Christopher D. Saunter

Direct measurement of the skew angle of the Poynting vector in a helically phased beam

We measure the local skew angle of the Poynting vector within a helically-phased, exp (il phi), beam using a Shack Hartmann wavefront sensor. It is the skew angle of the Poynting vector with respect to the beam axis that gives rise to the orbital angular momentum of a light beam. We confirm that this skew angle is l/kr, corresponding to an orbital angular momentum of l? per photon. Measurement of orbital angular momentum in this way is an alternative to interferometric techniques giving a non-ambiguous result to both the magnitude and sign of l from a single measurement, without any restriction on the optical bandwidth.

Mitochondrial Motility and Vascular Smooth Muscle Proliferation

Objective—Mitochondria are widely described as being highly dynamic and adaptable organelles, and their movement is thought to be vital for cell function. Yet, in various native cells, including those of heart and smooth muscle, mitochondria are stationary and rigidly structured. The significance of the differences in mitochondrial behavior to the physiological function of cells is unclear and was studied in single myocytes and intact resistance-sized cerebral arteries. We hypothesized that mitochondrial dynamics is controlled by the proliferative status of the cells. Methods and Results—High-speed fluorescence imaging of mitochondria in live vascular smooth muscle cells shows that the organelle undergoes significant reorganization as cells become proliferative. In nonproliferative cells, mitochondria are individual (≈2 &mgr;m by 0.5 &mgr;m), stationary, randomly dispersed, fixed structures. However, on entering the proliferative state, mitochondria take on a more diverse architecture and become small spheres, short rod-shaped structures, long filamentous entities, and networks. When cells proliferate, mitochondria also continuously move and change shape. In the intact pressurized resistance artery, mitochondria are largely immobile structures, except in a small number of cells in which motility occurred. When proliferation of smooth muscle was encouraged in the intact resistance artery, in organ culture, the majority of mitochondria became motile and the majority of smooth muscle cells contained moving mitochondria. Significantly, restriction of mitochondrial motility using the fission blocker mitochondrial division inhibitor prevented vascular smooth muscle proliferation in both single cells and the intact resistance artery. Conclusion—These results show that mitochondria are adaptable and exist in intact tissue as both stationary and highly dynamic entities. This mitochondrial plasticity is an essential mechanism for the development of smooth muscle proliferation and therefore presents a novel therapeutic target against vascular disease.

Real-time optical gating for three-dimensional beating heart imaging

We demonstrate real-time microscope image gating to an arbitrary position in the cycle of the beating heart of a zebrafish embryo. We show how this can be used for high-precision prospective gating of fluorescence image slices of the moving heart. We also present initial results demonstrating the application of this technique to 3-D structural imaging of the beating embryonic heart.

Dynamic lens and monovision 3D displays to improve viewer comfort.

Stereoscopic 3D (S3D) displays provide an additional sense of depth compared to non-stereoscopic displays by sending slightly different images to the two eyes. But conventional S3D displays do not reproduce all natural depth cues. In particular, focus cues are incorrect causing mismatches between accommodation and vergence: The eyes must accommodate to the display screen to create sharp retinal images even when binocular disparity drives the eyes to converge to other distances. This mismatch causes visual discomfort and reduces visual performance. We propose and assess two new techniques that are designed to reduce the vergence-accommodation conflict and thereby decrease discomfort and increase visual performance. These techniques are much simpler to implement than previous conflict-reducing techniques. The first proposed technique uses variable-focus lenses between the display and the viewers eyes. The power of the lenses is yoked to the expected vergence distance thereby reducing the mismatch between vergence and accommodation. The second proposed technique uses a fixed lens in front of one eye and relies on the binocularly fused percept being determined by one eye and then the other, depending on simulated distance. We conducted performance tests and discomfort assessments with both techniques and compared the results to those of a conventional S3D display. The first proposed technique, but not the second, yielded clear improvements in performance and reductions in discomfort. This dynamic-lens technique therefore offers an easily implemented technique for reducing the vergence-accommodation conflict and thereby improving viewer experience.

From structure to function: mitochondrial morphology, motion and shaping in vascular smooth muscle

The diversity of mitochondrial arrangements, which arise from the organelle being static or moving, or fusing and dividing in a dynamically reshaping network, is only beginning to be appreciated. While significant progress has been made in understanding the proteins that reorganise mitochondria, the physiological significance of the various arrangements is poorly understood. The lack of understanding may occur partly because mitochondrial morphology is studied most often in cultured cells. The simple anatomy of cultured cells presents an attractive model for visualizing mitochondrial behaviour but contrasts with the complexity of native cells in which elaborate mitochondrial movements and morphologies may not occur. Mitochondrial changes may take place in native cells (in response to stress and proliferation), but over a slow time-course and the cellular function contributed is unclear. To determine the role mitochondrial arrangements play in cell function, a crucial first step is characterisation of the interactions among mitochondrial components. Three aspects of mitochondrial behaviour are described in this review: (1) morphology, (2) motion and (3) rapid shape changes. The proposed physiological roles to which various mitochondrial arrangements contribute and difficulties in interpreting some of the physiological conclusions are also outlined.

