Automated cell viability assessment using a microfluidics based portable imaging flow analyzer

In this work, we report a system-level integration of portable microscopy and microfluidics for the realization of optofluidic imaging flow analyzer with a throughput of 450 cells/s. With the use of a cellphone augmented with off-the-shelf optical components and custom designed microfluidics, we demonstrate a portable optofluidic imaging flow analyzer. A multiple microfluidic channel geometry was employed to demonstrate the enhancement of throughput in the context of low frame-rate imaging systems. Using the cell-phone based digital imaging flow analyzer, we have imaged yeast cells present in a suspension. By digitally processing the recorded videos of the flow stream on the cellphone, we demonstrated an automated cell viability assessment of the yeast cell population. In addition, we also demonstrate the suitability of the system for blood cell counting. (C) 2015 AIP Publishing LLC.

Fermionic edge states and new physics

We investigate the properties of the Dirac operator on manifolds with boundaries in the presence of the Atiyah-Patodi-Singer boundary condition. An exact counting of the number of edge states for boundaries with isometry of a sphere is given. We show that the problem with the above boundary condition can be mapped to one where the manifold is extended beyond the boundary and the boundary condition is replaced by a delta function potential of suitable strength. We also briefly highlight how the problem of the self-adjointness of the operators in the presence of moving boundaries can be simplified by suitable transformations which render the boundary fixed and modify the Hamiltonian and the boundary condition to reflect the effect of moving boundary.

High-throughput miniaturized microfluidic microscopy with radially parallelized channel geometry

In this article, we present a novel approach to throughput enhancement in miniaturized microfluidic microscopy systems. Using the presented approach, we demonstrate an inexpensive yet high-throughput analytical instrument. Using the high-throughput analytical instrument, we have been able to achieve about 125,880 cells per minute (more than one hundred and twenty five thousand cells per minute), even while employing cost-effective low frame rate cameras (120 fps). The throughput achieved here is a notable progression in the field of diagnostics as it enables rapid quantitative testing and analysis. We demonstrate the applicability of the instrument to point-of-care diagnostics, by performing blood cell counting. We report a comparative analysis between the counts (in cells per mu l) obtained from our instrument, with that of a commercially available hematology analyzer.

Development of a spin polarized low energy electron diffraction system

We have designed and constructed a spin polarized low energy electron diffraction system working in the reflected electron pulse counting mode. This system is capable of measuring asymmetries due to spin-orbit and exchange interactions. Photoemission from a strained GaAs/GaAsP super lattice is used as the source of spin polarized electrons. Spin-orbit asymmetry is evaluated for Ir(100) single crystal at various energies. Subsequently, exchange asymmetry has been evaluated on 40 monolayer Fe deposited on Ir(100). This instrument proves to be useful in understanding structure and magnetism at surfaces. (C) 2016 AIP Publishing LLC.

Detection and Characterization of Long-Pulse Low-Velocity Impact Damage in Plastic Bonded Explosives

Damage not only degrades the mechanical properties of explosives, but also influences the shock sensitivity, combustion and even detonation behavior of explosives. The study of impact damage is crucial in the vulnerability evaluation of explosives. A long-pulse low-velocity gas gun with a gas buffer was developed and used to induce impact damage in a hot pressed plastic bonded explosive. Various methods were used to detect and characterize the impact damage of the explosive. The microstructure was examined by use of polarized light microscopy. Fractal analysis of the micrographs was conducted by use of box counting method. The correlation between the fractal dimensions and microstructures was analyzed. Ultrasonic testing was conducted using a pulse through-transmission method to obtain the ultrasonic velocity and ultrasonic attenuation. Spectra analyses were carried out for recorded ultrasonic signals using fast Fourier transform. The correlations between the impact damage and ultrasonic parameters including ultrasonic velocities and attenuation coefficients were also analyzed. To quantitatively assess the impact induced explosive crystal fractures, particle size distribution analyses of explosive crystals were conducted by using a thorough etching technique, in which the explosives samples were soaked in a solution for enough time that the binder was totally removed. Impact induces a large extent of explosive crystal fractures and a large number of microcracks. The ultrasonic velocity decreases and attenuation coefficients increase with the presence of impact damage. Both ultrasonic parameters and fractal dimension can be used to quantitatively assess the impact damage of plastic bonded explosives.

