Demonstration of an Unusual Thermal Effect in the Casimir Force from Graphene.

We report precision measurements of the gradient of the Casimir force between an Au-coated sphere and graphene sheet deposited on a silica plate. The measurement data are compared with exact theory using the polarization tensor found in the framework of the Dirac model including effects of the nonzero chemical potential and energy gap of the graphene sample with no fitting parameters. The very good agreement between experiment and theory demonstrates the unusually big thermal effect at separations below 1 µm which has never been observed for conventional 3D materials. Thus, it is confirmed experimentally that for graphene the effective temperature is determined by the Fermi velocity rather than by the speed of light.

Demonstration of angle-dependent Casimir force between corrugations.

The normal Casimir force between a sinusoidally corrugated gold coated plate and a sphere was measured at various angles between the corrugations using an atomic force microscope. A strong dependence on the orientation angle of the corrugation is found. The measured forces were found to deviate from the proximity force approximation and are in agreement with the theory based on the gradient expansion including correlation effects of geometry and material properties. We analyze the role of temperature. The obtained results open new opportunities for control of the Casimir effect in micromechanical systems.

Demonstration of the Casimir force between ferromagnetic surfaces of a Ni-coated sphere and a Ni-coated plate.

We demonstrate the Casimir interaction between two ferromagnetic boundary surfaces using the dynamic atomic force microscope. The experimental data are found to be in excellent agreement with the predictions of the Lifshitz theory for magnetic boundary surfaces combined with the plasma model approach. It is shown that for magnetic materials the role of hypothetical patch potentials is opposite to that required for reconciliation of the data with the Drude model.

Variable-temperature device for precision Casimir-force-gradient measurement.

We present the design and use of an instrument that is based on a microcantilever to perform precision force gradient measurements. We demonstrate its performance through measurements of the Casimir pressure at various temperatures. The instrument can operate in high vacuum environments and temperatures between 5 K and 300 K. It uses an all-fiber optical interferometer to detect the resonant-frequency shift of a customized microcantilever due to the presence of a force gradient. To measure this shift we use both, a technique of frequency-modulation atomic force microscopy and the direct recording of the thermomechanical resonant frequency.

How to modify the van der Waals and Casimir forces without change of the dielectric permittivity.

We propose a new experiment on the measurement of the Casimir force and its gradient between a Au-coated sphere and two different plates made of doped semiconductors. The concentrations of charge carriers in the plates are chosen slightly below and above the critical density at which the Mott-Anderson insulator-metal transition occurs. We calculate changes in the Casimir force and the Casimir pressure due to the insulator-metal transition using the standard Lifshitz theory and the phenomenological approach neglecting the contribution of free charge carriers in the dielectric permittivity of insulator materials (this approach was recently supported by the measurement data of several experiments). It is demonstrated that for the special selection of semiconductor materials (S- or Se-doped Si, B-doped diamond) the calculation results using the two theoretical approaches differ significantly and the predicted effects are easily detectable using the existing laboratory setups. In the case that the prediction of the phenomenological approach is confirmed, this would open opportunities to modify the van der Waals and Casimir forces with almost no change of room temperature dielectric permittivity.

Single-molecule measurements of dissociation rates and energy landscapes of binary trans snare complexes in parallel versus antiparallel orientation.

Interactions between synaptobrevin 2 (Sb2) and syntaxin 1A (Sx1A) can be readily isolated and studied with the use of force spectroscopy single-molecule measurements. We studied interactions between Sx1A and Sb2 in two different orientations (parallel and antiparallel) using four different terminus configurations of these proteins. Force-loading experiments indicated that protein pairs in any configuration/orientation are zippered. We measured the extension and force for disassembly of these interactions, calculated the spontaneous dissociation lifetimes, and determined their free energies, enthalpies, and entropies. Although the free energies were very similar for all four configurations (∼28 k(B)T (Eyring model) and ∼20 k(B)T (Kramers model)), the enthalpy changes of binary Sx1A-Sb2 interactions varied between 24.7 k(B)T and 33.1 k(B)T. This variation is consistent with the conformation changes that occur during disassembly of the various protein terminus configurations, as verified by alterations in the extension. The parallel interactions appear to be energetically somewhat advantageous over antiparallel configurations/orientation, especially when the N-termini of Sx1A-Sb2 are left to interact freely.

