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Collier, C. Patrick

Publications and source records attributed to Collier, C. Patrick.

33 records · Page 2
All sourcesNASA NTRSDOE OSTI
DOE OSTI2017

Capacitive Detection of Low-Enthalpy, Higher-Order Phase Transitions in Synthetic and Natural Composition Lipid Membranes

In-plane lipid organization and phase separation in natural membranes play key roles in regulating many cellular processes. Highly cooperative, first-order phase transitions in model membranes consisting of few lipid components are well understood and readily detectable via calorimetry, densitometry, and fluorescence. However, far less is known about natural membranes containing numerous lipid species and high concentrations of cholesterol, for which thermotropic transitions are undetectable by the above-mentioned techniques. We demonstrate that membrane capacitance is highly sensitive to low-enthalpy thermotropic transitions taking place in complex lipid membranes. Specifically, we measured the electrical capacitance as a function of temperature for droplet interface bilayer model membranes of increasing compositional complexity, namely, (a) a single lipid species, (b) domain-forming ternary mixtures, and (c) natural brain total lipid extract (bTLE). We observed that, for single-species lipid bilayers and some ternary compositions, capacitance exhibited an abrupt, temperature-dependent change that coincided with the transition detected by other techniques. In addition, capacitance measurements revealed transitions in mixed-lipid membranes that were not detected by the other techniques. Most notably, capacitance measurements of bTLE bilayers indicated a transition at ~38 °C not seen with any other method. Likewise, capacitance measurements detected transitions in some well-studied ternary mixtures that, while known to yield coexisting lipid phases, are not detected with calorimetry or densitometry. These results indicate that capacitance is exquisitely sensitive to low-enthalpy membrane transitions because of its sensitivity to changes in bilayer thickness that occur when lipids and excess solvent undergo subtle rearrangements near a phase transition. Our findings also suggest that heterogeneity confers stability to natural membranes that function near transition temperatures by preventing unwanted defects and macroscopic demixing associated with high-enthalpy transitions commonly found in simpler mixtures.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2017

Real-Time Sensing of Single-Ligand Delivery with Nanoaperture-Integrated Microfluidic Devices

The measurement of biological events on the surface of live cells at the single-molecule level is complicated by several factors including high protein densities that are incompatible with single-molecule imaging, cellular autofluorescence, and protein mobility on the cell surface. Here, we fabricated a device composed of an array of nanoscale apertures coupled with a microfluidic delivery system to quantify single-ligand interactions with proteins on the cell surface. We cultured live cells directly on the device and isolated individual epidermal growth factor receptors (EGFRs) in the apertures while delivering fluorescently labeled epidermal growth factor. We observed single ligands binding to EGFRs, allowing us to quantify the ligand turnover in real time. These results demonstrate that this nanoaperture-coupled microfluidic device allows for the spatial isolation of individual membrane proteins while maintaining them in their cellular environment, providing the capability to monitor single-ligand binding events while maintaining receptors in their physiological environment. Lastly, these methods should be applicable to a wide range of membrane proteins.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2017

Tuning Superhydrophobic Nanostructures To Enhance Jumping-Droplet Condensation

It was recently discovered that condensation growing on a nanostructured superhydrophobic surface can spontaneously jump off the surface, triggered by naturally occurring coalescence events. Many reports have observed that droplets must grow to a size of order 10 μm before jumping is enabled upon coalescence; however, it remains unknown how the critical jumping size relates to the topography of the underlying nanostructure. Here, we characterize the dynamic behavior of condensation growing on six different superhydrophobic nanostructures, where the topography of the nanopillars was systematically varied. The critical jumping diameter was observed to be highly dependent upon the height, diameter, and pitch of the nanopillars: tall and slender nanopillars promoted 2 μm jumping droplets while short and stout nanopillars increased the critical size to over 20 μm. The topology of each surface is successfully correlated to the critical jumping diameter by constructing an energetic model that predicts how large a nucleating embryo needs to grow before it can inflate into the air with an apparent contact angle large enough for jumping. Furthermore, by extending our model to consider any possible surface, it is revealed that properly designed nanostructures should enable nanometric jumping droplets, which would further enhance jumping droplet condensers for heat transfer, anti-fogging, and anti-frosting applications.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2017

