Diurnal fluctuation in the number of hypocretin/orexin and histamine producing: Implication for understanding and treating neuronal loss.

The loss of specific neuronal phenotypes, as determined by immunohistochemistry, has become a powerful tool for identifying the nature and cause of neurological diseases. Here we show that the number of neurons identified and quantified using this method misses a substantial percentage of extant neurons in a phenotype specific manner. In mice, 24% more hypocretin/orexin (Hcrt) neurons are seen in the night compared to the day, and an additional 17% are seen after inhibiting microtubule polymerization with colchicine. We see no such difference between the number of MCH (melanin concentrating hormone) neurons in dark, light or colchicine conditions, despite MCH and Hcrt both being hypothalamic peptide transmitters. Although the size of Hcrt neurons did not differ between light and dark, the size of MCH neurons was increased by 15% in the light phase. The number of neurons containing histidine decarboxylase (HDC), the histamine synthesizing enzyme, was 34% greater in the dark than in the light, but, like Hcrt, cell size did not differ. We did not find a significant difference in the number or the size of neurons expressing choline acetyltransferase (ChAT), the acetylcholine synthesizing enzyme, in the horizontal diagonal band (HBD) during the dark and light conditions. As expected, colchicine treatment did not increase the number of these neurons. Understanding the function and dynamics of transmitter production within "non-visible" phenotypically defined cells has fundamental implications for our understanding of brain plasticity

Holographic Measurements of Anisotropic Three-Dimensional Diffusion of Colloidal Clusters

We measure all nonzero elements of the three-dimensional diffusion tensor D for clusters of colloidal spheres to a precision of 1% or better using digital holographic microscopy. We study both dimers and triangular trimers of spheres, for which no analytical calculations of the diffusion tensor exist. We observe anisotropic rotational and translational diffusion arising from the asymmetries of the clusters. In the case of the three-particle triangular cluster, we also detect a small but statistically significant difference in the rotational diffusion about the two in-plane axes. We attribute this difference to weak breaking of threefold rotational symmetry due to a small amount of particle polydispersity. Our experimental measurements agree well with numerical calculations and show how diffusion constants can be measured under conditions relevant to colloidal self-assembly, where theoretical and even numerical prediction is difficult.Engineering and Applied SciencesPhysic

Fast High Resolution Echelle Spectroscopy Of A Laboratory Plasma

An echelle diffraction grating and a multianode photomultiplier tube are paired to construct a high resolution (R=lambda/delta lambda approximate to 2.5x10(4)) spectrograph with fast time response for use from the UV through the visible. This instrument has analyzed the line shape of C III impurity ion emission at 229.687 nm over the lifetime (approximate to 100 mu s) of the hydrogen plasmas produced at SSX. The ion temperature and line of sight average velocity are inferred from the observed thermal broadening and Doppler shift of the line. The time resolution of these measurements is about 1 mu s, sufficient to observe the fastest magnetohydrodynamic activity

Measuring the 3D Dynamics of Multiple Colloidal Particles with Digital Holographic Microscopy

We discuss digital holographic microscopy (DHM), a 3D imaging technique capable of measuring the positions of micron-sized colloidal particles with nanometer precision and sub-millisecond temporal resolution. We use exact electromagnetic scattering solutions to model holograms of multiple colloidal spheres. While the Lorenz-Mie solution for scattering by isolated spheres has previously been used to model digital holograms, we apply for the first time an exact multisphere superposition scattering model that is capable of modeling holograms from spheres that are sufficiently close together to exhibit optical coupling.Physic

Using the Discrete Dipole Approximation and Holographic Microscopy to Measure Rotational Dynamics of Non-spherical Colloidal Particles

