Micron-sized Objects Research Articles

Averaging fractional Fourier domains for background noise removal applied to digital lensless holographic microscopy

A background noise removal method based on averaging fractional Fourier domains is presented. The method is applied to Digital Lensless Holographic Microscopy (DLHM) intensity reconstructions, where the background is perturbed by the weak yet detrimental presence of information of the twin image. A set of fractional Fourier domains of a DLHM intensity reconstruction is computed and thereafter averaged leading to a sensible reduction of the background noise and, therefore, an increase in the overall contrast of the resulting image. The maximum reach of the fractional rotations used in the method is determined by measuring the spatial resolution in a regular star test target such that the spatial resolution is kept within the span of interest for a given application. The set of images to be averaged is composed of fractional rotations of the original intensity reconstruction that are smaller than the previously determined maximum reach. The number of fractional rotations that are finally averaged is determined by the sought increase in the contrast of the resulting image. Experimental samples of micrometer-sized objects and an intricate biological specimen have been used to validate the proposal.

The evolution of organic material on Asteroid 162173 Ryugu and its delivery to Earth

The recent return of samples from asteroid 162173 Ryugu provides a first insight into early Solar System prebiotic evolution from known planetary bodies. Ryugu’s samples are CI chondrite-like, rich in water and organic material, and primarily composed of phyllosilicate. This phyllosilicate surrounds micron to submicron macromolecular organic particles known as insoluble organic matter. Using advanced microscopy techniques on Hayabusa-2 samples, we find that aqueous alteration on Ryugu produced organic particles richer in aromatics compared to less altered carbonaceous chondrites. This challenges the view that aromatic-rich organic matter formed pre-accretion. Additionally, widespread diffuse organic material occurs in phyllosilicate more aliphatic-, carboxylic-rich, and aromatic-poor than the discrete organic particles, likely preserving the soluble organic material. Some organic particles evolved to encapsulate phyllosilicate, indicating that aqueous alteration on Ryugu led to the containment of soluble organic matter within these particles. Earth therefore has been, and continues to be, delivered micron-sized polymeric organic objects containing biologically relevant molecules.

Multifunctional Hydrogel with 3D Printability, Fluorescence, Biodegradability, and Biocompatibility for Biomedical Microrobots.

As micron-sized objects, mobile microrobots have shown significant potential for future biomedical applications, such as targeted drug delivery and minimally invasive surgery. However, to make these microrobots viable for clinical applications, several crucial aspects should be implemented, including customizability, motion-controllability, imageability, biodegradability, and biocompatibility. Developing materials to meet these requirements is of utmost importance. Here, a gelatin methacryloyl (GelMA) and (2-(4-vinylphenyl)ethene-1,1,2-triyl)tribenzene (TPEMA)-based multifunctional hydrogel with 3D printability, fluorescence imageability, biodegradability, and biocompatibility is demonstrated. By using 3D direct laser writing method, the hydrogel exhibits its versatility in the customization and fabrication of 3D microstructures. Spherical hydrogel microrobots were fabricated and decorated with magnetic nanoparticles on their surface to render them magnetically responsive, and have demonstrated excellent movement performance and motion controllability. The hydrogel microstructures also represented excellent drug loading/release capacity and degradability by using collagenase, along with stable fluorescence properties. Moreover, cytotoxicity assays showed that the hydrogel was non-toxic, as well as able to support cell attachment and growth, indicating excellent biocompatibility of the hydrogel. The developed multifunctional hydrogel exhibits great potential for biomedical microrobots that are integrated with customizability, 3D printability, motion controllability, drug delivery capacity, fluorescence imageability, degradability, and biocompatibility, thus being able to realize the real in vivo biomedical applications of microrobots.

Tunable viscoelastic size-based particle separation in straight microchannels with triangular cross-sections

In recent years, viscoelastic particle manipulation within microfluidic systems has received much attention due to the ease with which micron-sized objects may be maneuvered and isolated on the basis of size. While several factors, including both fluid and particle properties, regulate the precise locations and trajectories of micron-sized species flowing along a microchannel, the role of channel cross-section shape is critical, since it directly influences the fluid velocity profile and thus the direction and magnitude of hydrodynamic forces. It is therefore surprising that this parameter has not been comprehensively investigated for cell-based separations, especially since most viscoelastic microfluidic systems are only able to efficiently sperate cells over limited size ranges. To address this shortcoming, we present a viscoelastic microfluidic system integrating a triangular cross-section microchannel, for efficient and tunable size-based separations of micron-sized species. We find that particle focusing patterns can be controlled by simple variation of volumetric flow rates, which allows for the efficient separation of particles and cells of variable size. To showcase the efficacy of the approach, we present a size-based separation of various blood components, including white blood cells, platelets, and rare cells. By quantifying the number of particles collected at the outlets, we achieve recovery efficiencies of over 98%.

