Recently, a bio-inspired, synthetic membrane-based hair cell sensor was fabricated and characterized. This sensor
generates current in response to mechanical stimuli, such as airflow or free vibration, which perturb the sensor’s hair.
Vibration transferred from the hair to a lipid membrane (lipid bilayer) causes a voltage-dependent time rate of change in
electrical capacitance of the membrane, which produces measurable current. Studies to date have been performed on
systems containing only two droplets and a single bilayer, even though an array of multiple bilayers can be formed with
more than 2 droplets. Thus, it is yet to be determined how multiple lipid bilayers affect the sensing response of a
membrane-based hair cell sensor. In this work, we assemble serial droplet arrays with more than 1 bilayer to
experimentally study the current generated by each membrane in response to perturbation of a single hair element. Two
serial array configurations are studied: The first consists of a serial array of 3 bilayers formed using 4 droplets with the
hair positioned in an end droplet. The second configuration consists of 3 droplets and 2 bilayers in series with the hair
positioned in the central droplet. In serial arrays of up to four droplets, we observe that mechanotransduction of the
hair’s motion into a capacitive current occurs at every membrane, with bilayers positioned adjacent to the droplet
containing the hair generating the largest sensing current. The measured currents suggest the total current generated by
all bilayers in a 4-droplet, 3-bilaye array is greater than the current produced by a single-membrane sensor and similar in
magnitude to the sum of currents output by 3, single-bilayer sensors operated independently. Moreover, we learned that
bilayers positioned on the same side of the hair produce sensing currents that are in-phase, whereas bilayers positioned
on opposite sides of the droplet containing the hair generate out-of-phase responses.
Hair cell structures are one of the most common forms of sensing elements found in nature. In nearly all
vertebrates hair cells are used for auditory and vestibular sensing. In humans, approximately 16,000
auditory hair cells can be found in the cochlea of the ear. Each hair cell contains a stereocilia, which is the
primary structure for sound transduction. This study looks to develop and characterize an artificial hair
cell that resembles the stereocilia of the human ear. Recently our research group has shown that a single
artificial hair cell can be formed in an open substrate using a single aqueous droplet and a hydrogel. In
this study, air was blown across the hair and analyzed using spectral analysis. The results of this study
provided the foundation for our current work toward an artificial hair cell that uses two aqueous droplets.
In the current study a test fixture was created in order to consistently measure various properties of the
encapsulated hair cell. The response of the hair cell was measured with an impulse input at various
locations on the test fixture. A frequency response function was then created using the impulse input and
the output of the sensor. It was found that the vibration of the hair was only detectable if the test fixture
was struck at the correct location. By changing the physical parameters of the hair sensor, such as hair
length, we were able to alter the response of the sensor. It was also found that the sensitivity of the sensor
was reliant on the size of the lipid bilayer.
Recent research has shown that a new class of mechanical sensor, assembled from biomolecules and which features an
artificial cell membrane as the sensing element, can be used to mimic basic hair cell mechanotransduction in vertebrates.
The work presented in this paper is motivated by the need to increase sensor performance and stability by refining the
methods used to fabricate and connect lipid-encapsulated hydrogels. Inspired by superficial neuromasts found on fish,
three hydrogel materials are compared for their ability to be readily shaped into neuromast-inspired geometries and
enable lipid bilayer formation using self-assembly at an oil/water interface. Agarose, polyethylene glycol (PEG,
6kg/mole), and hydroxyethyl methacrylate (HEMA) gel materials are compared. The results of this initial study
determined that UV-curable gel materials such as PEG and HEMA enable more accurate shaping of the gel-needed for
developing a sensor that uses a gel material both for mechanical support and membrane formation-compared to
agarose. However, the lower hydrophobicity of agarose and PEG materials provide a more fluid, water-like environment
for membrane formation-unlike HEMA. In working toward a neuromast-inspired design, a final experiment demonstrates that a bilayer can also be formed directly between two lipid-covered PEG surfaces. These initial results suggest that candidate gel materials with a low hydrophobicity, high fluidity, and a low modulus can be used to provide membrane support.
The formation of lipid bilayers between ionic liquid droplets is presented as a new means of forming functional
bimolecular networks. Ionic liquids are molten salts that have a number of interesting properties, such as the ability
to be a liquid at room temperature and exceedingly low vapor pressure. Our research demonstrates that it is possible
to consistently and repeatable form lipid bilayers on droplets of ionic liquid solutions. Characterization of the bilayers
interfaces shows that the ionic liquids have negligible effects on the stability and electrical properties of the bilayer. It
is also shown that the conductance levels in the gating events of Alamethicin peptide are affected by some ionic liquids.