Initial development of microdamage under impact loading

In this paper, the initial development of microdamage in material subjected to impulsive loading was investigated experimentally and analytically with controllable short-load duration. Based on a general solution to the statistical evolution of a one-dimensional system of ideal microcracks, a prerequisite to experimental investigation of nucleation of microcracks was derived. By counting the number of microcracks, the distribution of nucleation of microcracks was studied. The law of the nucleation rate of microcracks can be expressed as a separable function of stress and cracksize. It is roughly linear dependence on loading stress. The normalized number density of microcracks is in agreement with that of a second-phase particle.

Use of aerial survey and aerophotogrammetry methods in monitoring manatee populations

We evaluated the use of strip-transect survey methods for manatees through a series of replicate aerial surveys in the Banana River, Brevard County, Florida, during summer 1993 and summer 1994. Transect methods sample a representative portion of the total study area, thus allowing for statistical extrapolation to the total area. Other advantages of transect methods are less flight time and less cost than total coverage, ease of navigation, and reduced likelihood of double-counting. Our objectives were: (1) to identify visibility biases associated with the transect survey method and to adjust the counts accordingly; (2) to derive a population estimate with known variance for the Banana River during summer; and (3) to evaluate the potential value of this survey method for monitoring trends in manatee population size over time. (51 page document)

Short-Term Nonlinear Response of Tension Leg Platform in Random Sea Waves

Most of the existing mathematical models for analyzing the dynamic response of TLP are based on explicit or implicit assumptions that motions (translations and rotations) are small magnitude. However, when TLP works in severe adverse conditions, the a priori assumption on small displacements may be inadequate. In such situation, the motions should be regarded as finite magnitude. This paper will study stochastic nonlinear dynamic responses of TLP with finite displacements in random waves. The nonlinearities considered are: large amplitude motions, coupling the six degrees-of-freedom, instantaneous position, instantaneous wet surface, free surface effects and viscous drag force. The nonlinear dynamic responses are calculated by using numerical integration procedure in the time domain. After the time histories of the dynamic responses are obtained, we carry out cycle counting of the stress histories of the tethers with rain-flow counting method to get the stress range distribution.

Characterization of the benthos, marine debris and bottom fish at Gray’s Reef National Marine Sanctuary

