This invention relates to smart sensors in the field of nano-technology, and in particular to, methods, devices and systems for electrically deflected nano-mechanical sensors to measure and characterize physical properties of materials from ambient temperatures and conditions to temperatures up to 400° C.
In the 21st century, people are developing smart materials and smart sensors. The smart sensors, made of smart materials, provide in association with other components like actuators and control systems, the functional capability to react to internal and external environments and achieve adaptability. Examples include, but are not limited to, life science research involving the study of interaction between biological and other molecules, in-vitro diagnostics, food safety whereby bacteria and toxins are detected without the need for time-consuming growth experiments, fresh water control involving the detection of heavy metal ions in fresh water or terror-related compounds, such as, ricin in fresh water supplies and gas detection, detection of explosives, chemical warfare agents, narcotics and the like.
The realization that many molecular phenomena result in mechanical responses at the nanoscale level promises to bring about a revolution in the field of chemical, physical, and biological sensor development. Exploiting nanoscale mechanics for molecular recognition is a paradigm shift in sensor technology. In a quest for smaller, faster, better, smarter sensors, the micro-electromechanical systems (MEMS) have been scaled to the submicron range, leading to the new category of nanoelectromechanical systems (NEMS). The nanomechanical sensor opens additional potential for low-power high-frequency devices for mobile communications, or bio-sensors for the detection of single molecules by sensitized NEMS surfaces.
For example, the resonance frequency of a cantilever beam varies sensitively as a function of molecular adsorption. In addition, when the adsorption is confined to one side of the cantilever, the cantilever undergoes deflection due to adsorption-induced variation in surface free energy. Chemical selectivity can be achieved by coating the cantilevers with selective molecules. It is postulated that the cantilever bending depends on the changes in surface free energy while the resonance frequency variation is entirely due to mass loading (both specific and nonspecific adsorption). The minimum detectable adsorbed mass on a cantilever sensor can be increased by orders of magnitude by changing the dimensions of the device; smaller and thicker cantilevers offer higher resonance frequency and therefore better mass detection sensitivity.
Thus, a fast developing area of smart sensors is evolving, and an advanced combination of materials, sensors, actuators, control and processing are blended suitably to achieve devices with a huge potential for applications in many different fields.
Highly sensitive and selective detection of chemicals and biomolecules using integrated micro- and nanodevices have attracted much attention due to the importance in national security, clinical diagnostics, and industrial processes. Among these devices, nanomechanical cantilever sensors can detect surface stress or resonant frequency of a thin solid device induced by subtle molecular interactions. These cantilever nanomechanical sensors are robust and reliable in the detection and can be mass-produced using microfabrication techniques. Furthermore, their label-free and real-time detection abilities can be combined with microfluidics and microelectronics to create portable lab-on-chip systems. After the specific receptors are immobilized on one side of the cantilever, the interaction with certain molecules changes the surface stresses and leads to the bending of the cantilever.
Alternatively, the cantilever sensor can detect some physical changes such as temperature, relative humidity, viscosity, magnetic field, radiation and light intensity. These sensors can work in a dynamic mode in which the resonant frequency is monitored to reflect the change in the effective mass induced by molecule adsorption. Due to their small size and mass, nanomechanical cantilever sensors are sensitive to the mass changes induced by molecular scale recognition or adsorption, and have been used to detect nucleic acids, gaseous molecules, proteins, explosives, and other substances. The bending and the vibration of the cantilever can be measured using different approaches, including piezoresistive and piezoelectric method, optical interferometry and laser reflection methods. The laser reflection method is frequently utilized wherein a laser beam is used to illuminate the free end of a one-end fixed cantilever and changes in position are detected by a position sensitive detector.
In spite of the many advantages of the nanomechanical cantilever sensor, there are limitations and disadvantages of nanomechanical cantilever sensors regarding their adaptability and reconfigurablility. The first limitation is that for a certain magnitude of bending, it is necessary to determine whether it is due to the interaction with target species or not. This occurs because the specificity and selectivity of nanomechanical sensors comes only from immobilized molecules or receptors. The mechanical deformations of the cantilevers do not directly relate to molecular structure or property, even if the peak shape (ascending side and descending side) contains some specific molecular information. Although spectroscopic techniques (infrared and visible spectrometry, mass spectrometry, and nuclear magnetic resonance) could provide detailed information for molecular identification, adding scanning wavelength spectroscopic accessories to portable sensors increases their size, complexity, and expense, while reducing adaptability of the devices. Because one of the most important driving forces of creating a small sensor is to achieve wide and easy deployment as the result of compact size, simple structure and low price, combining the device with one spectroscopic method is thus not preferred.
Secondly, for most of the biomolecular detection applications, cantilever sensors have to take an extra step (e.g., heat treatment or stringent washing) to remove attached molecular species for the next detection. In other words, the sensor needs to recognize and detect molecules with adjustable sensitivity over a large concentration range call for an adaptive and reconfigurable sensing strategy.