Enhancement of optical levitation with hyperbolic metamaterials

The tightly focused laser beam in an optical trap has become a useful tool for many recent research areas. The momentum change in the photon-stream path of incident laser beam induces radiation force that enables trapping and manipulating mesoscopic micron-sized objects. In this study, we report the first analytical demonstration of optical trapping and levitation with radiation pressure on a transparent micron-sized spherical object made of hyperbolic metamaterial (HMM). The optical radial and axial forces acting on dielectric and HMM spherical particles are calculated using ray-optics approximation, assuming an optical levitation trapping setup. We compared the net force acting on the two objects, finding that the net radiation force exerted towards HMM particle is enhanced in the axial direction: The optical force enhancement in the HMM particle is more than ~ 8 times stronger compared to the induced force on the conventional dielectric particle with the corresponding material parameters. Besides, a better performance in the radial stabilization is observed for the HMM particle in comparison with the dielectric case, at which some oscillations and unstable saturation locations for the radial stabilization is monitored for TEM00 beam incidence. Furthermore, “zero-force” paths where radial stabilization of the HMM particle exists are also obtained for both TEM00 and TEM01∗\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$TEM_{01}^{*}$$\\end{document} laser beam incidences. Such phenomenon does not occur for particles of only dielectric and only metal material, which can be considered as another superiority of the proposed HMM particle.

Co-Expression of type 1 fimbriae and flagella in Escherichia coli: consequences for adhesion at interfaces.

Escherichia coli expresses surface appendages including fimbriae, flagella, and curli, at various levels in response to environmental conditions and external stimuli. Previous studies have revealed an interplay between expression of fimbriae and flagella in several E. coli strains, but how this regulation between fimbrial and flagellar expression affects adhesion to interfaces is incompletely understood. Here, we investigate how the concurrent expression of fimbriae and flagella by engineered strains of E. coli MG1655 affects their adhesion at liquid-solid and liquid-liquid interfaces. We tune fimbrial and flagellar expression on the cell surface through plasmid-based inducible expression of the fim operon and fliC-flhDC genes. We show that increased fimbrial expression increases interfacial adhesion as well as bacteria-driven actuation of micron-sized objects. Co-expression of flagella in fimbriated bacteria, however, does not greatly affect either of these properties. Together, these results suggest that interfacial adhesion as well as motion actuated by adherent bacteria can be altered by controlling the expression of surface appendages.

Motion of an active bent rod with an articulating hinge: exploring mechanical and chemical modes of swimming

Swimming at the microscale typically involves two modes of motion: mechanical propulsion and propulsion due to field interactions. During mechanical propulsion, particles swim by reconfiguring their geometry. When propelled by field interactions, body forces such as phoretic interactions drive mobility. In this work, we employ slender-body theory to explore how a bent rod actuator propels due to a mechanical mode of swimming via hinge articulations and due to a chemical mode of swimming via diffusiophoretic interactions with a solute field. Although previous theoretical studies have examined mechanical and chemical modes of swimming in isolation, the simultaneous investigation of both modes has remained unexplored. For the mechanical mode of swimming, our calculations, both numerical and analytical, recover Purcell’s scallop theorem and show that the bent rod actuator experiences zero net displacement during reciprocal motion. Additionally, we calculate the trajectories traced by a bent rod actuator under a non-reciprocal hinge articulation, revealing that these trajectories are influenced by the amplitude of the hinge articulation, geometric asymmetry, and the angular velocity distribution between the two arms of the bent rod actuator. We provide intuitive explanations for these effects using free-body diagrams. Furthermore, we explore the motion induced by simultaneous hinge articulations and self-diffusiophoresis. We observe that hinge articulations can modify the effective phoretic forces and torques acting on the bent rod actuator, either supporting or impeding propulsion. Additionally, during self-diffusiophoretic propulsion, reciprocal hinge articulations no longer result in zero net displacement. In summary, our findings chart a new direction for designing micron-sized objects that harness both mechanical and chemical modes of propulsion synchronously, offering a mechanism to enact control over trajectories.

Photothermal heterodyne imaging of micron-sized objects.