Receptors known as hair cells give many animals this ability to sense a wide range of stimuli, such as sound, orientation,
vibration, and flow. Previous researchers have mimicked natural hair cells by building electromechanical sensor systems
that produce an electric response due to the bending of artificial hairs. Inspired by the roles of sensory hairs in fish, this
work builds on previous research by investigating the flow dependent electrical response of a 'skin'-encapsulated
artificial hair cell in an aqueous flow. This study presents the design, fabrication, and characterization of a flow sensor
that will help close the loop between the sensing mechanisms and control strategies that aquatic organisms employ for
functions such as locomotion regulation, prey capture, and particulate capture. The system is fabricated with a durable,
artificial bilayer that forms at the interface between lipid-encased aqueous volumes contained in a flexible encapsulated
polyurethane substrate. Flow experiments are conducted by placing the bio-inspired sensor in a flow chamber and
subjecting it to pulse-like flows. Specifically, through temporal responses of the measured current and power spectral
density (PSD) analysis, our results show that the amplitude and frequency of the current response are related to the flow
over the hair. This preliminary study demonstrates that the encapsulated artificial hair cell flow sensor is capable of
sensing changes in flow through a mechanoelectrical response and that its sensing capabilities may be altered by varying
its surface morphology.
In this paper, we discuss a new form of electroactive material that consists of both synthetic polymers and
biological molecules. This modular material system is inspired by the compartmentalized and hierarchical organization
of cellular systems and features an artificial cell membrane, or lipid bilayer, which acts as the primary
transduction element in the material. Building on recent developments by our group, the lipid bilayer is formed
at the interface between phospholipid-encased hydrogel volumes surrounded by oil and contained in a solid substrate.
Results are presented that demonstrate how the electromechanical properties of the lipid bilayer can be
used for both static and dynamic sensing and actuation. Specifically, a relative change in length of the outer
substrate of 10-15% due to an applied force yields large changes in capacitance (> 90% reduction) or resistance
(20-30% increase) depending on the composition of the bilayer. The capacitive nature of the membrane is also
used in a dynamic sensing application, whereby the perturbation of an artificial hair structure induces bending
in a bilayer formed at the base of the hair. This oscillation results in a time-varying membrane capacitance that
in turn produces an electrical current on the order of 1 - 100pA. The ability to actuate the amount of contact
between neighboring modules is also discussed and a concept for fabricating higher-order biomolecular arrays
that connect internally to form networks of lipid bilayers is also presented.
A gel-supported lipid bilayer formed at the base of an artificial hair is used as the transduction element in a
membrane-based artificial haircell sensor inspired by the structure and function of mammalian outer hair cells.
This paper describes the initial fabrication and characterization of a bioderived, soft-material alternative to
previous artificial haircells that used the transduction properties of synthetic materials for flow and touch sensing.
Under an applied air flow, the artificial hair structure vibrates, triggering a picoamp-level electrical current across
the bilayer. Experimental analysis of this mechanoelectrical transduction process supports the hypothesis that
the oscillating current is produced by a time-varying change in the capacitance of the membrane caused by the
vibration of the hair. Specifically, frequency analysis of both the motion of the hair and the measured current
show that both phenomena occur at similar frequencies, which suggests that changes in capacitance occur as a
result of membrane bending during excitation.
Biomolecular networks formed from droplet interface bilayers (DIB) use principles of phase separation and
molecular self-assembly to create a new type of functional material. The original DIB embodiment consists
of lipid-encased aqueous droplets surrounding by a large volume of oil contained in a shallow well. However,
recent results have shown that, by reducing the amount of oil that separates the droplets from the supporting
substrate, physically-encapsulated DIBs display increased durability and portability. In this paper we extend
the concept of encapsulated biomolecular networks to one in which phase separation and molecular self-assembly
occur entirely within internally-structured reservoirs of a solid material. Flexible substrates with 200μm wideby-
200μm deep internal microchannels for holding the aqueous and oil phases are fabricated from Sylgard 184 polydimethylsiloxane (PDMS) using soft-lithography microfabrication techniques. Narrowed apertures along the
microchannels enable the use of the regulated attachment method (RAM) to subdivide and reattach lipid-encased
aqueous volumes contained within the material with an applied external force. The use of perfluorodecalin, a
fluorocarbon oil, instead of hexadecane eliminates absorption of the oil phase into the PDMS bulk while a silanization surface treatment of the internal channel walls maximizes wetting by the oil phase to retain a thin layer of oil within the channels to provide a fluid oil/water interface around the aqueous volumes. High-quality 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPHPC) lipid bilayers formed within the prototype substrates have electrical resistance between 1-100GΩ, enabling the measurement of single and few-channel recordings of alpha-hemolysin (αHL) and alamethicin proteins incorporated into the bilayers.