Another limitation of nanomechanical cantilever sensors is the structural configuration wherein the sensor must have rectangular cross sections with thickness at sub-micrometer scale, and require that the top surface and the bottom surface be made of different material or be chemically modified. Usually a gold thin film is deposited on one side of the silicon substrate to reflect a laser beam and immobilize receptors. Such asymmetric bimorph thin film structures are sensitive to temperature change. The temperature must be controlled precisely using a thermoelectrically stabilized cell and stabilized for hours before detection, otherwise the specific binding signal cannot be differentiated from thermal noise.
Another limitation of the nanomechanical cantilever sensor is that during laser deflection detection, accurate bending measurements depend on the position of laser beam shining on the cantilever. However, 5-20% misplacement is normal because the size of the laser spot is larger than the actual width of the cantilever. A slight positional change of the laser spot on the cantilever changes the detection sensitivity. Therefore, an arbitrary bending of the sensor cannot be determined if the laser illumination position is changed due to a system perturbation. Such uncertainty makes it difficult to cross-compare the bending results.
It is important to note most surface modification based sensing devices, such as the nanomechanical cantilever sensors, do not possess the desired adaptability, reconfigurability and identification ability. These sensors have narrow detection or operation ranges, cannot detect target species at ultra low concentrations, and will be saturated at ultra high concentrations. Most importantly, all the surface modified sensors have to depend on either highly selective molecules or a large number of sensors in an array to determine the nature of molecules, making these sensors unsuitable for highly specific detection of species in uncontrolled field environments.
To place each sensing element in specific locations various techniques have been explored, but the deposition of sensing elements on contact electrodes with good registry is a low yield process and usually requires sophisticated instruments for simultaneous observation and electrode formation. It turns out that the manipulation of different sensing units is hard and time-consuming. The fabrication costs are prohibitively high for daily manufacture. When compared with instruments that are used in traditional chemical and biological analysis, micro- or nanoscale sensing devices are sensitive, integratable, relatively inexpensive, easy to be deployed and fast in response. However, unless a significant advancement in mechanism is realized, such devices are continuously challenged by the miniaturization of traditional instruments such as micro-mass spectrometers, optical sensors, and on-chip microanalysis systems.
The nanomechanical sensors of the present invention solve many problems and overcome many limitations in the prior art.
The first objective of the present invention is to provide an electrically induced deflective amplification (E-IDEA) sensor for the adaptive and reconfigurable detection of chemical and biological molecules using optical fiber transducers.
The second objective of the present invention is to provide an electrically induced deflective amplification sensor for the adaptive and reconfigurable detection of chemical and biological molecules using waveguide transducers.
The third objective of the present invention is to provide an electrically induced deflective amplification sensor for the adaptive and reconfigurable detection of chemical and biological molecules using microfabricated cantilever transducers.
The fourth objective of the present invention is to provide an electrically induced deflective amplification sensor for the adaptive and reconfigurable detection of chemical and biological molecules using nanowire transducers.
The fifth objective of this invention is to provide an electrically induced deflective amplification sensor for the adaptive and reconfigurable detection of chemical and biological molecules using nanotube transducers.
The sixth objective of this invention is to provide an electrically induced deflective amplification sensor for the adaptive and reconfigurable detection of chemical and biological molecules using piezoelectric or piezoresistive methods.
The seventh objective of this invention is to provide an electrically induced deflective amplification sensor for the adaptive and reconfigurable detection of chemical and biological molecules using optical interfermetry.
The eighth objective of this invention is to provide an electrically induced deflective amplification sensor for the adaptive and reconfigurable detection of chemical and biological molecules for accurate, on-line and in-situ monitoring.
The ninth objective of the invention is to provide an electrically induced deflective amplification sensor for the adaptive and reconfigurable detection of chemical and biological molecules at a temperature range between ambient temperatures to approximately 400° C.
The tenth objective of the invention is to provide an electrically induced deflective amplification (E-IDEA) sensor for the adaptive and reconfigurable detection of chemical and biological molecules that is not sensitive to temperature change.
A first embodiment of the present invention provides an electrically deflected nanomechanical sensor including a first and a second parallel electrode plates separated by a gap for applying a voltage difference to the first and second parallel electrode plates to generate an electric field between the first and second parallel electrode plates, an adaptive and reconfigurable transducer positioned within the gap between the first and second parallel electrode plates, the adaptive and reconfigurable transducer bending in response to detecting a physical property of a material within range and a detecting device for detecting a position of the adaptive and reconfigurable transducer, the detected material identified by a change in position of the adaptive and reconfigurable transducer. In an embodiment, the adaptive and reconfigurable transducer includes a micro- or nano-scale optical fiber and a coupling device for guiding and confining a light beam for transmitting through the optical fiber, the change in position of the optical fiber output detected by the detector to detect the bending of the optical fiber in response to the detected material. The detecting device includes a position sensitive detector for detecting a position of the transmitted light for determining the bending of the optical fiber, a piezoelectric or piezosensitive device for detecting a position of the transmitted light beam for determining the bending of the optical fiber. In another embodiment, the transducer is at least one of an optical fiber, a waveguide, a microfabricated cantilever, a nanowire, a nanotube, and a glass nanofiber and the detecting device is used to detect at least one of a gaseous molecule, a chemical molecule and a biological molecule.