Micron-sized dye-doped polymer beads were imaged using transmitted/reflected light microscopy and photothermal heterodyne imaging (PHI) measurements. The transmitted/reflected light images show distinct ring patterns that are attributed to diffraction effects and/or internal reflections within the beads. In the PHI experiments pump laser induced heating changes the refractive index and size of the bead, which causes changes in the diffraction pattern and internal reflections. This creates an analogous ring pattern in the PHI images. The ring pattern disappears in both the reflected light and PHI experiments when an incoherent light source is used as a probe. When the beads are imaged in an organic medium heat transfer changes the refractive index of the environment, and gives rise to a ring pattern external to the beads in the PHI images. This causes the beads to appear larger than their physical dimensions in PHI experiments. This external signal does not appear when the beads are imaged in air because the refractive index changes in air are very small.

Designing Ordered Organic Small-Molecule Domains for Ultraviolet Detection and Micrometer-Sized Flexible Imaging.

Photodetectors displaying an ultraviolet (UV) spectral response window are typically based on wide-bandgap semiconductors that have long been dominated by inorganic materials that suffer from bottlenecks of low flexibility and a limited material family. Here, we synthesized a novel organic small molecule and controlled its crystallization to suppress leakage currents and facilitate separation of the carriers, and the relationship between the nanoscale phase separation morphology and the optoelectrical performance of the photodetectors is disclosed. Our optimized organic photodetector (OPD) presents a UV spectral response window, with superior self-powered responsivities of 45 mA/W (under 250 nm light) and 70 mA/W (under 300 nm light), outperforming the Si photodiode and rivaling other reported UV self-powered photodetectors. Finally, an imaging system was constructed to demonstrate the application potential of the OPD in UV flexible imaging with high-resolution arrays of 400 pixels × 400 pixels (5 μm × 5 μm per pixel), which could work in bent states and successfully output images of micrometer-sized objects.

Endocytosis-Like Vesicle Fission Mediated by a Membrane-Expanding Molecular Machine Enables Virus Encapsulation for In Vivo Delivery.

Biological membranes are functionalized by membrane-associated protein machinery. Membrane-associated transport processes, such as endocytosis, represent a fundamental and universal function mediated by membrane-deforming protein machines, by which small biomolecules and even micrometer-size substances can be transported via encapsulation into membrane vesicles. Although synthetic molecules that induce dynamic membrane deformation have been reported, a molecular approach enabling membrane transport in which membrane deformation is coupled with substance binding and transport remains critically lacking. Here, we developed an amphiphilic molecular machine containing a photoresponsive diazocine core (AzoMEx) that localizes in a phospholipid membrane. Upon photoirradiation, AzoMEx expands the liposomal membrane to bias vesicles toward outside-in fission in the membrane deformation process. Cargo components, including micrometer-size M13 bacteriophages that interact with AzoMEx, are efficiently incorporated into the vesicles through the outside-in fission. Encapsulated M13 bacteriophages are transiently protected from the external environment and therefore retain biological activity during distribution throughout the body via the blood following administration. This research developed a molecular approach using synthetic molecular machinery for membrane functionalization to transport micrometer-size substances and objects via vesicle encapsulation. The molecular design demonstrated in this study to expand the membrane for deformation and binding to a cargo component can lead to the development of drug delivery materials and chemical tools for controlling cellular activities.

Acoustic radiation forces and torques on elastic micro rings in standing waves

Acoustic radiation forces and torques are very effective in manipulating micron-sized objects such as cells, droplets, particles, and organisms. In this work, we present analysis of acoustic radiation forces and torques on ring-shaped slender microstructures under a standing wave in an inviscid fluid using on a three-dimensional finite-element-method (FEM) model. The influence of geometric and physical parameters on the radiation forces and torques is characterized. The radiation force tends to push the rings towards the pressure nodes or anti-nodes depending on the contrast factor and exhibits a volumetric dependence in magnitude on geometric parameters. Moreover, a nonzero net torque develops when the ring is not co-planar with plane waves and varies in magnitude and direction depending on the position, orientation, and material properties of the ring. Large variations are observed only in torque values for specific combinations of geometric and physical parameters as an indicative of resonance. Furthermore, the FEM results are compared with a reduced-order model called chain-of-spheres, which works well in estimating the radiation forces at a fraction of the computational cost but deviates significantly in torque evaluations. Lastly, a segmented ring is used to understand the relative effect of secondary forces due to self-scattering. The findings of the study are applicable to development of acoustic manipulation systems for ring-like elastic microfilaments and slender bodies with arbitrary shapes and orientations. These results also can be used in directional reinforcing of ring-shaped composites.