Dynamic Defrosting on Scalable Superhydrophobic Surfaces

Recent studies have shown that frost can grow in a suspended Cassie state on nanostructured superhydrophobic surfaces. During defrosting, the melting sheet of Cassie frost spontaneously dewets into quasi-spherical slush droplets that are highly mobile. Promoting Cassie frost would therefore seem advantageous from a defrosting standpoint; however, nobody has systematically compared the efficiency of defrosting Cassie ice versus defrosting conventional surfaces. Here, we characterize the defrosting of an aluminum plate, one-half of which exhibits a superhydrophobic nanostructure while the other half is smooth and hydrophobic. For thick frost sheets (>1 mm), the superhydrophobic surface was able to dynamically shed the meltwater, even at very low tilt angles. In contrast, the hydrophobic surface was unable to shed any appreciable meltwater even at a 90° tilt angle. For thin frost layers (≲1 mm), not even the superhydrophobic surface could mobilize the meltwater. We attribute this to the large apparent contact angle of the meltwater, which for small amounts of frost serves to minimize coalescence events and prevent droplets from approaching the capillary length. Finally, we demonstrate a new mode of dynamic defrosting using an upside-down surface orientation, where the melting frost was able to uniformly detach from the superhydrophobic side and subsequently pull the frost from the hydrophobic side in a chain reaction. Treating surfaces to enable Cassie frost is therefore very desirable for enabling rapid and low-energy thermal defrosting, but only for frost sheets that are sufficiently thick.

36 MATERIALS SCIENCE↗
DOE OSTI2016

Resource Sharing Controls Gene Expression Bursting

Episodic gene expression, with periods of high expression separated by periods of no expression, is a pervasive biological phenomenon. This bursty pattern of expression draws from a finite reservoir of expression machinery in a highly time variant way, i.e., requiring no resources most of the time but drawing heavily on them during short intense bursts, that intimately links expression bursting and resource sharing. Yet, most recent investigations have focused on specific molecular mechanisms intrinsic to the bursty behavior of individual genes, while little is known about the interplay between resource sharing and global expression bursting behavior. In this paper, we confine Escherichia coli cell extract in both cell-sized microfluidic chambers and lipid-based vesicles to explore how resource sharing influences expression bursting. Interestingly, expression burst size, but not burst frequency, is highly sensitive to the size of the shared transcription and translation resource pools. The intriguing implication of these results is that expression bursts are more readily amplified than initiated, suggesting that burst formation occurs through positive feedback or cooperativity. When extrapolated to prokaryotic cells, these results suggest that large translational bursts may be correlated with large transcriptional bursts. This correlation is supported by recently reported transcription and translation bursting studies in E. coli. Finally, the results reported here demonstrate a strong intimate link between global expression burst patterns and resource sharing, and they suggest that bursting plays an important role in optimizing the use of limited, shared expression resources.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2016

A Comparison of Single-Molecule Emission in Aluminum and Gold Zero-Mode Waveguides

We characterized the effect of gold and aluminum zero-mode waveguides (ZMWs) on the brightness of immobilized single emitters by probing fluorophores that absorb in the green and red regions of the visible spectrum. Aluminum ZMWs enhance the emission of Atto565 fluorophores upon green excitation, but they do not enhance the emission of Atto647N fluorophores upon red excitation. Gold ZMWs increase emission of both fluorophores with Atto647N showing enhancement that is threefold higher than that observed for Atto565. Our work indicates that 200 nm gold ZMWs are better suited for single-molecule fluorescence studies in the red region of the visible spectrum, while aluminum appears more suited for the green region of the visible spectrum.

36 MATERIALS SCIENCE↗
DOE OSTI2016

Hydrodynamic trapping for rapid assembly and in situ electrical characterization of droplet interface bilayer arrays

The droplet interface bilayer (DIB) is a modular technique for assembling planar lipid membranes between water droplets in oil. The DIB method thus provides a unique capability for developing digital, droplet-based membrane platforms for rapid membrane characterization, drug screening and ion channel recordings. This paper demonstrates a new, low-volume microfluidic system that automates droplet generation, sorting, and sequential trapping in designated locations to enable the rapid assembly of arrays of DIBs. The channel layout of the device is guided by an equivalent circuit model, which predicts that a serial arrangement of hydrodynamic DIB traps enables sequential droplet placement and minimizes the hydrodynamic pressure developed across filled traps to prevent squeeze-through of trapped droplets. Furthermore, the incorporation of thin-film electrodes fabricated via evaporation metal deposition onto the glass substrate beneath the channels allows for the first time in situ, simultaneous electrical interrogation of multiple DIBs within a sealed device. Combining electrical measurements with imaging enables measurements of membrane capacitance and resistance and bilayer area, and our data show that DIBs formed in different trap locations within the device exhibit similar sizes and transport properties. Simultaneous, single channel recordings of ion channel gating in multiple membranes are obtained when alamethicin peptides are incorporated into the captured droplets, qualifying the thin-film electrodes as a means for measuring stimuli-responsive functions of membrane-bound biomolecules. Furthermore, this novel microfluidic-electrophysiology platform provides a reproducible, high throughput method for performing electrical measurements to study transmembrane proteins and biomembranes in low-volume, droplet-based membranes.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2016