A second embodiment provides a method for fabricating an electrically deflected nanomechanical sensor by forming a first conductive layer as a first electrode, depositing a first photoresist layer on the first conductive layer, placing a transducer on the photoresist layer, depositing a second photoresist layer on the first photoresist layer on the first photoresist layer over the transducer, forming a second conductive layer on the second photoresist layer and exposing the formed layers to an ultra violet light to remove the exposed first and second photoresist layers leaving a partially exposed transducer between the first and second conductive layers. Placing a transducer on the photoresist layer is accomplished by fabricating an optical fiber for transmitting a laser beam. The method further includes designing the optical fiber to achieve a predetermined mechanical and optical property. The step of placing a transducer on the photoresist layer includes placing a cantilever on the first photoresist layer and forming a cantilever having a porous structure or forming plural nanostructured glass arrays having an ordered arrangement on an external surface of the cantilever.
A third embodiment provides a system for detecting a physical property of a material including a first and a second parallel electrode plate separated by a gap, a power source connected between the first and second parallel electrode plates for generating an electric potential difference in the gap between the first and second parallel electrode plates, an adaptive and reconfigurable transducer positioned within the gap between the first and second parallel electrode plates and a detection device for detecting a bending of the adaptive and reconfigurable transducer in response to the detected a physical property of a material within range of the sensor. In an embodiment, the adaptive and reconfigurable transducer includes a micro- or nano-scale optical fiber, a laser source for providing a laser beam, a waveguide coupled with the optical fiber for guiding and confining the laser beam for transmitting through the optical fiber and an amplifier for amplifying the transmitted fiber beam, the detection device detecting a position of the amplified laser beam for determining a physical characteristic of the material. The power source includes a variable voltage source for generating a variable voltage for applying to the first and a second parallel electrode.
Further objects and advantages of this invention will be apparent from the following detailed description of a presently preferred embodiment that is illustrated schematically in the accompanying drawings.
a shows an electrically-induced deflective amplification transducer in cantilever form before molecular detection.
b shows an electrically-induced deflective amplification transducer in cantilever form after molecular detection, as shown by bending.
c is a schematic drawing of an electrically-induced deflective amplification transducer in optic fiber form before molecular detection.
d is a schematic drawing of an electrically-induced deflective amplification transducer in optic fiber form after molecular detection.
a is a photograph showing how a localized electric field breaks small organic molecules, and the fragment reacts with each other and forms condensed polymers of polythiophene.
b is a photograph showing how a localized electric field breaks small organic molecules, and the fragment reacts with each other and forms condensed polymers of polypyrrole.
a is an illustration of how the bending of E-IDEA microfiber and nanowire transducers are measured.
b is an illustration of how the bending of E-IDEA cantilever transducers are measured.
a illustrates E-IDEA based electrophoretic analysis of DNA mixture on a nanostructured transducer.
b shows two graphs showing the bending of the transducer at different electric field strengths and the corresponding graph converting to DNA amount verses length.
Before explaining the disclosed embodiment of the present invention in detail it is to be understood that the invention is not limited in its application to the details of the particular arrangement shown since the invention is capable of other embodiments. Also, the terminology used herein is for the purpose of description and not of limitation.
The following is a listing of reference numerals used throughout the specification and the drawings:
110 bottom conductive surface
115 top conductive surface
120 bottom photoresist layer
125 top photoresist layer
130 fiber
140 laser beam
150 lens
160 detector
The present invention is a nanomechanical sensor having electrically induced deflective amplification (E-IDEA) for the adaptive and reconfigurable detection of chemical and biological molecules using various transducers including optical fibers, microfabricated cantilevers, wave-guide and nanowires/nanotubes or using piezoelectric or piezoresistive methods, optical interfemetry or other methods that will be obvious to those skilled in the art.
The bending of the transducer changes the position of an emitted or reflected laser beam that is monitored by using a position sensitive photo-detector as shown in
By monitoring the deflection magnitude, the amount of molecules bound to the transducer is determined in real-time and without any label. Based on this completely different bending mechanism, E-IDEA sensors have the same advantages as traditional cantilever sensors, yet, overcome the limitations of surface modification sensors and provide both adaptivity and reconfigurability for detection, recognition and system integration.
In addition, the transducer bending is actively controlled by external electric fields, and the surface molecules are removed when the magnitude of electrostatic attraction force acting on the molecules exceeds a threshold value, desorption free energy change. The electric field induced desorption reflects the nature of adsorbed molecules using a novel spectroscopic method named as electric field programmed desorption (EFPD). Compared to asymmetric structured transducers, E-IDEA transducers with symmetric shape (fiber and wire) operate at high temperature where the emitted laser is used for detection to completely eliminate the problem of laser beam misalignment in prior art cantilever sensors.
