency region. In this manuscript, we explore a different situation where the cell is applied with the drug Latrunculin-B which inhibits actin polymerization and find the effect on membrane fluctuations. We find that even as the normal fluctuations show a power law (frequency)-5/3as is the case for a free membrane, the slope fluctuations PSD remains (frequency)-1, with exactly the same coefficient as the case when the drug was not applied. Moreover, while sometimes, when the normal fluctuations at high frequency yield a power law of (frequency)-4/3, the pitch PSD still yields (frequency)-1. Thus, this presents a convenient opportunity to study membrane parameters like bending rigidity as a function of time after application of the drug, while the membrane softens. We also investigate the active athermal fluctuations of the membrane appearing in the PSD at low frequencies and find active timescales of slower than 1 s.
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Compuestos Bicíclicos Heterocíclicos con Puentes , Membrana Celular , Tiazolidinas , Compuestos Bicíclicos Heterocíclicos con Puentes/química , Tiazolidinas/químicaRESUMEN
Particles can be assembled at the air-water interface due to optically induced local heating. This induces convection currents in the water which brings particles to the surface. We improve the technique by employing an upconverting particle (UCP), which, when illuminated with 975 nm light, not only emits visible emission but also generates heat owing to the poor efficiency of the upconversion process. This induces strong convection currents which makes particles dispersed in the suspension assemble at the interface and immediately under the UCP. We show assembly of polystyrene particles of 1 µm diameter and diamonds of 500 nm diameter bearing Nitrogen-Vacancy (NV) centers around the UCP. We also show, for the first time, that the microdiamonds are assembled within about 30 nm at the bottom of the UCP by utilizing non-radiative energy transfer that reduces the lifetime of the 550 nm emission from about 90 µs to about 50 µs.
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A colloidal particle placed inside the cell cytoplasm is enmeshed within a network of cytoskeletal fibres immersed in the cytosolic fluid. The translational mode is believed to yield different rheological parameters than the rotational mode, given that these modes stretch the fibers differently. We compare the parameters for Michigan Cancer Foundation-7 (MCF-7) cells in this manuscript and find that the results are well comparable to each other. At low values of 0 Hz viscosity, the rotational and translational viscoelasticity matches well. However, discrepancies appear at higher values which may indicate that the cytoskeletal modes involved in rotation and translation of the particle are getting invoked. We also show that the 0 Hz viscosity increases as the cell ages under the conditions of constant room temperature of 25°C on the sample chamber.
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Tumor-associated collagen signature-3 (TACS-3) is a prognostic indicator for breast cancer survival. It is characterized by highly organized, parallel bundles of collagen fibers oriented perpendicular to the tumor boundary, serving as directional, confining channels for cancer cell invasion. Here we design a TACS-3-mimetic anisotropic, confined collagen I matrix and examine the relation between anisotropy of matrix, directed cellular migration, and anisotropy of cell membrane-the first direct contact between TACS-3 and cell-using Michigan Cancer Foundation-7 (MCF-7) cells as cancer-model. Using unidirectional freezing, we generated â¼50µm-wide channels filled with collagen I. Optical tweezer (OT) microrheology shows that anisotropic confinement increases collagen viscoelasticity by two orders of magnitude, and the elastic modulus is significantly greater along the direction of anisotropic confinement compared to that along the orthogonal direction, thus establishing matrix anisotropy. Furthermore, MCF-7 cells embedded in anisotropic collagen I, exhibit directionality in cellular morphology and migration. Finally, using customized OT to trap polystyrene probes bound to cell-membrane (and not to ECM) of either free cells or cells under anisotropic confinement, we quantified the effect of matrix anisotropy on membrane viscoelasticity, both in-plane and out-of-plane, vis-à-vis the membrane. Both bulk and viscous modulus of cell-membrane of MCF-7 cells exhibit significant anisotropy under anisotropic confinement. Moreover, the cell membrane of MCF-7 cells under anisotropic confinement is significantly softer (both in-plane and out-of-plane moduli) despite their local environment being five times stiffer than free cells. In order to test if the coupling between anisotropy of extracellular matrix and anisotropy of cell-membrane is regulated by cell-cytoskeleton, actin cytoskeleton was depolymerized for both free and confined cells. Results show that cell membrane viscoelasticity of confined MCF-7 cells is unaffected by actin de-polymerization, in contrast to free cells. Together, these findings suggest that anisotropy of ECM induces directed migration and correlates with anisotropy of cell-membrane viscoelasticity of the MCF-7 cells in an actin-independent manner.