We present a new, high-speed technique to track the three-dimensional translation and rotation of non-spherical colloidal particles. We capture digital holograms of micrometer-scale silica rods and sub-micrometer-scale Janus particles freely diffusing in water, and then fit numerical scattering models based on the discrete dipole approximation to the measured holograms. This inverse-scattering approach allows us to extract the the position and orientation of the particles as a function of time, along with static parameters including the size, shape, and refractive index. The best-fit sizes and refractive indices of both particles agree well with expected values. The technique is able to track the center of mass of the rod to a precision of 35 nm and its orientation to a precision of 1.5$^\circ$, comparable to or better than the precision of other 3D diffusion measurements on non-spherical particles. Furthermore, the measured translational and rotational diffusion coefficients for the silica rods agree with hydrodynamic predictions for a spherocylinder to within 0.3%. We also show that although the Janus particles have only weak optical asymmetry, the technique can track their 2D translation and azimuthal rotation over a depth of field of several micrometers, yielding independent measurements of the effective hydrodynamic radius that agree to within 0.2%. The internal and external consistency of these measurements validate the technique. Because the discrete dipole approximation can model scattering from arbitrarily shaped particles, our technique could be used in a range of applications, including particle tracking, microrheology, and fundamental studies of colloidal self-assembly or microbial motion.Comment: 11 pages, 9 figures, 2 table

Two Fluid Effects On Three-Dimensional Reconnection In The Swarthmore Spheromak Experiment With Comparisons To Space Data

Several new experimental results are reported from spheromak merging studies at the Swarthmore SpheromakExperiment[M. R. Brown, Phys. Plasmas6, 1717 (1999)] with relevance to three-dimensional (3D) reconnection in laboratory and space plasmas. First, recent velocity measurements of impurity ions using ion Doppler spectroscopy are reported. Bidirectional outflow at nearly the Alfvén speed is clearly observed. Second, experimental measurements of the out-of-plane magnetic field in a reconnection volume showing a quadrupolar structure at the ion inertial scale are discussed. Third, a measurement of in-plane Hall electric field and nonideal terms of the generalized Ohm’s law in a reconnection volume of a weakly collisional laboratory plasma is presented. Time resolved vector magnetic field measurements on a 3D lattice [B(r,t)] enables evaluation of the various terms. Results show that the Hall electric field dominates everywhere (J×B∕ne) and also exhibits a quadrupolar structure at the ion inertial scale; resistive and electron inertia terms are small. Each of these is related to and compared with similar measurements in a solar or space context

Random-subset fitting of digital holograms for fast three-dimensional particle tracking

Fitting scattering solutions to time series of digital holograms is a precise way to measure three-dimensional dynamics of microscale objects such as colloidal particles. However, this inverse-problem approach is computationally expensive. We show that the computational time can be reduced by an order of magnitude or more by fitting to a random subset of the pixels in a hologram. We demonstrate our algorithm on experimentally measured holograms of micrometer-scale colloidal particles, and we show that 20-fold increases in speed, relative to fitting full frames, can be attained while introducing errors in the particle positions of 10 nm or less. The method is straightforward to implement and works for any scattering model. It also enables a parallelization strategy wherein random-subset fitting is used to quickly determine initial guesses that are subsequently used to fit full frames in parallel. This approach may prove particularly useful for studying rare events, such as nucleation, that can only be captured with high frame rates over long times.Engineering and Applied SciencesPhysic

Measuring Dynamics and Interactions of Colloidal Particles with Digital Holographic Microscopy

We investigate how colloidal particles self-assemble in confined and nonequilibrium systems, including particles trapped at liquid-liquid interfaces (e.g. emulsion droplets) and inside spherical containers. Although common in industrial formulations and fundamental condensed matter studies, these systems remain poorly understood, primarily because no existing experimental probes, including confocal microscopy, can yield real-space data with sufficiently fast acquisition times to resolve 3D dynamics. We use a powerful interferometric technique, Digital Holographic Microscopy (DHM), in concert with particle synthesis and algorithm development to overcome these limitations. Preliminary data show that the technique is capable of tracking several micrometer-sized colloidal particles with 30 nm spatial precision in all three dimensions on millisecond time scales. DHM may be able to yield the most complete physical picture to date of dynamics, interactions, and assembly in colloidal suspensions.Engineering and Applied SciencesPhysic