Media Compositions for Three Dimensional Mammalian Tissue Growth Under Microgravity Culture Conditions

Science.gov (United States)

Goodwin, Thomas J. (Inventor)

1998-01-01

Normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural and blood tissue. The cells are grown in vitro under microgravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel.

Microengineering methods for cell-based microarrays and high-throughput drug-screening applications

International Nuclear Information System (INIS)

Xu Feng; Wu Jinhui; Wang Shuqi; Gurkan, Umut Atakan; Demirci, Utkan; Durmus, Naside Gozde

2011-01-01

Screening for effective therapeutic agents from millions of drug candidates is costly, time consuming, and often faces concerns due to the extensive use of animals. To improve cost effectiveness, and to minimize animal testing in pharmaceutical research, in vitro monolayer cell microarrays with multiwell plate assays have been developed. Integration of cell microarrays with microfluidic systems has facilitated automated and controlled component loading, significantly reducing the consumption of the candidate compounds and the target cells. Even though these methods significantly increased the throughput compared to conventional in vitro testing systems and in vivo animal models, the cost associated with these platforms remains prohibitively high. Besides, there is a need for three-dimensional (3D) cell-based drug-screening models which can mimic the in vivo microenvironment and the functionality of the native tissues. Here, we present the state-of-the-art microengineering approaches that can be used to develop 3D cell-based drug-screening assays. We highlight the 3D in vitro cell culture systems with live cell-based arrays, microfluidic cell culture systems, and their application to high-throughput drug screening. We conclude that among the emerging microengineering approaches, bioprinting holds great potential to provide repeatable 3D cell-based constructs with high temporal, spatial control and versatility.

Microengineering methods for cell-based microarrays and high-throughput drug-screening applications

Energy Technology Data Exchange (ETDEWEB)

Xu Feng; Wu Jinhui; Wang Shuqi; Gurkan, Umut Atakan; Demirci, Utkan [Department of Medicine, Demirci Bio-Acoustic-MEMS in Medicine (BAMM) Laboratory, Center for Biomedical Engineering, Brigham and Women' s Hospital, Harvard Medical School, Boston, MA (United States); Durmus, Naside Gozde, E-mail: udemirci@rics.bwh.harvard.edu [School of Engineering and Division of Biology and Medicine, Brown University, Providence, RI (United States)

2011-09-15

Screening for effective therapeutic agents from millions of drug candidates is costly, time consuming, and often faces concerns due to the extensive use of animals. To improve cost effectiveness, and to minimize animal testing in pharmaceutical research, in vitro monolayer cell microarrays with multiwell plate assays have been developed. Integration of cell microarrays with microfluidic systems has facilitated automated and controlled component loading, significantly reducing the consumption of the candidate compounds and the target cells. Even though these methods significantly increased the throughput compared to conventional in vitro testing systems and in vivo animal models, the cost associated with these platforms remains prohibitively high. Besides, there is a need for three-dimensional (3D) cell-based drug-screening models which can mimic the in vivo microenvironment and the functionality of the native tissues. Here, we present the state-of-the-art microengineering approaches that can be used to develop 3D cell-based drug-screening assays. We highlight the 3D in vitro cell culture systems with live cell-based arrays, microfluidic cell culture systems, and their application to high-throughput drug screening. We conclude that among the emerging microengineering approaches, bioprinting holds great potential to provide repeatable 3D cell-based constructs with high temporal, spatial control and versatility.

Entrapment of cultured pancreas islets in three-dimensional collagen matrices.

Science.gov (United States)

Chao, S H; Peshwa, M V; Sutherland, D E; Hu, W S

1992-01-01

In vitro culture of islets of Langerhans decreases their immunogenicity, presumably by eliminating passenger leukocytes and other Ia+ presenting cells within the islets. Islets cultivated in petri dishes either at 37 degrees C or at 25 degrees C gradually disintegrate during culture in a time-dependent manner which is related to the free-floating condition of the islets. Also, a fraction of the islets disperse as single cells and beta-cell aggregates or adhere to the bottom of the culture dishes. Thus, the retrieval rate of transplantable islets is dampened due to their disintegration and spontaneous dispersion in conventional petri dish cultures. Entrapment of freshly harvested islets of Langerhans in a three-dimensional collagen matrix was studied as an alternative method for islet cultivation. The contraction of collagen fibrils during in vitro culture counteracts the dispersion of islets and helps in maintaining their integrity while in culture. It was observed that the entrapped islets maintain satisfactory morphology, viability, and capability of glucose-dependent insulin secretion for over 2 wk. The oxygen consumption rate and glucose metabolism of these islets was not deranged when entrapped in collagen. Also, the retrieval of islets is easier and more efficient than that observed in conventional culture systems. Our results indicate that culture of islets in three-dimensional collagen gels can potentially develop into an ideal system applicable to clinical transplantation of cultured islets or beta-cell aggregates.

Apical polarity in three-dimensional culture systems: where to now?

Energy Technology Data Exchange (ETDEWEB)

Inman, J.L.; Bissell, Mina

2010-01-21

Delineation of the mechanisms that establish and maintain the polarity of epithelial tissues is essential to understanding morphogenesis, tissue specificity and cancer. Three-dimensional culture assays provide a useful platform for dissecting these processes but, as discussed in a recent study in BMC Biology on the culture of mammary gland epithelial cells, multiple parameters that influence the model must be taken into account.

High Throughput Neuro-Imaging Informatics

Directory of Open Access Journals (Sweden)

Michael I Miller

2013-12-01

Full Text Available This paper describes neuroinformatics technologies at 1 mm anatomical scale based on high throughput 3D functional and structural imaging technologies of the human brain. The core is an abstract pipeline for converting functional and structural imagery into their high dimensional neuroinformatic representations index containing O(E3-E4 discriminating dimensions. The pipeline is based on advanced image analysis coupled to digital knowledge representations in the form of dense atlases of the human brain at gross anatomical scale. We demonstrate the integration of these high-dimensional representations with machine learning methods, which have become the mainstay of other fields of science including genomics as well as social networks. Such high throughput facilities have the potential to alter the way medical images are stored and utilized in radiological workflows. The neuroinformatics pipeline is used to examine cross-sectional and personalized analyses of neuropsychiatric illnesses in clinical applications as well as longitudinal studies. We demonstrate the use of high throughput machine learning methods for supporting (i cross-sectional image analysis to evaluate the health status of individual subjects with respect to the population data, (ii integration of image and non-image information for diagnosis and prognosis.

Cell culture for three-dimensional modeling in rotating-wall vessels: an application of simulated microgravity

Science.gov (United States)

Schwarz, R. P.; Goodwin, T. J.; Wolf, D. A.

1992-01-01

High-density, three-dimensional cell cultures are difficult to grow in vitro. The rotating-wall vessel (RWV) described here has cultured BHK-21 cells to a density of 1.1 X 10(7) cells/ml. Cells on microcarriers were observed to grow with enhanced bridging in this batch culture system. The RWV is a horizontally rotated tissue culture vessel with silicon membrane oxygenation. This design results in a low-turbulence, low-shear cell culture environment with abundant oxygenation. The RWV has the potential to culture a wide variety of normal and neoplastic cells.

Growing B Lymphocytes in a Three-Dimensional Culture System

Science.gov (United States)

Wu, J. H. David; Bottaro, Andrea

2010-01-01

A three-dimensional (3D) culture system for growing long-lived B lymphocytes has been invented. The capabilities afforded by the system can be expected to expand the range of options for immunological research and related activities, including testing of immunogenicity of vaccine candidates in vitro, generation of human monoclonal antibodies, and immunotherapy. Mature lymphocytes, which are the effectors of adaptive immune responses in vertebrates, are extremely susceptible to apoptotic death, and depend on continuous reception of survival-inducing stimulation (in the forms of cytokines, cell-to-cell contacts, and antigen receptor signaling) from the microenvironment. For this reason, efforts to develop systems for long-term culture of functional, non-transformed and non-activated mature lymphocytes have been unsuccessful until now. The bone-marrow microenvironment supports the growth and differentiation of many hematopoietic lineages, in addition to B-lymphocytes. Primary bone-marrow cell cultures designed to promote the development of specific cell types in vitro are highly desirable experimental systems, amenable to manipulation under controlled conditions. However, the dynamic and complex network of stromal cells and insoluble matrix proteins is disrupted in prior plate- and flask-based culture systems, wherein the microenvironments have a predominantly two-dimensional (2D) character. In 2D bone-marrow cultures, normal B-lymphoid cells become progressively skewed toward precursor B-cell populations that do not retain a normal immunophenotype, and such mature B-lymphocytes as those harvested from the spleen or lymph nodes do not survive beyond several days ex vivo in the absence of mitogenic stimulation. The present 3D culture system is a bioreactor that contains highly porous artificial scaffolding that supports the long-term culture of bone marrow, spleen, and lymph-node samples. In this system, unlike in 2D culture systems, B-cell subpopulations developing

AELAS: Automatic ELAStic property derivations via high-throughput first-principles computation

Science.gov (United States)

Zhang, S. H.; Zhang, R. F.

2017-11-01

The elastic properties are fundamental and important for crystalline materials as they relate to other mechanical properties, various thermodynamic qualities as well as some critical physical properties. However, a complete set of experimentally determined elastic properties is only available for a small subset of known materials, and an automatic scheme for the derivations of elastic properties that is adapted to high-throughput computation is much demanding. In this paper, we present the AELAS code, an automated program for calculating second-order elastic constants of both two-dimensional and three-dimensional single crystal materials with any symmetry, which is designed mainly for high-throughput first-principles computation. Other derivations of general elastic properties such as Young's, bulk and shear moduli as well as Poisson's ratio of polycrystal materials, Pugh ratio, Cauchy pressure, elastic anisotropy and elastic stability criterion, are also implemented in this code. The implementation of the code has been critically validated by a lot of evaluations and tests on a broad class of materials including two-dimensional and three-dimensional materials, providing its efficiency and capability for high-throughput screening of specific materials with targeted mechanical properties. Program Files doi:http://dx.doi.org/10.17632/f8fwg4j9tw.1 Licensing provisions: BSD 3-Clause Programming language: Fortran Nature of problem: To automate the calculations of second-order elastic constants and the derivations of other elastic properties for two-dimensional and three-dimensional materials with any symmetry via high-throughput first-principles computation. Solution method: The space-group number is firstly determined by the SPGLIB code [1] and the structure is then redefined to unit cell with IEEE-format [2]. Secondly, based on the determined space group number, a set of distortion modes is automatically specified and the distorted structure files are generated

Three-dimensional neural cultures produce networks that mimic native brain activity.

Science.gov (United States)

Bourke, Justin L; Quigley, Anita F; Duchi, Serena; O'Connell, Cathal D; Crook, Jeremy M; Wallace, Gordon G; Cook, Mark J; Kapsa, Robert M I

2018-02-01

Development of brain function is critically dependent on neuronal networks organized through three dimensions. Culture of central nervous system neurons has traditionally been limited to two dimensions, restricting growth patterns and network formation to a single plane. Here, with the use of multichannel extracellular microelectrode arrays, we demonstrate that neurons cultured in a true three-dimensional environment recapitulate native neuronal network formation and produce functional outcomes more akin to in vivo neuronal network activity. Copyright © 2017 John Wiley & Sons, Ltd.

Quantitative volumetric Raman imaging of three dimensional cell cultures

KAUST Repository

Kallepitis, Charalambos

2017-03-22

The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell–material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy.

Quantitative volumetric Raman imaging of three dimensional cell cultures

Science.gov (United States)

Kallepitis, Charalambos; Bergholt, Mads S.; Mazo, Manuel M.; Leonardo, Vincent; Skaalure, Stacey C.; Maynard, Stephanie A.; Stevens, Molly M.

2017-03-01

The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell-material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy.

High-resolution two-dimensional and three-dimensional modeling of wire grid polarizers and micropolarizer arrays

Science.gov (United States)

Vorobiev, Dmitry; Ninkov, Zoran

2017-11-01

Recent advances in photolithography allowed the fabrication of high-quality wire grid polarizers for the visible and near-infrared regimes. In turn, micropolarizer arrays (MPAs) based on wire grid polarizers have been developed and used to construct compact, versatile imaging polarimeters. However, the contrast and throughput of these polarimeters are significantly worse than one might expect based on the performance of large area wire grid polarizers or MPAs, alone. We investigate the parameters that affect the performance of wire grid polarizers and MPAs, using high-resolution two-dimensional and three-dimensional (3-D) finite-difference time-domain simulations. We pay special attention to numerical errors and other challenges that arise in models of these and other subwavelength optical devices. Our tests show that simulations of these structures in the visible and near-IR begin to converge numerically when the mesh size is smaller than ˜4 nm. The performance of wire grid polarizers is very sensitive to the shape, spacing, and conductivity of the metal wires. Using 3-D simulations of micropolarizer "superpixels," we directly study the cross talk due to diffraction at the edges of each micropolarizer, which decreases the contrast of MPAs to ˜200∶1.

Advancing three-dimensional MEMS by complimentary laser micro manufacturing

Science.gov (United States)

Palmer, Jeremy A.; Williams, John D.; Lemp, Tom; Lehecka, Tom M.; Medina, Francisco; Wicker, Ryan B.

2006-01-01

This paper describes improvements that enable engineers to create three-dimensional MEMS in a variety of materials. It also provides a means for selectively adding three-dimensional, high aspect ratio features to pre-existing PMMA micro molds for subsequent LIGA processing. This complimentary method involves in situ construction of three-dimensional micro molds in a stand-alone configuration or directly adjacent to features formed by x-ray lithography. Three-dimensional micro molds are created by micro stereolithography (MSL), an additive rapid prototyping technology. Alternatively, three-dimensional features may be added by direct femtosecond laser micro machining. Parameters for optimal femtosecond laser micro machining of PMMA at 800 nanometers are presented. The technical discussion also includes strategies for enhancements in the context of material selection and post-process surface finish. This approach may lead to practical, cost-effective 3-D MEMS with the surface finish and throughput advantages of x-ray lithography. Accurate three-dimensional metal microstructures are demonstrated. Challenges remain in process planning for micro stereolithography and development of buried features following femtosecond laser micro machining.

Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres.

Directory of Open Access Journals (Sweden)

Delyan P Ivanov

Full Text Available Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money.

Evaluation of the osteogenic differentiation of gingiva-derived stem cells grown on culture plates or in stem cell spheroids: Comparison of two- and three-dimensional cultures.

Science.gov (United States)

Lee, Sung-Il; Ko, Youngkyung; Park, Jun-Beom

2017-09-01

Three-dimensional cell culture systems provide a convenient in vitro model for the study of complex cell-cell and cell-matrix interactions in the absence of exogenous substrates. The current study aimed to evaluate the osteogenic differentiation potential of gingiva-derived stem cells cultured in two-dimensional or three-dimensional systems. To the best of our knowledge, the present study is the first to compare the growth of gingiva-derived stem cells in monolayer culture to a three-dimensional culture system with microwells. For three-dimensional culture, gingiva-derived stem cells were isolated and seeded into polydimethylsiloxane-based concave micromolds. Alkaline phosphatase activity and alizarin red S staining assays were then performed to evaluate osteogenesis and the degree of mineralization, respectively. Stem cell spheroids had a significantly increased level of alkaline phosphatase activity and mineralization compared with cells from the two-dimensional culture. In addition, an increase in mineralized deposits was observed with an increase in the loading cell number. The results of present study indicate that gingiva-derived stem cell spheroids exhibit an increased osteogenic potential compared with stem cells from two-dimensional culture. This highlights the potential of three-dimensional culture systems using gingiva-derived stem cells for regenerative medicine applications requiring stem cells with osteogenic potential.

Three-dimensional optofluidic device for isolating microbes

Science.gov (United States)

Keloth, A.; Paterson, L.; Markx, G. H.; Kar, A. K.

2015-03-01

Development of efficient methods for isolation and manipulation of microorganisms is essential to study unidentified and yet-to-be cultured microbes originating from a variety of environments. The discovery of novel microbes and their products have the potential to contribute to the development of new medicines and other industrially important bioactive compounds. In this paper we describe the design, fabrication and validation of an optofluidic device capable of redirecting microbes within a flow using optical forces. The device holds promise to enable the high throughput isolation of single microbes for downstream culture and analysis. Optofluidic devices are widely used in clinical research, cell biology and biomedical engineering as they are capable of performing analytical functions such as controlled transportation, compact and rapid processing of nanolitres to millilitres of clinical or biological samples. We have designed and fabricated a three dimensional optofluidic device to control and manipulate microorganisms within a microfluidic channel. The device was fabricated in fused silica by ultrafast laser inscription (ULI) followed by selective chemical etching. The unique three-dimensional capability of ULI is utilized to integrate microfluidic channels and waveguides within the same substrate. The main microfluidic channel in the device constitutes the path of the sample. Optical waveguides are fabricated at right angles to the main microfluidic channel. The potential of the optical scattering force to control and manipulate microorganisms is discussed in this paper. A 980 nm continuous wave (CW) laser source, coupled to the waveguide, is used to exert radiation pressure on the particle and particle migrations at different flow velocities are recorded. As a first demonstration, device functionality is validated using fluorescent microbeads and initial trials with microalgae are presented.

Increased Paracrine Immunomodulatory Potential of Mesenchymal Stromal Cells in Three-Dimensional Culture

DEFF Research Database (Denmark)

Follin, Bjarke; Juhl, Morten; Cohen, Smadar

2016-01-01

Mesenchymal stromal/stem cells (MSCs) have been investigated extensively through the past years, proving to have great clinical therapeutic potential. In vitro cultivation of MSCs in three-dimensional (3D) culture systems, such as scaffolds, hydrogels, or spheroids, have recently gained attention...... for tissue engineering applications. Studies on MSC spheroids demonstrated that such cultivation increased the paracrine immunomodulatory potential of the MSCs, accompanied by phenotypic alterations. In this review, we gather results from recent experimental studies on the immunomodulatory abilities of MSCs...... when cultured as spheroids or in biomaterials like scaffolds or hydrogels compared to regular two-dimensional (2D) culture and show that alterations occurring to MSCs in spheroids also occur in MSCs in biomaterials. We provide a brief description of known mechanisms of MSC immunomodulatory capacity...

Fabrication of cell-benign inverse opal hydrogels for three-dimensional cell culture.

Science.gov (United States)

Im, Pilseon; Ji, Dong Hwan; Kim, Min Kyung; Kim, Jaeyun

2017-05-15

Inverse opal hydrogels (IOHs) for cell culture were fabricated and optimized using calcium-crosslinked alginate microbeads as sacrificial template and gelatin as a matrix. In contrast to traditional three-dimensional (3D) scaffolds, the gelatin IOHs allowed the utilization of both the macropore surface and inner matrix for cell co-culture. In order to remove templates efficiently for the construction of 3D interconnected macropores and to maintain high cell viability during the template removal process using EDTA solution, various factors in fabrication, including alginate viscosity, alginate concentration, alginate microbeads size, crosslinking calcium concentration, and gelatin network density were investigated. Low viscosity alginate, lower crosslinking calcium ion concentration, and lower concentration of alginate and gelatin were found to obtain high viability of cells encapsulated in the gelatin matrix after removal of the alginate template by EDTA treatment by allowing rapid dissociation and diffusion of alginate polymers. Based on the optimized fabrication conditions, gelatin IOHs showed good potential as a cell co-culture system, applicable to tissue engineering and cancer research. Copyright © 2017 Elsevier Inc. All rights reserved.

Cytokeratin expression of engrafted three-dimensional culture tissues using epithelial cells derived from porcine periodontal ligaments.

Science.gov (United States)

Yamada, Rie; Kitajima, Kayoko; Arai, Kyoko; Igarashi, Masaru

2014-09-01

This study investigated the differentiation and proliferation of epithelial cells derived from periodontal ligaments after three-dimensional culture using collagen gel with fibroblasts in vitro and in vivo. Epithelial cells and fibroblasts were derived from porcine periodontal ligaments. Epithelial cells were labeled using a fluorescent red membrane marker (PKH-26GL) and were seeded onto collagen gel with fibroblasts, followed by incubation in an air-liquid interface for 7 days. Three-dimensional cultures were grafted onto the backs of nude mice and removed at 1, 7, and 14 days after surgery (in vivo model). Unfixed sections (5 μm) were used to detect the presence of red fluorescent cells. Paraffin sections were analyzed histologically and immunohistochemically. Specimens were compared with three-dimensional culture tissues at 8, 14 and 21 days (in vitro model). Grafted three-dimensional cultures formed a stratified epithelial structure similar to skin in vivo. Epithelial cells were sequenced in basal-layer-like structures at 14 days in vivo. Immunohistochemical findings showed that the expression of cytokeratin was detected in the epithelial layer in in vitro and in vivo models. Ck8 + 18 + 19 was expressed in the upper epithelial layer in the in vitro model at 14 and 21 days, but not in vivo. Involucrin was expressed in the certified layers in vitro at 14 days, but not in vivo. Laminin was detected at the dermo-epidermal junction in vivo at 7 and 14 days, but not in vitro. These results suggest that differentiation of three-dimensional culture tissues differs in vivo and in vitro. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Influence of flow conditions and matrix coatings on growth and differentiation of three-dimensionally cultured rat hepatocytes.

Science.gov (United States)

Fiegel, Henning C; Havers, Joerg; Kneser, Ulrich; Smith, Molly K; Moeller, Tim; Kluth, Dietrich; Mooney, David J; Rogiers, Xavier; Kaufmann, Peter M

2004-01-01

Maintenance of liver-specific function of hepatocytes in culture is still difficult. Improved culture conditions may enhance the cell growth and function of cultured cells. We investigated the effect of three-dimensional culture under flow conditions, and the influence of surface modifications in hepatocyte cultures. Hepatocytes were harvested from Lewis rats. Cells were cultured on three-dimensional polymeric poly-lactic-co-glycolic acid (PLGA) matrices in static culture, or in a pulsatile flow-bioreactor system. Different surface modifications of matrices were investigated: coating with collagen I, collagen IV, laminin, or fibronectin; or uncoated matrix. Hepatocyte numbers, DNA content, and albumin secretion rate were assessed over the observation period. Culture under flow condition significantly enhanced cell numbers. An additional improvement of this effect was observed, when matrix coating was used. Cellular function also showed a significant increase (4- to 5-fold) under flow conditions when compared with static culture. Our data showed that culture under flow conditions improves cell number, and strongly enhances cellular function. Matrix modification by coating with extracellular matrix showed overall an additive stimulatory effect. Our conclusion is that combining three-dimensional culture under flow conditions and using matrix modification significantly improves culture conditions and is therefore attractive for the development of successful culture systems for hepatocytes.

BioSig3D: High Content Screening of Three-Dimensional Cell Culture Models.

Directory of Open Access Journals (Sweden)

Cemal Cagatay Bilgin

Full Text Available BioSig3D is a computational platform for high-content screening of three-dimensional (3D cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony. BioSig3D also enables visualization of raw and processed 3D volumetric data for quality control, and integrates advanced bioinformatics analysis. The system consists of multiple computational and annotation modules that are coupled together with a strong use of controlled vocabularies to reduce ambiguities between different users. It is a web-based system that allows users to: design an experiment by defining experimental variables, upload a large set of volumetric images into the system, analyze and visualize the dataset, and either display computed indices as a heatmap, or phenotypic subtypes for heterogeneity analysis, or download computed indices for statistical analysis or integrative biology. BioSig3D has been used to profile baseline colony formations with two experiments: (i morphogenesis of a panel of human mammary epithelial cell lines (HMEC, and (ii heterogeneity in colony formation using an immortalized non-transformed cell line. These experiments reveal intrinsic growth properties of well-characterized cell lines that are routinely used for biological studies. BioSig3D is being released with seed datasets and video-based documentation.

Three-dimensional printing of stem cell-laden hydrogels submerged in a hydrophobic high-density fluid

International Nuclear Information System (INIS)

Duarte Campos, Daniela F; Blaeser, Andreas; Weber, Michael; Fischer, Horst; Jäkel, Jörg; Neuss, Sabine; Jahnen-Dechent, Wilhelm

2013-01-01

Over the last decade, bioprinting technologies have begun providing important tissue engineering strategies for regenerative medicine and organ transplantation. The major drawback of past approaches has been poor or inadequate material-printing device and substrate combinations, as well as the relatively small size of the printed construct. Here, we hypothesise that cell-laden hydrogels can be printed when submerged in perfluorotributylamine (C 12 F 27 N), a hydrophobic high-density fluid, and that these cells placed within three-dimensional constructs remain viable allowing for cell proliferation and production of extracellular matrix. Human mesenchymal stem cells and MG-63 cells were encapsulated into agarose hydrogels, and subsequently printed in high aspect ratio in three dimensional structures that were supported in high density fluorocarbon. Three-dimensional structures with various shapes and sizes were manufactured and remained stable for more than six months. Live/dead and DAPI stainings showed viable cells 24 h after the printing process, as well as after 21 days in culture. Histological and immunohistochemical analyses after 14 and 21 days revealed viable cells with marked matrix production and signs of proliferation. The compressive strength values of the printed gels consequently increased during the two weeks in culture, revealing encouraging results for future applications in regenerative medicine. (paper)

Three-dimensional HepaRG model as an attractive tool for toxicity testing.

Science.gov (United States)

Leite, Sofia B; Wilk-Zasadna, Iwona; Zaldivar, Jose M; Airola, Elodie; Reis-Fernandes, Marcos A; Mennecozzi, Milena; Guguen-Guillouzo, Christiane; Chesne, Christopher; Guillou, Claude; Alves, Paula M; Coecke, Sandra

2012-11-01

The culture of HepaRG cells as three dimensional (3D) structures in the spinner-bioreactor may represent added value as a hepatic system for toxicological purposes. The use of a cost-effective commercially available bioreactor, which is compatible with high-throughput cell analysis, constitutes an attractive approach for routine use in the drug testing industry. In order to assess specific aspects of the biotransformation capacity of the bioreactor-based HepaRG system, the induction of CYP450 enzymes (i.e., CYP1A2, 2B6, 2C9, and 3A4) and the activity of the phase II enzyme, uridine diphosphate glucuronoltransferase (UGT), were tested. The long-term functionality of the system was demonstrated by 7-week stable profiles of albumin secretion, CYP3A4 induction, and UGT activities. Immunofluorescence-based staining showed formation of tissue-like arrangements including bile canaliculi-like structures and polar distribution of transporters. The use of in silico models to analyze the in vitro data related to hepatotoxic activity of acetaminophen (APAP) demonstrated the advantage of the integration of kinetic and dynamic aspects for a better understanding of the in vitro cell behavior. The bioactivation of APAP and its related cytotoxicity was assessed in a system compatible to high-throughput screening. The approach also proved to be a good strategy to reduce the time necessary to obtain fully differentiated cell cultures. In conclusion, HepaRG cells cultured in 3D spinner-bioreactors are an attractive tool for toxicological studies, showing a liver-like performance and demonstrating a practical applicability for toxicodynamic approaches.

