Sangjin Ryu

Chemical control of Vorticella bioactuator using microfluidics

In this report, we demonstrate a microfluidic platform to control the stalk contraction and extension of Vorticella convallaria by changing concentration of Ca2+ with pneumatically-actuated elastomeric microvalves. Habitation, extraction and control of V. convallaria were carried out in a PDMS-based microfluidic device. By treating the cells with the permeant saponin, external actuation of cell-anchoring stalk between an extended and contracted state was achieved by cyclic exposure of the cells to a Ca2+ buffer (10(-6) M) and a rinse buffer containing EGTA as a chelation agent. When solutions were switched, the stalk contracted and extended responding to the ambient Ca2+ concentration change. The length of the stalk changed between 20 and 60 microm, resulting in a working distance of about 40 microm.

Nearby boundaries create eddies near microscopic filter feeders.

We show through calculations, simulations and experiments that the eddies often observed near sessile filter feeders are frequently due to the presence of nearby boundaries. We model the common filter feeder Vorticella, which is approximately 50 µm across and which feeds by removing bacteria from ocean or pond water that it draws towards itself. We use both an analytical stokeslet model and a Brinkman flow approximation that exploits the narrow-gap geometry to predict the size of the eddy caused by two parallel no-slip boundaries that represent the slides between which experimental observations are often made. We also use three-dimensional finite-element simulations to fully solve for the flow around a model Vorticella and analyse the influence of multiple nearby boundaries. Additionally, we track particles around live feeding Vorticella in order to determine the experimental flow field. Our models are in good agreement both with each other and with experiments. We also provide approximate equations to predict the experimental eddy sizes owing to boundaries both for the case of a filter feeder between two slides and for the case of a filter feeder attached to a perpendicular surface between two slides.

Three-Dimensional Confocal Microscopy Indentation Method for Hydrogel Elasticity Measurement

The stiffness of the extracellular matrix (ECM) plays an important role in controlling cell functions. As an alternative to the ECM, hydrogels of tunable elasticity are widely used for in vitro cell mechanobiology studies. Therefore, characterizing the Youngs modulus of the hydrogel substrate is crucial. In this paper, we propose a confocal microscopy indentation method for measuring the elasticity of polyacrylamide gel as a model hydrogel. Our new indentation method is based on three-dimensional imaging of the indented gel using confocal microscopy and automated image processing to measure indentation depth from the three-dimensional image stack. We tested and validated our method by indenting polyacrylamide gels of different rigidities with various sphere indentors and by comparing it with the rheometric method. Our measurements show consistent results regardless of the type of the indentors and agree with rheometric measurements. Therefore, the proposed confocal microscopy indentation method can accurately measure the stiffness of hydrogels.

A New Angle on Microscopic Suspension Feeders near Boundaries

Microscopic sessile suspension feeders are a critical component in aquatic ecosystems, acting as an intermediate trophic stage between bacteria and higher eukaryotic taxa. Because they live attached to boundaries, it has long been thought that recirculation of the feeding currents produced by sessile suspension feeders inhibits their ability to access fresh fluid. However, previous models for the feeding flows of these organisms assume that they feed by pushing fluid perpendicular to surfaces they live upon, whereas we observe that sessile suspension feeders often feed at an angle to these boundaries. Using experiments and calculations, we show that living suspension feeders (Vorticella) likely actively regulate the angle that they feed relative to a substratum. We then use theory and simulations to show that angled feeding increases nutrient and particle uptake by reducing the reprocessing of depleted water. This work resolves an open question of how a key class of suspension-feeding organisms escapes physical limitations associated with their sessile lifestyle.

In Situ Hydrodynamic Lateral Force Calibration of AFM Colloidal Probes

Lateral force microscopy (LFM) is an application of atomic force microscopy (AFM) to sense lateral forces applied to the AFM probe tip. Recent advances in tissue engineering and functional biomaterials have shown a need for the surface characterization of their material and biochemical properties under the application of lateral forces. LFM equipped with colloidal probes of well-defined tip geometries has been a natural fit to address these needs but has remained limited to provide primarily qualitative results. For quantitative measurements, LFM requires the successful determination of the lateral force or torque conversion factor of the probe. Usually, force calibration results obtained in air are used for force measurements in liquids, but refractive index differences between air and liquids induce changes in the conversion factor. Furthermore, in the case of biochemically functionalized tips, damage can occur during calibration because tip-surface contact is inevitable in most calibration methods. Therefore, a nondestructive in situ lateral force calibration is desirable for LFM applications in liquids. Here we present an in situ hydrodynamic lateral force calibration method for AFM colloidal probes. In this method, the laterally scanned substrate surface generated a creeping Couette flow, which deformed the probe under torsion. The spherical geometry of the tip enabled the calculation of tip drag forces, and the lateral torque conversion factor was calibrated from the lateral voltage change and estimated torque. Comparisons with lateral force calibrations performed in air show that the hydrodynamic lateral force calibration method enables quantitative lateral force measurements in liquid using colloidal probes.