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Actinas , Colágeno , Humanos , Anisotropía , Células MCF-7 , Membrana CelularRESUMEN
Typically a rigid body can have three degrees of rotational freedom. Among these, there can be two types of out-of-plane rotational modes, called the pitch and the roll. The pitch motion is typically to turn the particle along an axis orthogonal to the axis of symmetry. However, rotation about the axis of symmetry (called the roll motion) has so far not been shown in optical tweezers. It is here that we use a hexagonal shaped particle (NaYF4) which prefers to align side on with the optical tweezers [Rodriguez-Sevilla et al., Nano Letters 16, 8005 (2016)]. In this work, we find that the stable configuration of the hexagonal particle changes while using one beam and two beams, so that when one of the tweezers beams is switched on and off, the particle tends to switch between the different configurations. Thus we get a controlled roll motion. This is the first time that controlled partial roll motions have been generated in optical tweezers.
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Optical tweezers have revolutionised micromanipulation from physics and biology to material science. However, the high laser power involved in optical trapping can damage biological samples. In this context, indirect trapping of microparticles and objects using fluid flow fields has assumed great importance. It has recently been shown that cells and particles can be turned in the pitch sense by opto-plasmonic heating of a gold surface constituting one side of a sample chamber. We extend that work to place two such hotspots in close proximity to each other to form a very unique configuration of flow fields forming an effective quasi-three-dimensional 'trap', assisted by thermophoresis. This is effectively a harmonic trap confining particles in all three dimensions without relying on other factors to confine the particles close to the surface. We use this to show indirect trapping of different types of upconverting particles and cells, and also show that we can approach a trap stiffness of 40 fN µm-1 indicating a weak confinement regime without relying on feedback.
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Mechanical properties of particle laden interfaces is crucial for various applications. For water droplets containing soft microgel particles, passive microrheology studies have revealed that the dynamically varying surface area of the evaporating drop results in a viscous to viscoelastic transition along the plane of the interface. However, the behaviour of the medium orthogonal to the interface has been elusive to study using passive microrheology techniques. In this work, we employ optical tweezers and birefringent probe particles to extract the direction-resolved viscoelastic properties of the particle-laden interface. By using special types of birefringent tracer particles, we detect not only the in-plane translational mode but also the out-of-plane translational (perpendicular to the interface) and rotational modes. We first compare different passive methods of probing the viscoelasticity of the microgel laden interface of sessile drop and then study the modes perpendicular to the interface and the out-of-plane rotational mode using optical tweezers based passive microrheology. The viscoelasticity of the interface using two different methods, i.e., multiple-particle tracking passive microrheology using video microscopy and by trapping birefringent tracer particles in optical tweezers, relying on different models are studied and found to exhibit comparable trends. Interestingly, the mode orthogonal to the interface and the rotational mode also show the viscous to viscoelastic transition as the droplet evaporates, but with lesser viscoelasticity during the same evaporation time than the in-plane mode.
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Upconverting particles like Yb and Er-doped NaYF4 are known to heat up after illumination with light at pump wavelength due to inefficient upconversion processes. Here we show that NaYF4 particles which have been co-doped not only with Yb and Er but also Fe improves the photothermal conversion efficiency. In addition, we show for the first time that alternating magnetic fields also heat up the ferromagnetic particles. Thereafter we show that a combination of optical and magnetic stimuli significantly increases the heat generated by the particles.
