Untethered capsules are capable of entering the gastrointestinal (GI) tract and collecting fluid samples containing microbial communities from specific locations, facilitating the study of chronic diseases. However, existing sampling capsules are designed for single-site sampling, making it challenging to gather samples from multiple targets. This paper reports a magnetic-driven capsule for multiple sampling within the GI tract and an on-demand magnetic-triggered fluid sampling strategy. The capsule consists of a body, a magnetic-triggered negative pressure unit, and a reservoir unit. Composed of an elastic membrane and Magnet I, the negative pressure unit controls pressure change inside the capsule cavity on demand to pump the sample by switching the magnetic field, while the embedded Magnet I also enables real-time magnetic localization for regional targeting and position tracking. The reservoir unit integrates three sampling papers for fluid absorption, two waterproof layers that maintain contamination levels below 25% to ensure reliable multi-site sampling, and a rotating arm embedded with Magnet II for posture adjustment of the sampling paper. The pumping and storage performance of the capsule was systematically evaluated and optimized. Meanwhile, the capsule, actuated by an external magnetic field, was evaluated for its active locomotion performance. Finally, the feasibility of using the capsule to perform active navigation and multi-target sampling in a porcine intestine was validated viaex vivoexperiments.

Soft tissue cutting is used for incision, separation and removal of tissues or cells. Due to high deformation of soft tissues resulting from their viscosity and elasticity, it is challenging to accurately cut the tissue along a desired path and control the force applied to the tissue for reducing invasiveness, especially at the microscale. This paper presents a robotic biopsy system for cutting and collecting trophectoderm cells from a highly deformable blastocyst. The system, for the first time, enables TE cell junction detection for laser ablation throughout the blastocyst biopsy process by using a convolutional neural network. The overall detection error was 2.13% in every 1,000 cell junctions with position RMSE of$1.63\ \mu \mathrm{m}\pm 0.29\ \mu \mathrm{m}$. A dynamics model was developed to describe the motion of the trophectoderm cells inside a biopsy micropipette. Based on this model, an adaptive control method was developed for trophectoderm cell aspiration and positioning inside the biopsy micropipette. Experimental results revealed that the controller was capable of effectively compensating for the cell positioning error by updating the varying system parameters according to the adaptation law. The success rate was 100%, the cell aggregate positioning accuracy was$\pm 1\ \mu \mathrm{m}$, the average settling time was 2 s, and the largest overshoot was$4.3\ \mu \mathrm{m}$. Compared to manual blastocyst biopsy, the robotic biopsy system shortened the blastocyst's recovery time (35 min vs. 50 min) which indicates lower invasiveness.

In biology, fluorescent dyes are routinely used for biochemical measurements such as pH and ion concentrations. They, especially when used for detecting a low concentration of ions, suffer from low signal-to-noise ratios (SNR); and increasing the concentration of fluorescent dyes causes more sever cytotoxicity. We invented a new approach that uses a low amount of fluorescent dye-coated magnetic nanoparticles for on-demand, accurately aggregating the nanoparticles and thus fluorescent dyes in a local region inside a cell for intracellular measurement. Experiments proved this approach is capable of achieving a significantly higher SNR and lower cytotoxicity. Different from existing magnetic micromanipulation systems that generate large swarms (several microns and above) or cannot move the generated swarm to an arbitrary position, we developed a five-pole magnetic micromanipulation system and technique for generating a small swarm (e.g., 1 μm; capable of generating a magnetic swarm from 0.52 μm to 52.7 μm with an error <; 7.5 %) and accurately positioning the small swarm (position control accuracy: 0.76 μm). As an example, the system performed intracellular pH mapping using a 1 μm swarm of pH sensitive fluorescent dye-coated magnetic nanoparticles. The swarm had an SNR inside a cell 10 times that by the traditional method, i.e., global dye treatment, with both cases using the same fluorescent dye concentration. Our intracellular measurement results, for the first time, quantitatively revealed the existence of pH gradient and polarized pH distribution in live migrating cells.

Existing approaches for robotic control of magnetic swarms are not capable of generating magnetic aggregates precisely in an arbitrarily specified target region in a fluidic flow environment. Such a swarm control capability is demanded by medical applications such as clinical embolization (i.e., localized clogging of blood vessels). This paper presents a new magnetic swarm control strategy to generate aggregates only in a specified target region under fluidic flow. Within the target region, the magnetic field generates sufficiently large magnetic forces among magnetic particles to maintain the aggregates' integrity at the junctions of blood vessels. In contrast, unintended aggregates outside the target region are disassembled by fluidic shear. The aggregation control approach achieved a mean absolute error of 0.15 mm in positioning a target region and a mean absolute error of 0.30 mm in controlling the target region's radius. With thrombin coating, 1 μm magnetic particles were controlled to perform embolization both in vitro (using microfluidic channel networks) and ex vivo (using porcine tissue). Experiments proved the effectiveness of the swarm control technique for on-demand, targeted embolization.