Liarozole inhibits transforming growth factor-β3–mediated extracellular matrix formation in human three-dimensional leiomyoma cultures

Science.gov (United States)

Levy, Gary; Malik, Minnie; Britten, Joy; Gilden, Melissa; Segars, James; Catherino, William H.

2014-01-01

Objective To investigate the impact of liarozole on transforming growth factor-β3 (TGF-β3) expression, TGF-β3 controlled profibrotic cytokines, and extracellular matrix formation in a three-dimensional (3D) leiomyoma model system. Design Molecular and immunohistochemical analysis in a cell line evaluated in a three-dimensional culture. Setting Laboratory study. Patient(s) None. Intervention(s) Treatment of leiomyoma and myometrial cells with liarozole and TGF-β3 in a three-dimensional culture system. Main Outcome Measure(s) Quantitative real-time reverse-transcriptase polymerase chain reaction and Western blotting to assess fold gene and protein expression of TGF-β3 and TGF-β3 regulated fibrotic cytokines: collagen 1A1 (COL1A1), fibronectin, and versican before and after treatment with liarozole, and confirmatory immunohistochemical stains of treated three-dimensional cultures. Result(s) Both TGF-β3 gene and protein expression were elevated in leiomyoma cells compared with myometrium in two-dimensional and 3D cultures. Treatment with liarozole decreased TGF-β3 gene and protein expression. Extracellular matrix components versican, COL1A1, and fibronectin were also decreased by liarozole treatment in 3D cultures. Treatment of 3D cultures with TGF-β3 increased gene expression and protein production of COL1A1, fibronectin, and versican. Conclusion(s) Liarozole decreased TGF-β3 and TGF-β3–mediated extracellular matrix expression in a 3D uterine leiomyoma culture system. PMID:24825427

Three-dimensional spheroid culture promotes odonto/osteoblastic differentiation of dental pulp cells.

Science.gov (United States)

Yamamoto, Mioko; Kawashima, Nobuyuki; Takashino, Nami; Koizumi, Yu; Takimoto, Koyo; Suzuki, Noriyuki; Saito, Masahiro; Suda, Hideaki

2014-03-01

Three-dimensional (3D) spheroid culture is a method for creating 3D aggregations of cells and their extracellular matrix without a scaffold mimicking the actual tissues. The aim of this study was to evaluate the effects of 3D spheroid culture on the phenotype of immortalized mouse dental papilla cells (MDPs) that have the ability to differentiate into odontoblasts. We cultured MDPs for 1, 3, 7, and 14 days in 96-well low-attachment culture plates for 3D spheroid culture or flat-bottomed plates for two-dimensional (2D) monolayer culture. Cell proliferation and apoptosis were detected by immunohistochemical staining of Ki67 and cleaved caspase-3, respectively. Hypoxia was measured by the hypoxia probe LOX-1. Odonto/osteoblastic differentiation marker gene expression was evaluated by quantitative PCR. We also determined mineralized nodule formation, alkaline phosphatase (ALP) activity, and dentine matrix protein-1 (DMP1) expression. Vinculin and integrin signalling-related proteins were detected immunohistochemically. Odonto/osteoblastic marker gene expression and mineralized nodule formation were significantly up-regulated in 3D spheroid-cultured MDPs compared with those in 2D monolayer-cultured MDPs (podonto/osteoblastic differentiation of MDPs, which may be mediated by integrin signalling. Copyright © 2013 Elsevier Ltd. All rights reserved.

Attempt to develop taste bud models in three-dimensional culture.

Science.gov (United States)

Nishiyama, Miyako; Yuki, Saori; Fukano, Chiharu; Sako, Hideyuki; Miyamoto, Takenori; Tomooka, Yasuhiro

2011-09-01

Taste buds are the end organs of taste located in the gustatory papillae, which occur on the surface of the oral cavity. The goal of the present study was to establish a culture model mimicking the lingual taste bud of the mouse. To this end, three cell lines were employed: taste bud-derived cell lines (TBD cell lines), a lingual epithelial cell-derived cell line (20A cell line), and a mesenchymal cell-derived cell line (TMD cell line). TBD cells embedded in collagen gel formed three-dimensional clusters, which had an internal cavity equipped with a tight junction-like structure, a microvilluslike structure, and a laminin-positive layer surrounding the cluster. The cells with this epitheliumlike morphology expressed marker proteins of taste cells: gustducin and NCAM. TBD cells formed a monolayer on collagen gel when they were co-cultured with TMD cells. TBD, 20A, and TMD cell lines were maintained in a triple cell co-culture, in which TBD cells were pre-seeded as aggregates or in suspension on the collagen gel containing TMD cells, and 20A cells were laid over the TBD cells. TBD cells in the triple cell co-culture expressed NCAM. This result suggests that co-cultured TBD cells exhibited a characteristic of Type III taste cells. The culture model would be useful to study morphogenesis and functions of the gustatory organ.

A Bioengineered Three-Dimensional Cell Culture Platform Integrated with Microfluidics To Address Antimicrobial Resistance in Tuberculosis

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Magdalena K. Bielecka

2017-02-01

Full Text Available Antimicrobial resistance presents one of the most significant threats to human health, with the emergence of totally drug-resistant organisms. We have combined bioengineering, genetically modified bacteria, longitudinal readouts, and fluidics to develop a transformative platform to address the drug development bottleneck, utilizing Mycobacterium tuberculosis as the model organism. We generated microspheres incorporating virulent reporter bacilli, primary human cells, and an extracellular matrix by using bioelectrospray methodology. Granulomas form within the three-dimensional matrix, and mycobacterial stress genes are upregulated. Pyrazinamide, a vital first-line antibiotic for treating human tuberculosis, kills M. tuberculosis in a three-dimensional culture but not in a standard two-dimensional culture or Middlebrook 7H9 broth, demonstrating that antibiotic sensitivity within microspheres reflects conditions in patients. We then performed pharmacokinetic modeling by combining the microsphere system with a microfluidic plate and demonstrated that we can model the effect of dynamic antibiotic concentrations on mycobacterial killing. The microsphere system is highly tractable, permitting variation of cell content, the extracellular matrix, sphere size, the infectious dose, and the surrounding medium with the potential to address a wide array of human infections and the threat of antimicrobial resistance.

Three-dimensional hydrogel cell culture systems for modeling neural tissue

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Frampton, John

Two-dimensional (2-D) neural cell culture systems have served as physiological models for understanding the cellular and molecular events that underlie responses to physical and chemical stimuli, control sensory and motor function, and lead to the development of neurological diseases. However, the development of three-dimensional (3-D) cell culture systems will be essential for the advancement of experimental research in a variety of fields including tissue engineering, chemical transport and delivery, cell growth, and cell-cell communication. In 3-D cell culture, cells are provided with an environment similar to tissue, in which they are surrounded on all sides by other cells, structural molecules and adhesion ligands. Cells grown in 3-D culture systems display morphologies and functions more similar to those observed in vivo, and can be cultured in such a way as to recapitulate the structural organization and biological properties of tissue. This thesis describes a hydrogel-based culture system, capable of supporting the growth and function of several neural cell types in 3-D. Alginate hydrogels were characterized in terms of their biomechanical and biochemical properties and were functionalized by covalent attachment of whole proteins and peptide epitopes. Methods were developed for rapid cross-linking of alginate hydrogels, thus permitting the incorporation of cells into 3-D scaffolds without adversely affecting cell viability or function. A variety of neural cell types were tested including astrocytes, microglia, and neurons. Cells remained viable and functional for longer than two weeks in culture and displayed process outgrowth in 3-D. Cell constructs were created that varied in cell density, type and organization, providing experimental flexibility for studying cell interactions and behavior. In one set of experiments, 3-D glial-endothelial cell co-cultures were used to model blood-brain barrier (BBB) structure and function. This co-culture system was

Culture of three-dimensional tissue model and its application in bystander-effect research

International Nuclear Information System (INIS)

Wu Ruqun; Xu An; Wu Lijun; Hu Burong

2012-01-01

Compared with the cultured monolayer (2D) cells, three-dimensional (3D) tissue could be more similar to the environment in vivo including the physical support, chemical factors, cell-cell and cell-matrix interaction and so on. With the development of three-dimensional cell culture techniques (TDCC), 3D tissue is widely used in the areas of bystander effect research. This review focuses on introducing the TDCC method and its application in bystander-effect research. First, the development process of 3D tissue culture method was introduced. Secondly, the induction of radiation induced bystander effects both in 2D cell and 3D tissue and its mechanisms were reviewed. Finally, because heavy ion (carbon ion beam) has been developed as a useful tool to cure solid cancer, and the 3D tissue model is an ideal material to study the damages on body after being irradiated and to understand the underlying mechanisms, future study about heavy ion radiation inducing bystander effect in 3D tissue was discussed. (authors)

CyTOF workflow: differential discovery in high-throughput high-dimensional cytometry datasets [version 1; referees: 2 approved

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Malgorzata Nowicka

2017-05-01

Full Text Available High dimensional mass and flow cytometry (HDCyto experiments have become a method of choice for high throughput interrogation and characterization of cell populations.Here, we present an R-based pipeline for differential analyses of HDCyto data, largely based on Bioconductor packages. We computationally define cell populations using FlowSOM clustering, and facilitate an optional but reproducible strategy for manual merging of algorithm-generated clusters. Our workflow offers different analysis paths, including association of cell type abundance with a phenotype or changes in signaling markers within specific subpopulations, or differential analyses of aggregated signals. Importantly, the differential analyses we show are based on regression frameworks where the HDCyto data is the response; thus, we are able to model arbitrary experimental designs, such as those with batch effects, paired designs and so on. In particular, we apply generalized linear mixed models to analyses of cell population abundance or cell-population-specific analyses of signaling markers, allowing overdispersion in cell count or aggregated signals across samples to be appropriately modeled. To support the formal statistical analyses, we encourage exploratory data analysis at every step, including quality control (e.g. multi-dimensional scaling plots, reporting of clustering results (dimensionality reduction, heatmaps with dendrograms and differential analyses (e.g. plots of aggregated signals.

Three-dimensional alginate spheroid culture system of murine osteosarcoma.

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Akeda, Koji; Nishimura, Akinobu; Satonaka, Haruhiko; Shintani, Ken; Kusuzaki, Katsuyuki; Matsumine, Akihiko; Kasai, Yuichi; Masuda, Koichi; Uchida, Atsumasa

2009-11-01

Osteosarcoma (OS) is the most common primary malignant tumor of the bone and often forms pulmonary metastases, which are the most important prognostic factor. For further elucidation of the mechanism underlying the progression and metastasis of human OS, a culture system mimicking the microenvironment of the tumor in vivo is needed. We report a novel three-dimensional (3D) alginate spheroid culture system of murine osteosarcoma. Two different metastatic clones, the parental Dunn and its derivative line LM8, which has a higher metastatic potential to the lungs, were encapsulated in alginate beads to develop the 3D culture system. The beads containing murine OS cells were also transplanted into mice to determine their metastatic potential in vivo. In this culture system, murine OS cells encapsulated in alginate beads were able to grow in a 3D structure with cells detaching from the alginate environment. The number of detaching cells was higher in the LM8 cell line than the Dunn cell line. In the in vivo alginate bead transplantation model, the rate of pulmonary metastasis was higher with LM8 cells compared with that of Dunn cells. The cell characteristics and kinetics in this culture system closely reflect the original malignant potential of the cells in vivo.

Cell-free DNA in a three-dimensional spheroid cell culture model

DEFF Research Database (Denmark)

Aucamp, Janine; Calitz, Carlemi; Bronkhorst, Abel J.

2017-01-01

Background Investigating the biological functions of cell-free DNA (cfDNA) is limited by the interference of vast numbers of putative sources and causes of DNA release into circulation. Utilization of three-dimensional (3D) spheroid cell cultures, models with characteristics closer to the in vivo...... cultures can serve as effective, simplified in vivo-simulating “closed-circuit” models since putative sources of cfDNA are limited to only the targeted cells. In addition, cfDNA can also serve as an alternative or auxiliary marker for tracking spheroid growth, development and culture stability. Biological...... significance 3D cell cultures can be used to translate “closed-circuit” in vitro model research into data that is relevant for in vivo studies and clinical applications. In turn, the utilization of cfDNA during 3D culture research can optimize sample collection without affecting the stability of the growth...

Three-Dimensional Cell Culture Models for Infectious Disease and Drug Development

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Nickerson, Cheryl A.; Honer zu Bentrup, Kerstin; Ott, C. Mark

2005-01-01

Three-dimensional (3-D) cell cultures hold enormous potential to advance our understanding of infectious disease and to effectively translate basic cellular research into clinical applications. Using novel NASA bioreactor technology, the rotating wall vessel (RWV), we have engineered physiologically relevant 3-D human tissue culture models for infectious disease studies. The design of the RWV is based on the understanding that organs and tissues function in a 3-D environment, and that this 3-D architecture is critical for the differentiated form and function of tissues in vivo. The RWV provides large numbers of cells which are amenable to a wide variety of experimental manipulations and provides an easy, reproducible, and cost-effective approach to enhance differentiated features of cell culture models.

The fabrication and cell culture of three-dimensional rolled scaffolds with complex micro-architectures

International Nuclear Information System (INIS)

Liu Yaxiong; Li Xiao; Qu Xiaoli; Zhu Lin; He Jiankang; Zhao Qian; Wu Wanquan; Li Dichen

2012-01-01

Cell cultures for tissue engineering are traditionally prepared on two-dimensional or three-dimensional scaffolds with simple pores; however, this limits mass transportation, which is necessary for cell viability and function. In this paper, an innovative method is proposed for fabricating porous scaffolds with designed complex micro-architectures. Channels devised by computer-aided design were used to simulate features of blood vessels in native rat liver. Rapid prototyping and microreplication were used to produce a negative polydimethylsiloxane mold, and then a planar porous scaffold with predefined microchannel parameters was obtained by freeze-drying a silk fibroin/gelatin solution of an optimized concentration. After seeding with rat primary hepatocytes, the planar scaffold was rolled up to build spatial channels. By reconstructing the three-dimensional channel model in the scaffold in the form of micro-computed topography data and observing the cross-sections of the scroll, we confirmed that the bent channels were still interconnected, with restricted deviations. A comparison of the primary hepatocyte culture in the scaffolds with and without the devised channels proved that our design influenced cell organization and improved cell survival and proliferation. This method can be used for the construction of complex tissues for implantation and for culturing cells in vitro for biological tests and observations.

Airway branching morphogenesis in three dimensional culture

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Gudjonsson Thorarinn

2010-11-01

Full Text Available Abstract Background Lungs develop from the fetal digestive tract where epithelium invades the vascular rich stroma in a process called branching morphogenesis. In organogenesis, endothelial cells have been shown to be important for morphogenesis and the maintenance of organ structure. The aim of this study was to recapitulate human lung morphogenesis in vitro by establishing a three dimensional (3D co-culture model where lung epithelial cells were cultured in endothelial-rich stroma. Methods We used a human bronchial epithelial cell line (VA10 recently developed in our laboratory. This cell line cell line maintains a predominant basal cell phenotype, expressing p63 and other basal markers such as cytokeratin-5 and -14. Here, we cultured VA10 with human umbilical vein endothelial cells (HUVECs, to mimic the close interaction between these cell types during lung development. Morphogenesis and differentiation was monitored by phase contrast microscopy, immunostainings and confocal imaging. Results We found that in co-culture with endothelial cells, the VA10 cells generated bronchioalveolar like structures, suggesting that lung epithelial branching is facilitated by the presence of endothelial cells. The VA10 derived epithelial structures display various complex patterns of branching and show partial alveolar type-II differentiation with pro-Surfactant-C expression. The epithelial origin of the branching VA10 colonies was confirmed by immunostaining. These bronchioalveolar-like structures were polarized with respect to integrin expression at the cell-matrix interface. The endothelial-induced branching was mediated by soluble factors. Furthermore, fibroblast growth factor receptor-2 (FGFR-2 and sprouty-2 were expressed at the growing tips of the branching structures and the branching was inhibited by the FGFR-small molecule inhibitor SU5402. Discussion In this study we show that a human lung epithelial cell line can be induced by endothelial cells to

Three-dimensional Huh7 cell culture system for the study of Hepatitis C virus infection

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Uprichard Susan L

2009-07-01

Full Text Available Abstract Background In order to elucidate how Hepatitis C Virus (HCV interacts with polarized hepatocytes in vivo and how HCV-induced alterations in cellular function contribute to HCV-associated liver disease, a more physiologically relevant hepatocyte culture model is needed. As such, NASA-engineered three-dimensional (3-D rotating wall vessel (RWV bioreactors were used in effort to promote differentiation of HCV-permissive Huh7 hepatoma cells. Results When cultured in the RWV, Huh7 cells became morphologically and transcriptionally distinct from more standard Huh7 two-dimensional (2-D monolayers. Specifically, RWV-cultured Huh7 cells formed complex, multilayered 3-D aggregates in which Phase I and Phase II xenobiotic drug metabolism genes, as well as hepatocyte-specific transcripts (HNF4α, Albumin, TTR and α1AT, were upregulated compared to 2-D cultured Huh7 cells. Immunofluorescence analysis revealed that these HCV-permissive 3-D cultured Huh7 cells were more polarized than their 2D counterparts with the expression of HCV receptors, cell adhesion and tight junction markers (CD81, scavenger receptor class B member 1, claudin-1, occludin, ZO-1, β-Catenin and E-Cadherin significantly increased and exhibiting apical, lateral and/or basolateral localization. Conclusion These findings show that when cultured in 3-D, Huh7 cells acquire a more differentiated hepatocyte-like phenotype. Importantly, we show that these 3D cultures are highly permissive for HCV infection, thus providing an opportunity to study HCV entry and the effects of HCV infection on host cell function in a more physiologically relevant cell culture system.

Three-dimensional organoid culture reveals involvement of Wnt/β-catenin pathway in proliferation of bladder cancer cells.

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Yoshida, Takahiro; Sopko, Nikolai A; Kates, Max; Liu, Xiaopu; Joice, Gregory; McConkey, David J; Bivalacqua, Trinity J

2018-02-16

There has been increasing awareness of the importance of three-dimensional culture of cancer cells. Tumor cells growing as multicellular spheroids in three-dimensional culture, alternatively called organoids, are widely believed to more closely mimic solid tumors in situ . Previous studies concluded that the Wnt/β-catenin pathway is required for regeneration of the normal urothelium after injury and that β-catenin is upregulated in human bladder cancers, but no clear evidence has been advanced to support the idea that the Wnt/β-catenin pathway is directly involved in deregulated proliferation and the other malignant characteristics of bladder cancer cells. Here we report that the Wnt/β-catenin pathway activator, CHIR99021, promoted proliferation of established human bladder cancer cell lines when they were grown in organoid culture but not when they were grown in conventional adherent cultures. CHIR99021 activated Wnt/β-catenin pathway in bladder cancer cell lines in organoid culture. CHIR99021 also stimulated proliferation and the Wnt/b-catenin pathway in primary human bladder cancer organoids. RNAi-mediated knockdown of β-catenin blocked growth of organoids. The effects of CHIR99021 were associated with decreased expression of the urothelial terminal differentiation marker, cytokeratin 20. Our data suggest that the Wnt/β-catenin pathway is required for the proliferation of bladder cancer cells in three-dimensional organoid culture and provide a concrete example of why organoid culture is important for cancer research.

Three-dimensional imaging and scanning: Current and future applications for pathology

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Navid Farahani

2017-01-01

Full Text Available Imaging is vital for the assessment of physiologic and phenotypic details. In the past, biomedical imaging was heavily reliant on analog, low-throughput methods, which would produce two-dimensional images. However, newer, digital, and high-throughput three-dimensional (3D imaging methods, which rely on computer vision and computer graphics, are transforming the way biomedical professionals practice. 3D imaging has been useful in diagnostic, prognostic, and therapeutic decision-making for the medical and biomedical professions. Herein, we summarize current imaging methods that enable optimal 3D histopathologic reconstruction: Scanning, 3D scanning, and whole slide imaging. Briefly mentioned are emerging platforms, which combine robotics, sectioning, and imaging in their pursuit to digitize and automate the entire microscopy workflow. Finally, both current and emerging 3D imaging methods are discussed in relation to current and future applications within the context of pathology.

Three-dimensional (3D) culture in sarcoma research and the clinical significance.

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Gao, Songtao; Shen, Jacson; Hornicek, Francis; Duan, Zhenfeng

2017-08-03

Sarcomas are rare malignant tumors that arise from transformed cells of mesenchymal origin. Despite the progress in diagnosis and treatment, sarcomas have a high mortality rate due to local recurrence, metastasis, and the development of drug resistance to chemotherapy. New models for sarcoma research are required to further understand the disease and to develop new therapies. In vitro sarcoma modeling is challenging because of significant genetic heterogeneities, diverse pathological, and overlapping clinical characteristics. Studies on the mechanisms of recurrence, metastasis, and drug resistance in sarcoma have resulted in the generation of novel three-dimensional (3D) culture models for sarcoma research. 3D culture models aim to recapitulate the tumor microenvironment that plays a critical role in the pathogenesis of sarcoma using biomaterial scaffolds of natural biological materials and artificial polymers. An ideal 3D culture model can properly mimic not only the microenvironment, oncogenesis, and maintenance of sarcoma cell growth, but also imitate the interactions between cells and to the extracellular matrix. More recently, 3D cell culture has been used to research the biological behavior and mechanism of chemotherapy and radiotherapy resistance in different sarcoma models. Ultimately, findings using 3D models that more accurately reflect human sarcoma biology are likely to translate into improved clinical outcomes. In this review, we discuss the most recent advances of 3D culture technologies in sarcoma research and emerging clinical applications.

Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model.

Science.gov (United States)

Chang, Robert; Emami, Kamal; Wu, Honglu; Sun, Wei

2010-12-01

In their normal in vivo matrix milieu, tissues assume complex well-organized three-dimensional architectures. Therefore, the primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario. This challenge can be addressed by applying emerging layered biofabrication approaches in which the precise configuration and composition of cells and bioactive matrix components can recapitulate the well-defined three-dimensional biomimetic microenvironments that promote cell-cell and cell-matrix interactions. Furthermore, the advent of and refinements in microfabricated systems can present physical and chemical cues to cells in a controllable and reproducible fashion unmatched with conventional cultures, resulting in the precise construction of engineered biomimetic microenvironments on the cellular length scale in geometries that are readily parallelized for high throughput in vitro models. As such, the convergence of layered solid freeform fabrication (SFF) technologies along with microfabrication techniques enables the creation of a three-dimensional micro-organ device to serve as an in vitro platform for cell culture, drug screening or to elicit further biological insights, particularly for NASA's interest in a flight-suitable high-fidelity microscale platform to study drug metabolism in space and planetary environments. The proposed model in this paper involves the combinatorial setup of an automated syringe-based, layered direct cell writing bioprinting process with micro-patterning techniques to fabricate a microscale in vitro device housing a chamber of bioprinted three-dimensional liver cell-encapsulated hydrogel-based tissue constructs in defined design patterns that biomimic the cell's natural microenvironment for enhanced biological functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, reproducibly fabricated tissue constructs were biologically characterized for

Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model

International Nuclear Information System (INIS)

Chang, Robert; Sun Wei; Emami, Kamal; Wu Honglu

2010-01-01

In their normal in vivo matrix milieu, tissues assume complex well-organized three-dimensional architectures. Therefore, the primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario. This challenge can be addressed by applying emerging layered biofabrication approaches in which the precise configuration and composition of cells and bioactive matrix components can recapitulate the well-defined three-dimensional biomimetic microenvironments that promote cell-cell and cell-matrix interactions. Furthermore, the advent of and refinements in microfabricated systems can present physical and chemical cues to cells in a controllable and reproducible fashion unmatched with conventional cultures, resulting in the precise construction of engineered biomimetic microenvironments on the cellular length scale in geometries that are readily parallelized for high throughput in vitro models. As such, the convergence of layered solid freeform fabrication (SFF) technologies along with microfabrication techniques enables the creation of a three-dimensional micro-organ device to serve as an in vitro platform for cell culture, drug screening or to elicit further biological insights, particularly for NASA's interest in a flight-suitable high-fidelity microscale platform to study drug metabolism in space and planetary environments. The proposed model in this paper involves the combinatorial setup of an automated syringe-based, layered direct cell writing bioprinting process with micro-patterning techniques to fabricate a microscale in vitro device housing a chamber of bioprinted three-dimensional liver cell-encapsulated hydrogel-based tissue constructs in defined design patterns that biomimic the cell's natural microenvironment for enhanced biological functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, reproducibly fabricated tissue constructs were biologically characterized for

Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model

Energy Technology Data Exchange (ETDEWEB)

Chang, Robert; Sun Wei [Department of Mechanical Engineering and Mechanics, Drexel University, Philadelphia, PA (United States); Emami, Kamal; Wu Honglu, E-mail: rcc34@drexel.ed, E-mail: sunwei@drexel.ed, E-mail: kamal.emami-1@nasa.go, E-mail: honglu.wu-1@nasa.go [Radiation Biophysics Laboratory, Human Adaptation and Countermeasures Office, NASA Johnson Space Center, Houston, TX (United States)

2010-12-15

In their normal in vivo matrix milieu, tissues assume complex well-organized three-dimensional architectures. Therefore, the primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario. This challenge can be addressed by applying emerging layered biofabrication approaches in which the precise configuration and composition of cells and bioactive matrix components can recapitulate the well-defined three-dimensional biomimetic microenvironments that promote cell-cell and cell-matrix interactions. Furthermore, the advent of and refinements in microfabricated systems can present physical and chemical cues to cells in a controllable and reproducible fashion unmatched with conventional cultures, resulting in the precise construction of engineered biomimetic microenvironments on the cellular length scale in geometries that are readily parallelized for high throughput in vitro models. As such, the convergence of layered solid freeform fabrication (SFF) technologies along with microfabrication techniques enables the creation of a three-dimensional micro-organ device to serve as an in vitro platform for cell culture, drug screening or to elicit further biological insights, particularly for NASA's interest in a flight-suitable high-fidelity microscale platform to study drug metabolism in space and planetary environments. The proposed model in this paper involves the combinatorial setup of an automated syringe-based, layered direct cell writing bioprinting process with micro-patterning techniques to fabricate a microscale in vitro device housing a chamber of bioprinted three-dimensional liver cell-encapsulated hydrogel-based tissue constructs in defined design patterns that biomimic the cell's natural microenvironment for enhanced biological functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, reproducibly fabricated tissue constructs were biologically

Improving Cardiac Action Potential Measurements: 2D and 3D Cell Culture.