Biological molecules including phospholipids and proteins offer scientists and engineers a diverse selection of
materials to develop new types of active materials and smart systems based on ion conduction. The inherent
energy-coupling abilities of these components create novel kinds of transduction elements. Networks formed from
droplet-interface bilayers (DIB) are a promising construct for creating cell mimics that allow for the assembly
and study of these active biological molecules. The current-voltage relationship of symmetric, "lipid-in" dropletinterface
bilayers are characterized using electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV).
"Lipid-in" diphytanoyl phosphatidylcholine (DPhPC) droplet-interface bilayers have specific resistances of nearly
10MΩ•cm2 and rupture at applied potentials greater than 300mV, indicating the "lipid-in" approach produces
higher quality interfacial membranes than created using the original "lipid-out" method. The incorporation
of phospholipids into the droplet interior allows for faster monolayer formation but does not inhibit the selfinsertion
of transmembrane proteins into bilayer interfaces that separate adjacent droplets. Alamethicin proteins
inserted into single and multi-DIB networks produce a voltage-dependent membrane conductance and current
measurements on bilayers containing this type of protein exhibit a reversible, 3-4 order-of-magnitude conductance
increase upon application of voltage.
Bimolecules have demonstrated the potential to function as active components in energy harvesting devices,
biosensors and bioinspired actuators. The bilayer lipid membrane (BLM) formed from lipid molecules and
supported in the pores of porous substrates is the standard platform for fabricating the biomolecule based
devices. The techniques for forming BLM in an in-vitro environment like lipid painting, Lagmuir-Blodgett,
Langmuir-Schaffer and lipid folding methods were developed by researchers in the biophysical community to
investigate the properties of membrane bound proteins. While all of these methods can form a BLM and has
been used in laboratory research for few decades, they are not equally well-suited for fabricating an engineering
device. Of the different methods, the lipid deposition technique for BLM self-assembly and protein insertion is
the closest in its qualities to an engineering prototyping method. This article presents a detailed electrical model
of the substrates and the BLM formed in the pores from SOPC, POPS:POPE and DPhPC lipids using lipid
deposition technique. The equivalent circuits of the substrates and the BLM are used to interrogate the quality
of the BLM by impedance spectroscopy. The deviations of the prepared BLMs from desirable parameters are
traced to the preparation procedure that could be used as a feedback information for fabricating a single BLM in
the pores of the substrate. The impedance response is also used to understand the change in electrical properties
of BLMs formed in an array of pores of a multi-porous substrate.
This study expands the number of novel synthetic ionomers specifically designed for performance as ionic polymer
transducers (IPT) membranes, specifically employing a highly branched sulfonated polysulfone. Control of the synthetic
design, characterization, and application of the novel ionomer is intended to allow fundamental study of the effect of
polymer branching on electromechanical transduction in IPTs. Fabrication methods were developed based upon the
direct application process (DAP) to construct a series of stand-alone electrodes as well as full IPTs with corresponding
electrode compositions. Specifically, the volumetric ratio of RuO2 conducting particles to the novel ionomeric matrix
was varied from 0 - 45 vol % in the electrodes. Electrical impedance spectroscopy was employed to determine the
electrical properties and their variation with electrode composition separate from and in the IPT. A percolation threshold
was detected for increased ionic conductivity of the stand-alone electrodes and the full IPTs based on increased loading
of conducting particles in the electrodes. An equivalent electrical circuit model was applied to fit the impedance data and
implicated interfacial and bulk effects contributing differently to the electrical properties of the electrodes and IPT as a
whole. The fabricated IPT series was further tested for bending actuation in response to applied step voltages and
represents the first demonstration of IPTs constructed with the DAP process using 100 % novel ionomer in all
components. The percolation behavior extended to the bending actuation responses for strain and voltage-normalized
strain rate and is useful in optimizing IPT components for maximum performance regardless of the ionomer employed.