Executive Summary: Baseline characterization of resources is an essential part of marine protected area (MPA) management and is critical to inform adaptive management. Gray’s Reef National Marine Sanctuary (GRNMS) currently lacks adequate characterization of several key resources as identified in the 2006 Final Management Plan. The objectives of this characterization were to fulfill this need by characterizing the bottom fish, benthic features, marine debris, and the relationships among them for the different bottom types within the sanctuary: ledges, sparse live bottom, rippled sand, and flat sand. Particular attention was given to characterizing the different ledge types, their fish communities, and the marine debris associated with them given the importance of this bottom type to the sanctuary. The characterization has been divided into four sections. Section 1 provides a brief overview of the project, its relevance to sanctuary needs, methods of site selection, and general field procedures. Section 2 provides the survey methods, results, discussion, and recommendations for monitoring specific to the benthic characterization. Section 3 describes the characterization of marine debris. Section 4 is specific to the characterization of bottom fish. Field surveys were conducted during August 2004, May 2005, and August 2005. A total of 179 surveys were completed over ledge bottom (n=92), sparse live bottom (n=51), flat sand (n=20), and rippled sand (n=16). There were three components to each field survey: fish counting, benthic assessment, and quantification of marine debris. All components occurred within a 25 x 4 m belt transect. Two divers performed the transect at each survey site. One diver was responsible for identification of fish species, size, and abundance using a visual survey. The second diver was responsible for characterization of benthic features using five randomly placed 1 m2 quadrats, measuring ledge height and other benthic structures, and quantifying marine debris within the entire transect. GRNMS is composed of four main bottom types: flat sand, rippled sand, sparsely colonized live bottom, and densely colonized live bottom (ledges). Independent evaluation of the thematic accuracy of the GRNMS benthic map produced by Kendall et al. (2005) revealed high overall accuracy (93%). Most discrepancies between map and diver classification occurred during August 2004 and likely can be attributed to several factors, including actual map or diver errors, and changes in the bottom type due to physical forces. The four bottom types have distinct physical and biological characteristics. Flat and rippled sand bottom types were composed primarily of sand substrate and secondarily shell rubble. Flat sand and rippled sand bottom types were characterized by low percent cover (0-2%) of benthic organisms at all sites. Although the sand bottom types were largely devoid of epifauna, numerous burrows indicate the presence of infaunal organisms. Sparse live bottom and ledges were colonized by macroalgae and numerous invertebrates, including coral, gorgonians, sponges, and “other” benthic species (such as tunicates, anemones, and bryozoans). Ledges and sparse live bottom were similar in terms of diversity (H’) given the level of classification used here. However, percent cover of benthic species, with the exception of gorgonians, was significantly greater on ledge than on sparse live bottom. Percent biotic cover at sparse live bottom ranged from 0.7-26.3%, but was greater than 10% at only 7 out of 51 sites. Colonization on sparse live bottom is likely inhibited by shifting sands, as most sites were covered in a layer of sediment up to several centimeters thick. On ledge bottom type, percent cover ranged from 0.42-100%, with the highest percent cover at ledges in the central and south-central region of GRNMS. Biotic cover on ledges is influenced by local ledge characteristics. Cluster analysis of ledge dimensions (total height, undercut height, undercut width) resulted in three main categories of ledges, which were classified as short, medium, and tall. Median total percent cover was 97.6%, 75.1%, and 17.7% on tall, medium, and short ledges, respectively. Total percent cover and cover of macroalgae, sponges, and other organisms was significantly lower on short ledges compared to medium and tall ledges, but did not vary significantly between medium and tall ledges. Like sparse live bottom, short ledges may be susceptible to burial by sand, however the results indicate that ledge height may only be important to a certain threshold. There are likely other factors not considered here that also influence spatial distribution and community structure (e.g., small scale complexity, ocean currents, differential settlement patterns, and biological interactions). GRNMS is a popular site for recreational fishing and boating, and there has been increased concern about the accumulation of debris in the sanctuary and potential effects on sanctuary resources. Understanding the types, abundance, and distribution of debris is essential to improving debris removal and education efforts. Approximately two-thirds of all observed debris items found during the field surveys were fishing gear, and about half of the fishing related debris was monofilament fishing line. Other fishing related debris included leaders and spear gun parts, and non-gear debris included cans, bottles, and rope. The