Guillaume Tresset

Highly coupled ATP synthesis by F1-ATPase single molecules

F1-ATPase is the smallest known rotary motor, and it rotates in an anticlockwise direction as it hydrolyses ATP. Single-molecule experiments point towards three catalytic events per turn, in agreement with the molecular structure of the complex. The physiological function of F1 is ATP synthesis. In the ubiquitous F0F1 complex, this energetically uphill reaction is driven by F0, the partner motor of F1, which forces the backward (clockwise) rotation of F1, leading to ATP synthesis. Here, we have devised an experiment combining single-molecule manipulation and microfabrication techniques to measure the yield of this mechanochemical transformation. Single F1 molecules were enclosed in femtolitre-sized hermetic chambers and rotated in a clockwise direction using magnetic tweezers. When the magnetic field was switched off, the F1 molecule underwent anticlockwise rotation at a speed proportional to the amount of synthesized ATP. At 10 Hz, the mechanochemical coupling efficiency was low for the α3β3γ subcomplex (F1-ɛ), but reached up to 77% after reconstitution with the ɛ-subunit (F1+ɛ). We provide here direct evidence that F1 is designed to tightly couple its catalytic reactions with the mechanical rotation. Our results suggest that the ɛ-subunit has an essential function during ATP synthesis.

Microfabricated arrays of femtoliter chambers allow single molecule enzymology

Precise understanding of biological functions requires tools comparable in size to the basic components of life. Single molecule studies have revealed molecular behaviors usually hidden in the ensemble- and time-averaging of bulk experiments. Although most such approaches rely on sophisticated optical strategies to limit the detection volume, another attractive approach is to perform the assay inside very small containers. We have developed a silicone device presenting a large array of micrometer-sized cavities. We used it to tightly enclose volumes of solution, as low as femtoliters, over long periods of time. The microchip insures that the chambers are uniform and precisely positioned. We demonstrated the feasibility of our approach by measuring the activity of single molecules of β-galactosidase and horseradish peroxidase. The approach should be of interest for many ultrasensitive bioassays at the single-molecule level.

The multiple faces of self-assembled lipidic systems

Lipids, the building blocks of cells, common to every living organisms, have the propensity to self-assemble into well-defined structures over short and long-range spatial scales. The driving forces have their roots mainly in the hydrophobic effect and electrostatic interactions. Membranes in lamellar phase are ubiquitous in cellular compartments and can phase-separate upon mixing lipids in different liquid-crystalline states. Hexagonal phases and especially cubic phases can be synthesized and observed in vivo as well. Membrane often closes up into a vesicle whose shape is determined by the interplay of curvature, area difference elasticity and line tension energies, and can adopt the form of a sphere, a tube, a prolate, a starfish and many more. Complexes made of lipids and polyelectrolytes or inorganic materials exhibit a rich diversity of structural morphologies due to additional interactions which become increasingly hard to track without the aid of suitable computer models. From the plasma membrane of archaebacteria to gene delivery, self-assembled lipidic systems have left their mark in cell biology and nanobiotechnology; however, the underlying physics is yet to be fully unraveled. PACS Codes: 87.14.Cc, 82.70.Uv

Real-time control of the q-profile in JET for steady state advanced tokamak operation

In order to simultaneously control the current and pressure profiles in high performance tokamak plasmas with internal transport barriers (ITB), a multi-variable model-based technique has been proposed. New algorithms using a truncated singular value decomposition (TSVD) of a linearized model operator and retaining the distributed nature of the system have been implemented in the JET control system. Their simplest versions have been applied to the control of the current density profile in reversed shear plasmas using three heating and current drive actuators (neutral beam injection, ion cyclotron resonant frequency heating and lower hybrid current drive). Successful control of the safety factor profile has been achieved in the quasi-steady-state, on a timescale of the order of the current redistribution time. How the TSVD algorithm will be used in the forthcoming campaigns for the simultaneous control of the current profile and of the ITB temperature gradient is discussed in some detail, but this has not yet been attempted in the present pioneering experiments.

Electroformation of giant liposomes in microfluidic channels

This paper describes a method to produce giant liposomes in microfluidic channels. To prepare the liposomes, we have investigated two different methods: electroformation and conventional gentle-hydration. We found that the liposomes produced by the conventional method were relatively small size, and several liposomes were enclosed inside another liposome. On the other hand, the liposomes formed by electroformation were mostly giant liposomes and did not enclose liposomes inside. Using microfluidic channels with electroformation method, we have succeeded in enclosing different types of the nano/micro materials into giant liposomes in the same microfluidic device simultaneously.