Bilayer lipid membranes (BLMs) have been studied extensively due to functional and structural similarities
to cell membranes, fostering research to understand ion-channel protein functions, measure bilayer mechanical
properties, and identify self-assembly mechanisms. BLMs have traditionally been formed across single pores in
substrates such as PTFE (Teflon). The incorporation of ion-channel proteins into the lipid bilayer enables the
selective transfer of ions and fluid through the BLM. Processes of this nature have led to the measurement of
ion current flowing across the lipid membrane and have been used to develop sensors that signal the presence of
a particular reactant (glucose, urea, penicillin), improve drug recognition in cells, and develop materials capable
of creating chemical energy from light. Recent research at Virginia Tech has shown that the incorporation of
proton transporters in a supported BLM formed across an array of pores can convert chemical energy available
in the adenosine triphosphate (ATP) into electricity. Experimental results from this work show that the
system-named Biocell-is capable of developing 2µW/cm2 of membrane area with 15μl of ATPase. Efforts to increase
the power output and conversion efficiency of this process while moving toward a packaged device present a
unique engineering problem. The bilayer, as host to the active proton transporters, must therefore be formed
evenly across a porous substrate, remain stable and yet fluid-like for protein interaction, and exhibit a large seal
resistance. This article presents the ongoing work to characterize the Biocell using impedance analysis. Electrical
impedance spectroscopy (EIS) is used to study the effect of adding ATPase proteins to POPS:POPE bilayer lipid
membranes and correlate structural changes evident in the impedance data to the energy-conversion capability
of various partial and whole Biocell assemblies. The specific membrane resistance of a pure BLM drops from
40-120kΩ•cm2 to only a few hundred Ω•cm2 upon reconstitution of ATPase proteins. Power characterization
indicates that ATP hydrolysis may result in charging of the
silver-silver chloride electrodes.
Plant and animal cell membranes transport charged species, neutral molecules and water through ion pumps
and channels. The energy required for moving species against established concentration and charge gradients
is provided by the biological fuel - adenosine triphosphate (ATP) -synthesized within the cell. The adenosine
triphosphatase (ATPases) in a plant cell membrane hydrolyze ATP in the cell cytoplasm to pump protons across
the cell membrane. This establishes a proton gradient across the membrane from the cell exterior into the cell
cytoplasm. This proton motive force stimulates ion channels that transport nutrients and other species into
the cell. This article discusses a device that converts the chemical energy stored in adenosine triphosphate into
electrical power using a transporter protein, ATPase. The V-type ATPase proteins used in our prototype are
extracted from red beet(Beta vulgaris) tonoplast membranes and reconstituted in a bilayer lipid membrane or
BLM formed from POPC and POPS lipids. A pH7 medium that can support ATP hydrolysis is provided on both
sides of the membrane and ATP is dissolved in the pH7 buffer on one side of the membrane. Hydrolysis of ATP
results in the formation of a phosphate ion and adenosine diphosphate. The energy from the reaction activates
ATPase in the BLM and moves a proton across the membrane. The charge gradient established across the BLM
due to the reaction and ion transport is converted into electrical current by half-cell reference electrodes. The
prototype ATPase cell with an effective BLM area of 4.15 mm2 carrying 15 &mgr;l of ATPase proteins was observed
to develop a steady state peak power output of 70 nW, which corresponds to a specific power of 1.69 &mgr;W/cm2
and a current density of 43.4 &mgr;A/cm2 of membrane area.
An active approach for initiating rigidization in carbon-fiber reinforced polymer (CFRP) thermosets links controllable mechanical stiffening to inherent electrical resistivity. With direct applications toward the rigidization of ultra-lightweight, inflatable space structures, temperature-controlled resistive heating is used to create oncommand rigidization. As required by the on-orbit conditions in space, flexible, rigidizable structures demand stable and space-survivable materials that incorporate techniques for providing shape control and structural stiffening. Methods currently employed to achieve a mechanical hardening include many passive techniques:
UV curing, sub-Tg hardening, and hydro-gel evaporation. The benefits of a passive system (simplicity, energy efficiency) are offset by their inherent lack of control, which can lead to long curing times and weak spots due to uneven curing. In efforts to significantly reduce the transition time of the composite from a structurally-vulnerable state to a fully-rigidized shape and to increase control of the curing process, an active approach is taken. Specifically, temperature-controlled internal resistive heating initiates thermoset curing in a coated carbon fiber composite to form an electrically-controlled, thermally-activated material. Through controlled heating,
this research examines how selective temperature control can be used to prescribe matrix consolidation and material rigidization on two different thermosetting resins, U-Nyte Set 201A and 201B. Feedback temperature control, based on a PID control algorithm, was applied to the process of resistive heating. Precise temperature tracking (less than 1.1°C RMS or ±3.3% error) was achieved for controlled sample heating. Using samples of the thermoset-coated carbon-fiber tow, composite hardening through resistive heating occurred in 24 minutes and required roughly 1 W-hr/inch of electrical energy. The rigidized material was measured to be 14-21 times stiffer in bending than the uncured material. In addition, the cure completion of the resin was measured through differential scanning calorimetry (DSC).
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