Robotic manipulation of deformable objects has been a classic topic in robotics. Compared to synthetic deformable objects such as rubber balls and clothes, biological cells are highly deformable and more prone to damage. This article presents robotic manipulation of deformable cells for orientation control (both out-of-plane and in-plane), which is required in both clinical (e.g., in vitro fertilization) and biomedical (e.g., clone) applications. Compared to manual cell orientation control based on empirical experience, the robotic approach, based on modeling and path planning, effectively rotates a cell, while consistently maintaining minimal cell deformation to avoid cell damage. A force model is established to determine the minimal force applied by the micropipette to rotate a spherical or, more generally, ellipsoidal oocyte. The force information is translated into indentation through a contact mechanics model, and the manipulation path of the micropipette is formed by connecting the indentation positions on the oocyte. An optimal controller is designed to compensate for the variations of mechanical properties across oocytes. The polar body of an oocyte is detected by deep neural networks with robustness to shape and size differences. In experiments, the system achieved an accuracy of 97.6% in polar body detection and an accuracy of 0.7° in oocyte orientation control with maximum oocyte deformation of 2.70 μm throughout the orientation control process.

Robotic manipulation of deformable objects (vs. rigid objects) has been a classic topic in robotics. Compared to deformable synthetic objects such as rubber balls and clothes, biological cells are highly deformable and more prone to damage. This paper presents robotic manipulation of deformable cells for orientation control (both out-of-plane and in-plane), which is required in both clinical (e.g., in vitro fertilization) and biomedical (e.g., clone) applications. Compared to manual cell rotation control based on empirical experience, the robotic approach, based on mathematical modeling and path planning, effectively rotates a cell while consistently maintaining minimal cell deformation to avoid cell damage. A force model is established to determine the minimal force applied by the micropipette to rotate a spherical or more generally, an ellipsoidal mouse oocyte. The force information is translated into indentation through a contact mechanics model, and the manipulation path of the micropipette is formed by connecting the indentation positions on the oocyte. A compensation controller is designed to compensate for the variations of mechanical properties across cells. The polar body of an oocyte is detected by deep neural networks with robustness to shape and size differences. Experimental results demonstrate that the system achieved an accuracy of 97.6% in polar body detection and an accuracy of 0.7° in oocyte orientation control with maximum oocyte deformation of 2.69 μm.

Measuring the motility and morphology parameters of motile cells is important for revealing their functional characteristics. This paper presents automation techniques that, for the first time, enable automated, non-invasive measurement of motility and morphology parameters of individual sperms. Compared to the status quo of qualitative estimation of single sperm's motility and morphology based on embryologists' empirical experience, the automation techniques provide quantitative data in nearly real time. An adapted joint probabilistic data association filter (JPDAF) was used for multi-sperm tracking and tackled challenges of identifying sperms that intersect or have small spatial distances. Since the standard differential interference contrast (DIC) imaging method has side illumination effect which causes inherent inhomogeneous image intensity and poses difficulties for accurate sperm morphology measurement, we integrated total variation norm into the quadratic cost function method, which together effectively removed inhomogeneous image intensity and retained sperm's subcellular structures after DIC image reconstruction. In order to relocate the same sperm of interest identified under low magnification after switching to high magnification, coordinate transformation was conducted to handle the changes in the field of view caused by magnification switch. Experimental results demonstrated an accuracy of 95.6% in sperm motility measurement and errors <;10% in morphology measurement.

Magnetic micromanipulation is an untethered technique and has enabled numerous applications in the scale of millimeters to micrometers from the tissue level to cell level. However, existing systems are not capable of maneuvering a sub-micrometer object for precise force control, preventing the realization of intracellular manipulation or `fantastic voyage' inside a single cell. The magnetic micromanipulation task achieved in this work is sub-micrometer position control and piconewton force control of a sub-micron (0.7 μm) magnetic bead inside a single human bladder cancer cell (RT4). The magnetic bead was 3D positioned in the cell using a generalized predictive controller that effectively tackled the control challenge caused by the slow visual feedback (1 Hz) from high-resolution confocal microscopy. The average positioning error was quantified to be 0.43 μm, which is slightly larger than Brownian motion-imposed constraint (0.31 μm). The system is capable of three-dimensionally applying a maximum force of 60 pN with a resolution of 4 pN. In experiments, a 0.7 μm magnetic bead was controlled to move from an initial position in a cell to target positions on the cell nucleus. Force-displacement data were obtained from multiple locations along the cell nucleus' major and minor axes. The results revealed, for the first time, significantly higher stiffness exists in the cell nucleus' major axis than the minor axis. This stiffness polarity was likely attributed to the aligned stress fibers of actin filament inside the cells.

