Minimally designed thermo-magnetic dual responsive soft robots for complex applications.

The fabrication of thermo-magnetic dual-responsive soft robots often requires intricate designs to implement complex locomotion patterns and utilize the implemented responsive behaviors. This work demonstrates a minimally designed soft robot based on poly-N-isopropylacrylamide (pNIPAM) and ferromagnetic particles, showcasing excellent control over both thermo- and magnetic responses. Free radical polymerization enables the magnetic particles to be entrapped homogeneously within the polymeric network. The integration of magnetic shape programming and temperature response allows the robot to perform various tasks including shaping, locomotion, pick-and-place, and release maneuvers of objects using independent triggers. The robot can be immobilized in a gripping state through magnetic actuation, and a subsequent increase in temperature transitions the robot from a swollen to a collapsed state. The temperature switch enables the robot to maintain a secured configuration while executing other movements via magnetic actuation. This approach offers a straightforward yet effective solution for achieving full control over both stimuli in dual-responsive soft robotics.

Two-photon microscopy for microrobotics: Visualization of micro-agents below fixed tissue.

Optical microscopy is frequently used to visualize microrobotic agents (i.e., micro-agents) and physical surroundings with a relatively high spatio-temporal resolution. However, the limited penetration depth of optical microscopy techniques used in microrobotics (in the order of 100 µm) reduces the capability of visualizing micro-agents below biological tissue. Two-photon microscopy is a technique that exploits the principle of two-photon absorption, permitting live tissue imaging with sub-micron resolution and optical penetration depths (over 500 µm). The two-photon absorption principle has been widely applied to fabricate sub-millimeter scale components via direct laser writing (DLW). Yet, its use as an imaging tool for microrobotics remains unexplored in the state-of-the-art. This study introduces and reports on two-photon microscopy as an alternative technique for visualizing micro-agents below biological tissue. In order to validate two-photon image acquisition for microrobotics, two-type micro-agents are fabricated and employed: (1) electrospun fibers stained with an exogenous fluorophore and (2) bio-inspired structure printed with autofluorescent resin via DLW. The experiments are devised and conducted to obtain three-dimensional reconstructions of both micro-agents, perform a qualitative study of laser-tissue interaction, and visualize micro-agents along with tissue using second-harmonic generation. We experimentally demonstrate two-photon microscopy of micro-agents below formalin-fixed tissue with a maximum penetration depth of 800 µm and continuous imaging of magnetic electrospun fibers with one frame per second acquisition rate (in a field of view of 135 × 135 µm2). Our results show that two-photon microscopy can be an alternative imaging technique for microrobotics by enabling visualization of micro-agents under in vitro and ex ovo conditions. Furthermore, bridging the gap between two-photon microscopy and the microrobotics field has the potential to facilitate in vivo visualization of micro-agents.

Locally Addressable Energy Efficient Actuation of Magnetic Soft Actuator Array Systems.

Advances in magnetoresponsive composites and (electro-)magnetic actuators have led to development of magnetic soft machines (MSMs) as building blocks for small-scale robotic devices. Near-field MSMs offer energy efficiency and compactness by bringing the field source and effectors in close proximity. Current challenges of near-field MSM are limited programmability of effector motion, dimensionality, ability to perform collaborative tasks, and structural flexibility. Herein, a new class of near-field MSMs is demonstrated that combines microscale thickness flexible planar coils with magnetoresponsive polymer effectors. Ultrathin manufacturing and magnetic programming of effectors is used to tailor their response to the nonhomogeneous near-field distribution on the coil surface. The MSMs are demonstrated to lift, tilt, pull, or grasp in close proximity to each other. These ultrathin (80 µm) and lightweight (100 gm-2 ) MSMs can operate at high frequency (25 Hz) and low energy consumption (0.5 W), required for the use of MSMs in portable electronics.

A Magnetic Bio-Inspired Soft Carrier as a Temperature-Controlled Gastrointestinal Drug Delivery System.