Electrically-induced deflective amplification (E-IDEA) is based on the Coulomb's law of electrostatic interaction. A charged object between two electrodes with a potential difference moves in one direction depending on the polarity of the object. The electric field induced motions was explored in Faraday's demonstration of electrostatic repulsion between charged gold particles, in Millikan's oil droplet experiment to measure the unit charge of electron, and also in electrophoresis to separate biomolecules. Millikan demonstrated that the charged oil droplet has constant velocity when the gravity is equilibrated by electrostatic interaction as shown in
There are many other examples of utilizing electric fields to control the motion of particles: for example, an ink-jet printer uses electric attraction to guide the motion of charged toner particles. However, implementing the fundamental effect as a novel-sensing concept into a functioning device that is capable of adaptive and reconfigurable detection and recognition is unexplored, even if electrostatic actuators have been used for many years to generate large motion and force. The effect of surface charge on the bending of E-IDEA transducers is calculated by using available electrophoretic data of 4.5 μm polystyrene microparticle. The electrophoretic force was measured using optical tweezers in an aqueous solution of sodium chloride, where an electric field of approximately 3.33×103 V/m generated a 2.5×10−13 N electrostatic force as discussed by K.-I. Wada et al. in “Electric charge measurement on a single microparticle using thermodynamic analysis of electrostatic forces, Appl. Phys. Lett. (2002), 81, 1768.
Providing the same surface charge density and electric field as those on the polystyrene particle, the electrostatic force acting on the free end of a cantilever transducer approximately 200 μm long, 20 μm wide, and spring constant of 0.05 N/m in this example, bends the transducer for 2.8 nm, which is the typical bending generated in stress-based detection. From an engineering perspective, the bending of the transducer depends on the strength of electric field rather than the absolute voltage. By reducing the electrode gap to micrometer scale, the required voltage can be dramatically decreased. For instance, approximately 1.92 V DC across a 10 μm electrode gap generates 1.92×105 V/m electric field strength which is the typical value in Millikan's experiment. If the charge screening is at the similar level as that of approximately 3.33×103 V/m, the electric field of 1.92×105 V/m produces approximately 160 nm bending at the end of the transducer.
For a solid object in dry conditions, all surface charges are neutralized due to adsorption of charged ions or particles from air. The charge balance is broken by various means such as friction, heat treatment, light illumination, and chemical modification. Two common examples are rubbing a glass rod with silk which produces a positive charge, and rubbing a plastic rod with fur which produces a negative charge. In solution, simple electrolytes and polyelectrolytes (e.g., DNA, protein, and peptides) hydrolyze to form charged ions, which attract oppositely charged ions and form electric double layer.
An important concept of E-IDEA is electric field induced desorption. Absorbed molecules can be removed from the transducer by increasing the electric field strength over the threshold. An example of this process comes from the gel electrophoresis of long DNA molecules. Some DNA chains are trapped on gel fibers at low voltage, and they extend as the electric field increases. Eventually the DNA chain is broken and the segments are carried away by electrophoretic force.
For E-IDEA based detection and desorption in liquid, especially aqueous condition, the voltage is carefully controlled to avoid the electrolysis of water into hydrogen and oxygen. The minimum hydrolysis voltage of water is approximately 1.24 volts. According to standard theory, no reaction should take place at room temperature below this voltage. At the same time, the electrode gap can be reduced to provide sufficient electric field in order to remove the adsorbed molecules. To facilitate desorption of absorbed gas molecules in dry conditions, extremely high electric field strengths are required which is incompatible with portable devices. It is important to note the field evaporation/desorption of strongly bonded surface atoms has been performed at the strength of 5×109 V/m.
However, the gas sensing applications usually require moderate adsorption, which means an optimal operation range between approximately 5×109 V/m and 1.92×105 V/m is used to remove absorbed gas molecules. In addition, the electric desorption is assisted by other techniques such as laser or thermal heating to reduce the energy barrier of desorption. E-IDEA transducer of the present invention is based on a different mechanism from electrostatic force microscopy or redox-molecule modified cantilever sensors. In both cases a conductive cantilever is used, and the charge density on both electrodes is determined by the voltage applied on the electrodes.
Three experiments were used to test the novel sensing technique of the present invention. The first experiment involved immobilization and hybridization of DNA on the cantilever. The gold-thiol interaction was utilized to immobilize thiolated single stranded DNA (ssDNA) onto a thin gold film deposited on one side of a cantilever sensor. The surface stress change was used to confirm the immobilization of captured ssDNA and the hybridization of target ssDNA. Both the immobilization and the hybridization generate changes in surface stresses that are measured using real-time in solution.
At the same time, when using a high DC voltage or even a pulsed voltage with a 20 V height and 10 ms width water electrolyses and bubbles form in aqueous solutions. Such problems are solved using low voltage less than approximately 1 V on a microfabricated electrode gap.
The second experiment involved electric attraction of charged microwires. In this example, short pieces of glass fibers with diameters from approximately 500 nm to approximately 200 μm were drawn using fiber drawing methods. The freshly drawn fibers stick strongly to a flat stainless steel plate. After staying in the ambient condition for few hours, the fibers can be removed from the plate easily. When an insulating collector is placed directly underneath the fiber outlet, the orientation of a fiber is determined by those already there, and the fibers are eventually distributed in a circular pattern in the round container.