Controlling condensation and frost growth with chemical micropatterns

Frost growth on chilled hydrophobic surfaces is an inter-droplet phenomenon, where frozen droplets harvest water from supercooled liquid droplets to grow ice bridges that propagate across the surface in a chain reaction. To date, no surface has been able to passively prevent the in-plane growth of frost across the population of supercooled condensate. Here, we demonstrate that when the nucleation sites for supercooled condensate are properly controlled with chemical micropatterns, the speed of frost growth can be slowed and even halted entirely. This stoppage of frost growth is attributed to the large interdroplet separation between condensate upon the onset of freezing, which was controlled by the pitch of the chemical patterns and by deliberately triggering an early freezing event. Lastly, these findings reveal that frost growth can be passively suppressed by designing surfaces to spatially control nucleation sites and/or temporally control the onset of freezing events.

36 MATERIALS SCIENCE↗
DOE OSTI2015

Adsorption kinetics dictate monolayer self-assembly for both lipid-in and lipid-out approaches to droplet interface bilayer formation

The droplet interface bilayer (DIB) is a method to assemble planar lipid bilayer membranes between lipid-coated aqueous droplets and has gained popularity among researchers in many fields. Well-packed lipid monolayer on aqueous droplet–oil interfaces is a prerequisite for successfully assembling DIBs. Such monolayers can be achieved by two different techniques: “lipid-in”, in which phospholipids in the form of liposomes are placed in water, and “lipid-out”, in which phospholipids are placed in oil as inverse micelles. While both approaches are capable of monolayer assembly needed for bilayer formation, droplet pairs assembled with these two techniques require significantly different incubation periods and exhibit different success rates for bilayer formation. In our study, we combine experimental interfacial tension measurements with molecular dynamics simulations of phospholipids (DPhPC and DOPC) assembled from water and oil origins to understand the differences in kinetics of monolayer formation. With the results from simulations and by using a simplified model to analyze dynamic interfacial tensions, we conclude that, at high lipid concentrations common to DIBs, monolayer formation is simple adsorption controlled for lipid-in technique, whereas it is predominantly adsorption-barrier controlled for the lipid-out technique due to the interaction of interface-bound lipids with lipid structures in the subsurface. The adsorption barrier established in lipid-out technique leads to a prolonged incubation time and lower bilayer formation success rate, proving a good correlation between interfacial tension measurements and bilayer formation. Also, we clarify that advective flow expedites monolayer formation and improves bilayer formation success rate by disrupting lipid structures, rather than enhancing diffusion, in the subsurface and at the interface for lipid-out technique. Additionally, electrical properties of DIBs formed with varying lipid placement and type are characterized.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2015

Direct in situ measurement of specific capacitance, monolayer tension, and bilayer tension in a droplet interface bilayer

In this study, thickness and tension are important physical parameters of model cell membranes. However, traditional methods to measure these quantities require multiple experiments using separate equipment. This work introduces a new multi-step procedure for directly accessing in situ multiple physical properties of droplet interface bilayers (DIB), including specific capacitance (related to thickness), lipid monolayer tension in the Plateau-Gibbs border, and bilayer tension. The procedure employs a combination of mechanical manipulation of bilayer area followed by electrowetting of the capacitive interface to examine the sensitivities of bilayer capacitance to area and contact angle to voltage, respectively. These data allow for determining the specific capacitance of the membrane and surface tension of the lipid monolayer, which are then used to compute bilayer thickness and tension, respectively. The use of DIBs affords accurate optical imaging of the connected droplets in addition to electrical measurements of bilayer capacitance, and it allows for reversibly varying bilayer area. After validating the accuracy of the technique with diphytanoyl phosphatidylcholine (DPhPC) DIBs in hexadecane, the method is applied herein to quantify separately the effects on membrane thickness and tension caused by varying the solvent in which the DIB is formed and introducing cholesterol into the bilayer. Because the technique relies only on capacitance measurements and optical images to determine both thickness and tension, this approach is specifically well-suited for studying the effects of peptides, biomolecules, natural and synthetic nanoparticles, and other species that accumulate within membranes without altering bilayer conductance.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2015