Reduction of the Casimir force from indium tin oxide film by UV treatment.

A significant decrease in the magnitude of the Casimir force (from 21% to 35%) was observed after an indium tin oxide sample interacting with an Au sphere was subjected to the UV treatment. Measurements were performed by using an atomic force microscope in high vacuum. The experimental results are compared with theory and a hypothetical explanation for the observed phenomenon is proposed.

Single molecule measurements of mechanical interactions within ternary SNARE complexes and dynamics of their disassembly: SNAP25 vs. SNAP23.

Regulated exocytosis is a crucial event for intercellular communication between neurons and astrocytes within the CNS. The soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE) complex, composed of synaptobrevin 2, syntaxin and synaptosome-associated protein of 25 kDa or 23 kDa (SNAP25 or SNAP23), is essential in this process. It was reported that SNAP25 and SNAP23 have distinct roles in exocytotic release, where SNAP25, but not SNAP23, supports an exocytotic burst. It is not clear, however, whether this is due to the intrinsic properties of the ternary SNARE complex, containing either SNAP25 or SNAP23, or perhaps due to the differential association of these proteins with ancillary proteins to the complex. Here, using force spectroscopy, we show from single molecule investigations of the SNARE complex, that SNAP23A created a local interaction at the ionic layer by cuffing syntaxin 1A and synaptobrevin 2, similar to the action of SNAP25B; thus either of the ternary complexes would allow positioning of vesicles at a maximal distance of approximately 13 nm from the plasma membrane. However, the stability of the ternary SNARE complex containing SNAP23A is less than half of that for the complex containing SNAP25B. Thus, differences in the stability of the two different ternary complexes could underlie some of the SNAP25/23 differential ability to control the exocytotic burst.

Single Molecule Measurements of Interaction Free Energies Between the Proteins Within Binary and Ternary SNARE Complexes.

We use an Atomic Force Microscope based single molecule measurements to evaluate the activation free energy in the interaction of SNARE proteins syntaxin 1A, SNAP25B and synaptobrevin 2 which regulate intracellular fusion of vesicles with target membranes. The dissociation rate of the binary syntaxin-synaptobrevin and the ternary syntaxin-SNAP25B-synaptobrevin complex was measured from the rupture force distribution as a function of the rate of applied force. The temperature dependence of the spontaneous dissociation rate was used to obtain the activation energy to the transition state of 19.8 +/- 3.5 kcal/mol = 33 +/- 6 k(B)T and 25.7 +/- 3.0 kcal/mol = 43 +/- 5 k(B)T for the binary and ternary complex, respectively. They are consistent with those measured previously for the ternary complex in lipid membranes and are of order expected for bilayer fusion and pore formation. The DeltaG was 12.4-16.6 kcal/mol = 21-28 k(B)T and 13.8-18.0 kcal/mol = 23-30 k(B)T for the binary and ternary complex, respectively. The ternary complex was more stable by 1.4 kcal/mol = 2.3 k(B)T, consistent with the spontaneous dissociation rates. The higher adhesion energies and smaller molecular extensions measured with SNAP25B point to its possible unique and important physiological role in tethering/docking the vesicle in closer proximity to the plasma membrane and increasing the probability for fusion completion.

Molecular form follows function: (un)snaring the SNAREs.

Exocytotic release of transmitters is mediated by the ternary SNARE complex. The form of this complex is consistent with its function in the positioning of vesicles to the plasma membrane and their fusion to it. Recent advances in single-molecule techniques, however, bring an additional layer of complexity to this process, implicating that there might be various modes of operation. For example, the binary syntaxin-synaptobrevin 2 complex, in addition to the ternary complex containing SNAP25, might enable vesicular docking. Single-molecule techniques allow direct measurements of the distance/extension, rupture force, spontaneous dissociation times and interaction energy for SNARE protein-protein interactions. These measurements are complementary to results and conclusions drawn from other techniques. Consequently, single-molecule techniques promise tremendous opportunities for in vitro investigations of SNARE proteins to improve our understanding of their role in exocytosis.

Comparative energy measurements in single molecule interactions.