spatial distribution of debris was concentrated in the center of the sanctuary and was most frequently associated with ledges rather than at other bottom types. Several factors may contribute to this observation. Ledges are often targeted by fishermen due to the association of recreationally important fish species with this bottom type. In addition, ledges are structurally complex and are often densely colonized by biota, providing numerous places for debris to become stuck or entangled. Analysis of observed boat locations indicated that higher boat activity, which is an indication of fishing, occurs in the center of the sanctuary. On ledges, the presence and abundance of debris was significantly related to observed boat density and physiographic features including ledge height, ledge area, and percent cover. While it is likely that most fishing related debris originates from boats inside the sanctuary, preliminary investigation of ocean current data indicate that currents may influence the distribution and local retention of more mobile items. Fish communities at GRNMS are closely linked to benthic habitats. A list of species encountered, probability of occurrence, abundance, and biomass by habitat is provided. Species richness, diversity, composition, abundance, and biomass of fish all showed striking differences depending on bottom type with ledges showing the highest values of nearly all metrics. Species membership was distinctly separated by bottom type as well, although very short, sparsely colonized ledges often had a similar community composition to that of sparse live bottom. Analysis of fish communities at ledges alone indicated that species richness and total abundance of fish were positively related to total percent cover of sessile invertebrates and ledge height. Either ledge attribute was sufficient to result in high abundance or species richness of fish. Fish diversity (H`) was negatively correlated with undercut height due to schools of fish species that utilize ledge undercuts such as Pareques species. Concurrent analysis of ledge types and fish communities indicated that there are five distinct combinations of ledge type and species assemblage. These include, 1) short ledges with little or no undercut that lacked many of the undercut associated species except Urophycis earlii ; 2) tall, heavily colonized, deeply undercut ledges typically with Archosargus probatocephalus, Mycteroperca sp., and Pareques sp.; 3) tall, heavily colonized but less undercut with high occurrence of Lagodon rhomboides and Balistes capriscus; 4) short, heavily colonized ledges typically with Centropristis ocyurus, Halichoeres caudalis, and Stenotomus sp.; and 5) tall, heavily colonized, less undercut typically with Archosargus probatocephalus, Caranx crysos and Seriola sp.. Higher levels of boating activity and presumably fishing pressure did not appear to influence species composition or abundance at the community level although individual species appeared affected. These results indicate that merely knowing the basic characteristics of a ledge such as total height, undercut width, and percent cover of sessile invertebrates would allow good prediction of not only species richness and abundance of fish but also which particular fish species assemblages are likely to occur there. Comparisons with prior studies indicate some major changes in the fish community at GRNMS over the last two decades although the causes of the changes are unknown. Species of interest to recreational fishermen including Centropristis striata, Mycteroperca microlepis, and Mycteroperca phenax were examined in relation to bottom features, areas of assumed high versus low fishing pressure, and spatial dispersion. Both Mycteroperca species were found more frequently when undercut height of ledges was taller. They often were found together in small mixed species groups at ledges in the north central and southwest central regions of the sanctuary. Both had lower mode size and proportion of fish above the fishery size limit in heavily fished areas of the sanctuary (i.e. high boat density) despite the presence of better habitat in that region. Black sea bass, C. striata, occurred at 98% of the ledges surveyed and appeared to be evenly distributed throughout the sanctuary. Abundance was best explained by a positive relationship with percent cover of sessile biota but was also negatively related to presence of either Mycteroperca species. This may be due to predation by the Mycteroperca species or avoidance of sites where they are present by C. striata. Suggestions for monitoring bottom features, marine debris, and bottom fish at GRNMS are provided at the end of each chapter. The present assessment has established quantitative baseline characteristics of many of the key resources and use issues at GRNMS. The methods can be used as a model for future assessments to track the trajectory of GRNMS resources. Belt transects are ideally suited to providing efficient and quantitative assessment of bottom features, debris, and fish at GRNMS. The limited visibility, sensitivity of sessile biota, and linear nature of ledge habitats greatly diminish the utility of other sampling techniques. Ledges should receive the bulk of future characterization effort due to their importance to the sanctuary and high variability in physical structure, benthic composition, and fish assemblages. (PDF contains 107 pages.)