Currently, gastrointestinal bleeding in the colon wall and the small bowel is diagnosed and treated with endoscopes. However, the locations of this condition are often problematic to treat using traditional flexible and tethered tools. New studies commonly consider untethered devices for solving this problem. However, there still exists a gap in the extant literature, and more research is needed to diagnose and deliver drugs in the lower gastrointestinal tract using soft robotic carriers. This paper discusses the development of an untethered, magnetically-responsive bio-inspired soft carrier. A molding process is utilized to produce prototypes from Diisopropylidene-1,6-diphenyl-1,6-hexanediol-based Polymer with Ethylene Glycol Dimethacrylate (DiAPLEX) MP-3510 - a shape memory polymer with a low transition temperature to enable the fabrication of these carriers. The soft carrier design is validated through simulation results of deformation caused by magnetic elements embedded in the carrier in response to an external field. The thermal responsiveness of the fabricated prototype carriers is assessed ex vivo and in a phantom. The results indicate a feasible design capable of administering drugs to a target inside a phantom of a large intestine. The soft carrier introduces a method for the controlled release of drugs by utilizing the rubbery modulus of the polymer and increasing the recovery force through magnetic actuation.

Locomotion of bovine spermatozoa during the transition from individual cells to bundles.

Various locomotion strategies employed by microorganisms are observed in complex biological environments. Spermatozoa assemble into bundles to improve their swimming efficiency compared to individual cells. However, the dynamic mechanisms for the formation of sperm bundles have not been fully characterized. In this study, we numerically and experimentally investigate the locomotion of spermatozoa during the transition from individual cells to bundles of two cells. Three consecutive dynamic behaviors are found across the course of the transition: hydrodynamic attraction/repulsion, alignment, and synchronization. The hydrodynamic attraction/repulsion depends on the relative orientation and distance between spermatozoa as well as their flagellar wave patterns and phase shift. Once the heads are attached, we find a stable equilibrium of the rotational hydrodynamics resulting in the alignment of the heads. The synchronization results from the combined influence of hydrodynamic and mechanical cell-to-cell interactions. Additionally, we find that the flagellar beat is regulated by the interactions during the bundle formation, whereby spermatozoa can synchronize their beats to enhance their swimming velocity.

Magnetic Soft Helical Manipulators with Local Dipole Interactions for Flexibility and Forces.

Magnetic continuum manipulators (MCMs) are a class of continuum robots that can be actuated without direct contact by an external magnetic field. MCMs operating in confined workspaces, such as those targeting medical applications, require flexible magnetic structures that contain combinations of magnetic components and polymers to navigate long and tortuous paths. In cylindrical MCM designs, a significant trade-off exists between magnetic moment and bending flexibility as the ratio between length and diameter decreases. In this study, we propose a new MCM design framework that enables increasing diameter without compromising on flexibility and magnetic moment. Magnetic soft composite helices constitute bending regions of the MCM and are separated by permanent ring magnets. Local dipole interactions between the permanent magnets can reduce bending stiffness, depending on their size and spacing. For the particular segment geometry presented herein, the local dipole interactions result in a 31% increase in angular deflection of composite helices inside an external magnetic field, compared to helices without local interactions. In addition, we demonstrate fabrication, maneuverability, and example applications of a multisegment MCM in a phantom of the abdominal aorta, such as passing contrast dye and guidewires.

Flagellar Propulsion of Sperm Cells Against a Time-Periodic Interaction Force.