Science.gov (United States)

Daily, Neil J; Yin, Yue; Kemanli, Pinar; Ip, Brian; Wakatsuki, Tetsuro

2015-11-01

Progress in the development of assays for measuring cardiac action potential is crucial for the discovery of drugs for treating cardiac disease and assessing cardiotoxicity. Recently, high-throughput methods for assessing action potential using induced pluripotent stem cell (iPSC) derived cardiomyocytes in both two-dimensional monolayer cultures and three-dimensional tissues have been developed. We describe an improved method for assessing cardiac action potential using an ultra-fast cost-effective plate reader with commercially available dyes. Our methods improve dramatically the detection of the fluorescence signal from these dyes and make way for the development of more high-throughput methods for cardiac drug discovery and cardiotoxicity.

Development of three-dimensional computed tomography system using TNRF2 of JRR-3M

Energy Technology Data Exchange (ETDEWEB)

Murata, Yutaka; Mochiki, Koh-ichi [Musashi Inst. of Tech., Tokyo (Japan); Matsubayashi, Masahito

1998-01-01

A three-dimensional filtering engine, a convolution engine, and a back projection engine were developed for real-time signal processing of three-dimensional computed tomography. The performance of the system was measured and through-put of 0.5 second per one cross sectional data processing was attained. (author)

High-throughput flow injection analysis mass spectroscopy with networked delivery of color-rendered results. 2. Three-dimensional spectral mapping of 96-well combinatorial chemistry racks.

Science.gov (United States)

Görlach, E; Richmond, R; Lewis, I

1998-08-01

For the last two years, the mass spectroscopy section of the Novartis Pharma Research Core Technology group has analyzed tens of thousands of multiple parallel synthesis samples from the Novartis Pharma Combinatorial Chemistry program, using an in-house developed automated high-throughput flow injection analysis electrospray ionization mass spectroscopy system. The electrospray spectra of these samples reflect the many structures present after the cleavage step from the solid support. The overall success of the sequential synthesis is mirrored in the purity of the expected end product, but the partial success of individual synthesis steps is evident in the impurities in the mass spectrum. However this latter reaction information, which is of considerable utility to the combinatorial chemist, is effectively hidden from view by the very large number of analyzed samples. This information is now revealed at the workbench of the combinatorial chemist by a novel three-dimensional display of each rack's complete mass spectral ion current using the in-house RackViewer Visual Basic application. Colorization of "forbidden loss" and "forbidden gas-adduct" zones, normalization to expected monoisotopic molecular weight, colorization of ionization intensity, and sorting by row or column were used in combination to highlight systematic patterns in the mass spectroscopy data.

Miniaturized microscope for high throughput screening of tumor spheroids in microfluidic devices

Science.gov (United States)

Uranga, Javier; Rodríguez-Pena, Alejandro; Gahigiro, Desiré; Ortiz-de-Solorzano, Carlos

2018-02-01

High-throughput in vitro screening of highly physiological three-dimensional cell cultures (3D-HTS) is rapidly gaining importance in preclinical studies, to study the effect of the microenvironment in tumor development, and to evaluate the efficacy of new anticancer drugs. Furthermore, it could also be envisioned the use of 3D-HTS systems in personalized anti-cancer treatment planning, based on tumor organoids or spheroids grown from tumor biopsies or isolated tumor circulating cells. Most commercial, multi-well plate based 3D-HTS systems are large, expensive, and are based on the use of multi-well plates that hardly provide a physiological environment and require the use of large amounts of biological material and reagents. In this paper we present a novel, miniaturized inverted microscope (hereinafter miniscospe), made up of low-cost, mass producible parts, that can be used to monitor the growth of living tumor cell spheroids within customized three-dimensional microfluidic platforms. Our 3D-HTS miniscope combines phase contrast imaging based on oblique back illumination technique with traditional widefield epi-fluorescence imaging, implemented using miniaturized electro-optical parts and gradient-index refraction lenses. This small (3x6x2cm), lightweight device can effectively image overtime the growth of (>200) tumor spheroids in a controlled and reproducible environment. Our miniscope can be used to acquire time-lapse images of cellular living spheroids over the course of several hours and captures their growth before and after drug treatment, to evaluate the effectiveness of the drug.

Three-dimensional spheroid culture targeting versatile tissue bioassays using a PDMS-based hanging drop array.

Science.gov (United States)

Kuo, Ching-Te; Wang, Jong-Yueh; Lin, Yu-Fen; Wo, Andrew M; Chen, Benjamin P C; Lee, Hsinyu

2017-06-29

Biomaterial-based tissue culture platforms have emerged as useful tools to mimic in vivo physiological microenvironments in experimental cell biology and clinical studies. We describe herein a three-dimensional (3D) tissue culture platform using a polydimethylsiloxane (PDMS)-based hanging drop array (PDMS-HDA) methodology. Multicellular spheroids can be achieved within 24 h and further boosted by incorporating collagen fibrils in PDMS-HDA. In addition, the spheroids generated from different human tumor cells exhibited distinct sensitivities toward drug chemotherapeutic agents and radiation as compared with two-dimensional (2D) cultures that often lack in vivo-like biological insights. We also demonstrated that multicellular spheroids may enable key hallmarks of tissue-based bioassays, including drug screening, tumor dissemination, cell co-culture, and tumor invasion. Taken together, these results offer new opportunities not only to achieve the active control of 3D multicellular spheroids on demand, but also to establish a rapid and cost-effective platform to study anti-cancer therapeutics and tumor microenvironments.

Adaptation of the three-dimensional wisdom scale (3D-WS) for the Korean cultural context.

Science.gov (United States)

Kim, Seungyoun; Knight, Bob G

2014-10-23

ABSTRACT Background: Previous research on wisdom has suggested that wisdom is comprised of cognitive, reflective, and affective components and has developed and validated wisdom measures based on samples from Western countries. To apply the measurement to Eastern cultures, the present study revised an existing wisdom scale, the three-dimensional wisdom scale (3D-WS, Ardelt, 2003) for the Korean cultural context. Methods: Participants included 189 Korean heritage adults (age range 19-96) living in Los Angeles. We added a culturally specific factor of wisdom to the 3D-WS: Modesty and Unobtrusiveness (Yang, 2001), which captures an Eastern aspect of wisdom. The structure and psychometrics of the scale were tested. By latent cluster analysis, we determined acculturation subgroups and examined group differences in the means of factors in the revised wisdom scale (3D-WS-K). Results: Three factors, Cognitive Flexibility, Viewpoint Relativism, and Empathic Modesty were found using confirmatory factor analysis. Respondents with high biculturalism were higher on Viewpoint Relativism and lower on Empathic Modesty. Conclusion: This study discovered that a revised wisdom scale had a distinct factor structure and item content in a Korean heritage sample. We also found acculturation influences on the meaning of wisdom.

High Throughput System for Plant Height and Hyperspectral Measurement

Science.gov (United States)

Zhao, H.; Xu, L.; Jiang, H.; Shi, S.; Chen, D.

2018-04-01

Hyperspectral and three-dimensional measurement can obtain the intrinsic physicochemical properties and external geometrical characteristics of objects, respectively. Currently, a variety of sensors are integrated into a system to collect spectral and morphological information in agriculture. However, previous experiments were usually performed with several commercial devices on a single platform. Inadequate registration and synchronization among instruments often resulted in mismatch between spectral and 3D information of the same target. And narrow field of view (FOV) extends the working hours in farms. Therefore, we propose a high throughput prototype that combines stereo vision and grating dispersion to simultaneously acquire hyperspectral and 3D information.

HIGH THROUGHPUT SYSTEM FOR PLANT HEIGHT AND HYPERSPECTRAL MEASUREMENT

Directory of Open Access Journals (Sweden)

H. Zhao

2018-04-01

Full Text Available Hyperspectral and three-dimensional measurement can obtain the intrinsic physicochemical properties and external geometrical characteristics of objects, respectively. Currently, a variety of sensors are integrated into a system to collect spectral and morphological information in agriculture. However, previous experiments were usually performed with several commercial devices on a single platform. Inadequate registration and synchronization among instruments often resulted in mismatch between spectral and 3D information of the same target. And narrow field of view (FOV extends the working hours in farms. Therefore, we propose a high throughput prototype that combines stereo vision and grating dispersion to simultaneously acquire hyperspectral and 3D information.

Alginate foam-based three-dimensional culture to investigate drug sensitivity in primary leukaemia cells.

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Karimpoor, Mahroo; Yebra-Fernandez, Eva; Parhizkar, Maryam; Orlu, Mine; Craig, Duncan; Khorashad, Jamshid S; Edirisinghe, Mohan

2018-04-01

The development of assays for evaluating the sensitivity of leukaemia cells to anti-cancer agents is becoming an important aspect of personalized medicine. Conventional cell cultures lack the three-dimensional (3D) structure of the bone marrow (BM), the extracellular matrix and stromal components which are crucial for the growth and survival of leukaemia stem cells. To accurately predict the sensitivity of the leukaemia cells in an in vitro assay a culturing system containing the essential components of BM is required. In this study, we developed a porous calcium alginate foam-based scaffold to be used for 3D culture. The new 3D culture was shown to be cell compatible as it supported the proliferation of both normal haematopoietic and leukaemia cells. Our cell differential assay for myeloid markers showed that the porous foam-based 3D culture enhanced myeloid differentiation in both leukaemia and normal haematopoietic cells compared to two-dimensional culture. The foam-based scaffold reduced the sensitivity of the leukaemia cells to the tested antileukaemia agents in K562 and HL60 leukaemia cell line model and also primary myeloid leukaemia cells. This observation supports the application of calcium alginate foams as scaffold components of the 3D cultures for investigation of sensitivity to antileukaemia agents in primary myeloid cells. © 2018 The Author(s).

Characterization of three human cell line models for high-throughput neuronal cytotoxicity screening.

Science.gov (United States)

Tong, Zhi-Bin; Hogberg, Helena; Kuo, David; Sakamuru, Srilatha; Xia, Menghang; Smirnova, Lena; Hartung, Thomas; Gerhold, David

2017-02-01

More than 75 000 man-made chemicals contaminate the environment; many of these have not been tested for toxicities. These chemicals demand quantitative high-throughput screening assays to assess them for causative roles in neurotoxicities, including Parkinson's disease and other neurodegenerative disorders. To facilitate high throughput screening for cytotoxicity to neurons, three human neuronal cellular models were compared: SH-SY5Y neuroblastoma cells, LUHMES conditionally-immortalized dopaminergic neurons, and Neural Stem Cells (NSC) derived from human fetal brain. These three cell lines were evaluated for rapidity and degree of differentiation, and sensitivity to 32 known or candidate neurotoxicants. First, expression of neural differentiation genes was assayed during a 7-day differentiation period. Of the three cell lines, LUHMES showed the highest gene expression of neuronal markers after differentiation. Both in the undifferentiated state and after 7 days of neuronal differentiation, LUHMES cells exhibited greater cytotoxic sensitivity to most of 32 suspected or known neurotoxicants than SH-SY5Y or NSCs. LUHMES cells were also unique in being more susceptible to several compounds in the differentiating state than in the undifferentiated state; including known neurotoxicants colchicine, methyl-mercury (II), and vincristine. Gene expression results suggest that differentiating LUHMES cells may be susceptible to apoptosis because they express low levels of anti-apoptotic genes BCL2 and BIRC5/survivin, whereas SH-SY5Y cells may be resistant to apoptosis because they express high levels of BCL2, BIRC5/survivin, and BIRC3 genes. Thus, LUHMES cells exhibited favorable characteristics for neuro-cytotoxicity screening: rapid differentiation into neurons that exhibit high level expression neuronal marker genes, and marked sensitivity of LUHMES cells to known neurotoxicants. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Two-dimensional materials from high-throughput computational exfoliation of experimentally known compounds

Science.gov (United States)

Mounet, Nicolas; Gibertini, Marco; Schwaller, Philippe; Campi, Davide; Merkys, Andrius; Marrazzo, Antimo; Sohier, Thibault; Castelli, Ivano Eligio; Cepellotti, Andrea; Pizzi, Giovanni; Marzari, Nicola

2018-02-01

Two-dimensional (2D) materials have emerged as promising candidates for next-generation electronic and optoelectronic applications. Yet, only a few dozen 2D materials have been successfully synthesized or exfoliated. Here, we search for 2D materials that can be easily exfoliated from their parent compounds. Starting from 108,423 unique, experimentally known 3D compounds, we identify a subset of 5,619 compounds that appear layered according to robust geometric and bonding criteria. High-throughput calculations using van der Waals density functional theory, validated against experimental structural data and calculated random phase approximation binding energies, further allowed the identification of 1,825 compounds that are either easily or potentially exfoliable. In particular, the subset of 1,036 easily exfoliable cases provides novel structural prototypes and simple ternary compounds as well as a large portfolio of materials to search from for optimal properties. For a subset of 258 compounds, we explore vibrational, electronic, magnetic and topological properties, identifying 56 ferromagnetic and antiferromagnetic systems, including half-metals and half-semiconductors.

A Novel Microgravity Simulator Applicable for Three-Dimensional Cell Culturing

Science.gov (United States)

Wuest, Simon L.; Richard, Stéphane; Walther, Isabelle; Furrer, Reinhard; Anderegg, Roland; Sekler, Jörg; Egli, Marcel

2014-10-01

Random Positioning Machines (RPM) were introduced decades ago to simulate microgravity. Since then numerous experiments have been carried out to study its influence on biological samples. The machine is valued by the scientific community involved in space relevant topics as an excellent experimental tool to conduct pre-studies, for example, before sending samples into space. We have developed a novel version of the traditional RPM to broaden its operative range. This novel version has now become interesting to researchers who are working in the field of tissue engineering, particularly those interested in alternative methods for three-dimensional (3D) cell culturing. The main modifications concern the cell culture condition and the algorithm that controls the movement of the frames for the nullification of gravity. An incubator was integrated into the inner frame of the RPM allowing precise control over the cell culture environment. Furthermore, several feed-throughs now allow a permanent supply of gas like CO 2. All these modifications substantially improve conditions to culture cells; furthermore, the rewritten software responsible for controlling the movement of the frames enhances the quality of the generated microgravity. Cell culture experiments were carried out with human lymphocytes on the novel RPM model to compare the obtained response to the results gathered on an older well-established RPM as well as to data from space flights. The overall outcome of the tests validates this novel RPM for cell cultivation under simulated microgravity conditions.

Preparation of three-dimensional macroporous chitosan-gelatin B microspheres and HepG2-cell culture.

Science.gov (United States)

Huang, Fang; Cui, Long; Peng, Cheng-Hong; Wu, Xu-Bo; Han, Bao-San; Dong, Ya-Dong

2016-12-01

Chitosan-gelatin B microspheres with an open, interconnected, highly macroporous (100-200 µm) structure were prepared via a three-step protocol combining freeze-drying with an electrostatic and ionic cross-linking method. Saturated tripolyphosphate ethanol solution (85% ethanol) was chosen as the crosslinking agent to prevent destruction of the porous structure and to improve the biostability of the chitosan-gelatin B microspheres, with N-(3-dimethylaminopropyl)-N'-ethyl-carbodiimide/N-hydroxysuccinimide as a second crosslinking agent to react with gelatin A and fixed chitosan-gelatin B microspheres to attain improved biocompatibility. Water absorption of the three-dimensional macroporous chitosan-gelatin B microspheres (3D-P-CGMs) was 12.84, with a porosity of 85.45%. In vitro lysozyme degradation after 1, 3, 5, 7, 10, 14, and 21 days showed improved biodegradation in the 3D-P-CGMs. The morphology of human hepatoma cell lines (HepG2 cells) cultured on the 3D-P-CGMs was spherical, unlike that of cells cultured under traditional two-dimensional conditions. Scanning electron microscopy and paraffin sections were used to confirm the porous structure of the 3D-P-CGMs. HepG2 cells were able to migrate inside through the pore. Cell proliferation and levels of albumin and lactate dehydrogenase suggested that the 3D-P-CGMs could provide a larger specific surface area and an appropriate microenvironment for cell growth and survival. Hence, the 3D-P-CGMs are eminently suitable as macroporous scaffolds for cell cultures in tissue engineering and cell carrier studies. Copyright © 2014 John Wiley & Sons, Ltd. Copyright © 2014 John Wiley & Sons, Ltd.

Three-dimensional co-culture facilitates the differentiation of embryonic stem cells into mature cardiomyocytes.

Science.gov (United States)

Ou, Dong-Bo; He, Yong; Chen, Rui; Teng, Ji-Wei; Wang, Hong-Tao; Zeng, Di; Liu, Xiong-Tao; Ding, Lu; Huang, Jin-Yan; Zheng, Qiang-Sun

2011-12-01

The cardiomyocyte (CM) differentiation of embryonic stem cells (ESCs) is routinely cultured as two-dimensional (2D) monolayer, which doesn't mimic in vivo physiological environment and may lead to low differentiated level of ESCs. Here, we develop a novel strategy that enhances CM differentiation of ESCs in collagen matrix three-dimensional (3D) culture combined with indirect cardiac fibroblasts co-culture. ESCs were cultured in hanging drops to form embryoid bodies (EBs) and then applied on collagen matrix. The EBs were indirectly co-cultured with cardiac fibroblasts by the hanging cell culture inserts (PET 1 µm). The molecular expressions and ultrastructural characteristics of ESC-derived CMs (ESCMs) were analyzed by real time RT-PCR, immunocytochemistry, and Transmission Electron Microscopy (TEM). We found that the percentage of beating EBs with cardiac fibroblasts co-culture was significantly higher than that without co-culture after differentiation period of 8 days. Type I collagen used as 3D substrates enhanced the late-stage CM differentiation of ESCs and had effect on ultrastructural mature of ESCMs in late-stage development. The combined effects of 3D and co-culture that mimic in vivo physiological environment further improved the efficiency of CM differentiation from ESCs, resulting in fiber-like structures of cardiac cells with organized sarcomeric structure in ESCMs. This novel 3D co-culture system emphasizes the fact that the ESC differentiation is actively responding to cues from their environment and those cues can drive phenotypic control, which provides a useful in vitro model to investigate CM differentiation of stem cells. Copyright © 2011 Wiley Periodicals, Inc.

Establishment of three-dimensional cultures of human pancreatic duct epithelial cells

International Nuclear Information System (INIS)

Gutierrez-Barrera, Angelica M.; Menter, David G.; Abbruzzese, James L.; Reddy, Shrikanth A.G.

2007-01-01

Three-dimensional (3D) cultures of epithelial cells offer singular advantages for studies of morphogenesis or the role of cancer genes in oncogenesis. In this study, as part of establishing a 3D culture system of pancreatic duct epithelial cells, we compared human pancreatic duct epithelial cells (HPDE-E6E7) with pancreatic cancer cell lines. Our results show, that in contrast to cancer cells, HPDE-E6E7 organized into spheroids with what appeared to be apical and basal membranes and a luminal space. Immunostaining experiments indicated that protein kinase Akt was phosphorylated (Ser473) and CTMP, a negative Akt regulator, was expressed in both HPDE-E6E7 and cancer cells. However, a nuclear pool of CTMP was detectable in HPDE-E6E7 cells that showed a dynamic concentrated expression pattern, a feature that further distinguished HPDE-E637 cells from cancer cells. Collectively, these data suggest that 3D cultures of HPDE-E6E7 cells are useful for investigating signaling and morphological abnormalities in pancreatic cancer cells

Preparation of wholemount mouse intestine for high-resolution three-dimensional imaging using two-photon microscopy.

Science.gov (United States)

Appleton, P L; Quyn, A J; Swift, S; Näthke, I

2009-05-01

Visualizing overall tissue architecture in three dimensions is fundamental for validating and integrating biochemical, cell biological and visual data from less complex systems such as cultured cells. Here, we describe a method to generate high-resolution three-dimensional image data of intact mouse gut tissue. Regions of highest interest lie between 50 and 200 mum within this tissue. The quality and usefulness of three-dimensional image data of tissue with such depth is limited owing to problems associated with scattered light, photobleaching and spherical aberration. Furthermore, the highest-quality oil-immersion lenses are designed to work at a maximum distance of image at high-resolution deep within tissue. We show that manipulating the refractive index of the mounting media and decreasing sample opacity greatly improves image quality such that the limiting factor for a standard, inverted multi-photon microscope is determined by the working distance of the objective as opposed to detectable fluorescence. This method negates the need for mechanical sectioning of tissue and enables the routine generation of high-quality, quantitative image data that can significantly advance our understanding of tissue architecture and physiology.

Three-Dimensional Messages for Interstellar Communication

Science.gov (United States)

Vakoch, Douglas A.

One of the challenges facing independently evolved civilizations separated by interstellar distances is to communicate information unique to one civilization. One commonly proposed solution is to begin with two-dimensional pictorial representations of mathematical concepts and physical objects, in the hope that this will provide a foundation for overcoming linguistic barriers. However, significant aspects of such representations are highly conventional, and may not be readily intelligible to a civilization with different conventions. The process of teaching conventions of representation may be facilitated by the use of three-dimensional representations redundantly encoded in multiple formats (e.g., as both vectors and as rasters). After having illustrated specific conventions for representing mathematical objects in a three-dimensional space, this method can be used to describe a physical environment shared by transmitter and receiver: a three-dimensional space defined by the transmitter--receiver axis, and containing stars within that space. This method can be extended to show three-dimensional representations varying over time. Having clarified conventions for representing objects potentially familiar to both sender and receiver, novel objects can subsequently be depicted. This is illustrated through sequences showing interactions between human beings, which provide information about human behavior and personality. Extensions of this method may allow the communication of such culture-specific features as aesthetic judgments and religious beliefs. Limitations of this approach will be noted, with specific reference to ETI who are not primarily visual.

Mouse embryonic stem cell culture for generation of three-dimensional retinal and cortical tissues.

Science.gov (United States)

Eiraku, Mototsugu; Sasai, Yoshiki

2011-12-15

Generation of compound tissues with complex structures is a major challenge in cell biology. In this article, we describe a protocol for mouse embryonic stem cell (ESC) culture for in vitro generation of three-dimensional retinal tissue, comparing it with the culture protocol for cortical tissue generation. Dissociated ESCs are reaggregated in a 96-well plate with reduced cell-plate adhesion and cultured as floating aggregates. Retinal epithelium is efficiently generated when ESC aggregates are cultured in serum-free medium containing extracellular matrix proteins, spontaneously forming hemispherical vesicles and then progressively transforming into a shape reminiscent of the embryonic optic cup in 9-10 d. In long-term culture, the ESC-derived optic cup generates a fully stratified retinal tissue consisting of all major neural retinal components. In contrast, the cortical differentiation culture can be started without exogenous extracellular matrix proteins, and it generates stratified cortical epithelia consisting of four distinct layers in 13 d.

Advanced three-dimensional culture of equine intestinal epithelial stem cells.

Science.gov (United States)

Stewart, A Stieler; Freund, J M; Gonzalez, L M

2018-03-01

Intestinal epithelial stem cells are critical to epithelial repair following gastrointestinal injury. The culture of intestinal stem cells has quickly become a cornerstone of a vast number of new research endeavours that range from determining tissue viability to testing drug efficacy for humans. This study aims to describe the methods of equine stem cell culture and highlights the future benefits of these techniques for the advancement of equine medicine. To describe the isolation and culture of small intestinal stem cells into three-dimensional (3D) enteroids in horses without clinical gastrointestinal abnormalities. Descriptive study. Intestinal samples were collected by sharp dissection immediately after euthanasia. Intestinal crypts containing intestinal stem cells were dissociated from the underlying tissue layers, plated in a 3D matrix and supplemented with growth factors. After several days, resultant 3D enteroids were prepared for immunofluorescent imaging and polymerase chain reaction (PCR) analysis to detect and characterise specific cell types present. Intestinal crypts were cryopreserved immediately following collection and viability assessed. Intestinal crypts were successfully cultured and matured into 3D enteroids containing a lumen and budding structures. Immunofluorescence and PCR were used to confirm the existence of stem cells and all post mitotic, mature cell types, described to exist in the horse intestinal epithelium. Previously frozen crypts were successfully cultured following a freeze-thaw cycle. Tissues were all derived from normal horses. Application of this technique for the study of specific disease was not performed at this time. The successful culture of equine intestinal crypts into 3D "mini-guts" allows for in vitro studies of the equine intestine. Additionally, these results have relevance to future development of novel therapies that harness the regenerative potential of equine intestine in horses with gastrointestinal disease

High throughput production of mouse monoclonal antibodies using antigen microarrays

DEFF Research Database (Denmark)

De Masi, Federico; Chiarella, P.; Wilhelm, H.

2005-01-01

Recent advances in proteomics research underscore the increasing need for high-affinity monoclonal antibodies, which are still generated with lengthy, low-throughput antibody production techniques. Here we present a semi-automated, high-throughput method of hybridoma generation and identification....... Monoclonal antibodies were raised to different targets in single batch runs of 6-10 wk using multiplexed immunisations, automated fusion and cell-culture, and a novel antigen-coated microarray-screening assay. In a large-scale experiment, where eight mice were immunized with ten antigens each, we generated...

High-resolution three-dimensional mapping of semiconductor dopant potentials

DEFF Research Database (Denmark)

Twitchett, AC; Yates, TJV; Newcomb, SB

2007-01-01

Semiconductor device structures are becoming increasingly three-dimensional at the nanometer scale. A key issue that must be addressed to enable future device development is the three-dimensional mapping of dopant distributions, ideally under "working conditions". Here we demonstrate how a combin......Semiconductor device structures are becoming increasingly three-dimensional at the nanometer scale. A key issue that must be addressed to enable future device development is the three-dimensional mapping of dopant distributions, ideally under "working conditions". Here we demonstrate how...... a combination of electron holography and electron tomography can be used to determine quantitatively the three-dimensional electrostatic potential in an electrically biased semiconductor device with nanometer spatial resolution....

Three-dimensional culture conditions lead to decreased radiation induced cytotoxicity in human mammary epithelial cells

International Nuclear Information System (INIS)

Sowa, Marianne B.; Chrisler, William B.; Zens, Kyra D.; Ashjian, Emily J.; Opresko, Lee K.

2010-01-01

For both targeted and non-targeted exposures, the cellular responses to ionizing radiation have predominantly been measured in two-dimensional monolayer cultures. Although convenient for biochemical analysis, the true interactions in vivo depend upon complex interactions between cells themselves and the surrounding extracellular matrix. This study directly compares the influence of culture conditions on radiation induced cytotoxicity following exposure to low-LET ionizing radiation. Using a three-dimensional (3D) human mammary epithelial tissue model, we have found a protective effect of 3D cell culture on cell survival after irradiation. The initial state of the cells (i.e., 2D versus 3D culture) at the time of irradiation does not alter survival, nor does the presence of extracellular matrix during and after exposure to dose, but long term culture in 3D which offers significant reduction in cytotoxicity at a given dose (e.g. ∼4-fold increased survival at 5 Gy). The cell cycle delay induced following exposure to 2 and 5 Gy was almost identical between 2D and 3D culture conditions and cannot account for the observed differences in radiation responses. However the amount of apoptosis following radiation exposure is significantly decreased in 3D culture relative to the 2D monolayer after the same dose. A likely mechanism of the cytoprotective effect afforded by 3D culture conditions is the down regulation of radiation induced apoptosis in 3D structures.