The highly charged glass fibers repel each other and are attracted to an oppositely charged object because the high temperature (e.g., 850° C.) drawing process produces surface oxygen sites with unsaturated dangling bonds. When pulled through a rubber puller, the glass fibers are charged. The adsorption of charged or dipolar species from air neutralizes the negative charges on the fibers, making them less repelling to each other. Actually there are many daily examples of electrostatic interactions that are similar to this one. The challenge is to implement electrostatic interaction into a small device without using high voltage.
A third experiment involving localized reaction in high electric fields was performed using microfabricated electrode gaps with sizes ranging from approximately 5 μm to approximately 50 μm which is similar to the designed electrode gaps in E-IDEA sensors. The three-dimensional structure of electrodes produced a localized high electric field greater than approximately 105 V/m at low voltage of approximately 10 V. When the silicon chip with electrodes was put into a chamber that was saturated with the vapor of organic molecules such as thiophene or pyrrole, a polymerization reaction occurred between the electrodes in the presence of water.
The experiments and analysis described above showed that the novel approach for adaptive and reconfigurable molecular sensing was viable. Two important questions were answered or confirmed. First, electrostatic interactions generate considerable force on micro- or nanoscale objects capable of changing their bending status and pathway. Second, localized electric fields break molecular bonds and create ionized species. Thus, E-IDEA provides a new technique for sensing using a small electrode gap with voltage drift control to produce a high strength electric field and avoid hydrolysis of water. The present invention significantly advances the fabrication of adaptive and reconfigurable sensors.
Fabrication of E-IDEA Fiber Transducer
After deriving the relation between the surface charge and the length and diameter of cylinder shaped E-IDEA transducers, the focus is on the operating principle which is to balance the elastic energy cost with the electrostatic energy gain. Providing two parallel plates with separation d and electric potential difference V, the bending of the transducer is determined by equating the elastic force with the electrostatic force. In this process y(x) is the deflected distance of the transducer from equilibrium position at point x, where x is the distance along a relaxed transducer from the fixed end. The static equilibrium shape y(x) at voltage V is obtained by minimizing the free energy
where D is charge density on the transducer; E, L, and R are the Young's modulus, the length and the diameter of the transducer, respectively. From the equation, a long and narrow fiber bends more, providing other conditions are identical. The bending of the E-IDEA transducer is dependent on charge density, electric field strength, and the mechanical property of material. In order to have high electric field strength, the electrodes are placed as close as possible. An approximately 1 V DC voltage generates 105 V/m field across a 10 μm gap. In this nanoscale exploratory research (NER), E-IDEA was proved and the first generation transducer was created with two electrodes 170 above and underneath an optical fiber 130 to detect the bending using a transmitted laser beam 140 as shown in
A fiber drawing tower was used, which makes micro- or nano-scale glass fibers by combining fiber drawing and selective chemical etching. In this method, the length, diameter and structure of the glass fiber is pre-designed to achieve desired mechanical and optical properties. Such fibers are incorporated in the fabrication of E-IDEA sensing devices using photolithography. The transducer can also be made using highly integrated photonics. Silicon waveguides are used to guide and confine light in micro or nanometer low-refractive-index materials. Two electrodes are fabricated across the suspended optical waveguide. The transducer bending is determined using a position sensitive detector. Alternately, either piezoelectric or piezoresistive approach can be substituted for the all-electronic detection of E-IDEA transducer bending.
Examples of Applications for E-IDEA
The sensitive and selective detection of charged ions in aqueous solution has significant implications for environmental protection, water treatment, national security, and health related issues. Current approaches depend mostly on electrochemical analysis, optical absorbance, and flame atomic absorption, which suffer from low sensitivity, bulky setup, and are inappropriate for on-site and real-time analysis. Micro- and nanodevices play an important role for such applications. However, most previous efforts were focused on the detection of certain molecules, media, metal ions, and chlorines, using established sensing platforms such as resistive/electric, optic and nanomechanical cantilever sensors. With the novel E-IDEA of the present invention, it is possible to build adaptive and reconfigurable sensing devices for the detection of charged non-biological species focusing on two common but important properties, pH value and chromium ion content. The purpose of the previous research is to establish the basic concept and methodology of E-IDEA sensing through a comparison with well-known detection systems.
In aqueous solution the electrolysis of water prevents the utilization of voltage high than approximately 1.24 V, otherwise the production of hydrogen and oxygen generates bubbles that disturb the bending status of the transducer. In order to have the same level of electric field as that in Millikan's experiment, the electrode gap is approximately 10 μm at voltage of approximately 1 V. Meanwhile the voltage drift in experiment is compensated relative to a reference electrode using a potentiostat. In this experiment self-assembled monolayers were used to modify the surfaces of E-IDEA transducers. To detect pH value, aminopropyltriethoxysilane that results in a surface terminating with —NH2 and —SiOH groups was used. These functional groups have different dissociation constants; thus the sensor can detect both low pH and high pH.
