Use of Optical Tweezers for Colloid Science

A space-borne optical tweezer apparatus for use with colloidal crystallization experiments has been characterized. The trapping force has been measured as a function of index mismatch between colloidal microspheres and the surrounding fluid and as a function of particle size. This work also presents a method to determine the refractive index of a colloidal microsphere, which is then used to calculate the applied trapping force for the case of an arbitrary background fluid. This is useful for work with dense colloidal suspensions when the usual (e.g., Stokes flow) trap force measurement methods do not apply, as well as microrheological studies of complex soft matter

Design and Construction of A Space-Borne Optical Tweezer Apparatus

A compact optical tweezer package has been developed for use on a microscope to be flown on the International Space Station as part of a series of experiments in colloid crystallization. A brief introduction to the principles of single-beam optical tweezer operation will be presented, after which a detailed system layout will be shown. Special design requirements due to the spaceflight nature of the hardware will also be discussed. The tweezer apparatus is capable of trapping many particles through use of a two-axis acousto-optical deflector. The trap strength is sufficient to perform the required science (50 pN at Δn=0.2). The trap beam behaves approximately as a diffraction limited single mode Gaussian beam of numerical aperture, NA=1.4, as shown through spot size measurements and confocal-type images of the focal region. This is the first time optical tweezers will be deployed in a microgravity environment

Use of Optical Tweezers to Probe Epithelial Mechanosensation

Cellular mechanosensation mechanisms have been implicated in a variety of disease states. Specifically in renal tubules, the primary cilium and associated mechanosensitive ion channels are hypothesized to play a role in water and salt homeostasis, with relevant disease states including polycystic kidney disease and hypertension. Previous experiments investigating ciliary-mediated cellular mechanosensation have used either fluid flow chambers or micropipetting to elicit a biological response. The interpretation of these experiments in terms of the ciliary hypothesis has been difficult due the spatially distributed nature of the mechanical disturbance-several competing hypotheses regarding possible roles of primary cilium, glycocalyx, microvilli, cell junctions, and actin cytoskeleton exist. I report initial data using optical tweezers to manipulate individual primary cilia in an attempt to elicit a mechanotransduction response-specifically, the release of intracellular calcium. The advantage of using laser tweezers over previous work is that the applied disturbance is highly localized. I find that stimulation of a primary cilium elicits a response, while stimulation of the apical surface membrane does not. These results lend support to the hypothesis that the primary cilium mediates transduction of mechanical strain into a biochemical response in renal epithelia. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3316378

Use of Optical Tweezers for Colloid Science

A space-borne optical tweezer apparatus for use with colloidal crystallization experiments has been characterized. The trapping force has been measured as a function of index mismatch between colloidal microspheres and the surrounding fluid and as a function of particle size. This work also presents a method to determine the refractive index of a colloidal microsphere, which is then used to calculate the applied trapping force for the case of an arbitrary background fluid. This is useful for work with dense colloidal suspensions when the usual (e.g., Stokes flow) trap force measurement methods do not apply, as well as microrheological studies of complex soft matter

Mechanical Properties of A Primary Cilium As Measured by Resonant Oscillation

Primary cilia are ubiquitous mammalian cellular substructures implicated in an ever-increasing number of regulatory pathways. The well-established ciliary hypothesis states that physical bending of the cilium (for example, due to fluid flow) initiates signaling cascades, yet the mechanical properties of the cilium remain incompletely measured, resulting in confusion regarding the biological significance of flow-induced ciliary mechanotransduction. In this work we measure the mechanical properties of a primary cilium by using an optical trap to induce resonant oscillation of the structure. Our data indicate 1) the primary cilium is not a simple cantilevered beam; 2) the base of the cilium may be modeled as a nonlinear rotatory spring, with the linear spring constant k of the cilium base calculated to be (4.6 ± 0.62) × 10−12 N/rad and nonlinear spring constant α to be (−1 ± 0.34) × 10−10 N/rad2; and 3) the ciliary base may be an essential regulator of mechanotransduction signaling. Our method is also particularly suited to measure mechanical properties of nodal cilia, stereocilia, and motile cilia—anatomically similar structures with very different physiological functions