A three-dimensional cell culture model to study the mechano-biological behavior in periodontal ligament regeneration

NARCIS (Netherlands)

Oortgiesen, D.A.W.; Yu, N.; Bronckers, A.L.J.J.; Yang, F.; Walboomers, X.F.; Jansen, J.A.

2012-01-01

Periodontitis is a disease affecting the supporting structures of the teeth, which can eventually result in tooth loss. A three-dimensional (3D) tissue culture model was developed that may serve to grow a 3D construct that not only transplants into defective periodontal sites, but also allows to

A Functional High-Throughput Assay of Myelination in Vitro

Science.gov (United States)

2014-07-01

Human induced pluripotent stem cells, hydrogels, 3D culture, electrophysiology, high-throughput assay 16. SECURITY CLASSIFICATION OF: 17...image the 3D rat dorsal root ganglion ( DRG ) cultures with sufficiently low background as to detect electrically-evoked depolarization events, as...of voltage-sensitive dyes. 8    We have made substantial progress in Task 4.1. We have fabricated neural fiber tracts from DRG explants and

Porcine spermatogonial stem cells self-renew effectively in a three dimensional culture microenvironment.

Science.gov (United States)

Park, Ji Eun; Park, Min Hee; Kim, Min Seong; Park, Yeo Reum; Yun, Jung Im; Cheong, Hee Tae; Kim, Minseok; Choi, Jung Hoon; Lee, Eunsong; Lee, Seung Tae

2017-12-01

Generally, self-renewal of spermatogonial stem cells (SSCs) is maintained in vivo in a three-dimensional (3D) microenvironment consisting of the seminiferous tubule basement membrane, indicating the importance of the 3D microenvironment for in vitro culture of SSCs. Here, we report a 3D culture microenvironment that effectively maintains porcine SSC self-renewal during culture. Porcine SSCs were cultured in an agarose-based 3D hydrogel and in 2D culture plates either with or without feeder cells. Subsequently, the effects of 3D culture on the maintenance of undifferentiated SSCs were identified by analyzing cell colony formation and morphology, AP activity, and transcriptional and translational regulation of self-renewal-related genes and the effects on proliferation by analyzing cell viability and single cell-derived colony number. The 3D culture microenvironment constructed using a 0.2% (w/v) agarose-based 3D hydrogel showed the strongest maintenance of porcine SSC self-renewal and induced significant improvements in proliferation compared with 2D culture microenvironments. These results demonstrate that self-renewal of porcine SSCs can be maintained more effectively in a 3D than in a 2D culture microenvironment. Moreover, this will play a significant role in developing novel culture systems for SSCs derived from diverse species in the future, which will contribute to SSC-related research. © 2017 International Federation for Cell Biology.

Direct Conversion of Equine Adipose-Derived Stem Cells into Induced Neuronal Cells Is Enhanced in Three-Dimensional Culture.

Science.gov (United States)

Petersen, Gayle F; Hilbert, Bryan J; Trope, Gareth D; Kalle, Wouter H J; Strappe, Padraig M

2015-12-01

The ability to culture neurons from horses may allow further investigation into equine neurological disorders. In this study, we demonstrate the generation of induced neuronal cells from equine adipose-derived stem cells (EADSCs) using a combination of lentiviral vector expression of the neuronal transcription factors Brn2, Ascl1, Myt1l (BAM) and NeuroD1 and a defined chemical induction medium, with βIII-tubulin-positive induced neuronal cells displaying a distinct neuronal morphology of rounded and compact cell bodies, extensive neurite outgrowth, and branching of processes. Furthermore, we investigated the effects of dimensionality on neuronal transdifferentiation, comparing conventional two-dimensional (2D) monolayer culture against three-dimensional (3D) culture on a porous polystyrene scaffold. Neuronal transdifferentiation was enhanced in 3D culture, with evenly distributed cells located on the surface and throughout the scaffold. Transdifferentiation efficiency was increased in 3D culture, with an increase in mean percent conversion of more than 100% compared to 2D culture. Additionally, induced neuronal cells were shown to transit through a Nestin-positive precursor state, with MAP2 and Synapsin 2 expression significantly increased in 3D culture. These findings will help to increase our understanding of equine neuropathogenesis, with prospective roles in disease modeling, drug screening, and cellular replacement for treatment of equine neurological disorders.

A gas trapping method for high-throughput metabolic experiments.

Science.gov (United States)

Krycer, James R; Diskin, Ciana; Nelson, Marin E; Zeng, Xiao-Yi; Fazakerley, Daniel J; James, David E

2018-01-01

Research into cellular metabolism has become more high-throughput, with typical cell-culture experiments being performed in multiwell plates (microplates). This format presents a challenge when trying to collect gaseous products, such as carbon dioxide (CO2), which requires a sealed environment and a vessel separate from the biological sample. To address this limitation, we developed a gas trapping protocol using perforated plastic lids in sealed cell-culture multiwell plates. We used this trap design to measure CO2 production from glucose and fatty acid metabolism, as well as hydrogen sulfide production from cysteine-treated cells. Our data clearly show that this gas trap can be applied to liquid and solid gas-collection media and can be used to study gaseous product generation by both adherent cells and cells in suspension. Since our gas traps can be adapted to multiwell plates of various sizes, they present a convenient, cost-effective solution that can accommodate the trend toward high-throughput measurements in metabolic research.

Three-dimensional bioprinting in tissue engineering and regenerative medicine.

Science.gov (United States)

Gao, Guifang; Cui, Xiaofeng

2016-02-01

With the advances of stem cell research, development of intelligent biomaterials and three-dimensional biofabrication strategies, highly mimicked tissue or organs can be engineered. Among all the biofabrication approaches, bioprinting based on inkjet printing technology has the promises to deliver and create biomimicked tissue with high throughput, digital control, and the capacity of single cell manipulation. Therefore, this enabling technology has great potential in regenerative medicine and translational applications. The most current advances in organ and tissue bioprinting based on the thermal inkjet printing technology are described in this review, including vasculature, muscle, cartilage, and bone. In addition, the benign side effect of bioprinting to the printed mammalian cells can be utilized for gene or drug delivery, which can be achieved conveniently during precise cell placement for tissue construction. With layer-by-layer assembly, three-dimensional tissues with complex structures can be printed using converted medical images. Therefore, bioprinting based on thermal inkjet is so far the most optimal solution to engineer vascular system to the thick and complex tissues. Collectively, bioprinting has great potential and broad applications in tissue engineering and regenerative medicine. The future advances of bioprinting include the integration of different printing mechanisms to engineer biphasic or triphasic tissues with optimized scaffolds and further understanding of stem cell biology.

Three-Dimensional Elastomeric Scaffolds Designed with Cardiac-Mimetic Structural and Mechanical Features

Science.gov (United States)

Neal, Rebekah A.; Jean, Aurélie; Park, Hyoungshin; Wu, Patrick B.; Hsiao, James; Engelmayr, George C.; Langer, Robert

2013-01-01

Tissue-engineered constructs, at the interface of material science, biology, engineering, and medicine, have the capacity to improve outcomes for cardiac patients by providing living cells and degradable biomaterials that can regenerate the native myocardium. With an ultimate goal of both delivering cells and providing mechanical support to the healing heart, we designed three-dimensional (3D) elastomeric scaffolds with (1) stiffnesses and anisotropy mimicking explanted myocardial specimens as predicted by finite-element (FE) modeling, (2) systematically varied combinations of rectangular pore pattern, pore aspect ratio, and strut width, and (3) structural features approaching tissue scale. Based on predicted mechanical properties, three scaffold designs were selected from eight candidates for fabrication from poly(glycerol sebacate) by micromolding from silicon wafers. Large 20×20 mm scaffolds with high aspect ratio features (5:1 strut height:strut width) were reproducibly cast, cured, and demolded at a relatively high throughput. Empirically measured mechanical properties demonstrated that scaffolds were cardiac mimetic and validated FE model predictions. Two-layered scaffolds providing fully interconnected pore networks were fabricated by layer-by-layer assembly. C2C12 myoblasts cultured on one-layered scaffolds exhibited specific patterns of cell elongation and interconnectivity that appeared to be guided by the scaffold pore pattern. Neonatal rat heart cells cultured on two-layered scaffolds for 1 week were contractile, both spontaneously and in response to electrical stimulation, and expressed sarcomeric α-actinin, a cardiac biomarker. This work not only demonstrated several scaffold designs that promoted functional assembly of rat heart cells, but also provided the foundation for further computational and empirical investigations of 3D elastomeric scaffolds for cardiac tissue engineering. PMID:23190320

Embryonic stem cells in scaffold-free three-dimensional cell culture: osteogenic differentiation and bone generation.

Science.gov (United States)

Handschel, Jörg; Naujoks, Christian; Depprich, Rita; Lammers, Lydia; Kübler, Norbert; Meyer, Ulrich; Wiesmann, Hans-Peter

2011-07-14

Extracorporeal formation of mineralized bone-like tissue is still an unsolved challenge in tissue engineering. Embryonic stem cells may open up new therapeutic options for the future and should be an interesting model for the analysis of fetal organogenesis. Here we describe a technique for culturing embryonic stem cells (ESCs) in the absence of artificial scaffolds which generated mineralized miromasses. Embryonic stem cells were harvested and osteogenic differentiation was stimulated by the addition of dexamethasone, ascorbic acid, and ß-glycerolphosphate (DAG). After three days of cultivation microspheres were formed. These spherical three-dimensional cell units showed a peripheral zone consisting of densely packed cell layers surrounded by minerals that were embedded in the extracellular matrix. Alizarine red staining confirmed evidence of mineralization after 10 days of DAG stimulation in the stimulated but not in the control group. Transmission electron microscopy demonstrated scorching crystallites and collagenous fibrils as early indication of bone formation. These extracellular structures resembled hydroxyl apatite-like crystals as demonstrated by distinct diffraction patterns using electron diffraction analysis. The micromass culture technique is an appropriate model to form three-dimensional bone-like micro-units without the need for an underlying scaffold. Further studies will have to show whether the technique is applicable also to pluripotent stem cells of different origin. © 2011 Handschel et al; licensee BioMed Central Ltd.

Embryonic stem cells in scaffold-free three-dimensional cell culture: osteogenic differentiation and bone generation

Directory of Open Access Journals (Sweden)

Meyer Ulrich

2011-07-01

Full Text Available Abstract Extracorporeal formation of mineralized bone-like tissue is still an unsolved challenge in tissue engineering. Embryonic stem cells may open up new therapeutic options for the future and should be an interesting model for the analysis of fetal organogenesis. Here we describe a technique for culturing embryonic stem cells (ESCs in the absence of artificial scaffolds which generated mineralized miromasses. Embryonic stem cells were harvested and osteogenic differentiation was stimulated by the addition of dexamethasone, ascorbic acid, and ß-glycerolphosphate (DAG. After three days of cultivation microspheres were formed. These spherical three-dimensional cell units showed a peripheral zone consisting of densely packed cell layers surrounded by minerals that were embedded in the extracellular matrix. Alizarine red staining confirmed evidence of mineralization after 10 days of DAG stimulation in the stimulated but not in the control group. Transmission electron microscopy demonstrated scorching crystallites and collagenous fibrils as early indication of bone formation. These extracellular structures resembled hydroxyl apatite-like crystals as demonstrated by distinct diffraction patterns using electron diffraction analysis. The micromass culture technique is an appropriate model to form three-dimensional bone-like micro-units without the need for an underlying scaffold. Further studies will have to show whether the technique is applicable also to pluripotent stem cells of different origin.

Highly ordered three-dimensional macroporous carbon spheres for determination of heavy metal ions

International Nuclear Information System (INIS)

Zhang, Yuxiao; Zhang, Jianming; Liu, Yang; Huang, Hui; Kang, Zhenhui

2012-01-01

Highlights: ► Highly ordered three dimensional macroporous carbon spheres (MPCSs) were prepared. ► MPCS was covalently modified by cysteine (MPCS–CO–Cys). ► MPCS–CO–Cys was first time used in electrochemical detection of heavy metal ions. ► Heavy metal ions such as Pb 2+ and Cd 2+ can be simultaneously determined. -- Abstract: An effective voltammetric method for detection of trace heavy metal ions using chemically modified highly ordered three dimensional macroporous carbon spheres electrode surfaces is described. The highly ordered three dimensional macroporous carbon spheres were prepared by carbonization of glucose in silica crystal bead template, followed by removal of the template. The highly ordered three dimensional macroporous carbon spheres were covalently modified by cysteine, an amino acid with high affinities towards some heavy metals. The materials were characterized by physical adsorption of nitrogen, scanning electron microscopy, and transmission electron microscopy techniques. While the Fourier-transform infrared spectroscopy was used to characterize the functional groups on the surface of carbon spheres. High sensitivity was exhibited when this material was used in electrochemical detection (square wave anodic stripping voltammetry) of heavy metal ions due to the porous structure. And the potential application for simultaneous detection of heavy metal ions was also investigated.

A multilayer microdevice for cell-based high-throughput drug screening

International Nuclear Information System (INIS)

Liu, Chong; Wang, Lei; Li, Jingmin; Ding, Xiping; Chunyu, Li; Xu, Zheng; Wang, Qi

2012-01-01

A multilayer polydimethylsiloxane microdevice for cell-based high-throughput drug screening is described in this paper. This established microdevice was based on a modularization method and it integrated a drug/medium concentration gradient generator (CGG), pneumatic microvalves and a cell culture microchamber array. The CGG was able to generate five steps of linear concentrations with the same outlet flow rate. The medium/drug flowed through CGG and then into the pear-shaped cell culture microchambers vertically. This vertical perfusion mode was used to reduce the impact of the shear stress on the physiology of cells induced by the fluid flow in the microchambers. Pear-shaped microchambers with two arrays of miropillars at each outlet were adopted in this microdevice, which were beneficial to cell distribution. The chemotherapeutics Cisplatin (DDP)-induced Cisplatin-resistant cell line A549/DDP apoptotic experiments were performed well on this platform. The results showed that this novel microdevice could not only provide well-defined and stable conditions for cell culture, but was also useful for cell-based high-throughput drug screening with less reagents and time consumption. (paper)

Parallel geometric classification of stem cells by their three-dimensional morphology

International Nuclear Information System (INIS)

Juba, Derek; Cardone, Antonio; Yiu Ip, Cheuk; Varshney, Amitabh; Simon Jr, Carl G; K Tison, Christopher; Kumar, Girish; Brady, Mary

2013-01-01

There is a need for tools to classify cells based on their three-dimensional (3D) shape. Cells exist in vivo in 3D, cells are frequently cultured within 3D scaffolds in vitro and 3D scaffolds are used for cell delivery in tissue engineering therapies. Recent work indicates that the physical structure of a tissue engineering scaffold can direct stem cell function by driving stem cells into morphologies that induce their differentiation. Thus, we have developed a rapid method for classifying cells based on their 3D shape. First, random lines are intersected with 3D Z-stacks of confocal images of stem cells. The intersection lengths are stored in histograms, which are then used to train a support vector machine (SVM) learning algorithm to distinguish between stem cells cultured on differentiation-inducing 3D scaffolds and those cultured on non-differentiating flat substrates. The trained SVM is able to properly classify the ‘new’ query cells over 80% of the time. The algorithm is easily parallelizable and we demonstrate its implementation on a commodity graphics processing unit (GPU). Use of a GPU to run the algorithm increases throughput by over 100-fold as compared to use of a CPU. The algorithm is also progressive, providing an approximate answer quickly and refining the answer over time. This allows further increase in the throughput of the algorithm by allowing the SVM classification scheme to terminate early if it becomes confident enough of the class of the cell being analyzed. These results demonstrate a rapid method for classifying stem cells based on their 3D shape that can be used by tissue engineers for identifying 3D tissue scaffold structures that drive stem cells into shapes that correlate with differentiation. (paper)

A physiologic three-dimensional cell culture system to investigate the role of decorin in matrix organisation and cell survival

International Nuclear Information System (INIS)

Seidler, Daniela G.; Schaefer, Liliana; Robenek, Horst; Iozzo, Renato V.; Kresse, Hans; Schoenherr, Elke

2005-01-01

In vivo cells exist in a three-dimensional environment generated and maintained by multiple cell-cell and cell-matrix interactions. Proteoglycans, like decorin, affect these complex interactions. Thus, we sought to investigate the role of decorin in a three-dimensional environment where the matrix was generated over time by decorin-deficient fibroblasts in the presence of L-ascorbic acid 2-phosphate. The cells were viable and proliferated in response to FGF2. Decorin was incorporated in the matrix and caused a ∼2 nm shift in the average diameter of the collagen fibrils, and the range and distribution of the fibrils became narrower and more uniform. Although there were no appreciable changes in collagen composition, we found that exogenous decorin induced the de novo synthesis of collagen I and V and cross-linked β (I). In the early phases of the three-dimensional culture, decorin reduced apoptosis. However, following the establishment of a three-dimensional matrix, the cells did not require decorin for their survival

Ho:YAG laser: intervertebral disk cell interaction using three-dimensional cell culture system

Science.gov (United States)

Sato, Masato; Ishihara, Miya; Arai, Tsunenori; Asazuma, Takashi; Kikuchi, Toshiyuki; Kikuchi, Makoto; Fujikawa, Kyosuke

2000-06-01

The purpose of this study is to evaluate the influence on the intervertebral disc cells after laser irradiation using three- dimensional culture system and to clarify the optimum Ho:YAG laser irradiation condition on percutaneous laser disc decompression (PLDD) for lumbar disc herniation. Since the Ho:YAG laser ablation is characterized by water-vapor bubble dynamics, not only thermal effect but also acoustic effect on cell metabolism might occur in the intervertebral disc. We studied the disc cell reaction from the metabolic point of view to investigate photothermal and photoacoustic effects on three-dimensional cultured disc cell. Intervertebral discs were obtained from female 30 Japanese white rabbits weighing about 1 kg. A pulsed Ho:YAG laser (wavelength: 2.1 micrometer, pulse width: about 200 microseconds) was delivered through a 200 micrometer-core diameter single silica glass fiber. We used the Ho:YAG laser irradiation fluence ranging from 60 to approximately 800 J/cm2 at the fiber end. To investigate acoustic effect, the acoustic transducer constructed with polyvinylidene fluoride (PVdF) film and acoustic absorber was used to detect the stress wave. Thermocouple and thermography were used to investigate thermal effect. Concerning damage of plasma membrane and ability of matrix synthesis, thermal effect might mainly affect cell reaction in total energy of 54 J (closed to practically used condition), but in 27 J, acoustic effect might contribute to it. We found that total energy was key parameter among the optimum condition, so that temperature and/or stress wave may influence Ho:YAG laser-disc cell interactions.

High-throughput miniaturized bioreactors for cell culture process development: reproducibility, scalability, and control.

Science.gov (United States)

Rameez, Shahid; Mostafa, Sigma S; Miller, Christopher; Shukla, Abhinav A

2014-01-01

Decreasing the timeframe for cell culture process development has been a key goal toward accelerating biopharmaceutical development. Advanced Microscale Bioreactors (ambr™) is an automated micro-bioreactor system with miniature single-use bioreactors with a 10-15 mL working volume controlled by an automated workstation. This system was compared to conventional bioreactor systems in terms of its performance for the production of a monoclonal antibody in a recombinant Chinese Hamster Ovary cell line. The miniaturized bioreactor system was found to produce cell culture profiles that matched across scales to 3 L, 15 L, and 200 L stirred tank bioreactors. The processes used in this article involve complex feed formulations, perturbations, and strict process control within the design space, which are in-line with processes used for commercial scale manufacturing of biopharmaceuticals. Changes to important process parameters in ambr™ resulted in predictable cell growth, viability and titer changes, which were in good agreement to data from the conventional larger scale bioreactors. ambr™ was found to successfully reproduce variations in temperature, dissolved oxygen (DO), and pH conditions similar to the larger bioreactor systems. Additionally, the miniature bioreactors were found to react well to perturbations in pH and DO through adjustments to the Proportional and Integral control loop. The data presented here demonstrates the utility of the ambr™ system as a high throughput system for cell culture process development. © 2014 American Institute of Chemical Engineers.

Scanning three-dimensional x-ray diffraction microscopy using a high-energy microbeam

International Nuclear Information System (INIS)

Hayashi, Y.; Hirose, Y.; Seno, Y.

2016-01-01

A scanning three-dimensional X-ray diffraction (3DXRD) microscope apparatus with a high-energy microbeam was installed at the BL33XU Toyota beamline at SPring-8. The size of the 50 keV beam focused using Kirkpatrick-Baez mirrors was 1.3 μm wide and 1.6 μm high in full width at half maximum. The scanning 3DXRD method was tested for a cold-rolled carbon steel sheet sample. A three-dimensional orientation map with 37 "3 voxels was obtained.

Scanning three-dimensional x-ray diffraction microscopy using a high-energy microbeam

Energy Technology Data Exchange (ETDEWEB)

Hayashi, Y., E-mail: y-hayashi@mosk.tytlabs.co.jp; Hirose, Y.; Seno, Y. [Toyota Central R& D Toyota Central R& D Labs., Inc., 41-1 Nagakute Aichi 480-1192 Japan (Japan)

2016-07-27

A scanning three-dimensional X-ray diffraction (3DXRD) microscope apparatus with a high-energy microbeam was installed at the BL33XU Toyota beamline at SPring-8. The size of the 50 keV beam focused using Kirkpatrick-Baez mirrors was 1.3 μm wide and 1.6 μm high in full width at half maximum. The scanning 3DXRD method was tested for a cold-rolled carbon steel sheet sample. A three-dimensional orientation map with 37 {sup 3} voxels was obtained.

On the Zeeman Effect in highly excited atoms: 2. Three-dimensional case

International Nuclear Information System (INIS)

Baseia, B.; Medeiros e Silva Filho, J.

1984-01-01

A previous result, found in two-dimensional hydrogen-atoms, is extended to the three-dimensional case. A mapping of a four-dimensional space R 4 onto R 3 , that establishes an equivalence between Coulomb and harmonic potentials, is used to show that the exact solution of the Zeeman effect in highly excited atoms, cannot be reached. (Author) [pt

Development of human nervous tissue upon differentiation of embryonic stem cells in three-dimensional culture.

Science.gov (United States)

Preynat-Seauve, Olivier; Suter, David M; Tirefort, Diderik; Turchi, Laurent; Virolle, Thierry; Chneiweiss, Herve; Foti, Michelangelo; Lobrinus, Johannes-Alexander; Stoppini, Luc; Feki, Anis; Dubois-Dauphin, Michel; Krause, Karl Heinz

2009-03-01

Researches on neural differentiation using embryonic stem cells (ESC) require analysis of neurogenesis in conditions mimicking physiological cellular interactions as closely as possible. In this study, we report an air-liquid interface-based culture of human ESC. This culture system allows three-dimensional cell expansion and neural differentiation in the absence of added growth factors. Over a 3-month period, a macroscopically visible, compact tissue developed. Histological coloration revealed a dense neural-like neural tissue including immature tubular structures. Electron microscopy, immunochemistry, and electrophysiological recordings demonstrated a dense network of neurons, astrocytes, and oligodendrocytes able to propagate signals. Within this tissue, tubular structures were niches of cells resembling germinal layers of human fetal brain. Indeed, the tissue contained abundant proliferating cells expressing markers of neural progenitors. Finally, the capacity to generate neural tissues on air-liquid interface differed for different ESC lines, confirming variations of their neurogenic potential. In conclusion, this study demonstrates in vitro engineering of a human neural-like tissue with an organization that bears resemblance to early developing brain. As opposed to previously described methods, this differentiation (a) allows three-dimensional organization, (b) yields dense interconnected neural tissue with structurally and functionally distinct areas, and (c) is spontaneously guided by endogenous developmental cues.

High-throughput determination of vancomycin in human plasma by a cost-effective system of two-dimensional liquid chromatography.

Science.gov (United States)

Sheng, Yanghao; Zhou, Boting

2017-05-26

Therapeutic drug monitoring (TDM) is one of the most important services of clinical laboratories. Two main techniques are commonly used: the immunoassay and chromatography method. We have developed a cost-effective system of two-dimensional liquid chromatography with ultraviolet detection (2D-LC-UV) for high-throughput determination of vancomycin in human plasma that combines the automation and low start-up costs of the immunoassay with the high selectivity and sensitivity of the liquid chromatography coupled with mass spectrometric detection without incurring their disadvantages, achieving high cost-effectiveness. This 2D-LC system offers a large volume injection to provide sufficient sensitivity and uses simulated gradient peak compression technology to control peak broadening and to improve peak shape. A middle column was added to reduce the analysis cycle time and make it suitable for high-throughput routine clinical assays. The analysis cycle time was 4min and the peak width was 0.8min. Compared with other chromatographic methods that have been developed, the analysis cycle time and peak width for vancomycin was reduced significantly. The lower limit of quantification was 0.20μg/mL for vancomycin, which is the same as certain LC-MS/MS methods that have been recently developed and validated. The method is rapid, automated, and low-cost and has high selectivity and sensitivity for the quantification of vancomycin in human plasma, thus making it well-suited for use in hospital clinical laboratories. Copyright © 2017 Elsevier B.V. All rights reserved.

Secure data storage by three-dimensional absorbers in highly scattering volume medium

International Nuclear Information System (INIS)

Matoba, Osamu; Matsuki, Shinichiro; Nitta, Kouichi

2008-01-01

A novel data storage in a volume medium with highly scattering coefficient is proposed for data security application. Three-dimensional absorbers are used as data. These absorbers can not be measured by interferometer when the scattering in a volume medium is strong enough. We present a method to reconstruct three-dimensional absorbers and present numerical results to show the effectiveness of the proposed data storage.

Crop 3D-a LiDAR based platform for 3D high-throughput crop phenotyping.

Science.gov (United States)

Guo, Qinghua; Wu, Fangfang; Pang, Shuxin; Zhao, Xiaoqian; Chen, Linhai; Liu, Jin; Xue, Baolin; Xu, Guangcai; Li, Le; Jing, Haichun; Chu, Chengcai

2018-03-01

With the growing population and the reducing arable land, breeding has been considered as an effective way to solve the food crisis. As an important part in breeding, high-throughput phenotyping can accelerate the breeding process effectively. Light detection and ranging (LiDAR) is an active remote sensing technology that is capable of acquiring three-dimensional (3D) data accurately, and has a great potential in crop phenotyping. Given that crop phenotyping based on LiDAR technology is not common in China, we developed a high-throughput crop phenotyping platform, named Crop 3D, which integrated LiDAR sensor, high-resolution camera, thermal camera and hyperspectral imager. Compared with traditional crop phenotyping techniques, Crop 3D can acquire multi-source phenotypic data in the whole crop growing period and extract plant height, plant width, leaf length, leaf width, leaf area, leaf inclination angle and other parameters for plant biology and genomics analysis. In this paper, we described the designs, functions and testing results of the Crop 3D platform, and briefly discussed the potential applications and future development of the platform in phenotyping. We concluded that platforms integrating LiDAR and traditional remote sensing techniques might be the future trend of crop high-throughput phenotyping.