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Cardiovascular and cancer illnesses often co-exist, share pathological pathways, and complicate therapy. In the context of the potential oncological role of cardiovascular-antihypertensive drugs (AHD), here we examine the role of calcium-channel blocking drugs on mechanics of extravasating cancer cells, choosing two clinically-approved calcium-channel blockers (CCB): Verapamil-hydrochloride and Nifedipine, as model AHD to simultaneously target cancer cells (MCF7 and or MDA231) and an underlying monolayer of endothelial cells (HUVEC). First, live-cell microscopy shows that exposure to Nifedipine increases the spreading-area, migration-distance, and frequency of transmigration of MCF-7 cells through the HUVEC monolayer, whereas Verapamil has the opposite effect. Next, impedance-spectroscopy shows that for monolayers of either endothelial or cancer cells, Nifedipine-treatment alone decreases the impedance of both cases, suggesting compromised cell-cell integrity. Furthermore, upon co-culturing MCF-7 on the HUVEC monolayers, Nifedipine-treated MCF-7 cells exhibit weaker impedance than Verapamil-treated MCF-7 cells. Following, fluorescent staining of CCB-treated cytoskeleton, focal adhesions, and cell-cell junction also indicated that Nifedipine treatment diminished the cell-cell integrity, whereas verapamil treatment preserved the integrity. Since CCBs regulate intracellular Ca2+, we next investigated if cancer cell's exposure to CCBs regulates calcium-dependent processes critical to extravasation, specifically traction and mechanics of plasma membrane. Towards this end, first, we quantified the 2D-cellular traction of cells in response to CCBs. Results show that exposure to F-actin depolymerizing drug decreases traction stress significantly only for Nifedipine-treated cells, suggesting an actin-independent mechanism of Verapamil activity. Next, using an optical tweezer to quantify the mechanics of plasma membrane (PM), we observe that under constant, externally-applied tensile strain, PM of Nifedipine-treated cells exhibits smaller relaxation-time than Verapamil and untreated cells. Finally, actin depolymerization significantly decreases MSD only for Verapamil treated cancer-cells and endothelial cells and not for Nifedipine-treated cells. Together, our results show that CCBs can have varied, mechanics-regulating effects on cancer-cell transmigration across endothelial monolayers. A judicious choice of CCBs is critical to minimizing the pro-metastatic effects of antihypertension therapy.
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Bloqueadores de los Canales de Calcio , Neoplasias , Calcio , Bloqueadores de los Canales de Calcio/farmacología , Bloqueadores de los Canales de Calcio/uso terapéutico , Diltiazem , Células Endoteliales , Neoplasias/tratamiento farmacológico , Nifedipino/farmacología , Nifedipino/uso terapéutico , Verapamilo/farmacología , Verapamilo/uso terapéuticoRESUMEN
3D Pitch (out-of-plane) rotational motion has been generated in spherical particles by maneuvering the laser spots of holographic optical tweezers. However, since the spherical particles, which are required to minimise drag are perfectly isotropic, a controllable torque cannot be applied with it. It remains free to spin about any axis even after moving the tweezers beams. It is here that we trap birefringent particles of about 3 µm diameter in two tweezers beams and then change the depth of one of the beam foci controllably to generate a pitch rotational torque-wrench and avoid the free spinning of the particle. We also detect the rotation with newly developed pitch motion detection technique and apply controlled torques on the particle.
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Conventionally, the work of adhesion at the nanoscale is estimated using an atomic force microscope with a tip of the size of 10 nm. It is pressed into a surface with nano-Newton forces and then retracted to ascertain when the tip breaks away from the surface. Thus this ensures "hard probing" of a surface. However there can be another configuration where the particle is barely placed into the surface when the work of adhesion attaches the particle to the surface and this can be called "soft probing". In this configuration, if a birefringent particle is confined in linearly polarized optical tweezers, and then the surface is moved in the direction tangential to the plane, a rolling motion can be induced. Study of this rolling motion can also show the work of adhesion. We use this configuration to find the rolling work of adhesion of a 3 µm diameter birefringent particle on a glass surface. We go on to study the effects of changing the surface to a hydrophobic slippery surface like polydimethyl siloxane (PDMS). Then we go on to show that even 500 nm diameter diamonds bearing nitrogen vacancy (NV) centers which are birefringent due to the stresses on the crystal could also be trapped and rolled to generate pitch (out-of-plane rotation) motion with 50 nm contact diameters. We find that this mode of soft probing yields a work of adhesion of about 1 mJ m-2 while the conventional nanoscale probing with atomic force microscopes (AFM) yields about 50 mJ m-2.