Electrostatic manipulation for saturated charging and orientating of recycled short-chopped carbon fibers based on dielectric polarization

Carbon fiber composites produce a large amount of scrap during their life cycle, to recycle and fully utilize these short-chopped carbon fibers (SCFs), present work proposes a method to disperse agglomerated SCFs using pure electrostatic forces, while using dielectric polarization to recycle and emit SCFs. SCFs are disorderly distributed or agglomerated into pellets in their natural state, and they need to be sufficiently charged and oriented for reliable collection. A novel plate-plate structure is proposed to maintain a higher load voltage to disperse and emit a larger amount of SCFs. The charging and orientation mechanisms of SCFs were investigated with high-speed camera. The dispersive characteristics of SCFs were statistically evaluated with Ripley's K function and Two-point spatial correlation. And SCFs were emitted by two different modes and the effects of embedment were studied. The results show that large-scale SCFs can be uniformly dispersed and sufficiently charged to 2 × 10−2C/kg in the electrostatic field and oriented to move under the driving of dielectric polarization. Moreover, the direction of embedded SCFs can be well controlled. This study describes the behavior pattern and motion characteristics of SCFs in the electrostatic field and provides a new method for the pollution-free, low-cost, efficient recycling of recycled fibers. It has broad application prospects in the field of carbon fiber reinforced polymer composites and is also significant for the pure electrostatic cross-scale control of micron-sized objects.

Optimal Cargo Size for Active Diffusion of Biohybrid Microcarriers

As society paves its way towards device miniaturization and precision medicine, microscale actuation and transport become increasingly prominent research fields with high impact in both technological and clinical contexts. In order to accomplish movement of micron-sized objects towards specific target sites, active biohybrid transport systems, such as motile living cells that act as smart biochemically powered microcarriers, have been suggested as an alternative to synthetic microrobots. Inspired by the motility of leukocytes, we propose the amoeboid crawling of eukaryotic cells as a promising mechanism for transport of micron-sized cargoes and present an in-depth study of this type of composite active matter. Its transport properties result from the interactions of an active element (cell) and a passive one (cargo) and reveal an optimal cargo size that enhances the locomotion of the load-carrying cells, even exceeding their motility in the absence of cargo. The experimental findings are rationalized in terms of a biohybrid active particle model that describes the emergent cell-cargo dynamics and enables us to derive the long-time diffusive transport of amoeboid microcarriers. As amoeboid locomotion is commonly observed for mammalian cells such as leukocytes, our results lay the foundations for the study of transport performance of other medically relevant cell types and for extending our findings to more advanced transport tasks in complex environments, such as tissues.

MiGriBot: A miniature parallel robot with integrated gripping for high-throughput micromanipulation.

Although robotic micromanipulation using microtweezers has been widely explored, the current manipulation throughput hardly exceeds one operation per second. Increasing the manipulation throughput is thus a key factor for the emergence of robotized microassembly industries. This article presents MiGriBot (Millimeter Gripper Robot), a miniaturized parallel robot with a configurable platform and soft joints, designed to perform pick-and-place operations at the microscale. MiGriBot combines in a single robot the benefits of a parallel kinematic architecture with a configurable platform and the use of soft joints at the millimeter scale. The configurable platform of the robot provides an internal degree of freedom that can be used to actuate microtweezers using piezoelectric bending actuators located at the base of the robot, which notably reduces the robot's inertia. The soft joints make it possible to miniaturize the mechanism and to avoid friction. These benefits enable MiGriBot to reach a throughput of 10 pick-and-place cycles per second of micrometer-sized objects, with a precision of 1 micrometer.

Optical diffraction tomography of 3D microstructures using a low coherence source.

Optical diffraction tomography (ODT) is a label-free technique for three dimensional imaging of micron-sized objects. Coherence and limited sampling of 3D Fourier space are often responsible for the appearance of artifacts. Here we present an ODT microscope that uses low temporal coherence light and spatial light modulators to retrieve reliable 3D maps of the refractive index. A common-path interferometer, based on a spatial light modulator, measures the complex fields transmitted by a sample. Measured fields, acquired while scanning the illumination direction using a digital micro-mirror device, are fed into a Rytov reconstruction algorithm to obtain refractive index maps whose accuracy is directly evaluated on microfabricated 3D test objects. Even for challenging shapes such as pyramids, bridges, and dumbbells, we obtain volumetric reconstructions that compare very well with electron microscopy images.

Molecular Diffusivity of Click Reaction Components: The Diffusion Enhancement Question.