A method for age determination of dolphins.

The use of growth layers in teeth as an indicator of age in odnotocetes and pinnipeds was suggested by Laws (1954) and since then the method has been used extensively in both marine and non-marine mammals. Dentinal growth layers are groups (growth layer groups) of repetitive alternating bands which in cross-section are similar to growth rings in trees. The most commonly used methods for counting growth layer groups (GLGs) are by undecalcified longitudinal thin sections (150 um) or decalcified and stained thin sections (10-30 um). In longitudinal sections viewed with light microscopy, GLGs appear as opaque and translucent cones nestled one inside another, with the oldest dentine Iying adjacent to the enamel, and the newest layer borderinq the pulp cavity.

Estimating age of spotted and spinner dolphins (Stenella attenuata and Stenella longirostris) from teeth

This paper is an account of preparation and examination techniques and criteria used to estimate age in decalcified and stained tooth thin sections from spinner and spotted dolphins. A dentinal growth layer group (GLG), composed of two thin light and two thicker dark-stained layers, is deposited annually. The GLG component layers are variably visible, but the "ideal" pattern and successive thinning of dentinal GLGs are used as a guide to determine GLG limits. Age-specific thicknesses of dentinal GLGs found in Hawaiian spinner dolphin teeth seem to be applicable to teeth of spotted dolphins and can be used as an aid in locating GLG boundaries. Cementa1 GLGs are composed of a dark-stained and alightly stained layer and usually are deposited at a rate of one per year, but may be deposited every other year or two or three times per year. Two slightly different methods of counting dentinal GLGs are presented, along with guidelines for determining whether dentinal or cementa1 GLG counts provide the best estimate of age for a specimen. (PDF contains 23 pages.)

KE Useage Statistics Guidelines

A Knowledge Exchange workshop took place in March 2010 to bring together technical experts working in partner projects collecting usage statistics including PIRUS2, OAstatistik and SURFsure projects. Experts from other related projects (RePec and NeeO) were also involved. Following the workshop, experts came to the conclusion that it would be valuable for institutions in all four Knowledge Exchange partner countries as well as countries outside of KE, to have an agreed set of guidelines which could be used when collecting and comparing data on usage statistics. This work connects with work already planned in projects already funded in three of the countries. Together they have set up a set of guidelines which will be tested and refined by Autumn 2010. An element in these guidelines is an agreed robotlist. This list extends on the COUNTER list of robots which should be excluded when counting downloads.

Metabolic efficiency with fast spiking in the squid axon

Fundamentally, action potentials in the squid axon are consequence of the entrance of sodium ions during the depolarization of the rising phase of the spike mediated by the outflow of potassium ions during the hyperpolarization of the falling phase. Perfect metabolic efficiency with a minimum charge needed for the change in voltage during the action potential would confine sodium entry to the rising phase and potassium efflux to the falling phase. However, because sodium channels remain open to a significant extent during the falling phase, a certain overlap of inward and outward currents is observed. In this work we investigate the impact of ion overlap on the number of the adenosine triphosphate (ATP) molecules and energy cost required per action potential as a function of the temperature in a Hodgkin–Huxley model. Based on a recent approach to computing the energy cost of neuronal action potential generation not based on ion counting, we show that increased firing frequencies induced by higher temperatures imply more efficient use of sodium entry, and then a decrease in the metabolic energy cost required to restore the concentration gradients after an action potential. Also, we determine values of sodium conductance at which the hydrolysis efficiency presents a clear minimum.

Growth of yellowfin tuna, Thunnus albacares, in the Eastern Pacific Ocean based on otolith increments