Self-propelled sweeping removal of dropwise condensate

Dropwise condensation can be enhanced by superhydrophobic surfaces, on which the condensate drops spontaneously jump upon coalescence. However, the self-propelled jumping in prior reports is mostly perpendicular to the substrate. Here, we propose a substrate design with regularly spaced micropillars. Coalescence on the sidewalls of the micropillars leads to self-propelled jumping in a direction nearly orthogonal to the pillars and therefore parallel to the substrate. This in- plane motion in turn produces sweeping removal of multiple neighboring drops. The spontaneous sweeping mechanism may greatly enhance dropwise condensation in a self-sustained manner.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗
DOE OSTI2014

Length scale selects directionality of droplets on vibrating pillar ratchet

Directional control of droplet motion at room temperature is of interest for applications such as microfluidic devices, self-cleaning coatings, and directional adhesives. Here, arrays of tilted pillars ranging in height from the nanoscale to the microscale are used as structural ratchets to directionally transport water at room temperature. Water droplets deposited on vibrating chips with a nanostructured ratchet move preferentially in the direction of the feature tilt while the opposite directionality is observed in the case of microstructured ratchets. This remarkable switch in directionality is consistent with changes in the contact angle hysteresis. To glean further insights into the length scale dependent asymmetric contact angle hysteresis, the contact lines formed by a nonvolatile room temperature ionic liquid placed onto the tilted pillar arrays were visualized and analyzed in situ in a scanning electron microscope. As a result, the ability to tune droplet directionality by merely changing the length scale of surface features all etched at the same tilt angle would be a versatile tool for manipulating multiphase flows and for selecting droplet directionality in other lap-on-chip applications.

42 ENGINEERING↗
DOE OSTI2013

Dynamic Defrosting on Nanostructured Superhydrophobic Surfaces

Water suspended on chilled superhydrophobic surfaces exhibits delayed freezing; however, the interdrop growth of frost through subcooled condensate forming on the surface seems unavoidable in humid environments. It is therefore of great practical importance to determine whether facile defrosting is possible on superhydrophobic surfaces. Here in this paper, we report that nanostructured superhydrophobic surfaces promote the growth of frost in a suspended Cassie state, enabling its dynamic removal upon partial melting at low tilt angles (<15°). The dynamic removal of the melting frost occurred in two stages: spontaneous dewetting followed by gravitational mobilization. This dynamic defrosting phenomenon is driven by the low contact angle hysteresis of the defrosted meltwater relative to frost on microstructured superhydrophobic surfaces, which forms in the impaled Wenzel state. Dynamic defrosting on nanostructured superhydrophobic surfaces minimizes the time, heat, and gravitational energy required to remove frost from the surface, and is of interest for a variety of systems in cold and humid environments.

36 MATERIALS SCIENCE↗
DOE OSTI2012

Monodisperse alginate microgel formation in a three-dimensional microfluidic droplet generator

Droplet based microfluidic systems provide an ideal platform for partitioning and manipulating aqueous samples for analysis. Identifying stable operating conditions under which droplets are generated is challenging yet crucial for real-world applications. A novel three-dimensional microfluidic platform that facilitates the consistent generation and gelation of alginate-calcium hydrogel microbeads for microbial encapsulation, over a broad range of backing pressures, in the absence of surfactants, is described. The unique three-dimensional design of the fluidic network utilizes a height difference at the junction between the aqueous sample injection and organic carrier channels to induce droplet formation via a surface tension enhanced self-shearing mechanism. Combined within a flow-focusing geometry, under constant pressure control, this arrangement facilitates predictable generation of droplets over a much broader range of operating conditions than conventional two-dimensional systems. The impact of operating pressures and geometry on droplet gelation, aqueous and organic material flow rates, microbead size and bead generation frequency are described. The system presented provides a robust platform for encapsulating single microbes in complex mixtures into individual hydrogel beads, and provides the foundation for the development of a complete system for sorting and analyzing microbes at the single cell level.

36 MATERIALS SCIENCE↗