Estimation of the proximal temperature rise of an excited upconversion particle by detecting the wavefront of emission.

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.

Comparison of thermal and athermal dynamics of the cell membrane slope fluctuations in the presence and absence of Latrunculin-B.

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.

High-resolution detection of pitch rotation in an optically confined hexagonal-shaped upconverting particle.

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.

Accelerated self assembly of particles at the air-water interface with optically assisted heating due to an upconverting particle.

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.

Anisotropic 3D confinement of MCF-7 cells induces directed cell-migration and viscoelastic anisotropy of cell-membrane.

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 ∼50µm-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.

Generation of partial roll rotation in a hexagonal NaYF4 particle by switching between different optical trapping configurations.

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.

A hydro-thermophoretic trap for microparticles near a gold-coated substrate.

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.

Directed Self-Assembly Driven Mesoscale Lithography Using Laser-Induced and Manipulated Microbubbles: Complex Architectures and Diverse Applications.

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.

Upconversion, MRI imaging and optical trapping studies of silver nanoparticle decorated multifunctional NaGdF4:Yb,Er nanocomposite.

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.

Detection of sub-degree angular fluctuations of the local cell membrane slope using optical tweezers.

Normal thermal fluctuations of the cell membrane have been studied extensively using high resolution microscopy and focused light, particularly at the peripheral regions of a cell. We use a single probe particle attached non-specifically to the cell-membrane to determine that the power spectral density is proportional to (frequency)-5/3 in the range of 5 Hz to 1 kHz. We also use a new technique to simultaneously ascertain the slope fluctuations of the membrane by relying upon the determination of pitch motion of the birefringent probe particle trapped in linearly polarized optical tweezers. In the process, we also develop the technique to identify pitch rotation to a high resolution using optical tweezers. We find that the power spectrum of slope fluctuations is proportional to (frequency)-1, which we also explain theoretically. We find that we can extract parameters like bending rigidity directly from the coefficient of the power spectrum particularly at high frequencies, instead of being convoluted with other parameters, thereby improving the accuracy of estimation. We anticipate this technique for determination of the pitch angle in spherical particles to high resolution as a starting point for many interesting studies using the optical tweezers.

Kinesin rotates unidirectionally and generates torque while walking on microtubules.

Cytoskeletal motors drive many essential cellular processes. For example, kinesin-1 transports cargo in a step-wise manner along microtubules. To resolve rotations during stepping, we used optical tweezers combined with an optical microprotractor and torsion balance using highly birefringent microspheres to directly and simultaneously measure the translocation, rotation, force, and torque generated by individual kinesin-1 motors. While, at low adenosine 5'-triphosphate (ATP) concentrations, motors did not generate torque, we found that motors translocating along microtubules at saturating ATP concentrations rotated unidirectionally, producing significant torque on the probes. Accounting for the rotational work makes kinesin a highly efficient machine. These results imply that the motor's gait follows a rotary hand-over-hand mechanism. Our method is generally applicable to study rotational and linear motion of molecular machines, and our findings have implications for kinesin-driven cellular processes.

Detection of self-generated nanowaves on the interface of an evaporating sessile water droplet.

Evaporating sessile droplets have been known to exhibit oscillations on the air-liquid interface. These are generally over millimeter scales. Using a novel approach, we are able to measure surface height changes of 500 nm amplitude using optical trapping of a set of microscopic particles at the interface, particularly when the vertical thickness of the droplet reduces to less than 50 µm. We find that at the later stages of the droplet evaporation, particularly when the convection currents become large, the top air-water interface starts to spontaneously oscillate vertically as a function of time in consistency with predictions. We also detect travelling wave trains moving in the azimuthal direction of the drop surface which are consistent with hydrothermal waves at a different combination of Reynolds, Prandtl and Evaporation numbers than previously observed. This is the first time that wave-trains have been observed in water, being extremely challenging to detect both interferometrically and with infra-red cameras. We also find that such waves apply a force parallel to the interface along the propagation direction.

Self-assembly and complex manipulation of colloidal mesoscopic particles by active thermocapillary stress.

We demonstrate that the active thermocapillary stresses induced by multiple microbubbles offer simple routes to directed self-assembly and complex but controllable micromanipulation of mesoscopic colloidal particles embedded in a liquid. The microbubbles are nucleated on a liquid-glass interface using optical tweezers. The flow around a single bubble causes self-assembly of the particles in rings at the bubble-base, while an asymmetric temperature profile generated across the bubble interface breaks the azimuthal symmetry of the flow, and induces simultaneous accumulation and repulsion of particles at different axial planes with respect to the bubble. The flow due to two adjacent bubbles leads to more diverse effects including the sorting of particles, and to local vorticity that causes radial and axial rotation of the particles - the latter being obtained for the first time using optical tweezers. The sorting is enabled by nucleating the bubbles on spatially discrete temperature profiles, while the vorticity is generated by nucleating them in the presence of a temperature gradient which once again causes a strong symmetry-breaking in the azimuthal flow. The flow profiles obtained in the experiments are explained by analytical solutions or qualitative explanations of the associated thermocapillary problem employing the Stokes and heat equations.