High-speed visualization of soap bubble blowing and image-processing-based analysis of pinch-off dynamics

Soap bubble blowing has long been an amusement for humans, and the process involves pinch-off similarly to liquid drops and gas bubbles. To visualize the pinch-off process of soap bubble blowing, we built an apparatus consisting of air jet flow and thin soap film on a circular ring, and replicated human soap bubbling. High-speed videography captured growing soap film tube and following pinch-off, and the minimal neck radius of the tube was measured based on image processing. Scaling law analyses show that regardless of the ring diameter, the scaling exponent of soap bubble pinch-off is about 2/3, which is similar to that of soap film bridge. Also, the speed of the airflow into the tube was evaluated based on volume calculation of the soap film tube, and the Reynolds number of the airflow was estimated to be 1060–2970, which suggests that soap bubbling may involve Bernoulli suction effect.Graphical abstract

Vorticella: A Protozoan for Bio-Inspired Engineering

In this review, we introduce Vorticella as a model biological micromachine for microscale engineering systems. Vorticella has two motile organelles: the oral cilia of the zooid and the contractile spasmoneme in the stalk. The oral cilia beat periodically, generating a water flow that translates food particles toward the animal at speeds in the order of 0.1–1 mm/s. The ciliary flow of Vorticella has been characterized by experimental measurement and theoretical modeling, and tested for flow control and mixing in microfluidic systems. The spasmoneme contracts in a few milliseconds, coiling the stalk and moving the zooid at 15–90 mm/s. Because the spasmoneme generates tension in the order of 10–100 nN, powered by calcium ion binding, it serves as a model system for biomimetic actuators in microscale engineering systems. The spasmonemal contraction of Vorticella has been characterized by experimental measurement of its dynamics and energetics, and both live and extracted Vorticellae have been tested for moving microscale objects. We describe past work to elucidate the contraction mechanism of the spasmoneme, recognizing that past and continuing efforts will increase the possibilities of using the spasmoneme as a microscale actuator as well as leading towards bioinspired actuators mimicking the spasmoneme.

Performance Characteristics of a V-type Probe Developed for Wall Vorticity Measurement

In order to investigate the relation between wall vorticity and streamwise velocity fluctuations in a turbulent boundary layer, a wall vorticity probe has been developed, which consists of two hot-wires on the wall aligned in V configuration. Although the measured intensity of spanwise wall vorticity fluctuations is somewhat lower than previous results, the intensity of streamwise wall vorticity fluctuations is in good agreement with them. It has been shown that the measured intensity of spanwise wall vorticity fluctuations is affected by transverse length of the wall vorticity probe. Instantaneous streamwise and spanwise wall vorticity fluctuations are compared with the results of DNS. Probability density function of spanwise wall vorticity fluctuations shows good agreement with previous results and is different from that of streamwise wall vorticity fluctuations. Energy spectrum of streamwisw wall vorticity fluctuations is lower than that of spanwise wall vorticity fluctuations in low frequency region.

Correlation of Wall Vorticity and Streamwise Velocity Fluctuations in a Turbulent Boundary Layer

A simultaneous measurement of wall vorticity and near-wall streamwise velocity fluctuations has been performed using a V-type wall vorticity probe and an I-type velocity probe to investigate the relation between them. Long-time averaged space-time correlations show that the wall vorticity is highly correlated with a turbulence structure which is tilted from the wall in the streamwise direction and that there is a streamwise vortex pair near the wall. It is shown that a structure correlated with the streamwise wall vorticity is smaller than and prior to a structure correlated with the spanwise wall vorticity. Tilting angles are obtained from the phase shift between the wall vorticity and streamwise velocity fluctuations. The tilting angle of the structure correlated with the streamwise wall vorticity is larger than that of the structure correlated with the spanwise wall vorticity. The convection velocity of the near-wall streamwise velocity fluctuations obtained from the space-time correlation is in good agreement with previous results.