Manipulation of motile cells such as bacteria and sperm is required in both cell biology and clinical applications. For immobilizing a motile sperm, the sperm head and tail positions must be accurately tracked, interference of proximal sperms on the target sperm must be tackled, and the orientation of the sperm must be properly aligned with the manipulation tool in order not to damage the sperm head where DNA is contained. Manual operation of sperm immobilization has stringent skill requirements, and both manual operation and existing robotic sperm immobilization suffer from inconsistent success rates and incapability of manipulating sperms swimming in all directions. This paper presents a robotic system for fully automated tracking, orientation control, and immobilization of motile sperms. Algorithms were developed for robustly tracking the sperm head and estimating the sperm tail positions under interfering conditions. A new visual servo control strategy was developed to enable the robotic system to actively adjust sperm orientation for immobilizing a sperm swimming in any direction. Experimental results from robotic immobilization of 400 sperms confirmed that the robotic system achieved a consistent success rate of 94.5 %, independent of sperm velocity or swimming direction.

Measuring cell motility and morphology is important for revealing their functional characteristics. This paper presents automation techniques that enable automated, non-invasive measurement of motility and morphology parameters of single sperm. Compared to the status quo of qualitative estimation of single sperm's motility and morphology manually, the automation techniques provide quantitative data for embryologists to select a single sperm for intracytoplasmic sperm injection. An adapted joint probabilistic data association filter was used for multi-sperm tracking and tackled challenges of identifying sperms that intersect or have small spatial distances. Since the standard differential interference contrast (DIC) imaging method has side illumination effect which causes inherent inhomogeneous image intensity and poses difficulties for accurate sperm morphology measurement, we integrated total variation norm into the quadratic cost function method, which together effectively removed inhomogeneous image intensity and retained sperm's subcellular structures after DIC image reconstruction. In order to relocate the same sperm of interest identified under low magnification after switching to high magnification, coordinate transformation was conducted to handle the changes in the field of view caused by magnification switch. The sperm's position after magnification switch was accurately predicted by accounting for the sperm's swimming motion during magnification switch. Experimental results demonstrated an accuracy of 95.6% in sperm motility measurement and an error <10% in morphology measurement.

Magnetic micromanipulation has the advantage of untethered control, high precision, and biocompatibility and has recently undergone great advances. The magnetic micromanipulation task to tackle in this paper is to three dimensionally navigate a 5-μm magnetic bead inside a mouse embryo and accurately apply forces to intraembryonic structures to perform mechanical measurements at multiple locations. Existing technologies are not able to achieve these navigation and measurement goals because of poor magnetic force scaling and/or lacking the capability of applying an accurately controlled force. This paper reports a three-dimensional magnetic tweezer system that enables, for the first time, intraembryonic magnetic navigation and force application. A single magnetic bead was introduced into a mouse embryo via robotic microinjection. The magnetic tweezer system accurately controlled the position of the magnetic bead via visually servoed magnetic control. By moving the magnetic bead with known forces inside the embryo, cytoplasm viscosity was measured, which is eight times the viscosity of water. For performing mechanical measurements on the cellular structures inside the mouse embryo, the system should be capable of applying forces up to 120 pN with a resolution of 4 pN. The results revealed that the middle region is significantly more deformable than the side regions of the inner cell mass.

Magnetic micromanipulation has the advantage of untethered control, high precision, and biocompatibility and has recently undergone great advances. The magnetic micromanipulation task to tackle in this work is to three-dimensionally navigate a 5-micrometer magnetic bead inside a mouse embryo and perform mechanical measurements at multiple locations. Existing technologies are not able to achieve these navigation and measurement goals because of poor magnetic force scaling and/or lacking the capability of applying an accurately controlled force. This paper reports a robotic magnetic tweezer system that enables, for the first time, intra- embryonic magnetic navigation and force application. A single magnetic bead was introduced into a mouse embryo via robotic microinjection. The robotic magnetic tweezer system accurately controls the position of the magnetic bead via visually servoed magnetic control. The system is also capable of applying forces up to 120 pN with a resolution of 1.78 pN for performing mechanical measurements on the cellular structures inside the mouse embryo, revealing that the middle region is more deformable than the side regions of the inner cell mass.