Highly ordered three-dimensional macroporous carbon spheres for determination of heavy metal ions

Energy Technology Data Exchange (ETDEWEB)

Zhang, Yuxiao; Zhang, Jianming [Institute of Functional Nano and Soft Materials (FUNSOM) and Jiangsu Key Laboratory for Carbon-Based Functional Materials and Devices, Soochow University, Suzhou 215123 (China); Liu, Yang, E-mail: yangl@suda.edu.cn [Institute of Functional Nano and Soft Materials (FUNSOM) and Jiangsu Key Laboratory for Carbon-Based Functional Materials and Devices, Soochow University, Suzhou 215123 (China); Huang, Hui [Institute of Functional Nano and Soft Materials (FUNSOM) and Jiangsu Key Laboratory for Carbon-Based Functional Materials and Devices, Soochow University, Suzhou 215123 (China); Kang, Zhenhui, E-mail: zhkang@suda.edu.cn [Institute of Functional Nano and Soft Materials (FUNSOM) and Jiangsu Key Laboratory for Carbon-Based Functional Materials and Devices, Soochow University, Suzhou 215123 (China)

2012-04-15

Highlights: Black-Right-Pointing-Pointer Highly ordered three dimensional macroporous carbon spheres (MPCSs) were prepared. Black-Right-Pointing-Pointer MPCS was covalently modified by cysteine (MPCS-CO-Cys). Black-Right-Pointing-Pointer MPCS-CO-Cys was first time used in electrochemical detection of heavy metal ions. Black-Right-Pointing-Pointer Heavy metal ions such as Pb{sup 2+} and Cd{sup 2+} can be simultaneously determined. -- Abstract: An effective voltammetric method for detection of trace heavy metal ions using chemically modified highly ordered three dimensional macroporous carbon spheres electrode surfaces is described. The highly ordered three dimensional macroporous carbon spheres were prepared by carbonization of glucose in silica crystal bead template, followed by removal of the template. The highly ordered three dimensional macroporous carbon spheres were covalently modified by cysteine, an amino acid with high affinities towards some heavy metals. The materials were characterized by physical adsorption of nitrogen, scanning electron microscopy, and transmission electron microscopy techniques. While the Fourier-transform infrared spectroscopy was used to characterize the functional groups on the surface of carbon spheres. High sensitivity was exhibited when this material was used in electrochemical detection (square wave anodic stripping voltammetry) of heavy metal ions due to the porous structure. And the potential application for simultaneous detection of heavy metal ions was also investigated.

The efficacy of cetuximab in a tissue-engineered three-dimensional in vitro model of colorectal cancer

Directory of Open Access Journals (Sweden)

Tarig Magdeldin

2014-07-01

Full Text Available The preclinical development process of chemotherapeutic drugs is often carried out in two-dimensional monolayer cultures. However, a considerable amount of evidence demonstrates that two-dimensional cell culture does not accurately reflect the three-dimensional in vivo tumour microenvironment, specifically with regard to gene expression profiles, oxygen and nutrient gradients and pharmacokinetics. With this objective in mind, we have developed and established a physiologically relevant three-dimensional in vitro model of colorectal cancer based on the removal of interstitial fluid from collagen type I hydrogels. We employed the RAFT™ (Real Architecture For 3D Tissue system for producing three-dimensional cultures to create a controlled reproducible, multiwell testing platform. Using the HT29 and HCT116 cell lines to model epidermal growth factor receptor expressing colorectal cancers, we characterized three-dimensional cell growth and morphology in addition to the anti-proliferative effects of the anti–epidermal growth factor receptor chemotherapeutic agent cetuximab in comparison to two-dimensional monolayer cultures. Cells proliferated well for 14 days in three-dimensional culture and formed well-defined cellular aggregates within the concentrated collagen matrix. Epidermal growth factor receptor expression levels revealed a twofold and threefold increase in three-dimensional cultures for both HT29 and HCT116 cells in comparison to two-dimensional monolayers, respectively (p < 0.05; p < 0.01. Cetuximab efficacy was significantly lower in HT29 three-dimensional cultures in comparison to two-dimensional monolayers, whereas HCT116 cells in both two-dimension and three-dimension were non-responsive to treatment in agreement with their KRAS mutant status. In summary, these results confirm the use of a three-dimensional in vitro cancer model as a suitable drug-screening platform for in vitro pharmacological testing.

High Throughput Facility

Data.gov (United States)

Federal Laboratory Consortium — Argonne?s high throughput facility provides highly automated and parallel approaches to material and materials chemistry development. The facility allows scientists...

Design and fabrication of human skin by three-dimensional bioprinting.

Science.gov (United States)

Lee, Vivian; Singh, Gurtej; Trasatti, John P; Bjornsson, Chris; Xu, Xiawei; Tran, Thanh Nga; Yoo, Seung-Schik; Dai, Guohao; Karande, Pankaj

2014-06-01

Three-dimensional (3D) bioprinting, a flexible automated on-demand platform for the free-form fabrication of complex living architectures, is a novel approach for the design and engineering of human organs and tissues. Here, we demonstrate the potential of 3D bioprinting for tissue engineering using human skin as a prototypical example. Keratinocytes and fibroblasts were used as constituent cells to represent the epidermis and dermis, and collagen was used to represent the dermal matrix of the skin. Preliminary studies were conducted to optimize printing parameters for maximum cell viability as well as for the optimization of cell densities in the epidermis and dermis to mimic physiologically relevant attributes of human skin. Printed 3D constructs were cultured in submerged media conditions followed by exposure of the epidermal layer to the air-liquid interface to promote maturation and stratification. Histology and immunofluorescence characterization demonstrated that 3D printed skin tissue was morphologically and biologically representative of in vivo human skin tissue. In comparison with traditional methods for skin engineering, 3D bioprinting offers several advantages in terms of shape- and form retention, flexibility, reproducibility, and high culture throughput. It has a broad range of applications in transdermal and topical formulation discovery, dermal toxicity studies, and in designing autologous grafts for wound healing. The proof-of-concept studies presented here can be further extended for enhancing the complexity of the skin model via the incorporation of secondary and adnexal structures or the inclusion of diseased cells to serve as a model for studying the pathophysiology of skin diseases.

A high throughput mass spectrometry screening analysis based on two-dimensional carbon microfiber fractionation system.

Science.gov (United States)

Ma, Biao; Zou, Yilin; Xie, Xuan; Zhao, Jinhua; Piao, Xiangfan; Piao, Jingyi; Yao, Zhongping; Quinto, Maurizio; Wang, Gang; Li, Donghao

2017-06-09

A novel high-throughput, solvent saving and versatile integrated two-dimensional microscale carbon fiber/active carbon fiber system (2DμCFs) that allows a simply and rapid separation of compounds in low-polar, medium-polar and high-polar fractions, has been coupled with ambient ionization-mass spectrometry (ESI-Q-TOF-MS and ESI-QqQ-MS) for screening and quantitative analyses of real samples. 2DμCFs led to a substantial interference reduction and minimization of ionization suppression effects, thus increasing the sensitivity and the screening capabilities of the subsequent MS analysis. The method has been applied to the analysis of Schisandra Chinensis extracts, obtaining with a single injection a simultaneous determination of 33 compounds presenting different polarities, such as organic acids, lignans, and flavonoids in less than 7min, at low pressures and using small solvent amounts. The method was also validated using 10 model compounds, giving limit of detections (LODs) ranging from 0.3 to 30ngmL -1 , satisfactory recoveries (from 75.8 to 93.2%) and reproducibilities (relative standard deviations, RSDs, from 1.40 to 8.06%). Copyright © 2017 Elsevier B.V. All rights reserved.

Three-dimensional true FISP for high-resolution imaging of the whole brain

International Nuclear Information System (INIS)

Schmitz, B.; Hagen, T.; Reith, W.

2003-01-01

While high-resolution T1-weighted sequences, such as three-dimensional magnetization-prepared rapid gradient-echo imaging, are widely available, there is a lack of an equivalent fast high-resolution sequence providing T2 contrast. Using fast high-performance gradient systems we show the feasibility of three-dimensional true fast imaging with steady-state precession (FISP) to fill this gap. We applied a three-dimensional true-FISP protocol with voxel sizes down to 0.5 x 0.5 x 0.5 mm and acquisition times of approximately 8 min on a 1.5-T Sonata (Siemens, Erlangen, Germany) magnetic resonance scanner. The sequence was included into routine brain imaging protocols for patients with cerebrospinal-fluid-related intracranial pathology. Images from 20 patients and 20 healthy volunteers were evaluated by two neuroradiologists with respect to diagnostic image quality and artifacts. All true-FISP scans showed excellent imaging quality free of artifacts in patients and volunteers. They were valuable for the assessment of anatomical and pathologic aspects of the included patients. High-resolution true-FISP imaging is a valuable adjunct for the exploration and neuronavigation of intracranial pathologies especially if cerebrospinal fluid is involved. (orig.)

Three-dimensional spheroid cell culture of umbilical cord tissue-derived mesenchymal stromal cells leads to enhanced paracrine induction of wound healing.

Science.gov (United States)

Santos, Jorge M; Camões, Sérgio P; Filipe, Elysse; Cipriano, Madalena; Barcia, Rita N; Filipe, Mariana; Teixeira, Mariana; Simões, Sandra; Gaspar, Manuela; Mosqueira, Diogo; Nascimento, Diana S; Pinto-do-Ó, Perpétua; Cruz, Pedro; Cruz, Helder; Castro, Matilde; Miranda, Joana P

2015-05-09

The secretion of trophic factors by mesenchymal stromal cells has gained increased interest given the benefits it may bring to the treatment of a variety of traumatic injuries such as skin wounds. Herein, we report on a three-dimensional culture-based method to improve the paracrine activity of a specific population of umbilical cord tissue-derived mesenchymal stromal cells (UCX®) towards the application of conditioned medium for the treatment of cutaneous wounds. A UCX® three-dimensional culture model was developed and characterized with respect to spheroid formation, cell phenotype and cell viability. The secretion by UCX® spheroids of extracellular matrix proteins and trophic factors involved in the wound-healing process was analysed. The skin regenerative potential of UCX® three-dimensional culture-derived conditioned medium (CM3D) was also assessed in vitro and in vivo against UCX® two-dimensional culture-derived conditioned medium (CM2D) using scratch and tubulogenesis assays and a rat wound splinting model, respectively. UCX® spheroids kept in our three-dimensional system remained viable and multipotent and secreted considerable amounts of vascular endothelial growth factor A, which was undetected in two-dimensional cultures, and higher amounts of matrix metalloproteinase-2, matrix metalloproteinase-9, hepatocyte growth factor, transforming growth factor β1, granulocyte-colony stimulating factor, fibroblast growth factor 2 and interleukin-6, when compared to CM2D. Furthermore, CM3D significantly enhanced elastin production and migration of keratinocytes and fibroblasts in vitro. In turn, tubulogenesis assays revealed increased capillary maturation in the presence of CM3D, as seen by a significant increase in capillary thickness and length when compared to CM2D, and increased branching points and capillary number when compared to basal medium. Finally, CM3D-treated wounds presented signs of faster and better resolution when compared to untreated and CM

High-throughput Cloning and Expression of Integral Membrane Proteins in Escherichia coli

Science.gov (United States)

Bruni, Renato

2014-01-01

Recently, several structural genomics centers have been established and a remarkable number of three-dimensional structures of soluble proteins have been solved. For membrane proteins, the number of structures solved has been significantly trailing those for their soluble counterparts, not least because over-expression and purification of membrane proteins is a much more arduous process. By using high throughput technologies, a large number of membrane protein targets can be screened simultaneously and a greater number of expression and purification conditions can be employed, leading to a higher probability of successfully determining the structure of membrane proteins. This unit describes the cloning, expression and screening of membrane proteins using high throughput methodologies developed in our laboratory. Basic Protocol 1 deals with the cloning of inserts into expression vectors by ligation-independent cloning. Basic Protocol 2 describes the expression and purification of the target proteins on a miniscale. Lastly, for the targets that express at the miniscale, basic protocols 3 and 4 outline the methods employed for the expression and purification of targets at the midi-scale, as well as a procedure for detergent screening and identification of detergent(s) in which the target protein is stable. PMID:24510647

Three-dimensional culture of buffalo granulosa cells in hanging drop mimics the preovulatory follicle stage.

Science.gov (United States)

Yadav, Monica; Agrawal, Himanshu; Pandey, Mamta; Singh, Dheer; Onteru, Suneel K

2018-03-01

Granulosa cell (GC) culture models mimicking the intrafollicular environment are limited. Such models have a great potential in reproductive toxicity studies. The buffalo, a monovulatory species like humans, could be a better model than polyovulatory rodents. Therefore, we targeted the development and characterization of three-dimensional (3D) culture systems for buffalo GCs. The GCs from small ovarian follicles (SF) maintained the CYP19 gene expression for 144 hr in a 2D culture system. Hence, GCs from SF were cultured directly in 3D using hanging drop and Poly-([2-hydroxyethyl methacrylate]) (polyHEMA) methods in the DMEM media containing 1 ng/ml FSH and 10 ng/ml IGF-1 for 144 hr. The expression profile of nine GC-specific transcripts; CYP19, TNFAIP6, AMH, PTI, NR4A1, FSHR, RUNX, LHR, and COX2/PTGS2; revealed that 3D-spheroids developed in hanging drop method maintained the GC phenotype of preovulatory follicles. Therefore, hanging drop method is a best method for culturing GCs to mimic the intrafollicular environment. © 2017 Wiley Periodicals, Inc.

High-throughput purification of recombinant proteins using self-cleaving intein tags.

Science.gov (United States)

Coolbaugh, M J; Shakalli Tang, M J; Wood, D W

2017-01-01

High throughput methods for recombinant protein production using E. coli typically involve the use of affinity tags for simple purification of the protein of interest. One drawback of these techniques is the occasional need for tag removal before study, which can be hard to predict. In this work, we demonstrate two high throughput purification methods for untagged protein targets based on simple and cost-effective self-cleaving intein tags. Two model proteins, E. coli beta-galactosidase (βGal) and superfolder green fluorescent protein (sfGFP), were purified using self-cleaving versions of the conventional chitin-binding domain (CBD) affinity tag and the nonchromatographic elastin-like-polypeptide (ELP) precipitation tag in a 96-well filter plate format. Initial tests with shake flask cultures confirmed that the intein purification scheme could be scaled down, with >90% pure product generated in a single step using both methods. The scheme was then validated in a high throughput expression platform using 24-well plate cultures followed by purification in 96-well plates. For both tags and with both target proteins, the purified product was consistently obtained in a single-step, with low well-to-well and plate-to-plate variability. This simple method thus allows the reproducible production of highly pure untagged recombinant proteins in a convenient microtiter plate format. Copyright © 2016 Elsevier Inc. All rights reserved.

High-throughput on-chip in vivo neural regeneration studies using femtosecond laser nano-surgery and microfluidics

Science.gov (United States)

Rohde, Christopher B.; Zeng, Fei; Gilleland, Cody; Samara, Chrysanthi; Yanik, Mehmet F.

2009-02-01

In recent years, the advantages of using small invertebrate animals as model systems for human disease have become increasingly apparent and have resulted in three Nobel Prizes in medicine or chemistry during the last six years for studies conducted on the nematode Caenorhabditis elegans (C. elegans). The availability of a wide array of species-specific genetic techniques, along with the transparency of the worm and its ability to grow in minute volumes make C. elegans an extremely powerful model organism. We present a suite of technologies for complex high-throughput whole-animal genetic and drug screens. We demonstrate a high-speed microfluidic sorter that can isolate and immobilize C. elegans in a well-defined geometry, an integrated chip containing individually addressable screening chambers for incubation and exposure of individual animals to biochemical compounds, and a device for delivery of compound libraries in standard multiwell plates to microfluidic devices. The immobilization stability obtained by these devices is comparable to that of chemical anesthesia and the immobilization process does not affect lifespan, progeny production, or other aspects of animal health. The high-stability enables the use of a variety of key optical techniques. We use this to demonstrate femtosecond-laser nanosurgery and three-dimensional multiphoton microscopy. Used alone or in various combinations these devices facilitate a variety of high-throughput assays using whole animals, including mutagenesis and RNAi and drug screens at subcellular resolution, as well as high-throughput high-precision manipulations such as femtosecond-laser nanosurgery for large-scale in vivo neural degeneration and regeneration studies.

High-resolution coherent three-dimensional spectroscopy of Br2.

Science.gov (United States)

Chen, Peter C; Wells, Thresa A; Strangfeld, Benjamin R

2013-07-25

In the past, high-resolution spectroscopy has been limited to small, simple molecules that yield relatively uncongested spectra. Larger and more complex molecules have a higher density of peaks and are susceptible to complications (e.g., effects from conical intersections) that can obscure the patterns needed to resolve and assign peaks. Recently, high-resolution coherent two-dimensional (2D) spectroscopy has been used to resolve and sort peaks into easily identifiable patterns for molecules where pattern-recognition has been difficult. For very highly congested spectra, however, the ability to resolve peaks using coherent 2D spectroscopy is limited by the bandwidth of instrumentation. In this article, we introduce and investigate high-resolution coherent three-dimensional spectroscopy (HRC3D) as a method for dealing with heavily congested systems. The resulting patterns are unlike those in high-resolution coherent 2D spectra. Analysis of HRC3D spectra could provide a means for exploring the spectroscopy of large and complex molecules that have previously been considered too difficult to study.

Macroporous Hydrogel Scaffolds for Three-Dimensional Cell Culture and Tissue Engineering.

Science.gov (United States)

Fan, Changjiang; Wang, Dong-An

2017-10-01

Hydrogels have been promising candidate scaffolds for cell delivery and tissue engineering due to their tissue-like physical properties and capability for homogeneous cell loading. However, the encapsulated cells are generally entrapped and constrained in the submicron- or nanosized gel networks, seriously limiting cell growth and tissue formation. Meanwhile, the spatially confined settlement inhibits attachment and spreading of anchorage-dependent cells, leading to their apoptosis. In recent years, macroporous hydrogels have attracted increasing attention in use as cell delivery vehicles and tissue engineering scaffolds. The introduction of macropores within gel scaffolds not only improves their permeability for better nutrient transport but also creates space/interface for cell adhesion, proliferation, and extracellular matrix deposition. Herein, we will first review the development of macroporous gel scaffolds and outline the impact of macropores on cell behaviors. In the first part, the advantages and challenges of hydrogels as three-dimensional (3D) cell culture scaffolds will be described. In the second part, the fabrication of various macroporous hydrogels will be presented. Third, the enhancement of cell activities within macroporous gel scaffolds will be discussed. Finally, several crucial factors that are envisaged to propel the improvement of macroporous gel scaffolds are proposed for 3D cell culture and tissue engineering.

Three dimensional illustrating - three-dimensional vision and deception of sensibility

Directory of Open Access Journals (Sweden)

Anita Gánóczy

2009-03-01

Full Text Available The wide-spread digital photography and computer use gave the opportunity for everyone to make three-dimensional pictures and to make them public. The new opportunities with three-dimensional techniques give chance for the birth of new artistic photographs. We present in detail the biological roots of three-dimensional visualization, the phenomena of movement parallax, which can be used efficiently in making three-dimensional graphics, the Zöllner- and Corridor-illusion. There are present in this paper the visual elements, which contribute to define a plane two-dimensional image in three-dimension: coherent lines, the covering, the measurement changes, the relative altitude state, the abatement of detail profusion, the shadings and the perspective effects of colors.

High-throughput continuous cryopump

International Nuclear Information System (INIS)

Foster, C.A.

1986-01-01

A cryopump with a unique method of regeneration which allows continuous operation at high throughput has been constructed and tested. Deuterium was pumped continuously at a throughput of 30 Torr.L/s at a speed of 2000 L/s and a compression ratio of 200. Argon was pumped at a throughput of 60 Torr.L/s at a speed of 1275 L/s. To produce continuous operation of the pump, a method of regeneration that does not thermally cycle the pump is employed. A small chamber (the ''snail'') passes over the pumping surface and removes the frost from it either by mechanical action with a scraper or by local heating. The material removed is topologically in a secondary vacuum system with low conductance into the primary vacuum; thus, the exhaust can be pumped at pressures up to an effective compression ratio determined by the ratio of the pumping speed to the leakage conductance of the snail. The pump, which is all-metal-sealed and dry and which regenerates every 60 s, would be an ideal system for pumping tritium. Potential fusion applications are for mpmp limiters, for repeating pneumatic pellet injection lines, and for the centrifuge pellet injector spin tank, all of which will require pumping tritium at high throughput. Industrial applications requiring ultraclean pumping of corrosive gases at high throughput, such as the reactive ion etch semiconductor process, may also be feasible

Reduced dimensionality (3,2)D NMR experiments and their automated analysis: implications to high-throughput structural studies on proteins.

Science.gov (United States)

Reddy, Jithender G; Kumar, Dinesh; Hosur, Ramakrishna V

2015-02-01

Protein NMR spectroscopy has expanded dramatically over the last decade into a powerful tool for the study of their structure, dynamics, and interactions. The primary requirement for all such investigations is sequence-specific resonance assignment. The demand now is to obtain this information as rapidly as possible and in all types of protein systems, stable/unstable, soluble/insoluble, small/big, structured/unstructured, and so on. In this context, we introduce here two reduced dimensionality experiments – (3,2)D-hNCOcanH and (3,2)D-hNcoCAnH – which enhance the previously described 2D NMR-based assignment methods quite significantly. Both the experiments can be recorded in just about 2-3 h each and hence would be of immense value for high-throughput structural proteomics and drug discovery research. The applicability of the method has been demonstrated using alpha-helical bovine apo calbindin-D9k P43M mutant (75 aa) protein. Automated assignment of this data using AUTOBA has been presented, which enhances the utility of these experiments. The backbone resonance assignments so derived are utilized to estimate secondary structures and the backbone fold using Web-based algorithms. Taken together, we believe that the method and the protocol proposed here can be used for routine high-throughput structural studies of proteins. Copyright © 2014 John Wiley & Sons, Ltd.

Accuracy Assessment for the Three-Dimensional Coordinates by High-Speed Videogrammetric Measurement

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Xianglei Liu

2018-01-01

Full Text Available High-speed CMOS camera is a new kind of transducer to make the videogrammetric measurement for monitoring the displacement of high-speed shaking table structure. The purpose of this paper is to validate the three-dimensional coordinate accuracy of the shaking table structure acquired from the presented high-speed videogrammetric measuring system. In the paper, all of the key intermediate links are discussed, including the high-speed CMOS videogrammetric measurement system, the layout of the control network, the elliptical target detection, and the accuracy validation of final 3D spatial results. Through the accuracy analysis, the submillimeter accuracy can be made for the final the three-dimensional spatial coordinates which certify that the proposed high-speed videogrammetric technique is a better alternative technique which can replace the traditional transducer technique for monitoring the dynamic response for the shaking table structure.

Spectrophotometric Enzyme Assays for High-Throughput Screening

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Jean-Louis Reymond

2004-01-01

Full Text Available This paper reviews high-throughput screening enzyme assays developed in our laboratory over the last ten years. These enzyme assays were initially developed for the purpose of discovering catalytic antibodies by screening cell culture supernatants, but have proved generally useful for testing enzyme activities. Examples include TLC-based screening using acridone-labeled substrates, fluorogenic assays based on the β-elimination of umbelliferone or nitrophenol, and indirect assays such as the back-titration method with adrenaline and the copper-calcein fluorescence assay for aminoacids.

Ultrasonic three-dimensional on-chip cell culture for dynamic studies of tumor immune surveillance by natural killer cells.

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Christakou, Athanasia E; Ohlin, Mathias; Önfelt, Björn; Wiklund, Martin

2015-08-07

We demonstrate a simple method for three-dimensional (3D) cell culture controlled by ultrasonic standing waves in a multi-well microplate. The method gently arranges cells in a suspension into a single aggregate in each well of the microplate and, by this, nucleates 3D tissue-like cell growth for culture times between two and seven days. The microplate device is compatible with both high-resolution optical microscopy and maintenance in a standard cell incubator. The result is a scaffold- and coating-free method for 3D cell culture that can be used for controlling the cellular architecture, as well as the cellular and molecular composition of the microenvironment in and around the formed cell structures. We demonstrate the parallel production of one hundred synthetic 3D solid tumors comprising up to thousands of human hepatocellular carcinoma (HCC) HepG2 cells, we characterize the tumor structure by high-resolution optical microscopy, and we monitor the functional behavior of natural killer (NK) cells migrating, docking and interacting with the tumor model during culture. Our results show that the method can be used for determining the collective ability of a given number of NK cells to defeat a solid tumor having a certain size, shape and composition. The ultrasound-based method itself is generic and can meet any demand from applications where it is advantageous to monitor cell culture from production to analysis of 3D tissue or tumor models using microscopy in one single microplate device.

Surface modified alginate microcapsules for 3D cell culture

Science.gov (United States)

Chen, Yi-Wen; Kuo, Chiung Wen; Chueh, Di-Yen; Chen, Peilin

2016-06-01

Culture as three dimensional cell aggregates or spheroids can offer an ideal platform for tissue engineering applications and for pharmaceutical screening. Such 3D culture models, however, may suffer from the problems such as immune response and ineffective and cumbersome culture. This paper describes a simple method for producing microcapsules with alginate cores and a thin shell of poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) to encapsulate mouse induced pluripotent stem (miPS) cells, generating a non-fouling surface as an effective immunoisolation barrier. We demonstrated the trapping of the alginate microcapsules in a microwell array for the continuous observation and culture of a large number of encapsulated miPS cells in parallel. miPS cells cultured in the microcapsules survived well and proliferated to form a single cell aggregate. Droplet formation of monodisperse microcapsules with controlled size combined with flow cytometry provided an efficient way to quantitatively analyze the growth of encapsulated cells in a high-throughput manner. The simple and cost-effective coating technique employed to produce the core-shell microcapsules could be used in the emerging field of cell therapy. The microwell array would provide a convenient, user friendly and high-throughput platform for long-term cell culture and monitoring.

Comprehensive analysis of signal transduction in three-dimensional ECM-based tumor cell cultures

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Iris Eke

2015-11-01

Full Text Available Analysis of signal transduction and protein phosphorylation is fundamental to understand physiological and pathological cell behavior as well as identification of novel therapeutic targets. Despite the fact that more physiological three-dimensional cell culture assays are increasingly used, particularly proteomics and phosphoproteomics remain challenging due to easy, robust and reproducible sample preparation. Here, we present an easy-to-perform, reliable and time-efficient method for the production of 3D cell lysates without compromising cell adhesion before cell lysis. The samples can be used for Western blotting as well as phosphoproteome array technology. This technique would be of interest for researchers working in all fields of biology and drug development.