Phragmoplast Orienting Kinesin 2 Is a Weak Motor Switching between Processive and Diffusive Modes.

Plant development and morphology relies on the accurate insertion of new cell walls during cytokinesis. However, how a plant cell correctly orients a new wall is poorly understood. Two kinesin class-12 members, phragmoplast orienting kinesin 1 (POK1) and POK2, are involved in the process, but how these molecular machines work is not known. Here, we used in vivo and single-molecule in vitro measurements to determine how Arabidopsis thaliana POK2 motors function mechanically. We found that POK2 is a very weak, on average plus-end-directed, moderately fast kinesin. Interestingly, POK2 switches between processive and diffusive modes characterized by an exclusive-state mean-squared-displacement analysis. Our results support a model that POK motors push against peripheral microtubules of the phragmoplast for its guidance. This pushing model may mechanically explain the conspicuous narrowing of the division site. Together, our findings provide mechanical insight into how active motors accurately position new cell walls in plants.

Using Brownian motion to measure shape asymmetry in mesoscopic matter using optical tweezers.

We propose a new method for quantifying shape asymmetry on the mesoscopic scale. The method takes advantage of the intrinsic coupling between rotational and translational Brownian motion (RBM and TBM, respectively) which happens in the case of asymmetric particles. We determine the coupling by measuring different correlation functions of the RBM and TBM for single, morphologically different, weakly trapped red blood cells in optical tweezers. The cells have different degrees of asymmetry that are controllably produced by varying the hypertonicity of their aqueous environment. We demonstrate a clear difference in the nature of the correlation functions both qualitatively and quantitatively for three types of cells having a varying degree of asymmetry. This method can have a variety of applications ranging from early stage disease diagnosis to quality control in microfabrication.

Generation of microswimmers from passive Brownian particles in a spherically aberrated optical trap.

We induce spontaneous motion that is both directed and complex in micron-sized asymmetric Brownian particles in a spherically aberrated optical trap to generate microswimmers. The aberrated optical trap is prepared in a slightly modified optical tweezers configuration where we use a refractive index mismatched cover slip leading to the formation of an annular intensity distribution near the trap focal plane. Asymmetric scattering from a micro-particle trapped in this annular trap gives rise to a net tangential force on the particle causing it to revolve spontaneously in the intensity ring. The rate of revolution can be controlled from sub-Hz to a few Hz by changing the intensity of the trapping light. Theoretical simulations performed using finite-difference time-domain method verify the experimental observations. We also experimentally demonstrate simultaneous spin and revolution of a micro-swimmer which shows that complex motion can be achieved by designing a suitable shape of a micro-swimmer in the optical potential.

Simultaneous detection of rotational and translational motion in optical tweezers by measurement of backscattered intensity.

We describe a simple yet powerful technique of simultaneously measuring both translational and rotational motion of mesoscopic particles in optical tweezers by measuring the backscattered intensity on a quadrant photodiode (QPD). While the measurement of translational motion by taking the difference of the backscattered intensity incident on adjacent quadrants of a QPD is well known, we demonstrate that rotational motion can be measured very precisely by taking the difference between the diagonal quadrants. The latter measurement eliminates the translational component entirely and leads to a detection sensitivity of around 50 mdeg at S/N of 2 for angular motion of a driven microrod. The technique is also able to resolve the translational and rotational Brownian motion components of the microrod in an unperturbed trap and can be very useful in measuring translation-rotation coupling of micro-objects induced by hydrodynamic interactions.

Self-assembly of mesoscopic materials to form controlled and continuous patterns by thermo-optically manipulated laser induced microbubbles.

The formation of continuous patterns of nanostructured materials using directed self-assembly under external fields has generated considerable current research interest. We demonstrate for the first time such continuous patterning by inducing irreversible self-assembly leading to nucleation in mesocopic materials (inorganic, organic, and nanoparticles) using a tightly focused laser beam in an optical tweezers apparatus. A dense aqueous dispersion or solution of the material which has high absorption at the laser wavelength is taken in a sample holder so that some material is adsorbed on the top surface. A hot spot is created on the top surface when the adsorbed material absorbs the high intensity at the focus of the laser beam (a submicrometer sized spot), due to which a water vapor bubble is formed. This causes self-assembly of material around the bubble due to Gibbs-Marangoni convection and capillary flow after which the material eventually nucleates into a crystalline state. The bubble is "trapped" at the hot spot due to the temperature gradient around it and can be manipulated by thermal forces generated optically, so that the system may be described as a "thermo-optical" tweezers. We translate the trapped bubble using the microscope sample holder stage of the apparatus so that the nucleation site of the material is simultaneously translated generating continuous patterns. We have demonstrated the technique using exotic inorganic materials such as soft oxometalates, an organic material such as glycine, and a fluorescent dye such as perylene as well as with carbon nanotubes. We have written patterns over lengths of nearly 1 mm at the rate of 1 Hz, with best resolution of about 4 µm. The technique has potential for a wide range of applications ranging from solution processed printable electronics to controlled catalysis.

Comparison of translational and rotational modes towards passive rheology of the cytoplasm of MCF-7 cells using optical tweezers.

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.

Controlled roll rotation of a microparticle in a hydro-thermophoretic trap.

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.