Progress towards steady-state operation and real-time control of internal transport barriers in JET

In JET, advanced tokamak research mainly focuses on plasmas with internal transport barriers (ITBs) that are strongly influenced by the current density profile. A previously developed optimized shear regime with low magnetic shear in the plasma centre has been extended to deeply negative magnetic shear configurations. High fusion performance with wide ITBs has been obtained transiently with negative central magnetic shear configuration: HIPB98(y,2) ~ 1.9, βN = 2.4 at Ip = 2.5 MA. At somewhat reduced performance, electron and ion ITBs have been sustained in full current drive operation with 1 MA of bootstrap current: HIPB98(y,2) ~ 1, βN = 1.7 at Ip = 2.0 MA. The ITBs were maintained for up to 11 s for the latter case. This duration, much larger than the energy confinement time (37 times larger), is already approaching a current resistive time. New real-time measurements and feedback control algorithms have been developed and implemented in JET for successfully controlling the ITB dynamics and the current density profile in the highly non-inductive current regime.

Towards fully non-inductive current drive operation in JET

Quasi-steady operation has been achieved at JET in the high-confinement regime with internal transport barriers (ITBs). The ITB has been maintained up to 11 s. This duration, much larger than the energy confinement time, is already approaching a current resistive time. The high-performance phase is limited only by plant constraints. The radial profiles of the thermal electron and ion pressures have steep gradients typically at mid-plasma radius. A large fraction of non-inductive current (above 80%) is sustained throughout the high-performance phase with a poloidal beta exceeding unity. The safety factor profile plays an important role in sustaining the ITB characteristics. In this regime where the self-generated bootstrap current (up to 1.0 MA) represents 50% of the total current, the resistive evolution of the non-monotonic q-profile is slowed down by using off-axis lower-hybrid current drive.

Influence of the q-profile shape on plasma performance in JET

The fusion performance of JET plasmas can be enhanced by the generation of internal transport barriers. The influence of the q-profile shape in the local and global plasma performance has been investigated in cases where the core magnetic shear ranges from small and positive to large and negative. Internal barriers extending to large plasma radii can be effective in raising the global performance of the plasma. It is found that such barriers tend to be generated more easily if the q-profile contains a region of negative magnetic shear. The formation is favoured by neutral beam injection compared with ion cyclotron resonance heating in scenarios where the two systems are used together. The minimum power level required to observe a local transport reduction is significantly lower than the value at which very steep pressure gradients can be achieved. This results in a practical threshold in the power to access a regime of high plasma performance that is sensitive to the q-profile shape.

Dielectrophoretic field-flow method for separating particle populations in a chip with asymmetric electrodes

This paper presents a field-flow method for separating particle populations in a dielectrophoretic (DEP) chip with asymmetric electrodes under continuous flow. The structure of the DEP device (with one thick electrode that defines the walls of the microfluidic channel and one thin electrode), as well as the fabrication and characterization of the device, was previously described. A characteristic of this structure is that it generates an increased gradient of electric field in the vertical plane that can levitate the particles experiencing negative DEP. The separation method consists of trapping one population to the bottom of the microfluidic channel using positive DEP, while the other population that exhibits negative DEP is levitated and flowed out. Viable and nonviable yeast cells were used for testing of the separation method.

Real-time control of internal transport barriers in JET

We present the results of recent experiments related to real-time control of internal transport barriers (ITBs) in JET. Using a simple criterion to characterize the ITB existence, location and strength, we have successfully controlled for the first time the radial electron temperature profile within the ITB. The dimensionless variable used in the real-time algorithm - ratio of the ion gyro-radius to the local gradient scale length of the electron temperature - is a measure of the normalized electron temperature gradient and characterizes satisfactorily the main ITB features with a relatively low computational cost. We show several examples of control of this variable in various experimental conditions of toroidal field and plasma current, using different heating systems as control actuators. We also present a double-loop feedback scheme where both the global neutron rate from D-D reactions and the ITB strength are controlled simultaneously. In this case the ITB is sustained in a fully non-inductive current drive regime during several seconds. With the proposed control method, disruptions are avoided by holding the plasma performance at a prescribed target and this opens the route towards stationary operation of tokamak plasmas with ITBs. Initial results suggest that the additional control of the current profile is an important issue for achieving steady-state operation, in particular in the triggering and the sustainment of the ITB.