Single molecule experiments have opened promising new avenues of investigations in biology, but the quantitative interpretation of results remains challenging. In particular, there is a need for a comparison of such experiments with theoretical methods. We experimentally determine the activation free energy for single molecule interactions between two synaptic proteins syntaxin 1A and synaptobrevin 2, using an atomic force microscope and the Jarzynski equality of nonequilibrium thermodynamics. The value obtained is shown to be reasonably consistent with that from single molecule reaction rate theory. The temperature dependence of the spontaneous dissociation lifetime along with different pulling speeds is used to confirm the approach to the adiabatic limit. This comparison of the Jarzynski equality for intermolecular interactions extends the procedure for calculation of activation energies in nonequilibrium processes.

Demonstration of optically modulated dispersion forces.

We report the first experiment on the optical modulation of dispersion forces through a change of the carrier density in a Si membrane. For this purpose a high-vacuum based atomic force microscope and excitation light pulses from an Ar laser are used. The experimental results are compared with two theoretical models. The modulation of the dispersion force will find applications in optomechanical micromachines.

Demonstration of the difference in the Casimir force for samples with different charge-carrier densities.

A measurement of the Casimir force between a gold coated sphere and two Si plates of different carrier densities is performed using a high vacuum based atomic force microscope. The results are compared with the Lifshitz theory and good agreement is found. Our experiment demonstrates that by changing the carrier density of the semiconductor plate by several orders of magnitude it is possible to modify the Casimir interaction. This result may find applications in nanotechnology.

Single molecule mechanical probing of the SNARE protein interactions.

Exocytotic release of neurotransmitters is mediated by the ternary soluble N-ethyl maleimide-sensitive fusion protein attachment protein receptors (SNAREs) complex, comprised of syntaxin (Sx), synaptosome-associated protein of 25 kDa (SNAP25), and synaptobrevin 2 (Sb2). Since exocytosis involves the nonequilibrium process of association and dissociation of bonds between molecules of the SNARE complex, dynamic measurements at the single molecule level are necessary for a detailed understanding of these interactions. To address this issue, we used the atomic force microscope in force spectroscopy mode to show from single molecule investigations of the SNARE complex, that Sx1A and Sb2 are zippered throughout their entire SNARE domains without the involvement of SNAP25. When SNAP25B is present in the complex, it creates a local interaction at the 0 (ionic) layer by cuffing Sx1A and Sb2. Force loading rate studies indicate that the ternary complex interaction is more stable than the Sx1A-Sb2 interaction.

Botulinum toxin type B micromechanosensor.

Botulinum neurotoxin (BoNT) types A, B, E, and F are toxic to humans; early and rapid detection is essential for adequate medical treatment. Presently available tests for detection of BoNTs, although sensitive, require hours to days. We report a BoNT-B sensor whose properties allow detection of BoNT-B within minutes. The technique relies on the detection of an agarose bead detachment from the tip of a micromachined cantilever resulting from BoNT-B action on its substratum, the synaptic protein synaptobrevin 2, attached to the beads. The mechanical resonance frequency of the cantilever is monitored for the detection. To suspend the bead off the cantilever we use synaptobrevin's molecular interaction with another synaptic protein, syntaxin 1A, that was deposited onto the cantilever tip. Additionally, this bead detachment technique is general and can be used in any displacement reaction, such as in receptor-ligand pairs, where the introduction of one chemical leads to the displacement of another. The technique is of broad interest and will find uses outside toxicology.

New features of the thermal Casimir force at small separations.

The difference of the thermal Casimir forces at different temperatures between real metals is shown to increase with a decrease of the separation distance. This opens new opportunities for the demonstration of the thermal dependence of the Casimir force. Both configurations of two parallel plates and a sphere above a plate are considered. Different approaches to the theoretical description of the thermal Casimir force are shown to lead to different measurable predictions.

Demonstration of the lateral casimir force.

The lateral Casimir force between a sinusoidally corrugated gold coated plate and large sphere was measured for surface separations between 0.2 to 0.3 microm using an atomic force microscope. The measured force shows the required periodicity corresponding to the corrugations. It also exhibits the necessary inverse fourth power distance dependence. The obtained results are shown to be in good agreement with a complete theory taking into account the imperfectness of the boundary metal. This demonstration opens new opportunities for the use of the Casimir effect for lateral translation in microelectromechanical systems.