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The multifunctional upconversion nanoparticles (UCNPs) are fascinating tool for biological applications. In the present work, photon upconverting NaGdF4:Yb,Er and Ag nanoparticles decorated NaGdF4:Yb,Er (NaGdF4:Yb,Er@Ag) nanoparticles were prepared using a simple polyol process. Rietveld refinement was performed for detailed crystal structural and phase fraction analysis. The morphology of the NaGdF4:Yb,Er@Ag was examined using high-resolution transmission electron microscope, which reveals silver nanoparticles of 8 nm in size were decorated over spherical shaped NaGdF4:Yb,Er nanoparticles with a mean particle size of 90 nm. The chemical compositions were confirmed by EDAX and inductively coupled plasma-optical emission spectrometry analyses. The upconversion luminescence (UCL) of NaGdF4:Yb,Er at 980 nm excitation showed an intense red emission. After incorporating the silver nanoparticles, the UCL intensity decreased due to weak scattering and surface plasmon resonance effect. The VSM magnetic measurement indicates both the UCNPs possess paramagnetic behaviour. The NaGdF4:Yb,Er@Ag showed computed tomography imaging. Magnetic resonance imaging study exhibited better T1 weighted relaxivity in the NaGdF4:Yb,Er than the commercial Gd-DOTA. For the first time, the optical trapping was successfully demonstrated for the upconversion NaGdF4:Yb,Er nanoparticle at near-infrared 980 nm light using an optical tweezer setup. The optically trapped UCNP possessing paramagnetic property exhibited a good optical trapping stiffness. The UCL of trapped single UCNP is recorded to explore the effect of the silver nanoparticles. The multifunctional properties for the NaGdF4:Yb,Er@Ag nanoparticle are demonstrated.
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We employ a single optically trapped upconverting nanoparticle (UCNP) of NaYF4:Yb,Er of diameter about 100 nm as a subdiffractive source to perform absorption spectroscopy. The experimentally expected mode volume of 100 nm of the backscatter profile of the nanoparticle matches well with a numerical simulation of the dominant backscattering modes to confirm our assertion of achieving a source dimension considerably lower than the diffraction limit set by the excitation wavelength of 975 nm for the UCNP. We perform absorption spectroscopy of several diverse entities such as the dye Rhodamine B in water, a thin gold film of thickness 30 nm, and crystalline soft oxometalates micro-patterned on a glass substrate using the UCNP as a source. The initial results lead to unambiguous utility of UCNPs as single nanoscopic sources for absorption spectroscopy of ultra-small sample volumes (femtolitres), and lead us to hypothesize a possible Resonance Energy Transfer mechanism between the UCNP and the molecules of the ambient medium, which may even lead to single molecule absorption spectroscopy applications.
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A microbubble nucleated due to the absorption of a tightly focused laser at the interface of a liquid-solid substrate enables directed and irreversible self-assembly of mesoscopic particles dispersed in the liquid at the bubble base. This phenomenon has facilitated a new microlithography technique which has grown rapidly over the past decade and can now reliably pattern a vast range of soft materials and colloids, ranging from polymers to metals to proteins. In this review, we discuss the science behind this technology and the present state-of-the-art. Thus, we describe the physics of the self-assembly driven by the bubble, the techniques for generating complex mesoarchitectures, both discrete and continuous, and their properties, and the various applications demonstrated in plastic electronics, site-specific catalysis, and biosensing. Finally, we describe a roadmap for the technique to achieve its potential of successfully patterning "everything" mesoscopic and the challenges that lie therein.
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3D pitch rotation of microparticles and cells assumes importance in a wide variety of applications in biology, physics, chemistry and medicine. Applications such as cell imaging and injection benefit from pitch-rotational manipulation. Generation of such motion in single beam optical tweezers has remained elusive due to the complexities of generating high enough ellipticity perpendicular to the direction of propagation. Further, trapping a perfectly spherical object at two locations and subsequent pitch rotation hasn't yet been demonstrated to be possible. Here, we use hexagonal-shaped upconverting particles and single cells trapped close to a gold-coated glass cover slip in a sample chamber to generate complete 360 degree and continuous pitch motion even with a single optical tweezer beam. The tweezers beam passing through the gold surface is partially absorbed and generates a hot-spot to produce circulatory convective flows in the vicinity which rotates the objects. The rotation rate can be controlled by the intensity of the laser light. Thus such a simple configuration can turn the particle in the pitch sense. The circulatory flows in this technique have a diameter of about 5 µm which is smaller than those reported using acousto-fluidic techniques.