Progress 03/01/99 to 02/28/04
Outputs
We report the electric-field influences on biomolecules and suspended particles in buffer solutions. Preliminary results demonstrated that 1)The translocation of biomolecules can be influenced by electric fields, 2)Careful electrode design can elicit various behavior of particle motion in AC fields, and 3)The rotary motion of the F1-ATPase enzyme is unaffected by such AC fields. The electrodes exerted limited negative DEP force on the biomolecules as particles were attracted towards attachment sites either by the induced fluidic flow from local temperature fluctuations or due to the repulsive effects of the intensified electric field regions. The smallest features of the electrode pattern the tips of the nickel attachment posts, were estimated to be less than 30 nm. Arrays of attachment sites were wired together to provide multiple assembly points for simultaneous observation of the rotary motion of the hybrid device. The nanoscale features therefore enabled the
application of greater electric field intensities on the assembly system. The biomolecules themselves, consisting of the biotinylated biomolecular motor F1-ATPase, are conjugated with Quantum Dots (QDOTS605 from Quantum Dots Corporation). The QDOTS have been labeled with streptavidin and therefore provide a reliable interface with the F1 motor. Furthermore, the 20 nm QDOTS do not photobleach, which is critical for their detection with a silicon intensified tube (SIT) camera (Hamamatsu Inc.). Infusion of these particles into the assembly area will be used to demonstrate directed attachment to the array. Initial results suggest that the particles seem to be influenced by negative DEP (nDEP) because they want to move away from field intense locations once they have been localized in the vicinity of electrodes. Further development of electrodes will utilize the trapping effect to localize assembled nano-devices at specific attachment points.
Impacts
The project established a pathway for other students at all ages to begin to experience the current efforts in nanotechnologies. As on of the main supporting projects encompassed in the CNSI program, the information learned from nano-assembly device development has been shared with others in the research community in an effort to refine the educational initiatives and extend opportunities for various individuals to participate in science.
Publications
- Soong, R.K., Neves, H.P., Schmidt, J., and Montemagno, C.D. 2001. Engineering Hybrid Nanoscale Devices Powered By Biomolecular Motors. Biomedical Microdevices, vol.3, p.69.
- Bachand, G.D., Soong, R.K., Neves, H.P., Olkhovets, A., Craighead, H.G. and Montemagno, C.D. 2001. Precision Attachment of Individual F1-ATPase Biomolecular Motors on Nanofabricated Substrate. Nano Letters, vol. 1, p. 42.
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Progress 01/01/02 to 12/31/02
Outputs
Current efforts are focused on directed assembly of hybrid nano-devices using dielectrophoresis (DEP). While previous work resulted in the engineering of functional rotary structures powered by F1-ATPase biomotors, the yield was less than 1 percent. The low device yield was attributed to poor control in the interfacing of components and the relatively low concentration of Ni rods (28 fM) compared with the motor proteins. The low concentration is inherent in the labor intensive electron beam (e-beam) lithography method of Ni bar synthesis. Furthermore, the low number of Ni rods suspended in solution coupled with the small contact area of the Ni post assembly points prevented effective diffusion of the metal bars to the motor proteins. Hence, DEP is utilized in an effort to implement control over the assembly process and more importantly, enforce greater reliability in the interfacing of components. Initial experimentation indicated that DEP forces might be used to
direct insulating 0.78 micron and 60 nm particles to electrode attachment sites through the application of a non-uniform electric field. Particles suspended in solution either collected at electrode tips (intense field areas) or remained in regions of low electrical potential upon utilization of an AC electric field. The latter describes the negative (nDEP) effect while the former is the positive (pDEP) effect. The conductivity of the media (typically 100 mM phosphate buffer in which the purified F1-ATPase is also suspended) and the frequency/amplitude of the sinusoidal AC field determine whether pDEP or nDEP is observed. Electrode geometry is critical in maintaining the proper electric field for the desired polarization and DEP effect on net neutral particles. A quadrupole configuration with nano-scale tips (minimum feature size of 20 nm) for maximum field gradients has been fabricated and tested for optimal, directed hybridizations. Although initial testing to demonstrate DEP
effects and the associated frequency responses were performed on insulating, fluorescent polystyrene (PS) microspheres, proteins such as F1-ATPase may also be modeled as having no net charge.
Impacts
Aside from DEP effects, electro-osmotic flow, and particle attraction/repulsion influenced by charge adsorption were observed as well. Frequency responses of these various effects due to the AC field will be explored to optimize the localization of biomolecules. It is also noted that Ni rods were attracted towards electrodes. Therefore once the mechanism for the controlled movement of F1-ATPase has been determined, full hybrid assembly may be achieved. Phosphorescent, eosin-labels on the F1 enzyme will enable visualization of the collection of the biomolecules on attachment sites. Ultimately, demonstration of the full capabilities of the new hybrid nano-device assembly system will be rendered by the visualization of an array of rotating Ni bars powered by F1-ATPase spaced evenly by the attachment sites.