Micrometer-sized objects are widely known to exhibit chemically driven motility in systems away from equilibrium. Experimental observation of reaction-induced motility or enhancement in diffusivity at the much shorter length scale of small molecules is, however, still a matter of debate. Here, we investigate the molecular diffusivity of reactants, catalyst, and product of a model reaction, the copper-catalyzed azide–alkyne cycloaddition click reaction, and develop new NMR diffusion approaches that allow the probing of reaction-induced diffusion enhancement in nanosized molecular systems with higher accuracy than the state of the art. Following two different approaches that enable the accounting of time-dependent concentration changes during NMR experiments, we closely monitored the diffusion coefficient of reaction components during the reaction. The reaction components showed distinct changes in the diffusivity: while the two reactants underwent a time-dependent decrease in their diffusivity, the diffusion coefficient of the product gradually increased and the catalyst showed only slight diffusion enhancement within the range expected for reaction-induced sample heating. The decrease in diffusion coefficient of the alkyne, one of the two reactants of click reaction, was not reproduced during its copper coordination when the second reactant, azide, was absent. Our results do not support the catalysis-induced diffusion enhancement of the components of the click reaction and, instead, point to the role of a relatively large intermediate species within the reaction cycle with diffusivity lower than that of both the reactants and product molecule.

Protein conformation and biomolecular condensates.

Protein conformation and cell compartmentalization are fundamental concepts and subjects of vast scientific endeavors. In the last two decades, we have witnessed exciting advances that unveiled the conjunction of these concepts. An avalanche of studies highlighted the central role of biomolecular condensates in membraneless subcellular compartmentalization that permits the spatiotemporal organization and regulation of myriads of simultaneous biochemical reactions and macromolecular interactions. These studies have also shown that biomolecular condensation, driven by multivalent intermolecular interactions, is mediated by order-disorder transitions of protein conformation and by protein domain architecture. Conceptually, protein condensation is a distinct level in protein conformational landscape in which collective folding of large collections of molecules takes place. Biomolecular condensates arise by the physical process of phase separation and comprise a variety of bodies ranging from membraneless organelles to liquid condensates to solid-like conglomerates, spanning lengths from mesoscopic clusters (nanometers) to micrometer-sized objects. In this review, we summarize and discuss recent work on the assembly, composition, conformation, material properties, thermodynamics, regulation, and functions of these bodies. We also review the conceptual framework for future studies on the conformational dynamics of condensed proteins in the regulation of cellular processes.

Exploiting the Structural Metamorphosis of Polymers to 'Wrap' Micron-Sized Spherical Objects.

There is growing interest in developing methods to 'wrap' nano- and micron-sized biological objects within films that may offer protection, enhance their stability or improve performance. We describe the successful 'wrapping' of lectin-decorated microspheres, which serve as appealing model micron-sized objects, within cross-linked polymer film. This approach utilizes polymer chains able to undergo a structural metamorphosis, from being intramolecularly cross-linked to intermolecularly cross-linked, a process that is triggered by polymer concentration upon the particle surface. Experiments demonstrate that both complementary molecular recognition and the dynamic covalent nature of the crosslinker are required for successful 'wrapping' to occur. This work is significant as it suggests that nano- and micron-sized biological objects such as virus-like particles, bacteria or mammalian cells-all of which may benefit from additional environmental protection or stabilization in emerging applications-may also be 'wrapped' by this approach.

Size control of shape switchable micronetworks by fast two-step microfluidic templating

Shape-memory polymer micronetworks (MN) are micrometer-sized objects that can switch their outer shape upon external command. This study aims to scale MN sizes to the low micrometer range at very narrow size distributions. In a two-step microfluidic strategy, the specific design of coaxial class capillary devices allowed stabilizing the thread of the dispersed phase to efficiently produce precursor particles in the tip-streaming regime at rates up to ~ 170 kHz and final sizes down to 4 µm. In a subsequent melt-based microfluidic photocrosslinking of the methacrylate-functionalized oligo(ɛ-caprolactone) precursor material, MN could be produced without particle aggregation. A comprehensive analysis of MN properties illustrated successful crosslinking, semi-crystalline morphology, and a shape-switching functionality for all investigated MN sizes (4, 6, 9, 12, 22 µm). Such functional micronetworks tailored to and below the dimension of cells can enable future applications in technology and medicine like controlling cell interaction.Graphic abstract

Designing a High-Resolution, LEGO-Based Microscope for an Educational Setting

ABSTRACT Microscopy is an essential tool in many fields of science. However, because of costs and fragility, the usage of microscopes is limited in classroom settings and nearly absent at home. In this article we present the construction of a microscope made of LEGO® bricks and low-cost, easily available lenses. We demonstrate that the obtained magnification and resolution are sufficient to resolve micrometer-sized objects and propose a series of experiments that explore various biophysical principles. Finally, a study with students in the age range of 9 to 13 y shows that the understanding of microscopy increases significantly after working with the LEGO microscope.