Title:
Micromachined membrane-based active probes for biomolecular force spectroscopy
Micromachined membrane-based active probes for biomolecular force spectroscopy
Author(s)
Torun, Hamdi
Advisor(s)
Degertekin, F. Levent
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Abstract
Atomic force microscope (AFM) is an invaluable tool for measurement of pico-Newton to nano-Newton levels of interaction forces in liquid. As such, it is widely used to measure single-molecular interaction forces through dynamic force spectroscopy. In this technique, the interaction force spectra between a specimen on the sharp tip of the cantilever and another specimen on the substrate is measured by repeatedly moving the cantilever in and out of contact with the substrate. By varying the loading rate and measuring the bond rupture force or bond lifetime give researchers information about the strength and dissociation rates of non-covalent bonds, which in turn determines the energy barriers to overcome. Commercially available cantilevers can resolve interaction forces as low as 5 pN with 1 kHz bandwidth in fluid. This resolution can be improved to 1 pN by using smaller cantilevers at the expense of microfabrication constraints and sophisticated detection systems. The pulling speed of the cantilever, which determines the loading rate of the bonds, is limited to the point where the hydrodynamic drag force becomes comparable to the level of the molecular interaction force. This level is around 10 um/s for most cantilevers while higher pulling speeds are required for complete understanding of force spectra. Thus, novel actuators that allow higher loading rates with minimal hydrodynamic drag forces on the cantilevers, and fast, sensitive force sensors with simple detection systems are highly desirable.
This dissertation presents the research efforts for the development of membrane-based active probe structures with electrostatic actuation and integrated diffraction-based optical interferometric force detection for single-molecular force measurements. Design, microfabrication and characterization of the probes are explained in detail. A setup including optics and electronics for experimental characterization and biological experiments with the probes membranes is also presented. Finally, biological experiments are included in this dissertation.
The "active" nature of the probe is because of the integrated, parallel-plate type electrostatic actuator. The actuation range of the membrane is controlled with the gap height between the membrane and the substrate. Within this range it is possible to actuate the membrane fast, with a speed limited by the membrane dynamics with negligible hydrodynamic drag. Actuating these membrane probes and using a cantilever coupled to the membrane, fast pulling experiments with an order of magnitude faster than achieved by regular AFM systems are demonstrated.
The displacement noise spectral density for the probe was measured to be below 10 fm/rtHz for frequencies as low as 3 Hz with differential readout scheme. This noise floor provides a force sensitivity of 0.3 - 3 pN with 1 kHz bandwidth using membranes with spring constants of 1 - 10 N/m. This low inherent noise has a potential to probe wide range of biomolecules. The probes have been demonstrated for fast-pulling and high-resolution force sensing. Feasibility for high throughput parallel operation has been explored. Unique capabilities of the probes such as electrostatic spring constant tuning and thermal drift cancellation in AFM are also presented in this dissertation.
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Date Issued
2010-01-04
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Dissertation