Publications
- No publications reported this period
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Progress 01/01/01 to 12/31/01
Outputs
During this period we have: Successfully engineered and expressed three different mutant F1-ATPase motor systems. These mutants have attachment handles on different portions of the protein and will be used to evaluate the effect of binding different portions of the protein to engineered substrates on the engineering performance of the motor. Imaged individual F1-ATPase motors on nanodots using Atomic Force Microcopy. This validated the hypothesis that we are positioning and attaching individual motors onto nanosized engineered features and established the investigative methodology to quantitatively determine the optimum feature size to ensure maximum site occupancy with a minimum of multiple motors on each site. Quantified the binding efficiency of different engineered surfaces to histidine peptide chains. This work establishes a direct measure of the binding strength of the motors to different substrates. Developed and demonstrated an instrument for measuring the
rotational displacement of the rotor of individual F1-ATPase motors. Using this device we have measured the eccentricity of the proteins conformation changes during rotation and the changes in angular velocity as a function of the rotor position during rotation. This is a significant accomplishment that was essential for quantitatively evaluating the performance of different engineered constructs of the F1-ATPase biomolecular motor. These results are currently being prepared for publication in either Science or Nature. Designed and successfully fabricated nano-propellers (60 nm) for attachment to F1-ATPase biomolecular motor. Produced and purified Bacteriorhodopsin and inserted the protein into 150 nm liposomes. This is the first step for producing a light powered biomolecular motor powered nanomechanical device. Begun the engineering evaluation of a porous membrane system to replace the organic liposome membrane system for producing fuel for the biomolecular motor powered NEMS
devices from light.
Impacts
The use of ATP-powered biomolecular motors opens the door for developing autonomous devices that may be maintained and fueled in a manner that is totally transparent to the user. The proposed research will create the world's first biomolecular motor-powered nanomechanical device, and establish prerequisite technologies for the further evolution of such devices.
Publications
- No publications reported this period
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Progress 01/01/00 to 12/31/00
Outputs
During this period we have: Successfully engineered and expressed three different mutant F1-ATPase motor systems. These mutants have attachment handles on different portions of the protein and will be used to evaluate the effect of binding different portions of the protein to engineered substrates on the engineering performance of the motor. Imaged individual F1-ATPase motors on nanodots using Atomic Force Microcopy. This validated the hypothesis that we are positioning and attaching individual motors onto nanosized engineered features and established the investigative methodology to quantitatively determine the optimum feature size to ensure maximum site occupancy with a minimum of multiple motors on each site. Quantified the binding efficiency of different engineered surfaces to histidine peptide chains. This work establishes a direct measure of the binding strength of the motors to different substrates. Developed and demonstrated an instrument for measuring the
rotational displacement of the rotor of individual F1-ATPase motors. Using this device we have measured the eccentricity of the proteins conformation changes during rotation and the changes in angular velocity as a function of the rotor position during rotation. This is a significant accomplishment that was essential for quantitatively evaluating the performance of different engineered constructs of the F1-ATPase biomolecular motor. These results are currently being prepared for publication in either Science or Nature. Designed and successfully fabricated nano-propellers (60 nm) for attachment to F1-ATPase biomolecular motor. Produced and purified Bacteriorhodopsin and inserted the protein into 150 nm liposomes. This is the first step for producing a light powered biomolecular motor powered nanomechanical device. Begun the engineering evaluation of a porous membrane system to replace the organic liposome membrane system for producing fuel for the biomolecular motor powered NEMS
devices from light.
Impacts
The use of ATP-powered biomolecular motors opens the door for developing autonomous devices that may be maintained and fueled in a manner that is totally transparent to the user. The proposed research will create the world's first biomolecular motor-powered nanomechanical device, and establish prerequisite technologies for the further evolution of such devices.
Publications
- Ma, Y., and C, D, Montemagno, '3D Image Analysis of 2-Phase Flow System in Porous Media', Proceedings of Geovision'99, Liege, Belgium, May, 1999.
- Montemagno, C. D., and G. Bachand, 'Constructing Biological Motor Powered Nano-mechanical Devices', Nanotechnology, 10(3), 225-231, 1999.
- Bachand G. and C. D. Montemagno, 'Constructing Biomolecular motor-powered , hybrid NEMS Devices', Proceedings of the International Symposium on Microelec-tronics and Micro-Electro-Mechanical Systems, SPIE #3892-14, Queensland, Australia, 1999.
- Soong, R. K., G. Bachand, and C. D. Montemagno, 'Precision Attachment of Engi-neered Biomolecular Motors to Nanofabricated Substrates', Proceedings of the Inter-national Conference on Modeling and Simulation of Microsystems, Puerto Rico, April, 1999.
- Montemagno, C.D., 'Integrating Life Processes into Engineered Nanofabricated Devices', Proceedings of the DARPA Workshop on Energy Harvesting I/Biofuel Cell I, Arlington, VA, March, 1999.
- Montemagno, C. D., G. Bachand, S. Stelick, and M. Bachand,'Constructing Biolog-ical Motor Powered Nanomechanical Devices', Sixth Foresight Conference on Molec-ular Nanotechnology, http://www.foresight.org/conference/MNT6/index.html, Santa Clara, CA, November, 1998.