At low pH, the —NH2 group is protonated to form —NH3+. At high pH, —SiOH is deprotonated to —SiO−, resulting a linear response in electric/resistive nanosensors. The bending direction and magnitude are dependent on polarity and electric field strength, and the bending is fully reversible when the solution changes between acidic and basic. In addition the effectiveness of E-IDEA transducers to detect chromium ions dissolved in water was studied. The transducer surface was modified with a self-assembled monolayer of L-cysteine that has high selectivity to chromium ions and the response of the E-IDEA transducer at different electric field strength was studied as a function chromium ion concentration.
Sensing of DNA Hybridization and Conformational Change
Another application for using the E-IDEA is sensing of DNA hybridization and conformational change. An important structural feature of DNA, as well as proteins and polypeptides, is that the molecule is highly charged under standard, physiological and pH conditions due to the existence of phosphate groups on backbone. A DNA chain has a net effective density of one fundamental negative charge each 0.17 nm of its length, which is approximately two times the charge density as a single stranded DNA (ssDNA) with the same length. The intrinsic charge difference is utilized to detect target DNA electronically or electrochemically. From another standpoint, both double and single stranded DNA move to the anode in solution, but with different attraction forces.
If ssDNA is first immobilized on an E-IDEA transducer, the hybridization to its complementary ssDNA increases the net negative charge. If the bending of the modified transducer is monitored in an electric field, the transducer bends more towards the anode due to the increased electrostatic attraction. In essence, a greater net negative charge and higher electric field strengths leads to a larger bending as shown in
The present invention focuses on the adaptive detection of target ssDNA, and the conformational change of DNA double helix using an E-IDEA transducer. Experiments have immobilized capture ssDNA on gold surface, hybridized to its complementary ssDNA, and studied the interaction of thus-formed double helix with intercalating drug (ethidium bromide) using cantilever sensors in stress-bending mode.
Other experiments have immobilized capture ssDNA on the glass fiber and detected the hybridization of target ssDNA using E-IDEA transducer at different field strength. Usually DNA double helix are treated as a semi-flexible rod with persistence length of approximately 50 nm under physiological conditions. Polycations neutralize the negative charges, and induce the condensation of DNA chains into compact globules, which are important for the delivery of gene drug and the packing of chromosomal DNA in cell nucleus. Experimentally, an E-IDEA is used transducer to monitor the real-time change in the electrostatic property of DNA. In addition, E-IDEA sensor are used to measure the mechanical properties of DNA in an electric field. Through extended activities, the applications of E-IDEA sensors is extended to various fields, and establish E-IDEA as the basis for the next generation platform with adjustable sensitivity, real-time and label-free detection ability, and portability.
Electric Field Programmed Molecule Desorption
E-IDEA provides an opportunity to identify the nature of surface adsorbed molecules by monitoring the bending behaviors under electric fields with programmed strength changes. It is possible to break the double helix DNA formed by two ssDNA chains, and obtain energy barriers related to separate different numbers of base pairs. The purpose of this is to determine whether E-IDEA can be used to sequence target DNA with high or single base pair precision by controlling the electric field strength. Previous works indicated the mechanical unzipping of DNA double helix with rough resolution as a way of sequencing.
For solution based E-IDEA, whether the DNA chain is removed or unzipped is strongly is dependent on the relative strengths of surface linkage and complementary basepairs. The covalent bonds between surface-molecules are broken at approximately 1.4 nN (sulfur-gold) and 2 nN (silicon-carbon) as shown in direct force measurements. For a DNA chain immobilized on an E-IDEA transducer, the forces acting on the surface linker and the complementary base pairs are:
where Eeff depends on the voltage, electrode gap, and charge screening in solution, a, b and c are the physical lengths of DNA chains shown in
By linearly increasing the electric potential with high resolution (mV) and stability using a potentiostat, the single base pair mismatch is distinguished using E-IDEA detected desorption. Not limited to DNA, the electric field induced desorption of adsorbed molecules in general determines how strong they are adsorbed, and identify their natures using characteristic desorption electric field strengths. This spectroscopy is called as electric field programmed desorption (EFPD) that is analogous to temperature programmed desorption (TPD). The temperature programmed desorption studies of adsorbed organic acids from several zeolites confirm that programmed desorption is an effective way to identify molecules. If the desorption strengths of the electric field in EFPD are too high for portable devices, other methods with constant energy inputs such as laser, UV, or heat to reduce the energy barrier are used.