Sperm cells undergo complex interactions with external environments, such as a solid-boundary, fluid flow, as well as other cells before arriving at the fertilization site. The interaction with the oviductal epithelium, as a site of sperm storage, is one type of cell-to-cell interaction that serves as a selection mechanism. Abnormal sperm cells with poor swimming performance, the major cause of male infertility, are filtered out by this selection mechanism. In this study, collinear bundles, consisting of two sperm cells, generate propulsive thrusts along opposite directions and allow to observe the influence of cell-to-cell interaction on flagellar wave-patterns. The developed elasto-hydrodynamic model demonstrates that steric and adhesive forces lead to highly symmetrical wave-pattern and reduce the bending amplitude of the propagating wave. It is measured that the free cells exhibit a mean flagellar curvature of 6.4 ± 3.5 rad mm-1 and a bending amplitude of 13.8 ± 2.8 rad mm-1 . After forming the collinear bundle, the mean flagellar curvature and bending amplitude are decreased to 1.8 ± 1.1 and 9.6 ± 1.4 rad mm-1 , respectively. This study presents consistent theoretical and experimental results important for understanding the adaptive behavior of sperm cells to the external time-periodic force encountered during sperm-egg interaction.

Embedded 3D printing of dilute particle suspensions into dense complex tissue fibers using shear thinning xanthan baths.

In order to fabricate functional organoids and microtissues, a high cell density is generally required. As such, the placement of cell suspensions in molds or microwells to allow for cell concentration by sedimentation is the current standard for the production of organoids and microtissues. Even though molds offer some level of control over the shape of the resulting microtissue, this control is limited as microtissues tend to compact towards a sphere after sedimentation of the cells. 3D bioprinting on the other hand offers complete control over the shape of the resulting structure. Even though the printing of dense cell suspensions in the ink has been reported, extruding dense cellular suspensions is challenging and generally results in high shear stresses on the cells and a poor shape fidelity of the print. As such, additional materials such as hydrogels are added in the bioink to limit shear stresses, and to improve shape fidelity and resolution. The maximum cell concentration that can be incorporated in a hydrogel-based ink before the ink's rheological properties are compromised, is significantly lower than the concentration in a tissue equivalent. Additionally, the hydrogel components often interfere with cellular self-assembly processes. To circumvent these limitations, we report a simple and inexpensive xanthan bath based embedded printing method to 3D print dense functional linear tissues using dilute particle suspensions consisting of cells, spheroids, hydrogel beads, or combinations thereof. Using this method, we demonstrated the self-organization of functional cardiac tissue fibers with a layer of epicardial cells surrounding a body of cardiomyocytes.

Visualization of micro-agents and surroundings by real-time multicolor fluorescence microscopy.

Optical microscopy techniques are a popular choice for visualizing micro-agents. They generate images with relatively high spatiotemporal resolution but do not reveal encoded information for distinguishing micro-agents and surroundings. This study presents multicolor fluorescence microscopy for rendering color-coded identification of mobile micro-agents and dynamic surroundings by spectral unmixing. We report multicolor microscopy performance by visualizing the attachment of single and cluster micro-agents to cancer spheroids formed with HeLa cells as a proof-of-concept for targeted drug delivery demonstration. A microfluidic chip is developed to immobilize a single spheroid for the attachment, provide a stable environment for multicolor microscopy, and create a 3D tumor model. In order to confirm that multicolor microscopy is able to visualize micro-agents in vascularized environments, in vitro vasculature network formed with endothelial cells and ex ovo chicken chorioallantoic membrane are employed as experimental models. Full visualization of our models is achieved by sequential excitation of the fluorophores in a round-robin manner and synchronous individual image acquisition from three-different spectrum bands. We experimentally demonstrate that multicolor microscopy spectrally decomposes micro-agents, organic bodies (cancer spheroids and vasculatures), and surrounding media utilizing fluorophores with well-separated spectrum characteristics and allows image acquisition with 1280 [Formula: see text] 1024 pixels up to 15 frames per second. Our results display that real-time multicolor microscopy provides increased understanding by color-coded visualization regarding the tracking of micro-agents, morphology of organic bodies, and clear distinction of surrounding media.

Intravascular Tracking of Micro-Agents Using Medical Ultrasound: Towards Clinical Applications.