Towards a high throughput droplet-based agglutination assay

KAUST Repository

Kodzius, Rimantas; Castro, David; Foulds, Ian G.

2013-01-01

This work demonstrates the detection method for a high throughput droplet based agglutination assay system. Using simple hydrodynamic forces to mix and aggregate functionalized microbeads we avoid the need to use magnetic assistance or mixing structures. The concentration of our target molecules was estimated by agglutination strength, obtained through optical image analysis. Agglutination in droplets was performed with flow rates of 150 µl/min and occurred in under a minute, with potential to perform high-throughput measurements. The lowest target concentration detected in droplet microfluidics was 0.17 nM, which is three orders of magnitude more sensitive than a conventional card based agglutination assay.

Towards a high throughput droplet-based agglutination assay

KAUST Repository

Kodzius, Rimantas

2013-10-22

This work demonstrates the detection method for a high throughput droplet based agglutination assay system. Using simple hydrodynamic forces to mix and aggregate functionalized microbeads we avoid the need to use magnetic assistance or mixing structures. The concentration of our target molecules was estimated by agglutination strength, obtained through optical image analysis. Agglutination in droplets was performed with flow rates of 150 µl/min and occurred in under a minute, with potential to perform high-throughput measurements. The lowest target concentration detected in droplet microfluidics was 0.17 nM, which is three orders of magnitude more sensitive than a conventional card based agglutination assay.

Three-Dimensional Electromagnetic High Frequency Axisymmetric Cavity Scars.

Energy Technology Data Exchange (ETDEWEB)

Warne, Larry Kevin; Jorgenson, Roy Eberhardt

2014-10-01

This report examines the localization of high frequency electromagnetic fi elds in three-dimensional axisymmetric cavities along periodic paths between opposing sides of the cavity. The cases where these orbits lead to unstable localized modes are known as scars. This report treats both the case where the opposing sides, or mirrors, are convex, where there are no interior foci, and the case where they are concave, leading to interior foci. The scalar problem is treated fi rst but the approximations required to treat the vector fi eld components are also examined. Particular att ention is focused on the normalization through the electromagnetic energy theorem. Both projections of the fi eld along the scarred orbit as well as point statistics are examined. Statistical comparisons are m ade with a numerical calculation of the scars run with an axisymmetric simulation. This axisymmetric cas eformstheoppositeextreme(wherethetwomirror radii at each end of the ray orbit are equal) from the two -dimensional solution examined previously (where one mirror radius is vastly di ff erent from the other). The enhancement of the fi eldontheorbitaxiscanbe larger here than in the two-dimensional case. Intentionally Left Blank

Hormonal regulation of epithelial organization in a three-dimensional breast tissue culture model.

Science.gov (United States)

Speroni, Lucia; Whitt, Gregory S; Xylas, Joanna; Quinn, Kyle P; Jondeau-Cabaton, Adeline; Barnes, Clifford; Georgakoudi, Irene; Sonnenschein, Carlos; Soto, Ana M

2014-01-01

The establishment of hormone target breast cells in the 1970's resulted in suitable models for the study of hormone control of cell proliferation and gene expression using two-dimensional (2D) cultures. However, to study mammogenesis and breast tumor development in vitro, cells must be able to organize in three-dimensional (3D) structures like in the tissue. We now report the development of a hormone-sensitive 3D culture model for the study of mammogenesis and neoplastic development. Hormone-sensitive T47D breast cancer cells respond to estradiol in a dose-dependent manner by forming complex epithelial structures. Treatment with the synthetic progestagen promegestone, in the presence of estradiol, results in flat epithelial structures that display cytoplasmic projections, a phenomenon reported to precede side-branching. Additionally, as in the mammary gland, treatment with prolactin in the presence of estradiol induces budding structures. These changes in epithelial organization are accompanied by collagen remodeling. Collagen is the major acellular component of the breast stroma and an important player in tumor development and progression. Quantitative analysis of second harmonic generation of collagen fibers revealed that collagen density was more variable surrounding budding and irregularly shaped structures when compared to more regular structures; suggesting that fiber organization in the former is more anisotropic than in the latter. In sum, this new 3D model recapitulates morphogenetic events modulated by mammogenic hormones in the breast, and is suitable for the evaluation of therapeutic agents.

Electron tomography, three-dimensional Fourier analysis and colour prediction of a three-dimensional amorphous biophotonic nanostructure

Science.gov (United States)

Shawkey, Matthew D.; Saranathan, Vinodkumar; Pálsdóttir, Hildur; Crum, John; Ellisman, Mark H.; Auer, Manfred; Prum, Richard O.

2009-01-01

Organismal colour can be created by selective absorption of light by pigments or light scattering by photonic nanostructures. Photonic nanostructures may vary in refractive index over one, two or three dimensions and may be periodic over large spatial scales or amorphous with short-range order. Theoretical optical analysis of three-dimensional amorphous nanostructures has been challenging because these structures are difficult to describe accurately from conventional two-dimensional electron microscopy alone. Intermediate voltage electron microscopy (IVEM) with tomographic reconstruction adds three-dimensional data by using a high-power electron beam to penetrate and image sections of material sufficiently thick to contain a significant portion of the structure. Here, we use IVEM tomography to characterize a non-iridescent, three-dimensional biophotonic nanostructure: the spongy medullary layer from eastern bluebird Sialia sialis feather barbs. Tomography and three-dimensional Fourier analysis reveal that it is an amorphous, interconnected bicontinuous matrix that is appropriately ordered at local spatial scales in all three dimensions to coherently scatter light. The predicted reflectance spectra from the three-dimensional Fourier analysis are more precise than those predicted by previous two-dimensional Fourier analysis of transmission electron microscopy sections. These results highlight the usefulness, and obstacles, of tomography in the description and analysis of three-dimensional photonic structures. PMID:19158016

High-Throughput Light Sheet Microscopy for the Automated Live Imaging of Larval Zebrafish

Science.gov (United States)

Baker, Ryan; Logan, Savannah; Dudley, Christopher; Parthasarathy, Raghuveer

The zebrafish is a model organism with a variety of useful properties; it is small and optically transparent, it reproduces quickly, it is a vertebrate, and there are a large variety of transgenic animals available. Because of these properties, the zebrafish is well suited to study using a variety of optical technologies including light sheet fluorescence microscopy (LSFM), which provides high-resolution three-dimensional imaging over large fields of view. Research progress, however, is often not limited by optical techniques but instead by the number of samples one can examine over the course of an experiment, which in the case of light sheet imaging has so far been severely limited. Here we present an integrated fluidic circuit and microscope which provides rapid, automated imaging of zebrafish using several imaging modes, including LSFM, Hyperspectral Imaging, and Differential Interference Contrast Microscopy. Using this system, we show that we can increase our imaging throughput by a factor of 10 compared to previous techniques. We also show preliminary results visualizing zebrafish immune response, which is sensitive to gut microbiota composition, and which shows a strong variability between individuals that highlights the utility of high throughput imaging. National Science Foundation, Award No. DBI-1427957.

The Influence of IL-10 and TNFα on Chondrogenesis of Human Mesenchymal Stromal Cells in Three-Dimensional Cultures

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Michal Jagielski

2014-09-01

Full Text Available Chondrogenic differentiated mesenchymal stromal cells (MSCs are a promising cell source for articular cartilage repair. This study was undertaken to determine the effectiveness of two three-dimensional (3D culture systems for chondrogenic MSC differentiation in comparison to primary chondrocytes and to assess the effect of Interleukin (IL-10 and Tumor Necrosis Factor (TNFα on chondrogenesis by MSCs in 3D high-density (H-D culture. MSCs were isolated from femur spongiosa, characterized using a set of typical markers and introduced in scaffold-free H-D cultures or non-woven polyglycolic acid (PGA scaffolds for chondrogenic differentiation. H-D cultures were stimulated with recombinant IL-10, TNFα, TNFα + IL-10 or remained untreated. Gene and protein expression of type II collagen, aggrecan, sox9 and TNFα were examined. MSCs expressed typical cell surface markers and revealed multipotency. Chondrogenic differentiated cells expressed cartilage-specific markers in both culture systems but to a lower extent when compared with articular chondrocytes. Chondrogenesis was more pronounced in PGA compared with H-D culture. IL-10 and/or TNFα did not impair the chondrogenic differentiation of MSCs. Moreover, in most of the investigated samples, despite not reaching significance level, IL-10 had a stimulatory effect on the type II collagen, aggrecan and TNFα expression when compared with the respective controls.

The influence of IL-10 and TNFα on chondrogenesis of human mesenchymal stromal cells in three-dimensional cultures.

Science.gov (United States)

Jagielski, Michal; Wolf, Johannes; Marzahn, Ulrike; Völker, Anna; Lemke, Marion; Meier, Carola; Ertel, Wolfgang; Godkin, Owen; Arens, Stephan; Schulze-Tanzil, Gundula

2014-09-09

Chondrogenic differentiated mesenchymal stromal cells (MSCs) are a promising cell source for articular cartilage repair. This study was undertaken to determine the effectiveness of two three-dimensional (3D) culture systems for chondrogenic MSC differentiation in comparison to primary chondrocytes and to assess the effect of Interleukin (IL)-10 and Tumor Necrosis Factor (TNF)α on chondrogenesis by MSCs in 3D high-density (H-D) culture. MSCs were isolated from femur spongiosa, characterized using a set of typical markers and introduced in scaffold-free H-D cultures or non-woven polyglycolic acid (PGA) scaffolds for chondrogenic differentiation. H-D cultures were stimulated with recombinant IL-10, TNFα, TNFα + IL-10 or remained untreated. Gene and protein expression of type II collagen, aggrecan, sox9 and TNFα were examined. MSCs expressed typical cell surface markers and revealed multipotency. Chondrogenic differentiated cells expressed cartilage-specific markers in both culture systems but to a lower extent when compared with articular chondrocytes. Chondrogenesis was more pronounced in PGA compared with H-D culture. IL-10 and/or TNFα did not impair the chondrogenic differentiation of MSCs. Moreover, in most of the investigated samples, despite not reaching significance level, IL-10 had a stimulatory effect on the type II collagen, aggrecan and TNFα expression when compared with the respective controls.

Tumor cell culture on collagen–chitosan scaffolds as three-dimensional tumor model: A suitable model for tumor studies

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Aziz Mahmoudzadeh

2016-07-01

Full Text Available Tumor cells naturally live in three-dimensional (3D microenvironments, while common laboratory tests and evaluations are done in two-dimensional (2D plates. This study examined the impact of cultured 4T1 cancer cells in a 3D collagen–chitosan scaffold compared with 2D plate cultures. Collagen–chitosan scaffolds were provided and passed confirmatory tests. 4T1 tumor cells were cultured on scaffolds and then tumor cells growth rate, resistance to X-ray radiation, and cyclophosphamide as a chemotherapy drug were analyzed. Furthermore, 4T1 cells were extracted from the scaffold model and were injected into the mice. Tumor growth rate, survival rate, and systemic immune responses were evaluated. Our results showed that 4T1 cells infiltrated the scaffolds pores and constructed a 3D microenvironment. Furthermore, 3D cultured tumor cells showed a slower proliferation rate, increased levels of survival to the X-ray irradiation, and enhanced resistance to chemotherapy drugs in comparison with 2D plate cultures. Transfer of extracted cells to the mice caused enhanced tumor volume and decreased life span. This study indicated that collagen–chitosan nanoscaffolds provide a suitable model of tumor that would be appropriate for tumor studies.

Tumor cell culture on collagen-chitosan scaffolds as three-dimensional tumor model: A suitable model for tumor studies.

Science.gov (United States)

Mahmoudzadeh, Aziz; Mohammadpour, Hemn

2016-07-01

Tumor cells naturally live in three-dimensional (3D) microenvironments, while common laboratory tests and evaluations are done in two-dimensional (2D) plates. This study examined the impact of cultured 4T1 cancer cells in a 3D collagen-chitosan scaffold compared with 2D plate cultures. Collagen-chitosan scaffolds were provided and passed confirmatory tests. 4T1 tumor cells were cultured on scaffolds and then tumor cells growth rate, resistance to X-ray radiation, and cyclophosphamide as a chemotherapy drug were analyzed. Furthermore, 4T1 cells were extracted from the scaffold model and were injected into the mice. Tumor growth rate, survival rate, and systemic immune responses were evaluated. Our results showed that 4T1 cells infiltrated the scaffolds pores and constructed a 3D microenvironment. Furthermore, 3D cultured tumor cells showed a slower proliferation rate, increased levels of survival to the X-ray irradiation, and enhanced resistance to chemotherapy drugs in comparison with 2D plate cultures. Transfer of extracted cells to the mice caused enhanced tumor volume and decreased life span. This study indicated that collagen-chitosan nanoscaffolds provide a suitable model of tumor that would be appropriate for tumor studies. Copyright © 2016. Published by Elsevier B.V.

Multi-channel counter-current chromatography for high-throughput fractionation of natural products for drug discovery.

Science.gov (United States)

Wu, Shihua; Yang, Lu; Gao, Yuan; Liu, Xiaoyue; Liu, Feiyan

2008-02-08

A multi-channel counter-current chromatography (CCC) method has been designed and fabricated for the high-throughput fractionation of natural products without complications sometimes encountered with other conventional chromatographic systems, such as irreversible adsorptive constituent losses and deactivation, tailing of solute peaks and contamination. It has multiple independent CCC channels and each channel connects independent separation column(s) by parallel flow tubes, and thus the multi-channel CCC apparatus can achieve simultaneously two or more independent chromatographic processes. Furthermore, a high-throughput CCC fractionation method for natural products has been developed by a combination of a new three-channel CCC apparatus and conventional parallel chromatographic devices including pumps, sample injectors, effluent detectors and collectors, and its performance has been displayed on the fractionation of ethyl acetate extracts of three natural materials Solidago canadensis, Suillus placidus, and Trichosanthes kirilowii, which are found to be potent cytotoxic to tumor cell lines in the course of screening the antitumor candidates. By combination of biological screening programs and preparative high-performance liquid chromatography (HPLC) purification, 22.8 mg 6 beta-angeloyloxykolavenic acid and 29.4 mg 6 beta-tigloyloxykolavenic acid for S. canadensis, 25.3mg suillin for S. placidus, and 6.8 mg 23,24-dihydrocucurbitacin B for T. Kirilowii as their major cytotoxic principles were isolated from each 1000 mg crude ethyl acetate extract. Their chemical structures were characterized by electrospray ionization mass spectrometry, one- and two-dimensional nuclear magnetic resonance. The overall results indicate the multi-channel CCC is very useful for high-throughput fractionation of natural products for drug discovery in spite of the solvent balancing requirement and the lower resolution of the shorter CCC columns.

High-throughput characterization methods for lithium batteries

Directory of Open Access Journals (Sweden)

Yingchun Lyu

2017-09-01

Full Text Available The development of high-performance lithium ion batteries requires the discovery of new materials and the optimization of key components. By contrast with traditional one-by-one method, high-throughput method can synthesize and characterize a large number of compositionally varying samples, which is able to accelerate the pace of discovery, development and optimization process of materials. Because of rapid progress in thin film and automatic control technologies, thousands of compounds with different compositions could be synthesized rapidly right now, even in a single experiment. However, the lack of rapid or combinatorial characterization technologies to match with high-throughput synthesis methods, limit the application of high-throughput technology. Here, we review a series of representative high-throughput characterization methods used in lithium batteries, including high-throughput structural and electrochemical characterization methods and rapid measuring technologies based on synchrotron light sources.

On the effect of long-term electrical stimulation on three-dimensional cell cultures: Hen embryo brain spheroids

OpenAIRE

Uroukov, Ivan S; Bull, Larry

2008-01-01

A comprehensive dataset of multielectrode array recordings was collected from three-dimensional hen embryo brain cell cultures, termed spheroids, under long-term electrical stimulation. The aim is to understand the ongoing changes in the spiking activity under electrical stimulation within the lifetime of 14–72DIV of the neuronal networks contained therein. The spiking dynamics were analyzed and behavioral characteristics derived. Some effects on spiking patterns and exhaustion were followed ...

A bioimage informatics platform for high-throughput embryo phenotyping.

Science.gov (United States)

Brown, James M; Horner, Neil R; Lawson, Thomas N; Fiegel, Tanja; Greenaway, Simon; Morgan, Hugh; Ring, Natalie; Santos, Luis; Sneddon, Duncan; Teboul, Lydia; Vibert, Jennifer; Yaikhom, Gagarine; Westerberg, Henrik; Mallon, Ann-Marie

2018-01-01

High-throughput phenotyping is a cornerstone of numerous functional genomics projects. In recent years, imaging screens have become increasingly important in understanding gene-phenotype relationships in studies of cells, tissues and whole organisms. Three-dimensional (3D) imaging has risen to prominence in the field of developmental biology for its ability to capture whole embryo morphology and gene expression, as exemplified by the International Mouse Phenotyping Consortium (IMPC). Large volumes of image data are being acquired by multiple institutions around the world that encompass a range of modalities, proprietary software and metadata. To facilitate robust downstream analysis, images and metadata must be standardized to account for these differences. As an open scientific enterprise, making the data readily accessible is essential so that members of biomedical and clinical research communities can study the images for themselves without the need for highly specialized software or technical expertise. In this article, we present a platform of software tools that facilitate the upload, analysis and dissemination of 3D images for the IMPC. Over 750 reconstructions from 80 embryonic lethal and subviable lines have been captured to date, all of which are openly accessible at mousephenotype.org. Although designed for the IMPC, all software is available under an open-source licence for others to use and develop further. Ongoing developments aim to increase throughput and improve the analysis and dissemination of image data. Furthermore, we aim to ensure that images are searchable so that users can locate relevant images associated with genes, phenotypes or human diseases of interest. © The Author 2016. Published by Oxford University Press.

High-throughput volumetric reconstruction for 3D wheat plant architecture studies

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Wei Fang

2016-09-01

Full Text Available For many tiller crops, the plant architecture (PA, including the plant fresh weight, plant height, number of tillers, tiller angle and stem diameter, significantly affects the grain yield. In this study, we propose a method based on volumetric reconstruction for high-throughput three-dimensional (3D wheat PA studies. The proposed methodology involves plant volumetric reconstruction from multiple images, plant model processing and phenotypic parameter estimation and analysis. This study was performed on 80 Triticum aestivum plants, and the results were analyzed. Comparing the automated measurements with manual measurements, the mean absolute percentage error (MAPE in the plant height and the plant fresh weight was 2.71% (1.08cm with an average plant height of 40.07cm and 10.06% (1.41g with an average plant fresh weight of 14.06g, respectively. The root mean square error (RMSE was 1.37cm and 1.79g for the plant height and plant fresh weight, respectively. The correlation coefficients were 0.95 and 0.96 for the plant height and plant fresh weight, respectively. Additionally, the proposed methodology, including plant reconstruction, model processing and trait extraction, required only approximately 20s on average per plant using parallel computing on a graphics processing unit (GPU, demonstrating that the methodology would be valuable for a high-throughput phenotyping platform.

Using DNase Hi-C techniques to map global and local three-dimensional genome architecture at high resolution.

Science.gov (United States)

Ma, Wenxiu; Ay, Ferhat; Lee, Choli; Gulsoy, Gunhan; Deng, Xinxian; Cook, Savannah; Hesson, Jennifer; Cavanaugh, Christopher; Ware, Carol B; Krumm, Anton; Shendure, Jay; Blau, C Anthony; Disteche, Christine M; Noble, William S; Duan, ZhiJun

2018-06-01

The folding and three-dimensional (3D) organization of chromatin in the nucleus critically impacts genome function. The past decade has witnessed rapid advances in genomic tools for delineating 3D genome architecture. Among them, chromosome conformation capture (3C)-based methods such as Hi-C are the most widely used techniques for mapping chromatin interactions. However, traditional Hi-C protocols rely on restriction enzymes (REs) to fragment chromatin and are therefore limited in resolution. We recently developed DNase Hi-C for mapping 3D genome organization, which uses DNase I for chromatin fragmentation. DNase Hi-C overcomes RE-related limitations associated with traditional Hi-C methods, leading to improved methodological resolution. Furthermore, combining this method with DNA capture technology provides a high-throughput approach (targeted DNase Hi-C) that allows for mapping fine-scale chromatin architecture at exceptionally high resolution. Hence, targeted DNase Hi-C will be valuable for delineating the physical landscapes of cis-regulatory networks that control gene expression and for characterizing phenotype-associated chromatin 3D signatures. Here, we provide a detailed description of method design and step-by-step working protocols for these two methods. Copyright © 2018 Elsevier Inc. All rights reserved.

Human umbilical cord mesenchymal stromal cells exhibit immature nucleus pulposus cell phenotype in a laminin-rich pseudo-three-dimensional culture system.

Science.gov (United States)

Chon, Brian H; Lee, Esther J; Jing, Liufang; Setton, Lori A; Chen, Jun

2013-10-02

Cell supplementation to the herniated or degenerated intervertebral disc (IVD) is a potential strategy to promote tissue regeneration and slow disc pathology. Human umbilical cord mesenchymal stromal cells (HUCMSCs) - originating from the Wharton's jelly - remain an attractive candidate for such endeavors with their ability to differentiate into multiple lineages. Previously, mesenchymal stem cells (MSCs) have been studied as a potential source for disc tissue regeneration. However, no studies have demonstrated that MSCs can regenerate matrix with unique characteristics matching that of immature nucleus pulposus (NP) tissues of the IVD. In our prior work, immature NP cells were found to express specific laminin isoforms and laminin-binding receptors that may serve as phenotypic markers for evaluating MSC differentiation to NP-like cells. The goal of this study is to evaluate these markers and matrix synthesis for HUCMSCs cultured in a laminin-rich pseudo-three-dimensional culture system. HUCMSCs were seeded on top of Transwell inserts pre-coated with Matrigel™, which contained mainly laminin-111. Cells were cultured under hypoxia environment with three differentiation conditions: NP differentiation media (containing 2.5% Matrigel™ solution to provide for a pseudo-three-dimensional laminin culture system) with no serum, or the same media supplemented with either insulin-like growth factor-1 (IGF-1) or transforming growth factor-β1 (TGF-β1). Cell clustering behavior, matrix production and the expression of NP-specific laminin and laminin-receptors were evaluated at days 1, 7, 13 and 21 of culture. Data show that a pseudo-three-dimensional culture condition (laminin-1 rich) promoted HUCMSC differentiation under no serum conditions. Starting at day 1, HUCMSCs demonstrated a cell clustering morphology similar to that of immature NP cells in situ and that observed for primary immature NP cells within the similar laminin-rich culture system (prior study

Three-dimensional spheroid culture of human umbilical cord mesenchymal stem cells promotes cell yield and stemness maintenance.

Science.gov (United States)

Li, Yi; Guo, Gang; Li, Li; Chen, Fei; Bao, Ji; Shi, Yu-Jun; Bu, Hong

2015-05-01

Mesenchymal stem cell (MSC) transplantation is a promising treatment of many diseases. However, conventional techniques with cells being cultured as a monolayer result in slow cell proliferation and insufficient yield to meet clinical demands. Three-dimensional (3D) culture systems are gaining attention with regard to recreating a complex microenvironment and to understanding the conditions experienced by cells. Our aim is to establish a novel 3D system for the culture of human umbilical cord MSCs (hUC-MSCs) within a real 3D microenvironment but with no digestion or passaging. Primary hUC-MSCs were isolated and grown in serum-free medium (SFM) on a suspension Rocker system. Cell characteristics including proliferation, phenotype and multipotency were recorded. The therapeutic effects of 3D-cultured hUC-MSCs on carbon tetrachloride (CCl4)-induced acute liver failure in mouse models were examined. In the 3D Rocker system, hUC-MSCs formed spheroids in SFM and maintained high viability and active proliferation. Compared with monolayer culture, the 3D-culture system yielded more hUC-MSCs cells within the same volume. The spheroids expressed higher levels of stem cell markers and displayed stronger multipotency. After transplantation into mouse, 3D hUC-MSCs significantly promoted the secretion of interferon-γ and interleukin-6 but inhibited that of tumor necrosis factor-α, thereby alleviating liver necrosis and promoting regeneration following CCl4 injury. The 3D culture of hUC-MSCs thus promotes cell yield and stemness maintenance and represents a promising strategy for hUC-MSCs expansion on an industrial scale with great potential for cell therapy and biotechnology.

Gene Expression Analysis of Escherichia Coli Grown in Miniaturized Bioreactor Platforms for High-Throughput Analysis of Growth and genomic Data

DEFF Research Database (Denmark)

Boccazzi, P.; Zanzotto, A.; Szita, Nicolas

2005-01-01

Combining high-throughput growth physiology and global gene expression data analysis is of significant value for integrating metabolism and genomics. We compared global gene expression using 500 ng of total RNA from Escherichia coli cultures grown in rich or defined minimal media in a miniaturize...... cultures using just 500 ng of total RNA indicate that high-throughput integration of growth physiology and genomics will be possible with novel biochemical platforms and improved detection technologies....

[Extraction of buildings three-dimensional information from high-resolution satellite imagery based on Barista software].

Science.gov (United States)

Zhang, Pei-feng; Hu, Yuan-man; He, Hong-shi

2010-05-01

The demand for accurate and up-to-date spatial information of urban buildings is becoming more and more important for urban planning, environmental protection, and other vocations. Today's commercial high-resolution satellite imagery offers the potential to extract the three-dimensional information of urban buildings. This paper extracted the three-dimensional information of urban buildings from QuickBird imagery, and validated the precision of the extraction based on Barista software. It was shown that the extraction of three-dimensional information of the buildings from high-resolution satellite imagery based on Barista software had the advantages of low professional level demand, powerful universality, simple operation, and high precision. One pixel level of point positioning and height determination accuracy could be achieved if the digital elevation model (DEM) and sensor orientation model had higher precision and the off-Nadir View Angle was relatively perfect.

Macrophage involvement affects matrix stiffness-related influences on cell osteogenesis under three-dimensional culture conditions.