In another experiment, it was shown that UV radiation removes organic species from a silicon nitride and titanium oxide cantilever operated in stress mode. In EFPD, the adsorbed species (e.g., gaseous molecules, biomolecules, or dissolved ions) are removed from the transducer surface evenly and the transducer bending is proportional to the number of the surface adsorbed molecules. The desorption rate of the surface species is then:
where Rdes is the desorption rate, x is the kinetic order of desorption, and Ea is the desorption activation energy. E is the external electric field strength and increased linearly with time from initial E0
E=E0+βt and dE=β·dt (4)
The bending of the transducer in terms of detector signal is proportional to desorbed molecule number, and the rate at which the surface concentration of adsorbed species is decreasing:
Electrophoresis Used I-IDEA Sensors
Gel electrophoresis identifies and separate DNA and protein, studies the interactions of biomolecules, and is critically important to genomics and proteomics. The separation depends on the charge and mass. The electric attraction drives the motion of molecule and the frictional force with the gel matrix separates the molecules by size or conformation. The gel is a cross-linked polymer matrix that is immersed in the buffer. Ions in the buffer carry the current and keep the pH at constant value. In capillary electrophoresis, friction arises from the interaction with the inner wall of the capillary tube. There are some limitations of current electrophoresis: (1) it takes large amounts of sample that may be difficult to prepare; (2) the sensitivity and resolution of the detection is low; (3) the analysis takes tens of hours to complete; (4) the microstructure of gel varies from sample to sample, making structure-performance correlation and cross-comparison difficult; (5) the trapping of long DNA molecules in the media will lead to band smearing and is one major challenge faced by the traditional gel electrophoresis; and (6) the high voltage used in capillary electrophoresis leads to sample heating and the possible denaturation of biomolecules.
Rather than pulling DNA off the transducer and doing traditional electrophoretic analysis, E-IDEA identifies the length of DNA in a mixture using a transducer with a porous structure as shown in
As a result, if an electric field is applied, the chains with different length experience different attraction per unit length, showing length-dependent mobility as shown in the graphs in
Integrated Nano-waveguide for E-IDEA Sensors
One-dimensional nanomaterials, such as, nanotubes, nanowires, and nanobelts, are highly promising sensing elements due to their small mass and large surface areas. Semiconductor nanowires have been used to make electric/resistive sensors or sensor arrays. However these sensors do not have good adaptability, reconfigurability, and molecular recognition ability. There are some technical challenges related to the fabrication as well. The first one is to manipulate the nanowire and control its position and orientation in a final device. The second is to address the response of each sensing element in an array. Alternative microfabrication can create integrated structure units that can be treated as nanomaterials with spatially defined locations and orientations.
Meanwhile, the effort to incorporate one-dimensional nanomaterials into nanomechanical devices is restricted by the lack of an integrated method to modify nanowires asymmetrically, and induce and monitor the bending of nanowires in real-time. Some semiconductor nanowire/belts are used as nano-waveguides that are capable of conducting light. Experiments were conducted to study the bending of nanowire/belt or glass waveguides in an electric field with an optical microscope or a position sensitive detector. Using E-IDEA according to the present invention, an asymmetric surface modification is not required and the bending magnitude is actively controlled by an electric field. Previously the bending and vibrating of individual carbon nanotubes have been observed using electron microscope, but not in a sensor format. Electrochemical processes can also induce the motion of carbon nanotube bundles in an aqueous solution. It is expected that the integrated E-IDEA nano-sensors would be able to detect antibodies or DNA at extremely low concentration or at the single molecule level.
Nanomechanical transducers work in a dynamic mode, where the resonant frequency is related to the effective mass of the transducer. One advantage of working in a dynamic mode is that it can detect small mass changes: single bacterial cells can be detected without damage. If the transducer surface is modified by some molecule or receptor, the change in concentration of target molecules is determined from the change in resonant frequency. By working in this mode, four response parameters, frequency, phase, amplitude, and Q-factor, are measured simultaneously to offer complementary information. As transducer sizes decrease to nanoscale or quantum scale, such devices approach gigahertz frequency, the sensitivity is higher, and many new phenomena may be discovered.
E-IDEA sensors work in dynamic mode by applying a time-dependent voltage to generate a time-dependent deflection. No piezoelectric crystal is required, which can be beneficial since the fabrication of piezoelectric crystals with high frequency and small size is difficult. E-IDEA in dynamic mode is tested using nanowires, or glass nanofibers, which are principally good sensing elements due to small size, large surface area, or superior mechanical properties.
During the duration of this experiment, the detection of mercury vapor was explored. First a thin layer of gold is deposited on the surface of a fiber or nanowire. An electric field is applied across the transducer to generate vibration. The transducer is then exposed to mercury vapor, the changes in response parameters before and after exposure are recorded. Mercury atoms form amalgam with gold film, increase the effective mass and shift the resonant frequency of the transducer. In the second experiment, aligned nanowire or glass nanofiber arrays were used as waveguide transducers. The bending of transducers in electric field was monitored using a fluorescence microscope or charge coupled device (CCD) camera as shown in
Detection of Gaseous Molecules at Elevated Temperatures
The adsorption or reaction of gaseous molecules on solid surfaces change the electronic state or the charge density of the surface. This is the basis of electric/resistive sensors for the detection of gaseous molecules at high temperature. Similar changes in surface charges are used for detection using E-IDEA transducer. By using an axially symmetric transducer, such as fiber, waveguide, and nanowire, the thermally induced bimorph bending of the traditional cantilever is eliminated. Thus E-IDEA transducers detect gaseous molecule between 200 and 700° C., below the softening temperature of the transducer material.