OBJECTIVE: This study demonstrates intravascular micro-agent visualization by utilizing robotic ultrasound-based tracking and visual servoing in clinically-relevant scenarios. METHODS: Visual servoing path is planned intraoperatively using a body surface point cloud acquired with a 3D camera and the vessel reconstructed from ultrasound (US) images, where both the camera and the US probe are attached to the robot end-effector. Developed machine vision algorithms are used for detection of micro-agents from minimal size of 250 µm inside the vessel contour and tracking with error recovery. Finally, real-time positions of the micro-agents are used for servoing of the robot with the attached US probe. Constant contact between the US probe and the surface of the body is accomplished by means of impedance control. RESULTS: Breathing motion is compensated to keep constant contact between the US probe and the body surface, with minimal measured force of 2.02 N. Anthropomorphic phantom vessels are segmented with an Intersection-Over-Union (IOU) score of 0.93 ± 0.05, while micro-agent tracking is performed with up to 99.8% success rate at 28-36 frames per second. Path planning, tracking and visual servoing are realized over 80 mm and 120 mm long surface paths. CONCLUSION: Experiments performed using anthropomorphic surfaces, biological tissue, simulation of physiological movement and simulation of fluid flow through the vessels indicate that robust visualization and tracking of micro-agents involving human patients is an achievable goal.

SonoTweezer: An Acoustically Powered End-Effector for Underwater Micromanipulation.

Recent advances in contactless micromanipulation strategies have revolutionized prospects of robotic manipulators as next-generation tools for minimally invasive surgeries. In particular, acoustically powered phased arrays offer dexterous means of manipulation both in air and water. Inspired by these phased arrays, we present SonoTweezer: a compact, low-power, and lightweight array of immersible ultrasonic transducers capable of trapping and manipulation of sub-mm sized agents underwater. Based on a parametric investigation with numerical pressure field simulations, we design and create a six-transducer configuration, which is small compared to other reported multi-transducer arrays (16-256 elements). Despite the small size of array, SonoTweezer can reach pressure magnitudes of 300 kPa at a low supply voltage of 25 V to the transducers, which is in the same order of absolute pressure as multi-transducer arrays. Subsequently, we exploit the compactness of our array as an end-effector tool for a robotic manipulator to demonstrate long-range actuation of sub-millimeter agents over a hundred times the agent's body length. Furthermore, a phase-modulation over its individual transducers allows our array to locally maneuver its target agents at sub-mm steps. The ability to manipulate agents underwater makes SonoTweezer suitable for clinical applications considering water's similarity to biological media, e.g., vitreous humor and blood plasma. Finally, we show trapping and manipulation of micro-agents under medical ultrasound (US) imaging modality. This application of our actuation strategy combines the usage of US waves for both imaging and micromanipulation.

CeFlowBot: A Biomimetic Flow-Driven Microrobot that Navigates under Magneto-Acoustic Fields.

Aquatic organisms within the Cephalopoda family (e.g., octopuses, squids, cuttlefish) exist that draw the surrounding fluid inside their bodies and expel it in a single jet thrust to swim forward. Like cephalopods, several acoustically powered microsystems share a similar process of fluid expulsion which makes them useful as microfluidic pumps in lab-on-a-chip devices. Herein, an array of acoustically resonant bubbles are employed to mimic this pumping phenomenon inside an untethered microrobot called CeFlowBot. CeFlowBot contains an array of vibrating bubbles that pump fluid through its inner body thereby boosting its propulsion. CeFlowBots are later functionalized with magnetic layers and steered under combined influence of magnetic and acoustic fields. Moreover, acoustic power modulation of CeFlowBots is used to grasp nearby objects and release it in the surrounding workspace. The ability of CeFlowBots to navigate remote environments under magneto-acoustic fields and perform targeted manipulation makes such microrobots useful for clinical applications such as targeted drug delivery. Lastly, an ultrasound imaging system is employed to visualize the motion of CeFlowBots which provides means to deploy such microrobots in hard-to-reach environments inaccessible to optical cameras.

Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.

Multicolor fluorescence microscopy is a powerful technique to fully visualize many biological phenomena by acquiring images from different spectrum channels. This study expands the scope of multicolor fluorescence microscopy by serial imaging of polystyrene micro-beads as surrogates for drug carriers, cancer spheroids formed using HeLa cells, and microfluidic channels. Three fluorophores with different spectral characteristics are utilized to perform multicolor microscopy. According to the spectrum analysis of the fluorophores, a multicolor widefield fluorescence microscope is developed. Spectral crosstalk is corrected by exciting the fluorophores in a round-robin manner and synchronous emitted light collection. To report the performance of the multicolor microscopy, a simplified 3D tumor model is created by placing beads and spheroids inside a channel filled with the cell culture medium is imaged at varying exposure times. As a representative case and a method for bio-hybrid drug carrier fabrication, a spheroid surface is coated with beads in a channel utilizing electrostatic forces under the guidance of multicolor microscopy. Our experiments show that multicolor fluorescence microscopy enables crosstalk-free and spectrally-different individual image acquisition of beads, spheroids, and channels with the minimum exposure time of 5.5 ms. The imaging technique has the potential to monitor drug carrier transportation to cancer cells in real-time.

A Recurrent Neural-Network-Based Real-Time Dynamic Model for Soft Continuum Manipulators.

This paper introduces and validates a real-time dynamic predictive model based on a neural network approach for soft continuum manipulators. The presented model provides a real-time prediction framework using neural-network-based strategies and continuum mechanics principles. A time-space integration scheme is employed to discretize the continuous dynamics and decouple the dynamic equations for translation and rotation for each node of a soft continuum manipulator. Then the resulting architecture is used to develop distributed prediction algorithms using recurrent neural networks. The proposed RNN-based parallel predictive scheme does not rely on computationally intensive algorithms; therefore, it is useful in real-time applications. Furthermore, simulations are shown to illustrate the approach performance on soft continuum elastica, and the approach is also validated through an experiment on a magnetically-actuated soft continuum manipulator. The results demonstrate that the presented model can outperform classical modeling approaches such as the Cosserat rod model while also shows possibilities for being used in practice.

Impact of Segmented Magnetization on the Flagellar Propulsion of Sperm-Templated Microrobots.

Technical design features for improving the way a passive elastic filament produces propulsive thrust can be understood by analyzing the deformation of sperm-templated microrobots with segmented magnetization. Magnetic nanoparticles are electrostatically self-assembled on bovine sperm cells with nonuniform surface charge, producing different categories of sperm-templated microrobots. Depending on the amount and location of the nanoparticles on each cellular segment, magnetoelastic and viscous forces determine the wave pattern of each category during flagellar motion. Passively propagating waves are induced along the length of these microrobots using external rotating magnetic fields and the resultant wave patterns are measured. The response of the microrobots to the external field reveals distinct flow fields, propulsive thrust, and frequency responses during flagellar propulsion. This work allows predictions for optimizing the design and propulsion of flexible magnetic microrobots with segmented magnetization.

Real-Time Multi-Modal Sensing and Feedback for Catheterization in Porcine Tissue.

OBJECTIVE: In this study, we introduce a multi-modal sensing and feedback framework aimed at assisting clinicians during endovascular surgeries and catheterization procedures. This framework utilizes state-of-the-art imaging and sensing sub-systems to produce a 3D visualization of an endovascular catheter and surrounding vasculature without the need for intra-operative X-rays. METHODS: The catheterization experiments within this study are conducted inside a porcine limb undergoing motions. A hybrid position-force controller of a robotically-actuated ultrasound (US) transducer for uneven porcine tissue surfaces is introduced. The tissue, vasculature, and catheter are visualized by integrated real-time US images, 3D surface imaging, and Fiber Bragg Grating (FBG) sensors. RESULTS: During externally-induced limb motions, the vasculature and catheter can be reliably reconstructed at mean accuracies of 1.9±0.3 mm and 0.82±0.21 mm, respectively. CONCLUSIONS: The conventional use of intra-operative X-ray imaging to visualize instruments and vasculature in the human body can be reduced by employing improved diagnostic technologies that do not operate via ionizing radiation or nephrotoxic contrast agents. SIGNIFICANCE: The presented multi-modal framework enables the radiation-free and accurate reconstruction of significant tissues and instruments involved in catheterization procedures.