Science.gov (United States)

He, Xiao-Tao; Wu, Rui-Xin; Xu, Xin-Yue; Wang, Jia; Yin, Yuan; Chen, Fa-Ming

2018-04-15

Accumulating evidence indicates that the physicochemical properties of biomaterials exert profound influences on stem cell fate decisions. However, matrix-based regulation selected through in vitro analyses based on a given cell population do not genuinely reflect the in vivo conditions, in which multiple cell types are involved and interact dynamically. This study constitutes the first investigation of how macrophages (Mφs) in stiffness-tunable transglutaminase cross-linked gelatin (TG-gel) affect the osteogenesis of bone marrow-derived mesenchymal stem cells (BMMSCs). When a single cell type was cultured, low-stiffness TG-gels promoted BMMSC proliferation, whereas high-stiffness TG-gels supported cell osteogenic differentiation. However, Mφs in high-stiffness TG-gels were more likely to polarize toward the pro-inflammatory M1 phenotype. Using either conditioned medium (CM)-based incubation or Transwell-based co-culture, we found that Mφs encapsulated in the low-stiffness matrix exerted a positive effect on the osteogenesis of co-cultured BMMSCs. Conversely, Mφs in high-stiffness TG-gels negatively affected cell osteogenic differentiation. When both cell types were cultured in the same TG-gel type and placed into the Transwell system, the stiffness-related influences of Mφs on BMMSCs were significantly altered; both the low- and high-stiffness matrix induced similar levels of BMMSC osteogenesis. Although the best material parameter for synergistically affecting Mφs and BMMSCs remains unknown, our data suggest that Mφ involvement in the co-culture system alters previously identified material-related influences on BMMSCs, such as matrix stiffness-related effects, which were identified based on a culture system involving a single cell type. Such Mφ-stem cell interactions should be considered when establishing proper matrix parameter-associated cell regulation in the development of biomimetic biomaterials for regenerative applications. The substrate stiffness

The high throughput biomedicine unit at the institute for molecular medicine Finland: high throughput screening meets precision medicine.

Science.gov (United States)

Pietiainen, Vilja; Saarela, Jani; von Schantz, Carina; Turunen, Laura; Ostling, Paivi; Wennerberg, Krister

2014-05-01

The High Throughput Biomedicine (HTB) unit at the Institute for Molecular Medicine Finland FIMM was established in 2010 to serve as a national and international academic screening unit providing access to state of the art instrumentation for chemical and RNAi-based high throughput screening. The initial focus of the unit was multiwell plate based chemical screening and high content microarray-based siRNA screening. However, over the first four years of operation, the unit has moved to a more flexible service platform where both chemical and siRNA screening is performed at different scales primarily in multiwell plate-based assays with a wide range of readout possibilities with a focus on ultraminiaturization to allow for affordable screening for the academic users. In addition to high throughput screening, the equipment of the unit is also used to support miniaturized, multiplexed and high throughput applications for other types of research such as genomics, sequencing and biobanking operations. Importantly, with the translational research goals at FIMM, an increasing part of the operations at the HTB unit is being focused on high throughput systems biological platforms for functional profiling of patient cells in personalized and precision medicine projects.

The in vitro biocompatibility of d-(+) raffinose modified chitosan: Two-dimensional and three-dimensional systems for culturing of horse articular chondrocytes.

Science.gov (United States)

De Angelis, Elena; Ravanetti, Francesca; Martelli, Paolo; Cacchioli, Antonio; Ivanovska, Ana; Corradi, Attilio; Nasi, Sonia; Bianchera, Annalisa; Passeri, Benedetta; Canelli, Elena; Bettini, Ruggero; Borghetti, Paolo

2017-12-01

The present study investigated the biocompatibility of chitosan films and scaffolds modified with d-(+)raffinose and their capability to support the growth and maintenance of the differentiation of articular chondrocytes in vitro. Primary equine articular chondrocytes were cultured on films and scaffolds of modified d-(+) raffinose chitosan. Their behavior was compared to that of chondrocytes grown in conventional bi- and three-dimensional culture systems, such as micromasses and alginate beads. Chitosan films maintained the phenotype of differentiated chondrocytes (typical round morphology) and sustained the synthesis of cartilaginous extracellular matrix (ECM), even at 4weeks of culture. Indeed, starting from 2weeks of culture, chondrocytes seeded on chitosan scaffolds were able to penetrate the surface pores and to colonize the internal matrix. Moreover they produced ECM expressing the genes of typical chondrocytes differentiation markers such as collagen II and aggrecan. In conclusion, chitosan modified with d-raffinose represents an ideal support for chondrocyte adhesion, proliferation and for the maintenance of cellular phenotypic and genotypic differentiation. This novel biomaterial could potentially be a reliable support for the re-differentiation of dedifferentiated chondrocytes. Copyright © 2017 Elsevier Ltd. All rights reserved.

High-Throughput Scoring of Seed Germination.

Science.gov (United States)

Ligterink, Wilco; Hilhorst, Henk W M

2017-01-01

High-throughput analysis of seed germination for phenotyping large genetic populations or mutant collections is very labor intensive and would highly benefit from an automated setup. Although very often used, the total germination percentage after a nominated period of time is not very informative as it lacks information about start, rate, and uniformity of germination, which are highly indicative of such traits as dormancy, stress tolerance, and seed longevity. The calculation of cumulative germination curves requires information about germination percentage at various time points. We developed the GERMINATOR package: a simple, highly cost-efficient, and flexible procedure for high-throughput automatic scoring and evaluation of germination that can be implemented without the use of complex robotics. The GERMINATOR package contains three modules: (I) design of experimental setup with various options to replicate and randomize samples; (II) automatic scoring of germination based on the color contrast between the protruding radicle and seed coat on a single image; and (III) curve fitting of cumulative germination data and the extraction, recap, and visualization of the various germination parameters. GERMINATOR is a freely available package that allows the monitoring and analysis of several thousands of germination tests, several times a day by a single person.

A cell-based biosensor for nanomaterials cytotoxicity assessment in three dimensional cell culture

International Nuclear Information System (INIS)

Dubiak-Szepietowska, Monika; Karczmarczyk, Aleksandra; Winckler, Thomas; Feller, Karl-Heinz

2016-01-01

Nanoparticles (NPs) are widely used in consumer and medicinal products. The high prevalence of nanoparticles in the environment raises concerns regarding their effects on human health, but there is limited knowledge about how NPs interact with cells or tissues. Because the European Union has called for a substantial reduction of animal experiments for scientific purposes (Directive 2010/63), increased efforts are required to develop in vitro models to evaluate potentially hazardous agents. Here, we describe a new cell-based biosensor for the evaluation of NPs cytotoxicity. The new biosensor is based on transgenic human hepatoblastoma cells (HepG2) that express a secreted form of alkaline phosphatase (SEAP) as a reporter protein whose expression is induced upon activation of a stress response pathway controlled by the transcription regulator nuclear factor-κB (NF-κB). The NF-κB-HepG2 sensor cells were cultured in a Matrigel-based three dimensional environment to simulate the in vivo situation. The new biosensor cells offer the advantage of generating fast and reproducible readout at lower concentrations and shorter incubation time than conventional viability assays, avoid possible interaction between nanomaterials and assay compounds, therefore, minimize generation of false positive or negative results and indicate mechanism of toxicity through NF-κB signaling.

High-Efficiency Dye-Sensitized Solar Cell with Three-Dimensional Photoanode

KAUST Repository

Tétreault, Nicolas

2011-11-09

Herein, we present a straightforward bottom-up synthesis of a high electron mobility and highly light scattering macroporous photoanode for dye-sensitized solar cells. The dense three-dimensional Al/ZnO, SnO2, or TiO 2 host integrates a conformal passivation thin film to reduce recombination and a large surface-area mesoporous anatase guest for high dye loading. This novel photoanode is designed to improve the charge extraction resulting in higher fill factor and photovoltage for DSCs. An increase in photovoltage of up to 110 mV over state-of-the-art DSC is demonstrated. © 2011 American Chemical Society.

High-Efficiency Dye-Sensitized Solar Cell with Three-Dimensional Photoanode

KAUST Repository

Té treault, Nicolas; Arsenault, É ric; Heiniger, Leo-Philipp; Soheilnia, Navid; Brillet, Jé ré mie; Moehl, Thomas; Zakeeruddin, Shaik; Ozin, Geoffrey A.; Grä tzel, Michael

2011-01-01

Herein, we present a straightforward bottom-up synthesis of a high electron mobility and highly light scattering macroporous photoanode for dye-sensitized solar cells. The dense three-dimensional Al/ZnO, SnO2, or TiO 2 host integrates a conformal passivation thin film to reduce recombination and a large surface-area mesoporous anatase guest for high dye loading. This novel photoanode is designed to improve the charge extraction resulting in higher fill factor and photovoltage for DSCs. An increase in photovoltage of up to 110 mV over state-of-the-art DSC is demonstrated. © 2011 American Chemical Society.

The self-crosslinking smart hyaluronic acid hydrogels as injectable three-dimensional scaffolds for cells culture.

Science.gov (United States)

Bian, Shaoquan; He, Mengmeng; Sui, Junhui; Cai, Hanxu; Sun, Yong; Liang, Jie; Fan, Yujiang; Zhang, Xingdong

2016-04-01

Although the disulfide bond crosslinked hyaluronic acid hydrogels have been reported by many research groups, the major researches were focused on effectively forming hydrogels. However, few researchers paid attention to the potential significance of controlling the hydrogel formation and degradation, improving biocompatibility, reducing the toxicity of exogenous and providing convenience to the clinical operations later on. In this research, the novel controllable self-crosslinking smart hydrogels with in-situ gelation property was prepared by a single component, the thiolated hyaluronic acid derivative (HA-SH), and applied as a three-dimensional scaffold to mimic native extracellular matrix (ECM) for the culture of fibroblasts cells (L929) and chondrocytes. A series of HA-SH hydrogels were prepared depending on different degrees of thiol substitution (ranging from 10 to 60%) and molecule weights of HA (0.1, 0.3 and 1.0 MDa). The gelation time, swelling property and smart degradation behavior of HA-SH hydrogel were evaluated. The results showed that the gelation and degradation time of hydrogels could be controlled by adjusting the component of HA-SH polymers. The storage modulus of HA-SH hydrogels obtained by dynamic modulus analysis (DMA) could be up to 44.6 kPa. In addition, HA-SH hydrogels were investigated as a three-dimensional scaffold for the culture of fibroblasts cells (L929) and chondrocytes cells in vitro and as an injectable hydrogel for delivering chondrocytes cells in vivo. These results illustrated that HA-SH hydrogels with controllable gelation process, intelligent degradation behavior, excellent biocompatibility and convenient operational characteristics supplied potential clinical application capacity for tissue engineering and regenerative medicine. Copyright © 2016 Elsevier B.V. All rights reserved.

Profiling stem cell states in three-dimensional biomaterial niches using high content image informatics.

Science.gov (United States)

Dhaliwal, Anandika; Brenner, Matthew; Wolujewicz, Paul; Zhang, Zheng; Mao, Yong; Batish, Mona; Kohn, Joachim; Moghe, Prabhas V

2016-11-01

materials relies on technologies that can sensitively discern cell response dynamics to biomaterials, while capturing cell-to-cell heterogeneity and preserving cellular native phenotypes. In this study, we illustrate the application of a novel high content image informatics platform to classify emergent human mesenchymal stem cell (hMSC) phenotypes in a diverse range of 3-D biomaterial scaffolds with high sensitivity and precision, and track cell responses to varied external stimuli. A major in silico innovation is the proposed image profiling technology based on unique three dimensional textural signatures of a mechanoreporter protein within the nuclei of stem cells cultured in 3-D scaffolds. This technology will accelerate the pace of high-fidelity biomaterial screening. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

The Figured Worlds of High School Science Teachers: Uncovering Three-Dimensional Assessment Decisions

Science.gov (United States)

Ewald, Megan

As a result of recent mandates of the Next Generation Science Standards, assessments are a "system of meaning" amidst a paradigm shift toward three-dimensional assessments. This study is motivated by two research questions: 1) how do high school science teachers describe their processes of decision-making in the development and use of three-dimensional assessments and 2) how do high school science teachers negotiate their identities as assessors in designing three-dimensional assessments. An important factor in teachers' assessment decision making is how they identify themselves as assessors. Therefore, this study investigated the teachers' roles as assessors through the Sociocultural Identity Theory. The most important contribution from this study is the emergent teacher assessment sub-identities: the modifier-recycler , the feeler-finder, and the creator. Using a qualitative phenomenological research design, focus groups, three-series interviews, think-alouds, and document analysis were utilized in this study. These qualitative methods were chosen to elicit rich conversations among teachers, make meaning of the teachers' experiences through in-depth interviews, amplify the thought processes of individual teachers while making assessment decisions, and analyze assessment documents in relation to teachers' perspectives. The findings from this study suggest that--of the 19 participants--only two teachers could consistently be identified as creators and aligned their assessment practices with NGSS. However, assessment sub-identities are not static and teachers may negotiate their identities from one moment to the next within socially constructed realms of interpretation known as figured worlds. Because teachers are positioned in less powerful figured worlds within the dominant discourse of standardization, this study raises awareness as to how the external pressures from more powerful figured worlds socially construct teachers' identities as assessors. For teachers

Migration ability and Toll-like receptor expression of human mesenchymal stem cells improves significantly after three-dimensional culture.

Science.gov (United States)

Zhou, Panpan; Liu, Zilin; Li, Xue; Zhang, Bing; Wang, Xiaoyuan; Lan, Jing; Shi, Qing; Li, Dong; Ju, Xiuli

2017-09-16

While the conventional two-dimensional (2D) culture protocol is well accepted for the culture of mesenchymal stem cells (MSCs), this method fails to recapitulate the in vivo native three-dimensional (3D) cellular microenvironment, and may result in phenotypic changes, and homing and migration capacity impairments. MSC preparation in 3D culture systems has been considered an attractive preparatory and delivery method recently. We seeded human umbilical cord-derived MSCs (hUCMSCs) in a 3D culture system with porcine acellular dermal matrix (PADM), and investigated the phenotypic changes, the expression changes of some important receptors, including Toll-like receptors (TLRs) and C-X-C chemokine receptor type 4 (CXCR4) when hUCMSCs were transferred from 2D to 3D systems, as well as the alterations in in vivo homing and migration potential. It was found that the percentage of CD105-positive cells decreased significantly, whereas that of CD34- and CD271-positive cells increased significantly in 3D culture, compared to that in 2D culture. The mRNA and protein expression levels of TLR2, TLR3, TLR4, TLR6, and CXCR4 in hUCMSCs were increased significantly upon culturing with PADM for 3 days, compared to the levels in 2D culture. The numbers of migratory 3D hUCMSCs in the heart, liver, spleen, and bone marrow were significantly greater than the numbers of 2D hUCMSCs, and the worst migration occurred in 3D + AMD3100 (CXCR4 antagonist) hUCMSCs. These results suggested that 3D culture of hUCMSCs with PADM could alter the phenotypic characteristics of hUCMSCs, increase their TLR and CXCR4 expression levels, and promote their migratory and homing capacity in which CXCR4 plays an important role. Copyright © 2017 Elsevier Inc. All rights reserved.

Data of a fluorescent imaging-based analysis of anti-cancer drug effects on three-dimensional cultures of breast cancer cells

Directory of Open Access Journals (Sweden)

Junji Itou

2015-12-01

Full Text Available Three-dimensional (3D cell culture is a powerful tool to study cell growth under 3D condition. To perform a simple test for anti-cancer drugs in 3D culture, visualization of non-proliferated cells is required. We propose a fluorescent imaging-based assay to analyze cancer cell proliferation in 3D culture. We used a pulse-labeling technique with a photoconvertible fluorescent protein Kaede to identify non-proliferated cells. This assay allows us to observe change in cell proliferation in 3D culture by simple imaging. Using this assay, we obtained the data of the effects of anti-cancer drugs, 5-fluorouracil and PD0332991 in a breast cancer cell line, MCF-7.

A robust robotic high-throughput antibody purification platform.

Science.gov (United States)

Schmidt, Peter M; Abdo, Michael; Butcher, Rebecca E; Yap, Min-Yin; Scotney, Pierre D; Ramunno, Melanie L; Martin-Roussety, Genevieve; Owczarek, Catherine; Hardy, Matthew P; Chen, Chao-Guang; Fabri, Louis J

2016-07-15

Monoclonal antibodies (mAbs) have become the fastest growing segment in the drug market with annual sales of more than 40 billion US$ in 2013. The selection of lead candidate molecules involves the generation of large repertoires of antibodies from which to choose a final therapeutic candidate. Improvements in the ability to rapidly produce and purify many antibodies in sufficient quantities reduces the lead time for selection which ultimately impacts on the speed with which an antibody may transition through the research stage and into product development. Miniaturization and automation of chromatography using micro columns (RoboColumns(®) from Atoll GmbH) coupled to an automated liquid handling instrument (ALH; Freedom EVO(®) from Tecan) has been a successful approach to establish high throughput process development platforms. Recent advances in transient gene expression (TGE) using the high-titre Expi293F™ system have enabled recombinant mAb titres of greater than 500mg/L. These relatively high protein titres reduce the volume required to generate several milligrams of individual antibodies for initial biochemical and biological downstream assays, making TGE in the Expi293F™ system ideally suited to high throughput chromatography on an ALH. The present publication describes a novel platform for purifying Expi293F™-expressed recombinant mAbs directly from cell-free culture supernatant on a Perkin Elmer JANUS-VariSpan ALH equipped with a plate shuttle device. The purification platform allows automated 2-step purification (Protein A-desalting/size exclusion chromatography) of several hundred mAbs per week. The new robotic method can purify mAbs with high recovery (>90%) at sub-milligram level with yields of up to 2mg from 4mL of cell-free culture supernatant. Copyright © 2016 Elsevier B.V. All rights reserved.

Management of High-Throughput DNA Sequencing Projects: Alpheus.

Science.gov (United States)

Miller, Neil A; Kingsmore, Stephen F; Farmer, Andrew; Langley, Raymond J; Mudge, Joann; Crow, John A; Gonzalez, Alvaro J; Schilkey, Faye D; Kim, Ryan J; van Velkinburgh, Jennifer; May, Gregory D; Black, C Forrest; Myers, M Kathy; Utsey, John P; Frost, Nicholas S; Sugarbaker, David J; Bueno, Raphael; Gullans, Stephen R; Baxter, Susan M; Day, Steve W; Retzel, Ernest F

2008-12-26

High-throughput DNA sequencing has enabled systems biology to begin to address areas in health, agricultural and basic biological research. Concomitant with the opportunities is an absolute necessity to manage significant volumes of high-dimensional and inter-related data and analysis. Alpheus is an analysis pipeline, database and visualization software for use with massively parallel DNA sequencing technologies that feature multi-gigabase throughput characterized by relatively short reads, such as Illumina-Solexa (sequencing-by-synthesis), Roche-454 (pyrosequencing) and Applied Biosystem's SOLiD (sequencing-by-ligation). Alpheus enables alignment to reference sequence(s), detection of variants and enumeration of sequence abundance, including expression levels in transcriptome sequence. Alpheus is able to detect several types of variants, including non-synonymous and synonymous single nucleotide polymorphisms (SNPs), insertions/deletions (indels), premature stop codons, and splice isoforms. Variant detection is aided by the ability to filter variant calls based on consistency, expected allele frequency, sequence quality, coverage, and variant type in order to minimize false positives while maximizing the identification of true positives. Alpheus also enables comparisons of genes with variants between cases and controls or bulk segregant pools. Sequence-based differential expression comparisons can be developed, with data export to SAS JMP Genomics for statistical analysis.

Quantifying the correlation between spatially defined oxygen gradients and cell fate in an engineered three-dimensional culture model

OpenAIRE

Ardakani, Amir G.; Cheema, Umber; Brown, Robert A.; Shipley, Rebecca J.

2014-01-01

A challenge in three-dimensional tissue culture remains the lack of quantitative information linking nutrient delivery and cellular distribution. Both in vivo and in vitro, oxygen is delivered by diffusion from its source (blood vessel or the construct margins). The oxygen level at a defined distance from its source depends critically on the balance of diffusion and cellular metabolism. Cells may respond to this oxygen environment through proliferation, death and chemotaxis, resulting in spat...

High throughput sample processing and automated scoring

Directory of Open Access Journals (Sweden)

Gunnar eBrunborg

2014-10-01

Full Text Available The comet assay is a sensitive and versatile method for assessing DNA damage in cells. In the traditional version of the assay, there are many manual steps involved and few samples can be treated in one experiment. High throughput modifications have been developed during recent years, and they are reviewed and discussed. These modifications include accelerated scoring of comets; other important elements that have been studied and adapted to high throughput are cultivation and manipulation of cells or tissues before and after exposure, and freezing of treated samples until comet analysis and scoring. High throughput methods save time and money but they are useful also for other reasons: large-scale experiments may be performed which are otherwise not practicable (e.g., analysis of many organs from exposed animals, and human biomonitoring studies, and automation gives more uniform sample treatment and less dependence on operator performance. The high throughput modifications now available vary largely in their versatility, capacity, complexity and costs. The bottleneck for further increase of throughput appears to be the scoring.

Three-dimensional porous graphene-Co{sub 3}O{sub 4} nanocomposites for high performance photocatalysts

Energy Technology Data Exchange (ETDEWEB)

Bin, Zeng, E-mail: 21467855@qq.com [College of Mechanical Engineering, Hunan University of Arts and Science, Changde 415000 (China); Hui, Long [Department of Applied Physics and Materials Research Center, The Hong Kong Polytechnic University, Hung Hom, Kowloon (Hong Kong)

2015-12-01

Highlights: • The three-dimensional porous graphene-Co{sub 3}O{sub 4} nanocomposites were synthesized. • Excellent photocatalytic performance. • Separated from the reaction medium by magnetic decantation. - Abstract: Novel three-dimensional porous graphene-Co{sub 3}O{sub 4} nanocomposites were synthesized by freeze-drying methods. Scanning and transmission electron microscopy revealed that the graphene formed a three-dimensional porous structure with Co{sub 3}O{sub 4} nanoparticles decorated surfaces. The as-obtained product showed high photocatalytic efficiency and could be easily separated from the reaction medium by magnetic decantation. This nanocomposite may be expected to have potential in water purification applications.

SAMNet: a network-based approach to integrate multi-dimensional high throughput datasets.

Science.gov (United States)

Gosline, Sara J C; Spencer, Sarah J; Ursu, Oana; Fraenkel, Ernest

2012-11-01

The rapid development of high throughput biotechnologies has led to an onslaught of data describing genetic perturbations and changes in mRNA and protein levels in the cell. Because each assay provides a one-dimensional snapshot of active signaling pathways, it has become desirable to perform multiple assays (e.g. mRNA expression and phospho-proteomics) to measure a single condition. However, as experiments expand to accommodate various cellular conditions, proper analysis and interpretation of these data have become more challenging. Here we introduce a novel approach called SAMNet, for Simultaneous Analysis of Multiple Networks, that is able to interpret diverse assays over multiple perturbations. The algorithm uses a constrained optimization approach to integrate mRNA expression data with upstream genes, selecting edges in the protein-protein interaction network that best explain the changes across all perturbations. The result is a putative set of protein interactions that succinctly summarizes the results from all experiments, highlighting the network elements unique to each perturbation. We evaluated SAMNet in both yeast and human datasets. The yeast dataset measured the cellular response to seven different transition metals, and the human dataset measured cellular changes in four different lung cancer models of Epithelial-Mesenchymal Transition (EMT), a crucial process in tumor metastasis. SAMNet was able to identify canonical yeast metal-processing genes unique to each commodity in the yeast dataset, as well as human genes such as β-catenin and TCF7L2/TCF4 that are required for EMT signaling but escaped detection in the mRNA and phospho-proteomic data. Moreover, SAMNet also highlighted drugs likely to modulate EMT, identifying a series of less canonical genes known to be affected by the BCR-ABL inhibitor imatinib (Gleevec), suggesting a possible influence of this drug on EMT.

Three-dimensional culture of human mesenchymal stem cells in a polyethylene terephthalate matrix

International Nuclear Information System (INIS)

Cao Yanfen; Li Ding; Shang Chunhua; Wang Jufang; Wang Xiaoning; Yang Shangtian

2010-01-01

Polyethylene terephthalate (PET) was used as the scaffold material to support the proliferation of human mesenchymal stem cells (hMSCs). The cells were cultured either statically in multi-wells or in a spinner flask agitated at 80 rpm for up to 20 days. To optimize the cell expansion condition, effects of the initial cell density and basic fibroblast growth factor (bFGF) were examined. During culture, cell growth and metabolism were tested. After 20 days, cells were harvested and surface markers were identified and quantified with flow cytometry. The results showed that hMSCs seeded at the lowest density gave the highest expansion fold. hMSCs grown in porous three-dimensional (3D) matrices displayed significantly different characteristics in terms of their proliferation and metabolism. PET matrices with 3D space could sustain cell proliferation for a long time. In addition, a low concentration (5 ng mL -1 ) of bFGF significantly enhanced the expansion of hMSCs in PET. Cell attachment and distribution in PET matrices were studied with confocal laser microscopy and scanning electron microscopy, which also confirmed cell proliferation. Furthermore, most of the cells in PET matrices were CD29, CD44 and CD105 positive, and CD34, CD45 and CD14 negative, confirming that hMSCs cultured in 3D PET matrices can be expanded and maintained in their undifferentiated state for at least 20 days without subculturing.

Magnetic resonance imaging-three-dimensional printing technology fabricates customized scaffolds for brain tissue engineering

Institute of Scientific and Technical Information of China (English)

Feng Fu; Chong Chen; Sai Zhang; Ming-liang Zhao; Xiao-hong Li; Zhe Qin; Chao Xu; Xu-yi Chen; Rui-xin Li; Li-na Wang; Ding-wei Peng; Hong-tao Sun; Yue Tu

2017-01-01

Conventional fabrication methods lack the ability to control both macro- and micro-structures of generated scaffolds. Three-dimensional printing is a solid free-form fabrication method that provides novel ways to create customized scaffolds with high precision and accuracy. In this study, an electrically controlled cortical impactor was used to induce randomized brain tissue defects. The overall shape of scaffolds was designed using rat-specific anatomical data obtained from magnetic resonance imaging, and the internal structure was created by computer- aided design. As the result of limitations arising from insufficient resolution of the manufacturing process, we magnified the size of the cavity model prototype five-fold to successfully fabricate customized collagen-chitosan scaffolds using three-dimensional printing. Results demonstrated that scaffolds have three-dimensional porous structures, high porosity, highly specific surface areas, pore connectivity and good internal characteristics. Neural stem cells co-cultured with scaffolds showed good viability, indicating good biocompatibility and biodegradability. This technique may be a promising new strategy for regenerating complex damaged brain tissues, and helps pave the way toward personalized medicine.