One advantage of using an E-IDEA high temperature sensor is that the sensitivity is increased at high electric field. As a preliminary confirmation of this application, the bending of a thin film modified E-IDEA transducer at different temperatures and different electric field strengths was measured. The titanium oxide thin film prepared by sol-gel method is reactive at high temperature. Either heat treatment or gas adsorption changes the surface charge density, resulting in the bending of E-IDEA transducer in electric field. Compared to non-catalytic surfaces, the reaction of titanium oxide is faster and generates more surface charges. Thus, even if symmetric E-IDEA sensors are immune to fluctuations of room temperature, they can still be used as high temperature sensors provided other factors, such as, heating, adsorption, reaction, and desorption, can change the surface charge density.
According to experimental results, the E-IDEA transducer is a novel high temperature sensor for the adaptive and reconfigurable detection of gaseous molecules including hydrogen, carbon monoxide and explosive vapors.
Fabrication of Fiber Waveguide Transducer
Although fabricating a cantilever shaped transducer is a routine task for microfabrication, putting two parallel electrodes with 10 μm separation across a cantilever is technically difficult and expensive, concerning the current aspect ratio in deep reactive ion etching (20:1). The present invention discloses a method to place optical micro-fiber or nano-fibers into E-IDEA based detection systems. The special fibers are made by scalable fiber drawing technique combined with the state-of-the-art selective etching. The fibers are cut into desired length ranging from millimeter to tens centimeter.
Surface Charge Density Change of Functional Monolayer in Solution
The self-assembled monolayer terminated with amine group hydrolyzde in an aqueous solution. On such surface, un-reacted surface hydroxyl groups coexist with amine groups. The two groups hydrolyze at different pH value: NH2 hydrolyzes to NH3+ at low pH, and OH hydrolyzes to O− at high pH. The combination of NH2 groups and OH groups extend the response to a wide range of pH as shown in
Temperature Programmed Desorption to Identify Organic Acids
In the study of basicity on zeolite catalysts, a preferred method to study the adsorption of different organic acids on the surface of zeolites is using temperature programmed desorption (TPD). The shape of TPD curves strongly depend on the molecules and the zeolites. For example,
UV Induced Desorption of Molecules from Surface
The exposure of a TiO2 coated cantilever to UV at a wavelength of approximately 365 nm bends the cantilever slightly. The exposure to UV at 254 nm radiation (5.1 eV) bends the cantilever significantly toward the light source, as shown in
When UV light at approximately 254 nm is periodically turned on and off, the cantilever shows bending corresponding to the on and off, and the magnitude is proportional to the length of the radiation time. The photo-induced bending is explained as the removal of surface adsorbed species (oxygen) and the generation of active surface sites with unsaturated coordination. The cantilever then experiences a compressive surface stress on the UV-exposed side and a tensile stress on the other side, and thus bends toward the radiation source as the result of the stress imbalance. When the UV light is turned off, the readsorption of atmospheric O2 which is usually faster than other adsorbates in ambient condition due to the high concentration, on active sites increases surface stress on the UV-exposed side and leads to a backward bending.
The partial recovery of bending confirms the effect of other adsorbates. The photodesorption follows the first-order kinetics: dn/dt=−σFn where n is the number of adsorbed O2, σ is the cross section of photodesorption, F is the photon flux, and t is radiation time. The bending of the TiO2 coated cantilever with UV light in
Glass Micro- and Nano-Fibers Made from Fiber Drawing Method
Using a fiber drawing tower, it is possible to make special nanostructured glass with various diameter ranging from hundreds of nanometer to hundreds of micrometers. These fibers have different geometry and structures to allow defined researches. For example,
Magnetic Force Induced Bending of Cantilever
The bending of nanomechanical transducers is a general phenomenon. Experiments on magnetic force induced bending of traditional cantilever sensor for the detection biomolecules using magnetic particle probes have been performed. In Sandwich immunoassay biotin modified cantilever and biotin labeled magnetic particles were used to detect antibodies in solution. The reaction between the biotin and streptavidin immobilizes the magnetic particles on the cantilever, which bends in the magnetic field of neodymium magnet. Other experiments were performed using 16-mercaptohexa-decanoic acid to capture iron oxide nanoparticles on the cantilever. Results showed that the magnitude of cantilever bending is not the same at different position over the magnet, indicating the effect of magnetic field strength on cantilever bending as shown in the graph of
The methods for fabricating optical fiber or waveguide nanomechanical sensors can be used to fabricate sensors commercially available for many diversified applications including national security, public health, and scientific and technological development. Applications for the E-IDEA sensor of the present invention includes, but are not limited to, detection of gaseous, chemical and biological species, molecular recognition, spectroscopy, electrophoretic analysis and sensing at high temperatures.
While the invention has been described, disclosed, illustrated and shown in various terms of certain embodiments or modifications which it has presumed in practice, the scope of the invention is not intended to be, nor should it be deemed to be, limited thereby and such other modifications or embodiments as may be suggested by the teachings herein are particularly reserved especially as they fall within the breadth and scope of the claims here appended.
This application claims the benefit of priority to U.S. Provisional Application No. 60/876,878 filed on Dec. 22, 2006.
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Number | Date | Country | |
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60876878 | Dec 2006 | US |