Contactless acoustic micro/nano manipulation: a paradigm for next generation applications in life sciences.

Acoustic actuation techniques offer a promising tool for contactless manipulation of both synthetic and biological micro/nano agents that encompass different length scales. The traditional usage of sound waves has steadily progressed from mid-air manipulation of salt grains to sophisticated techniques that employ nanoparticle flow in microfluidic networks. State-of-the-art in microfabrication and instrumentation have further expanded the outreach of these actuation techniques to autonomous propulsion of micro-agents. In this review article, we provide a universal perspective of the known acoustic micromanipulation technologies in terms of their applications and governing physics. Hereby, we survey these technologies and classify them with regards to passive and active manipulation of agents. These manipulation methods account for both intelligent devices adept at dexterous non-contact handling of micro-agents, and acoustically induced mechanisms for self-propulsion of micro-robots. Moreover, owing to the clinical compliance of ultrasound, we provide future considerations of acoustic manipulation techniques to be fruitfully employed in biological applications that range from label-free drug testing to minimally invasive clinical interventions.

Correction: Dynamic modeling of soft continuum manipulators using lie group variational integration.

[This corrects the article DOI: 10.1371/journal.pone.0236121.].

IRONSperm: Sperm-templated soft magnetic microrobots.

We develop biohybrid magnetic microrobots by electrostatic self-assembly of nonmotile sperm cells and magnetic nanoparticles. Incorporating a biological entity into microrobots entails many functional advantages beyond shape templating, such as the facile uptake of chemotherapeutic agents to achieve targeted drug delivery. We present a single-step electrostatic self-assembly technique to fabricate IRONSperms, soft magnetic microswimmers that emulate the motion of motile sperm cells. Our experiments and theoretical predictions show that the swimming speed of IRONSperms exceeds 0.2 body length/s (6.8 ± 4.1 µm/s) at an actuation frequency of 8 Hz and precision angle of 45°. We demonstrate that the nanoparticle coating increases the acoustic impedance of the sperm cells and enables localization of clusters of IRONSperm using ultrasound feedback. We also confirm the biocompatibility and drug loading ability of these microrobots, and their promise as biocompatible, controllable, and detectable biohybrid tools for in vivo targeted therapy.

Dynamic modeling of soft continuum manipulators using lie group variational integration.

This paper presents the derivation and experimental validation of algorithms for modeling and estimation of soft continuum manipulators using Lie group variational integration. Existing approaches are generally limited to static and quasi-static analyses, and are not sufficiently validated for dynamic motion. However, in several applications, models need to consider the dynamical behavior of the continuum manipulators. The proposed modeling and estimation formulation is obtained from a discrete variational principle, and therefore grants outstanding conservation properties to the continuum mechanical model. The main contribution of this article is the experimental validation of the dynamic model of soft continuum manipulators, including external torques and forces (e.g., generated by magnetic fields, friction, and the gravity), by carrying out different experiments with metal rods and polymer-based soft rods. To consider dissipative forces in the validation process, distributed estimation filters are proposed. The experimental and numerical tests also illustrate the algorithm's performance on a magnetically-actuated soft continuum manipulator. The model demonstrates good agreement with dynamic experiments in estimating the tip position of a Polydimethylsiloxane (PDMS) rod. The experimental results show an average absolute error and maximum error in tip position estimation of 0.13 mm and 0.58 mm, respectively, for a manipulator length of 60.55 mm.