Cell Mechanical and Physiological Behavior in the Regime of Rapid Mechanical Compressions that Lead to Cell Volume Change.

Cells respond to mechanical forces by deforming in accordance with viscoelastic solid behavior. Studies of microscale cell deformation observed by high speed video microscopy have elucidated a new cell behavior in which sufficiently rapid mechanical compression of cells can lead to transient cell volume loss and then recovery. This work has discovered that the resulting volume exchange between the cell interior and the surrounding fluid can be utilized for efficient, convective delivery of large macromolecules (2000 kDa) to the cell interior. However, many fundamental questions remain about this cell behavior, including the range of deformation time scales that result in cell volume loss and the physiological effects experienced by the cell. In this study, a relationship is established between cell viscoelastic properties and the inertial forces imposed on the cell that serves as a predictor of cell volume loss across human cell types. It is determined that cells maintain nuclear envelope integrity and demonstrate low protein loss after the volume exchange process. These results define a highly controlled cell volume exchange mechanism for intracellular delivery of large macromolecules that maintains cell viability and function for invaluable downstream research and clinical applications.

Label-free microfluidic enrichment of photoreceptor cells.

Inherited retinal degenerative disorders such as retinitis pigmentosa and Usher syndrome are characterized by progressive death of photoreceptor cells. To restore vision to patients blinded by these diseases, a stem cell-based photoreceptor cell replacement strategy will likely be required. Although retinal stem cell differentiation protocols suitable for generating photoreceptor cells exist, they often yield a rather heterogenous mixture of cell types. To enrich the donor cell population for one or a few cell types, scientists have traditionally relied upon the use of antibody-based selection approaches. However, these strategies are quite labor intensive and require animal derived reagents and equipment that are not well suited to current good manufacturing practices (cGMP). The purpose of this study was to develop and evaluate a microfluidic cell sorting device capable of exploiting the physical and mechanical differences between retinal cell types to enrich specific donor cell populations such as Retinal Pigment Epithelial (RPE) cells and photoreceptor cells. Using this device, we were able to separate a mixture of RPE and iPSC-derived photoreceptor precursor cell lines into two substantially enriched fractions. The enrichment factor of the RPE fraction was 2 and that of the photoreceptor precursor cell fraction was 2.7. Similarly, when human retina, obtained from 3 independent donors, was dissociated and passed through the sorting device, the heterogeneous mixture could be reliably sorted into RPE and photoreceptor cell rich fractions. In summary, microfluidic cell sorting is a promising approach for antibody free enrichment of retinal cell populations.

Metachronal motion of artificial magnetic cilia.

Organisms use hair-like cilia that beat in a metachronal fashion to actively transport fluid and suspended particles. Metachronal motion emerges due to a phase difference between beating cycles of neighboring cilia and appears as traveling waves propagating along ciliary carpet. In this work, we demonstrate biomimetic artificial cilia capable of metachronal motion. The cilia are micromachined magnetic thin filaments attached at one end to a substrate and actuated by a uniform rotating magnetic field. We show that the difference in magnetic cilium length controls the phase of the beating motion. We use this property to induce metachronal waves within a ciliary array and explore the effect of operation parameters on the wave motion. The metachronal motion in our artificial system is shown to depend on the magnetic and elastic properties of the filaments, unlike natural cilia, where metachronal motion arises due to fluid coupling. Our approach enables an easy integration of metachronal magnetic cilia in lab-on-a-chip devices for enhanced fluid and particle manipulations.

Metachronal Actuation of Microscale Magnetic Artificial Cilia.

Biological cells often interact with the environment through carpets of microscopic hair-like cilia. These elastic structures are known to beat in a synchronized wavy fashion called metachronal motion to produce fluid transport. Metachronal motion emerges due to a phase difference between beating cycles of neighboring cilia and appears as traveling waves propagating along the ciliary carpet. We demonstrate submerged in water microscale magnetic cilia that are externally actuated to beat in a metachronal fashion. Two approaches are used to induce coordinated phase differences among the beating cilia. In the first case, we fabricate cilia with an imposed gradient of geometrical properties that are subject to a rotating uniform magnetic field. In the second scenario, a ciliary array is composed of identical cilia that experience a magnetic field that varies spatiotemporally. We demonstrate that magnetic cilia can achieve symplectic, antiplectic, and leoplectic metachrony.

Microfluidic pumping using artificial magnetic cilia.

One of the vital functions of naturally occurring cilia is fluid transport. Biological cilia use spatially asymmetric strokes to generate a net fluid flow that can be utilized for feeding, swimming, and other functions. Biomimetic synthetic cilia with similar asymmetric beating can be useful for fluid manipulations in lab-on-chip devices. In this paper, we demonstrate the microfluidic pumping by magnetically actuated synthetic cilia arranged in multi-row arrays. We use a microchannel loop to visualize flow created by the ciliary array and to examine pumping for a range of cilia and microchannel parameters. We show that magnetic cilia can achieve flow rates of up to 11 µl/min with the pressure drop of ~1 Pa. Such magnetic ciliary array can be useful in microfluidic applications requiring rapid and controlled fluid transport.

Asymmetric motion of magnetically actuated artificial cilia.

Most microorganisms use hair-like cilia with asymmetric beating to perform vital bio-physical processes. In this paper, we demonstrate a novel fabrication method for creating magnetic artificial cilia capable of such a biologically inspired asymmetric beating pattern essential for inducing microfluidic transport at low Reynolds number. The cilia are fabricated using a lithographic process in conjunction with deposition of magnetic nickel-iron permalloy to create flexible filaments that can be manipulated by varying an external magnetic field. A rotating permanent magnet is used to actuate the cilia. We examine the kinematics of a cilium and demonstrate that the cilium motion is defined by an interplay among elastic, magnetic, and viscous forces. Specifically, the forward stroke is induced by the rotation of the magnet which bends the cilium, whereas the recovery stroke is defined by the straightening of the deformed cilium, releasing accumulated elastic potential energy. This difference in dominating forces acting during the forward stroke and the recovery stroke leads to an asymmetric beating pattern of the cilium. Such magnetic cilia can find applications in microfluidic pumping, mixing, and other fluid handling processes.

Vortex chain formation in regions of ion concentration polarization.

The local vortical flow generated inside an ion concentration polarization (ICP) region is evaluated experimentally. The ICP is induced by a patterned nanoporous self-assembling membrane integrated inside a single microchannel. A bottom-view image of the depletion region near the membrane revealed a primary vortex which results from the electric field amplification. A unique perspective of the flow is obtained by imaging the microchannel from its side. This visualization shows for the first time the formation of a chain of three vortices all rotating in the same direction in the depletion region. While observation of multiple vortices has been previously reported, it was in reference to counter rotating vortex pairs and not to the same direction of rotating vortex chain formation. A physical model is proposed which considers a two dimensionally varying concentration profile in the depletion region to account for the formation of multiple vortices rotating in the same direction. The fast rotating primary vortex changes the local concentration in regions adjacent to it, as the advection time scale is much higher than the diffusion time scale. Near the membrane, it moves the low concentration electrolyte from the bottom wall upwards into a higher concentration region. Away from the membrane, it moves the high concentration electrolyte from the middle of the channel downwards into a low concentration region. These local changes in the wall concentration result in a varying slip velocity capable of inducing a secondary vortex. Similarly, this secondary vortex can induce a tertiary one. A numerical simulation is performed using the proposed varying slip velocity model which showed excellent agreement with the experimental observations.