A Model for the Force Exerted on A Primary Cilium by An Optical Trap and The Resulting Deformation

Cilia are slender flexible structures extending from the cell body; genetically similar to flagella. Although their existence has been long known, the mechanical and functional properties of non-motile (“primary”) cilia are largely unknown. Optical traps are a non-contact method of applying a localized force to microscopic objects and an ideal tool for the study of ciliary mechanics. We present a method to measure the mechanical properties of a cilium using an analytic model of a flexible, anchored cylinder held within an optical trap. The force density is found using the discrete-dipole approximation. Utilizing Euler-Bernoulli beam theory, we then integrate this force density and numerically obtain the equilibrium deformation of the cilium in response to an optical trap. The presented results demonstrate that optical trapping can provide a great deal of information and insight about the properties and functions of the primary cilium

Physics of Martial Arts: Incorporation of Angular Momentum to Model Body Motion and Strikes

We develop a physics-based kinematic model of martial arts movements incorporating rotation and angular momentum, extending prior analyses. Here, our approach is designed for a classroom environment; we begin with a warm-up exercise introducing counter-intuitive aspects of rotational motion before proceeding to a set of model collision problems that are applied to martial arts movements. Finally, we develop a deformable solid-body mechanics model of a martial arts practitioner suitable for an intermediate mechanics course. We provide evidence for our improved model based on calculations from biomechanical data obtained from prior reports as well as time-lapse images of several different kicks. In addition to incorporating angular motion, our model explicitly makes reference to friction between foot and ground as an action-reaction pair, showing that this interaction provides the motive force/torque for nearly all martial arts movements. Moment-of-inertia tensors are developed to describe kicking movements and show that kicks aimed high, towards the head, transfer more momentum to the target than kicks aimed lower, e.g. towards the body

Force-Response Considerations in Ciliary Mechanosensation

Considerable experimental evidence indicates that the primary, nonmotile cilium is a mechanosensory organelle in several epithelial cell types. As the relationship between cellular responses and nature and magnitude of applied forces is not well understood, we have investigated the effects of exposure of monolayers of renal collecting duct chief cells to orbital shaking and quantified the forces incident on cilia. An exposure of 24 h of these cells to orbital shaking resulted in a decrease of amiloride-sensitive sodium current by ∼60% and ciliary length by ∼30%. The sensitivity of the sodium current to shaking was dependent on intact cilia. The drag force on cilia due to induced fluid flow during orbital shaking was estimated at maximally 5.2 × 10−3 pN at 2 Hz, ∼4 times that of thermal noise. The major structural feature of cilia contributing to their sensitivity appears to be ciliary length. As more than half of the total drag force is exerted on the ciliary cap, one function of the slender stalk may be to expose the cap to greater drag force. Regardless, the findings indicate that the cilium is a mechanosensory organelle with a sensitivity much lower than previously recognized

Stabilization of Hypoxia Inducible Factor by Cobalt Chloride Can Alter Renal Epithelial Transport

© 2017 The Authors. Given the importance of the transcriptional regulator hypoxia-inducible factor-1 (HIF-1) for adaptive hypoxia responses, we examined the effect of stabilized HIF-1α on renal epithelial permeability and directed sodium transport. This study was motivated by histological analysis of cystic kidneys showing increased expression levels of HIF-1α and HIF-2α. We hypothesize that compression induced localized ischemia-hypoxia of normal epithelia near a cyst leads to local stabilization of HIF-1α, leading to altered transepithelial transport that encourages cyst expansion. We found that stabilized HIF-1α alters both transcellular and paracellular transport through renal epithelial monolayers in a manner consistent with secretory behavior, indicating localized ischemia-hypoxia may lead to altered salt and water transport through kidney epithelial monolayers. A quantity of 100 µmol/L Cobalt chloride (CoCl2) was used acutely to stabilize HIF-1α in confluent cultures of mouse renal epithelia. We measured increased transepithelial permeability and decreased transepithelial resistance (TER) when HIF-1α was stabilized. Most interestingly, we measured a change in the direction of sodium current, most likely corresponding to abnormal secretory function, supporting our positive-feedback hypothesis