Micro-endoscope for in vivo widefield high spatial resolution fluorescent imaging

In this paper we report the design, testing and use of a scannerless probe specifically for minimally invasive imaging of deep tissue in vivo with an epi-fluorescence modality. The probe images a 500 μm diameter field of view through a 710 μm outer diameter probe with a maximum tissue penetration depth of 15 mm specifically configured for eGFP imaging. Example results are given from imaging the pituitary gland of rats and zebrafish hearts with lateral resolution of 2.5 μm.

Stochastically determined directed movement explains the dominant small-scale mitochondrial movements within non-neuronal tissue culture cells

The apparently stationary phase of mitochondrial motion was investigated in epithelial cells by spinning disk confocal light microscopy combined with image correlation based single particle tracking using custom software producing sub‐pixel accuracy measurements (∼5 nm) at 10–12 Hz frame‐rates. The analysis of these data suggests that the previously described stationary, or anchored phase, in mitochondrial movement actually comprise Brownian diffusion, interspersed with frequent and brief motor‐driven events whose duration are stochastically determined. We have therefore discovered a new aspect of mitochondrial behavior, which we call stochastically determined, directed movement.

Acceleration of adaptive optics simulations using programmable logic

Numerical simulation is an essential part of the design and optimization of astronomical adaptive optics (AO) systems. Simulations of AO are computationally expensive and the problem scales rapidly with telescope aperture size, as the required spatial order of the correcting system increases. Practical realistic simulations of AO systems for extremely large telescopes are beyond the capabilities of all but the largest of modern parallel supercomputers. Here, we describe a more cost-effective approach through the use of hardware acceleration using field programmable gate arrays. By transferring key parts of the simulation into programmable logic, large increases in computational bandwidth can be expected. We show that the calculation of wavefront sensor image centroids can be accelerated by a factor of 4 by transferring the algorithm into hardware. Implementing more demanding parts of the AO simulation in hardware will lead to much greater performance improvements of up to 1000 times. Ke yw ords: instrumentation: adaptive optics ‐ methods: numerical ‐ techniques: miscellaneous ‐ telescopes ‐ instrumentation: high angular resolution.

Development of a portable SLODAR turbulence profiler

We report on the development of a prototype portable monitor for profiling of the altitude and velocity of atmospheric optical turbulence. The instrument is based on the SLODAR Shack-Hartmann wave-front sensing technique, applied to a portable telescope and employing an electron-multiplication (EM) CCD camera as the wave-front sensor detector. Constructed for ESO by the astronomical instrumentation group at the University of Durham, the main applications of the monitor will be in support of the ESO multi-conjugate adaptive optics demonstrator (MAD) project, and for site characterization surveys for future extremely large telescopes. The monitor can profile the whole atmosphere or can be optimized for profiling of low altitude (0-1km) turbulence, with a maximum altitude resolution of approximately 150m. First tests of the system have been carried out at the La Palma observatory.

Entry into the nuclear pore complex is controlled by a cytoplasmic exclusion zone containing dynamic GLFG-repeat nucleoporin domains

ABSTRACT Nuclear pore complexes (NPCs) mediate nucleocytoplasmic movement. The central channel contains proteins with phenylalanine-glycine (FG) repeats, or variations (GLFG, glycine-leucine-phenylalanine-glycine). These are ‘intrinsically disordered’ and often represent weak interaction sites that become ordered upon interaction. We investigated this possibility during nuclear transport. Using electron microscopy of S. cerevisiae, we show that NPC cytoplasmic filaments form a dome-shaped structure enclosing GLFG domains. GLFG domains extend out of this structure and are part of an ‘exclusion zone’ that might act as a partial barrier to entry of transport-inert proteins. The anchor domain of a GLFG nucleoporin locates exclusively to the central channel. By contrast, the localisation of the GLFG domains varied between NPCs and could be cytoplasmic, central or nucleoplasmic and could stretch up to 80 nm. These results suggest a dynamic exchange between ordered and disordered states. In contrast to diffusion through the NPC, transport cargoes passed through the exclusion zone and accumulated near the central plane. We also show that movement of cargo through the NPC is accompanied by relocation of GLFG domains, suggesting that binding, restructuring and movement of these domains could be part of the translocation mechanism.