AMPA glutamate ion channels are tetrameric receptors in which activation to form the open channel depends on the binding of possibly multiple glutamate molecules. However, it is unclear whether AMPA receptors bound with a different number of glutamate molecules (i.e. one being the minimal and four being the maximal number of glutamate molecules) open the channels with different kinetic constants. Using a laser pulse photolysis technique that provides microsecond time resolution, we investigated the channel-opening kinetic mechanism of a nondesensitizing AMPA receptor, i.e. GluR1Q flip L497Y or a leucine-to-tyrosine substitution mutant, in the entire range of glutamate concentrations to ensure receptor saturation. We found that the minimal number of glutamate molecules required to bind to the receptor and to open the channel is two (or n ‫؍‬ 2), and that the entire channel-opening kinetics can be adequately described by just one channel-opening rate constant, k op , which correlates to n ‫؍‬ 2. This result suggests that higher receptor occupancy (n ‫؍‬ The ␣-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) 3 glutamate receptors are ligand-gated ion channels that are activated by binding of neurotransmitter glutamate (1, 2). An AMPA receptor is a tetrameric assembly with each subunit containing a glutamate binding site. The receptor can adopt multiple conductance levels, especially at high receptor occupancy, as observed in the single-channel records of wild-type and mutant recombinant receptors (3, 4) as well as native AMPA receptors (5). However, it remains unclear whether receptor occupancy plays a significant role in determining the kinetic constants for an ensemble rate process of channel opening as a function of glutamate concentration. The ensemble rate process is manifested in a whole-cell current response to the binding of glutamate in vitro and best represents the glutamatergic synaptic activity in vivo, such as excitatory postsynaptic current. Therefore, determining the number of glutamate molecules bound to a receptor or the percentage of the receptor occupancy pertinent to the rate of the channel opening is a basic question to be answered for understanding the function of AMPA receptors.To address this question, we investigated the channel-opening kinetics for a GluR1 AMPA receptor channel carrying a substitution of leucine (L) to tyrosine (Y) or L497Y. The discovery of this point mutation by Stern-Bach et al. (3) was a significant event in understanding the structure and function relationship of AMPA receptors in that (a) phenomenologically, the single leucine-to-tyrosine substitution renders the homomeric receptor channels virtually non-desensitizing (3), and (b) the phenotypic effect of this mutation is conserved at equivalent positions in all AMPA receptor subunits, i.e. GluR1-4 (6 -8). Furthermore, this mutation is thought to have no effect on either the main conductance level or the channel opening probability (3,7,9). From a crystallographic study, Sun et al. (6) revealed that this ...

For decades three-dimensional (3D) measurements of engineering components have been made using fixed metrology-room based coordinate measuring machines (CMMs) fitted most commonly with single point or to a much lesser extent, scanning tactile probes. Over the past decade there has been a rapid uptake in development and subsequent use of portable optical-based 3D coordinate measuring systems. These optical-based systems capture vast quantities of point data in a very short time, often permitting freeform surfaces to be digitized. Documented standards, for example ISO 10360, for the verification of fixed CMMs fitted with tactile probes are now widely available, whereas verification procedures and more specifically verification artefacts for optical-based systems are still in their infancy. Furthermore, the industry is seeking traceability in 3D measurements of high precision components. A recent requirement is the demand for highly accurate measurements of large gears with diameters up to 1000 mm as used in gear boxes of wind turbines. Up until now it has been impossible to ensure traceability of 3D measurements of big gears, since no traceable standards were available. This paper describes three different types of artefacts that were developed during the project, namely tetrahedron artefacts for testing the basic measurement capability of optical 3D devices, freeform verification artefacts for testing the capability of measuring complex geometry, and a large gear artefact for task related calibration of different types of CMMs. In addition, artefact calibration data and associated measurement uncertainties and international intercomparisons are presented. These developments will be of considerable value to end users, calibration laboratories and producers of optical and tactile CMMs.

Through its ability to measure otherwise inaccessible internal structures, x-ray computed tomography (CT) has become a useful tool in the field of dimensional metrology. However, the lack of international standards for performance verification and the determination of metrological characteristics has so far prevented it from becoming a standardized measurement technology. This paper considers how a test object comprising a dismantleable tetrahedral assembly of four calibrated alumina spheres can be employed to assess the metrological performance of an industrial-type CT system. The measurement accuracy was characterized in terms of sphere size, separation and form errors. In all cases, sub-voxel accuracy was achieved, with errors as small as 1/10 of a voxel being obtained. Good measurement repeatability was demonstrated even upon dismantling and reconstructing the reference object. The tetrahedral test object offers a simple robust solution for the verification of metrological performance with great versatility and adaptability.

The use of x-ray computed tomography for dimensional measurements associated with engineering applications has flourished in recent years. However, error sources associated with the technology are not well understood. In this paper, a novel two-sphere reference sample has been developed and used to investigate the stability of the imaging system that consists of an x-ray tube and a detector. In contrast with other research work reported, this work considered relative positional variation along the x-, y- and z-axes. This sample is a significant improvement over the one sphere sample proposed previously, which can only be used to observe the stability of the imaging system along x- and y-axes. Temperature variations of different parts of the system have been monitored and the relationship between temperature variations and x-ray image stability has been studied. Other effects that may also influence the stability of the imaging system have been discussed. The proposed reference sample and testing method are transferable to other types of x-ray computed tomography systems, for example, systems with transmission targets and systems with sub-micrometre focal spots.