ENGLISH: The growth of yellowfin tuna in the eastern Pacific is described in terms of several measurements taken from the fish and their otoliths (sagittae). Equations are also developed to predict age from the readily available dimensions of fork length and head length. The data for all of these relationships were obtained from a sample of 196 fish collected during 1977 through 1979 from purse seiners fishing north of the equator and east of 137°W. The fork-length range of the sample was 30-170 cm. The number of increments on a sagitta of each fish was used as a direct estimate of its age in days. The correspondence between increments and days has been validated for yellowfin in the length range of 40-110 cm. Circumstantial evidence indicates that the relationship also applies in the intervals of 0-40 cm and 110-170 cm. This circumstancial evidence was derived from: 1) literature on validated increments during early growth for other species, 2) knowledge that structures assumed to be daily increments on yellowfin otoliths have subsequently been validated in the corresponding zone on bluefin otoliths, and 3) a comparison of the growth curve based on increments to others obtained from length frequency modal analysis. Based on this information the age estimates over the entire size range of sampled fish are believed to be accurate. In addition to the general growth and age-predictive relationships, the major conclusions of the study are that: 1) Sexually dimorphic growth exists in terms of fork length, fish weight and the length of the otolith counting path for the entire data set. Examination of the data for 1977 and 1979 also revealed that the fork-length growth of each sex differed within years. 2) For combined sexes there were significant differences among the fork-length growth curves for yellowfin sampled in different years. 3) Yellowfin caught inshore (within 275 miles of the coast) were heavier than those caught offshore for fork lengths between 30 and 110 cm. The situation was reversed for lengths greater than 110 cm. 4) Back-calculated spawning months were distributed uniformly throughout the year in 1974 and 1977, but in 1975-1976 and 1978 spawning activity was apparently concentrated in the latter half of the year. SPANISH: El crecimiento del atún aleta amarilla en el Pacífico oriental se describe en términos de varias medidas obtenidas de peces y otolitos (sagita). Se formularon también ecuaciones para pronosticar la edad, según las dimensiones fácilmente disponibles de la longitud horquilla y longitud de la cabeza. Los datos de todas estas relaciones fueron obtenidos mediante una muestra de 196 peces recolectados desde 1977hasta 1979, en barcos cerqueros que estaban pescando al norte de la línea ecuatorial y al este de los 137°W. El intervalo de la longitud horquilla de la muestra fue de 30-170 cm. Se empleó el número de incrementos en la sagita de cada pez como un estimado directo de la edad en días. Se ha comprobado la relación entre los incrementos y los días en el intervalo de longitud de 40-110 cm del aleta amarilla. La evidencia circunstancial indica que se aplica también la relación a los intervalos de 0-40 cm y 110-170 cm. Esta evidencia circunstancial se dedujo: 1) de las publicaciones sobre incrementos comprobados de otras especies durante el primer crecimiento, 2) del conocimientoque las estructuras que se supone son incrementos diarios en los otolitos del aleta amarilla han sido comprobadas luego en la parte correspondiente de otolitos del aleta azul y 3) por una comparación de la curva de crecimiento, basada en incrementos relacionados a otras curvas obtenidas según el análisis modal frecuencia-talla. Se cree, basados en esta información, que las estimaciones de la edad sobre toda la amplitud de talla de los peces muestreados, es acertada. Además de la relación del crecimiento general y del pronóstico de la edad, las principales conclusiones de este estudio son: 1) En toda la serie de datos existe el crecimiento sexualmente dimórfico en términos de longitud horquilla, peso del pez y longitud del plano de conteo del otolito. El examen de los datos de 1977 y 1979, revelan también que el crecimiento longitud horquilla de cada sexo es diferente en los años. 2) En los sexos combinados hubo diferencias significativas entre las curvas de crecimiento longitud horquilla del aleta amarilla muestreado en diferentes años. 3) El aleta amarilla capturado cerca a la costa (en las primeras 275 millas) fue más pesado que el capturado en las aguas mar afuera, correspondiente a la longitud horquilla entre 30 y 110 cm. La situación fue inversa para tallas de más de 110 cm. 4) En 1974 y 1977, los meses retrocalculados del desove se distribuyeron uniformemente durante el año, pero en 1975-1976 y 1978, la actividad del desove se concentró aparentemente en el último semestre del año. (PDF contains 62 pages.)

Two-body photodisintegration of the deuteron at intermediate energy

We have measured the differential cross section for two-body deuteron photodisintegration at center-of-mass angles of 90°, 53° and 37° with photon energies from 1.6 Ge V to 2.8 Ge V. Additional data were taken at θ* = 37° and E_γ = 4.2 GeV. Invariant cross sections at θ* =90° and 53° appear to follow a simple scaling law predicted by constituent counting rules of perturbative QCD, while the cross section at θ* = 37° shows a slower fall-off with photon energy. Angular distributions show increasing forward peaking at higher energies. Agreement with various theoretical predictions based on pQCD and meson-exchange models is discussed.