- Soong, R.K., Bachand, G.D., Neves, H.P., Olkhovets, A.G.,Craighead, H.G., and Montemagno, C.D., Powering an inorganic nanodevices with a biomolecular motor. Science, 290: 1555-1558, 2000.
- Bachand, G.D., and Montemagno, C.D. Constructing organic/inorganic NEMS devices powered by biomolecular motors; Biomedical Microdevices, 2(3) 179-184, 2000.
- Nam, T. K., M. B. Timmons, C. D. Montemagno, and S. M. Tsukuda. Biofilm characteristics as affected by sand size and location in fluidized bed vessels, Aquacultural Engineering, 22, 213-224, 2000.
- Montemagno, C. D. and Yu Ma, 'Measurement of Interfacial Surface Areas for Two-Phase Flow in Porous Media from PVI Data', in Proc. of Characterization and Measurement of the Hydraulic Properties of Unsaturated Porous Media, M.Th. van Genuchten (ed.), Riverside, CA., 121-132, 1999.
- Montemagno, C. D. and L. J. Pyrak-Nolte, 'Fracture Network versus Single Fractures: Measurement of Fracture Geometry with X-ray Tomography', Physics and Chemistry of the Earth, 24(7), 575-579, 1999.
- Montemagno, C. D., 'Constructing Biological Motor Powered Nanomechanical Devices', Proceedings of the 10th International Conference Solid-State Sensors and Actuators, Sendai, Japan, 1999.
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Progress 01/01/99 to 12/31/99
Outputs
Successfully engineered and expressed three different mutant F1-ATPase motor systems. These mutants have attachment handles on different portions of the protein and will be used to evaluate the effect of binding different portions of the protein to engineered substrates on the engineering performance of the motor. Imaged individual F1-ATPase motors on nanodots using Atomic Force Microcopy. This validated the hypothesis that we are positioning and attaching individual motors onto nanosized engineered features and established the investigative methodology to quantitatively determine the optimum feature size to ensure maximum site occupancy with a minimum of multiple motors on each site. Quantified the binding efficiency of different engineered surfaces to histidine peptide chains. This work establishes a direct measure of the binding strength of the motors to different substrates. Developed and demonstrated an instrument for measuring the rotational displacement of the
rotor of individual F1-ATPase motors. Using this device we have measured the eccentricity of the proteins conformation changes during rotation and the changes in angular velocity as a function of the rotor position during rotation. This is a significant accomplishment that was essential for quantitatively evaluating the performance of different engineered constructs of the F1-ATPase biomolecular motor. These results are currently being prepared for publication in either Science or Nature. Designed and successfully fabricated nano-propellers (60 nm) for attachment to F1-ATPase biomolecular motor. Produced and purified Bacteriorhodopsin and inserted the protein into 150 nm liposomes. This is the first step for producing a light powered biomolecular motor powered nanomechanical device. Begun the engineering evaluation of a porous membrane system to replace the organic liposome membrane system for producing fuel for the biomolecular motor powered NEMS devices from light.
Impacts
The use of ATP-powered biomolecular motors opens the door for developing autonomous devices that may be maintained and fueled in a manner that is totally transparent to the user. The proposed research will create the world's first biomolecular motor-powered nanomechanical device, and establish prerequisite technologies for the further evolution of such devices.
Publications
- Montemagno, C. D., and G. Bachand, Constructing Biological Motor Powered Nano-mechanical Devices, Nanotechnology, 10(3), 225-231, 1999.
- Bachand G. and C. D. Montemagno, Constructing Biomolecular motor-powered , hybrid NEMS Devices, Proceedings of the International Symposium on Microelec-tronics and Micro-Electro-Mechanical Systems, SPIE #3892-14, Queensland, Australia, 1999.
- Soong, R. K., G. Bachand, and C. D. Montemagno, Precision Attachment of Engi-neered Biomolecular Motors to Nanofabricated Substrates, Proceedings of the Inter-national Conference on Modeling and Simulation of Microsystems, Puerto Rico, April, 1999.
- Montemagno, C.D., Integrating Life Processes into Engineered Nanofabricated Devices, Proceedings of the DARPA Workshop on Energy Harvesting I/Biofuel Cell I, Arlington, VA, March, 1999.
- Montemagno, C. D., G. Bachand, S. Stelick, and M. Bachand, Constructing Biolog-ical Motor Powered Nanomechanical Devices, Sixth Foresight Conference on Molec-ular Nanotechnology, http://www.foresight.org/conference/MNT6/index.html, Santa Clara, CA, November, 1998.
- Montemagno, C. D., Constructing Biological Motor Powered Nanomechanical Devices", Proceedings of the 10th International Conference Solid-State Sensors and Actuators, Sendai, Japan, 1999.
- Montemagno, C.D., Integrating Life Processes into Engineered Nanofabricated Devices, Proceedings of Biomedical Applications of Nanofabrication Workshop, New York, NY, March, 1999.
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