Three-dimensional scaffold-free fusion culture: the way to enhance chondrogenesis of in vitro propagated human articular chondrocytes

Directory of Open Access Journals (Sweden)

M. Lehmann

2013-11-01

Full Text Available Cartilage regeneration based on isolated and culture-expanded chondrocytes has been studied in various in vitro models, but the quality varies with respect to the morphology and the physiology of the synthesized tissues. The aim of our study was to promote in vitro chondrogenesis of human articular chondrocytes using a novel three-dimensional (3-D cultivation system in combination with the chondrogenic differentiation factors transforming growth factor beta 2 (TGF-b2 and L-ascorbic acid. Articular chondrocytes isolated from six elderly patients were expanded in monolayer culture. A single-cell suspension of the dedifferentiated chondrocytes was then added to agar-coated dishes without using any scaffold material, in the presence, or absence of TGF-b2 and/or L-ascorbic acid. Three-dimensional cartilage-like constructs, called single spheroids, and microtissues consisting of several spheroids fused together, named as fusions, were formed. Generated tissues were mainly characterized using histological and immunohistochemical techniques. The morphology of the in vitro tissues shared some similarities to native hyaline cartilage in regard to differentiated S100-positive chondrocytes within a cartilaginous matrix, with strong collagen type II expression and increased synthesis of proteoglycans. Finally, our innovative scaffold-free fusion culture technique supported enhanced chondrogenesis of human articular chondrocytes in vitro. These 3-D hyaline cartilage-like microtissues will be useful for in vitro studies of cartilage differentiation and regeneration, enabling optimization of functional tissue engineering and possibly contributing to the development of new approaches to treat traumatic cartilage defects or osteoarthritis.

Multiparallel Three-Dimensional Optical Microscopy

Science.gov (United States)

Nguyen, Lam K.; Price, Jeffrey H.; Kellner, Albert L.; Bravo-Zanoquera, Miguel

2010-01-01

Multiparallel three-dimensional optical microscopy is a method of forming an approximate three-dimensional image of a microscope sample as a collection of images from different depths through the sample. The imaging apparatus includes a single microscope plus an assembly of beam splitters and mirrors that divide the output of the microscope into multiple channels. An imaging array of photodetectors in each channel is located at a different distance along the optical path from the microscope, corresponding to a focal plane at a different depth within the sample. The optical path leading to each photodetector array also includes lenses to compensate for the variation of magnification with distance so that the images ultimately formed on all the photodetector arrays are of the same magnification. The use of optical components common to multiple channels in a simple geometry makes it possible to obtain high light-transmission efficiency with an optically and mechanically simple assembly. In addition, because images can be read out simultaneously from all the photodetector arrays, the apparatus can support three-dimensional imaging at a high scanning rate.

Quantitative imaging reveals heterogeneous growth dynamics and treatment-dependent residual tumor distributions in a three-dimensional ovarian cancer model

Science.gov (United States)

Celli, Jonathan P.; Rizvi, Imran; Evans, Conor L.; Abu-Yousif, Adnan O.; Hasan, Tayyaba

2010-09-01

Three-dimensional tumor models have emerged as valuable in vitro research tools, though the power of such systems as quantitative reporters of tumor growth and treatment response has not been adequately explored. We introduce an approach combining a 3-D model of disseminated ovarian cancer with high-throughput processing of image data for quantification of growth characteristics and cytotoxic response. We developed custom MATLAB routines to analyze longitudinally acquired dark-field microscopy images containing thousands of 3-D nodules. These data reveal a reproducible bimodal log-normal size distribution. Growth behavior is driven by migration and assembly, causing an exponential decay in spatial density concomitant with increasing mean size. At day 10, cultures are treated with either carboplatin or photodynamic therapy (PDT). We quantify size-dependent cytotoxic response for each treatment on a nodule by nodule basis using automated segmentation combined with ratiometric batch-processing of calcein and ethidium bromide fluorescence intensity data (indicating live and dead cells, respectively). Both treatments reduce viability, though carboplatin leaves micronodules largely structurally intact with a size distribution similar to untreated cultures. In contrast, PDT treatment disrupts micronodular structure, causing punctate regions of toxicity, shifting the distribution toward smaller sizes, and potentially increasing vulnerability to subsequent chemotherapeutic treatment.

Three-dimensional bio-printing.

Science.gov (United States)

Gu, Qi; Hao, Jie; Lu, YangJie; Wang, Liu; Wallace, Gordon G; Zhou, Qi

2015-05-01

Three-dimensional (3D) printing technology has been widely used in various manufacturing operations including automotive, defence and space industries. 3D printing has the advantages of personalization, flexibility and high resolution, and is therefore becoming increasingly visible in the high-tech fields. Three-dimensional bio-printing technology also holds promise for future use in medical applications. At present 3D bio-printing is mainly used for simulating and reconstructing some hard tissues or for preparing drug-delivery systems in the medical area. The fabrication of 3D structures with living cells and bioactive moieties spatially distributed throughout will be realisable. Fabrication of complex tissues and organs is still at the exploratory stage. This review summarize the development of 3D bio-printing and its potential in medical applications, as well as discussing the current challenges faced by 3D bio-printing.

Dual-wavelength optical-resolution photoacoustic microscopy for cells with gold nanoparticle bioconjugates in three-dimensional cultures

Science.gov (United States)

Lee, Po-Yi; Liu, Wei-Wen; Chen, Shu-Ching; Li, Pai-Chi

2016-03-01

Three-dimensional (3D) in vitro models bridge the gap between typical two-dimensional cultures and in vivo conditions. However, conventional optical imaging methods such as confocal microscopy and two-photon microscopy cannot accurately depict cellular processing in 3D models due to limited penetration of photons. We developed a dualwavelength optical-resolution photoacoustic microscopy (OR-PAM), which provides sufficient penetration depth and spatial resolution, for studying CD8+ cytotoxic T lymphocytes (CTLs) trafficking in an in vitro 3D tumor microenvironment. CTLs play a cardinal role in host defense against tumor. Efficient trafficking of CTLs to the tumor microenvironment is a critical step for cancer immunotherapy. For the proposed system, gold nanospheres and indocyanine green (ICG) have been remarkable choices for contrast agents for photoacoustic signals due to their excellent biocompatibility and high optical absorption. With distinct absorption spectrums, targeted cells with gold nanospheres and ICG respectively can be identified by switching 523-nm and 800-nm laser irradiation. Moreover, we use an x-y galvanometer scanner to obtain high scanning rate. In the developed system, lateral and axial resolutions were designed at 1.6 μm and 5 μm, respectively. We successfully showed that dual-spectral OR-PAM can map either the distribution of CTLs with gold nanospheres at a visible wavelength of 523 nm or the 3D structure of tumor spheres with ICG in an in vitro 3D microenvironment. Our OR-PAM can provide better biological relevant information in cellular interaction and is potential for preclinical screening of anti-cancer drugs.

Three-dimensional instability of standing waves

Science.gov (United States)

Zhu, Qiang; Liu, Yuming; Yue, Dick K. P.

2003-12-01

We investigate the three-dimensional instability of finite-amplitude standing surface waves under the influence of gravity. The analysis employs the transition matrix (TM) approach and uses a new high-order spectral element (HOSE) method for computation of the nonlinear wave dynamics. HOSE is an extension of the original high-order spectral method (HOS) wherein nonlinear wave wave and wave body interactions are retained up to high order in wave steepness. Instead of global basis functions in HOS, however, HOSE employs spectral elements to allow for complex free-surface geometries and surface-piercing bodies. Exponential convergence of HOS with respect to the total number of spectral modes (for a fixed number of elements) and interaction order is retained in HOSE. In this study, we use TM-HOSE to obtain the stability of general three-dimensional perturbations (on a two-dimensional surface) on two classes of standing waves: plane standing waves in a rectangular tank; and radial/azimuthal standing waves in a circular basin. For plane standing waves, we confirm the known result of two-dimensional side-bandlike instability. In addition, we find a novel three-dimensional instability for base flow of any amplitude. The dominant component of the unstable disturbance is an oblique (standing) wave oriented at an arbitrary angle whose frequency is close to the (nonlinear) frequency of the original standing wave. This finding is confirmed by direct long-time simulations using HOSE which show that the nonlinear evolution leads to classical Fermi Pasta Ulam recurrence. For the circular basin, we find that, beyond a threshold wave steepness, a standing wave (of nonlinear frequency Omega) is unstable to three-dimensional perturbations. The unstable perturbation contains two dominant (standing-wave) components, the sum of whose frequencies is close to 2Omega. From the cases we consider, the critical wave steepness is found to generally decrease/increase with increasing radial

Hanging Drop, A Best Three-Dimensional (3D) Culture Method for Primary Buffalo and Sheep Hepatocytes.

Science.gov (United States)

Shri, Meena; Agrawal, Himanshu; Rani, Payal; Singh, Dheer; Onteru, Suneel Kumar

2017-04-26

Livestock, having close resemblance to humans, could be a better source of primary hepatocytes than rodents. Herein, we successfully developed three-dimensional (3D) culturing system for primary sheep and buffalo hepatocytes. The 3D-structures of sheep hepatocytes were formed on the fifth-day and maintained until the tenth-day on polyHEMA-coated plates and in hanging drops with William's E media (HDW). Between the cultured and fresh cells, we observed a similar expression of GAPDH, HNF4α, ALB, CYP1A1, CK8 and CK18. Interestingly, a statistically significant increase was noted in the TAT, CPS, AFP, AAT, GSP and PCNA expression. In buffalo hepatocytes culture, 3D-like structures were formed on the third-day and maintained until the sixth-day on polyHEMA and HDW. The expression of HNF4α, GSP, CPS, AFP, AAT, PCNA and CK18 was similar between cultured and fresh cells. Further, a statistically significant increase in the TAT and CK8 expression, and a decrease in the GAPDH, CYP1A1 and ALB expression were noted. Among the culture systems, HDW maintained the liver transcript markers more or less similar to the fresh hepatocytes of the sheep and buffalo for ten and six days, respectively. Taken together, hanging drop is an efficient method for 3D culturing of primary sheep and buffalo hepatocytes.

A simple, specific high-throughput enzyme-linked immunosorbent assay (ELISA) for quantitative determination of melatonin in cell culture medium.

Science.gov (United States)

Li, Ye; Cassone, Vincent M

2015-09-01

A simple, specific, high-throughput enzyme-linked immunosorbent assay (ELISA) for quantitative determination of melatonin was developed for directly measuring melatonin in cell culture medium with 10% FBS. This assay adopts a commercial monoclonal melatonin antibody and melatonin-HRP conjugate, so it can be applied in multiple labs rapidly with low cost compared with commercial RIA and ELISA kits. In addition, the procedure is much simpler with only four steps: 1) sample/conjugate incubation, 2) plate washing, 3) TMB color reaction and 4) reading of results. The standards of the assay cover a wide working range from 100 pg/mL to 10 ng/mL. The sensitivity was 68 pg/mL in cell culture medium with 10% FBS and 26 pg/mL in PBS with as little as 25 μL sample volume. The recovery of melatonin from cell culture medium was 101.0%. The principal cross-reacting compound was 5-methoxytryptophol (0.1%). The variation coefficients of the assay, within and between runs, ranged between 6.68% and 15.76% in cell culture medium. The mean linearity of a series diluted cell culture medium sample was 105% (CV=5%), ranging between 98% and 111%, y=5.5263x+0.0646, R(2)=0.99. The assay enables small research and teaching labs to reliably measure this important neurohormone. Copyright © 2015 Elsevier B.V. All rights reserved.

Three-Dimensional Porous Nitrogen-Doped NiO Nanostructures as Highly Sensitive NO2 Sensors

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Van Hoang Luan

2017-10-01

Full Text Available Nickel oxide has been widely used in chemical sensing applications, because it has an excellent p-type semiconducting property with high chemical stability. Here, we present a novel technique of fabricating three-dimensional porous nitrogen-doped nickel oxide nanosheets as a highly sensitive NO2 sensor. The elaborate nanostructure was prepared by a simple and effective hydrothermal synthesis method. Subsequently, nitrogen doping was achieved by thermal treatment with ammonia gas. When the p-type dopant, i.e., nitrogen atoms, was introduced in the three-dimensional nanostructures, the nickel-oxide-nanosheet-based sensor showed considerable NO2 sensing ability with two-fold higher responsivity and sensitivity compared to non-doped nickel-oxide-based sensors.

Three-dimensional ICT reconstruction

International Nuclear Information System (INIS)

Zhang Aidong; Li Ju; Chen Fa; Sun Lingxia

2005-01-01

The three-dimensional ICT reconstruction method is the hot topic of recent ICT technology research. In the context, qualified visual three-dimensional ICT pictures are achieved through multi-piece two-dimensional images accumulation by, combining with thresholding method and linear interpolation. Different direction and different position images of the reconstructed pictures are got by rotation and interception respectively. The convenient and quick method is significantly instructive to more complicated three-dimensional reconstruction of ICT images. (authors)

Three-dimensional ICT reconstruction

International Nuclear Information System (INIS)

Zhang Aidong; Li Ju; Chen Fa; Sun Lingxia

2004-01-01

The three-dimensional ICT reconstruction method is the hot topic of recent ICT technology research. In the context qualified visual three-dimensional ICT pictures are achieved through multi-piece two-dimensional images accumulation by order, combining with thresholding method and linear interpolation. Different direction and different position images of the reconstructed pictures are got by rotation and interception respectively. The convenient and quick method is significantly instructive to more complicated three-dimensional reconstruction of ICT images. (authors)

Three-dimensional graphene/polyaniline composite material for high-performance supercapacitor applications

International Nuclear Information System (INIS)

Liu, Huili; Wang, Yi; Gou, Xinglong; Qi, Tao; Yang, Jun; Ding, Yulong

2013-01-01

Highlights: ► A novel 3D graphene showed high specific surface area and large mesopore volume. ► Aniline monomer was polymerized in the presence of 3D graphene at room temperature. ► The supercapacitive properties were studied by CV and charge–discharge tests. ► The composite show a high gravimetric capacitance and good cyclic stability. ► The 3D graphene/polyaniline has never been report before our work. -- Abstract: A novel three-dimensional (3D) graphene/polyaniline nanocomposite material which is synthesized using in situ polymerization of aniline monomer on the graphene surface is reported as an electrode for supercapacitors. The morphology and structure of the material are characterized by scanning electron microscopy (SEM), transmission electron microscope (TEM), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). The electrochemical properties of the resulting materials are systematically studied using cyclic voltammetry (CV) and constant current charge–discharge tests. A high gravimetric capacitance of 463 F g −1 at a scan rate of 1 mV s −1 is obtained by means of CVs with 3 mol L −1 KOH as the electrolyte. In addition, the composite material shows only 9.4% capacity loss after 500 cycles, indicating better cyclic stability for supercapacitor applications. The high specific surface area, large mesopore volume and three-dimensional nanoporous structure of 3D graphene could contribute to the high specific capacitance and good cyclic life

Three-dimensional neuroepithelial culture from human embryonic stem cells and its use for quantitative conversion to retinal pigment epithelium.

Directory of Open Access Journals (Sweden)

Yu Zhu

Full Text Available A goal in human embryonic stem cell (hESC research is the faithful differentiation to given cell types such as neural lineages. During embryonic development, a basement membrane surrounds the neural plate that forms a tight, apico-basolaterally polarized epithelium before closing to form a neural tube with a single lumen. Here we show that the three-dimensional epithelial cyst culture of hESCs in Matrigel combined with neural induction results in a quantitative conversion into neuroepithelial cysts containing a single lumen. Cells attain a defined neuroepithelial identity by 5 days. The neuroepithelial cysts naturally generate retinal epithelium, in part due to IGF-1/insulin signaling. We demonstrate the utility of this epithelial culture approach by achieving a quantitative production of retinal pigment epithelial (RPE cells from hESCs within 30 days. Direct transplantation of this RPE into a rat model of retinal degeneration without any selection or expansion of the cells results in the formation of a donor-derived RPE monolayer that rescues photoreceptor cells. The cyst method for neuroepithelial differentiation of pluripotent stem cells is not only of importance for RPE generation but will also be relevant to the production of other neuronal cell types and for reconstituting complex patterning events from three-dimensional neuroepithelia.

Bacterial diversity of the Colombian fermented milk "Suero Costeño" assessed by culturing and high-throughput sequencing and DGGE analysis of 16S rRNA gene amplicons.

Science.gov (United States)

Motato, Karina Edith; Milani, Christian; Ventura, Marco; Valencia, Francia Elena; Ruas-Madiedo, Patricia; Delgado, Susana

2017-12-01

"Suero Costeño" (SC) is a traditional soured cream elaborated from raw milk in the Northern-Caribbean coast of Colombia. The natural microbiota that characterizes this popular Colombian fermented milk is unknown, although several culturing studies have previously been attempted. In this work, the microbiota associated with SC from three manufacturers in two regions, "Planeta Rica" (Córdoba) and "Caucasia" (Antioquia), was analysed by means of culturing methods in combination with high-throughput sequencing and DGGE analysis of 16S rRNA gene amplicons. The bacterial ecosystem of SC samples was revealed to be composed of lactic acid bacteria belonging to the Streptococcaceae and Lactobacillaceae families; the proportions and genera varying among manufacturers and region of elaboration. Members of the Lactobacillus acidophilus group, Lactocococcus lactis, Streptococcus infantarius and Streptococcus salivarius characterized this artisanal product. In comparison with culturing, the use of molecular in deep culture-independent techniques provides a more realistic picture of the overall bacterial communities residing in SC. Besides the descriptive purpose, these approaches will facilitate a rational strategy to follow (culture media and growing conditions) for the isolation of indigenous strains that allow standardization in the manufacture of SC. Copyright © 2017 Elsevier Ltd. All rights reserved.

Three-dimensional analysis of craniofacial bones using three-dimensional computer tomography

Energy Technology Data Exchange (ETDEWEB)

Ono, Ichiro; Ohura, Takehiko; Kimura, Chu (Hokkaido Univ., Sapporo (Japan). School of Medicine) (and others)

1989-08-01

Three-dimensional computer tomography (3DCT) was performed in patients with various diseases to visualize stereoscopically the deformity of the craniofacial bones. The data obtained were analyzed by the 3DCT analyzing system. A new coordinate system was established using the median sagittal plane of the face (a plane passing through sella, nasion and basion) on the three-dimensional image. Three-dimensional profilograms were prepared for detailed analysis of the deformation of craniofacial bones for cleft lip and palate, mandibular prognathia and hemifacial microsomia. For patients, asymmetry in the frontal view and twist-formed complicated deformities were observed, as well as deformity of profiles in the anteroposterior and up-and-down directions. A newly developed technique allows three-dimensional visualization of changes in craniofacial deformity. It would aid in determining surgical strategy, including crani-facial surgery and maxillo-facial surgery, and in evaluating surgical outcome. (N.K.).

Three-dimensional analysis of craniofacial bones using three-dimensional computer tomography

International Nuclear Information System (INIS)

Ono, Ichiro; Ohura, Takehiko; Kimura, Chu

1989-01-01

Three-dimensional computer tomography (3DCT) was performed in patients with various diseases to visualize stereoscopically the deformity of the craniofacial bones. The data obtained were analyzed by the 3DCT analyzing system. A new coordinate system was established using the median sagittal plane of the face (a plane passing through sella, nasion and basion) on the three-dimensional image. Three-dimensional profilograms were prepared for detailed analysis of the deformation of craniofacial bones for cleft lip and palate, mandibular prognathia and hemifacial microsomia. For patients, asymmetry in the frontal view and twist-formed complicated deformities were observed, as well as deformity of profiles in the anteroposterior and up-and-down directions. A newly developed technique allows three-dimensional visualization of changes in craniofacial deformity. It would aid in determining surgical strategy, including crani-facial surgery and maxillo-facial surgery, and in evaluating surgical outcome. (N.K.)

Comprehensive optical and data management infrastructure for high-throughput light-sheet microscopy of whole mouse brains.

Science.gov (United States)

Müllenbroich, M Caroline; Silvestri, Ludovico; Onofri, Leonardo; Costantini, Irene; Hoff, Marcel Van't; Sacconi, Leonardo; Iannello, Giulio; Pavone, Francesco S

2015-10-01

Comprehensive mapping and quantification of neuronal projections in the central nervous system requires high-throughput imaging of large volumes with microscopic resolution. To this end, we have developed a confocal light-sheet microscope that has been optimized for three-dimensional (3-D) imaging of structurally intact clarified whole-mount mouse brains. We describe the optical and electromechanical arrangement of the microscope and give details on the organization of the microscope management software. The software orchestrates all components of the microscope, coordinates critical timing and synchronization, and has been written in a versatile and modular structure using the LabVIEW language. It can easily be adapted and integrated to other microscope systems and has been made freely available to the light-sheet community. The tremendous amount of data routinely generated by light-sheet microscopy further requires novel strategies for data handling and storage. To complete the full imaging pipeline of our high-throughput microscope, we further elaborate on big data management from streaming of raw images up to stitching of 3-D datasets. The mesoscale neuroanatomy imaged at micron-scale resolution in those datasets allows characterization and quantification of neuronal projections in unsectioned mouse brains.

High-resolution non-destructive three-dimensional imaging of integrated circuits.

Science.gov (United States)

Holler, Mirko; Guizar-Sicairos, Manuel; Tsai, Esther H R; Dinapoli, Roberto; Müller, Elisabeth; Bunk, Oliver; Raabe, Jörg; Aeppli, Gabriel

2017-03-15

Modern nanoelectronics has advanced to a point at which it is impossible to image entire devices and their interconnections non-destructively because of their small feature sizes and the complex three-dimensional structures resulting from their integration on a chip. This metrology gap implies a lack of direct feedback between design and manufacturing processes, and hampers quality control during production, shipment and use. Here we demonstrate that X-ray ptychography-a high-resolution coherent diffractive imaging technique-can create three-dimensional images of integrated circuits of known and unknown designs with a lateral resolution in all directions down to 14.6 nanometres. We obtained detailed device geometries and corresponding elemental maps, and show how the devices are integrated with each other to form the chip. Our experiments represent a major advance in chip inspection and reverse engineering over the traditional destructive electron microscopy and ion milling techniques. Foreseeable developments in X-ray sources, optics and detectors, as well as adoption of an instrument geometry optimized for planar rather than cylindrical samples, could lead to a thousand-fold increase in efficiency, with concomitant reductions in scan times and voxel sizes.

High-resolution non-destructive three-dimensional imaging of integrated circuits

Science.gov (United States)

Holler, Mirko; Guizar-Sicairos, Manuel; Tsai, Esther H. R.; Dinapoli, Roberto; Müller, Elisabeth; Bunk, Oliver; Raabe, Jörg; Aeppli, Gabriel

2017-03-01

Modern nanoelectronics has advanced to a point at which it is impossible to image entire devices and their interconnections non-destructively because of their small feature sizes and the complex three-dimensional structures resulting from their integration on a chip. This metrology gap implies a lack of direct feedback between design and manufacturing processes, and hampers quality control during production, shipment and use. Here we demonstrate that X-ray ptychography—a high-resolution coherent diffractive imaging technique—can create three-dimensional images of integrated circuits of known and unknown designs with a lateral resolution in all directions down to 14.6 nanometres. We obtained detailed device geometries and corresponding elemental maps, and show how the devices are integrated with each other to form the chip. Our experiments represent a major advance in chip inspection and reverse engineering over the traditional destructive electron microscopy and ion milling techniques. Foreseeable developments in X-ray sources, optics and detectors, as well as adoption of an instrument geometry optimized for planar rather than cylindrical samples, could lead to a thousand-fold increase in efficiency, with concomitant reductions in scan times and voxel sizes.

Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.

Science.gov (United States)

Kaneko, Ai; Sankai, Yoshiyuki

2014-01-01

The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months) primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4) cells/mL (8.9×10(3) cells/cm2) without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm), greater cell viability (≥30%) for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks).

Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.

Directory of Open Access Journals (Sweden)

Ai Kaneko

Full Text Available The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4 cells/mL (8.9×10(3 cells/cm2 without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm, greater cell viability (≥30% for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks.

THREE-DIMENSIONAL OBSERVATIONS ON THICK BIOLOGICAL SPECIMENS BY HIGH VOLTAGE ELECTRON MICROSCOPY

Directory of Open Access Journals (Sweden)

Tetsuji Nagata

2011-05-01

Full Text Available Thick biological specimens prepared as whole mount cultured cells or thick sections from embedded tissues were stained with histochemical reactions, such as thiamine pyrophosphatase, glucose-6-phosphatase, cytochrome oxidase, acid phosphatase, DAB reactions and radioautography, to observe 3-D ultrastructures of cell organelles producing stereo-pairs by high voltage electron microscopy at accerelating voltages of 400-1000 kV. The organelles demonstrated were Golgi apparatus, endoplasmic reticulum, mitochondria, lysosomes, peroxisomes, pinocytotic vesicles and incorporations of radioactive compounds. As the results, those cell organelles were observed 3- dimensionally and the relative relationships between these organelles were demonstrated.

Cell fiber-based three-dimensional culture system for highly efficient expansion of human induced pluripotent stem cells.

Science.gov (United States)

Ikeda, Kazuhiro; Nagata, Shogo; Okitsu, Teru; Takeuchi, Shoji

2017-06-06

Human pluripotent stem cells are a potentially powerful cellular resource for application in regenerative medicine. Because such applications require large numbers of human pluripotent stem cell-derived cells, a scalable culture system of human pluripotent stem cell needs to be developed. Several suspension culture systems for human pluripotent stem cell expansion exist; however, it is difficult to control the thickness of cell aggregations in these systems, leading to increased cell death likely caused by limited diffusion of gases and nutrients into the aggregations. Here, we describe a scalable culture system using the cell fiber technology for the expansion of human induced pluripotent stem (iPS) cells. The cells were encapsulated and cultured within the core region of core-shell hydrogel microfibers, resulting in the formation of rod-shaped or fiber-shaped cell aggregations with sustained thickness and high viability. By encapsulating the cells with type I collagen, we demonstrated a long-term culture of the cells by serial passaging at a high expansion rate (14-fold in four days) while retaining its pluripotency. Therefore, our culture system could be used for large-scale expansion of human pluripotent stem cells for use in regenerative medicine.

In-Field High-Throughput Phenotyping of Cotton Plant Height Using LiDAR

Directory of Open Access Journals (Sweden)

Shangpeng Sun

2017-04-01

Full Text Available A LiDAR-based high-throughput phenotyping (HTP system was developed for cotton plant phenotyping in the field. The HTP system consists of a 2D LiDAR and an RTK-GPS mounted on a high clearance tractor. The LiDAR scanned three rows of cotton plots simultaneously from the top and the RTK-GPS was used to provide the spatial coordinates of the point cloud during data collection. Configuration parameters of the system were optimized to ensure the best data quality. A height profile for each plot was extracted from the dense three dimensional point clouds; then the maximum height and height distribution of each plot were derived. In lab tests, single plants were scanned by LiDAR using 0.5° angular resolution and results showed an R2 value of 1.00 (RMSE = 3.46 mm in comparison to manual measurements. In field tests using the same angular resolution; the LiDAR-based HTP system achieved average R2 values of 0.98 (RMSE = 65 mm for cotton plot height estimation; compared to manual measurements. This HTP system is particularly useful for large field application because it provides highly accurate measurements; and the efficiency is greatly improved compared to similar studies using the side view scan.

Three dimensional strained semiconductors

Science.gov (United States)

Voss, Lars; Conway, Adam; Nikolic, Rebecca J.; Leao, Cedric Rocha; Shao, Qinghui

2016-11-08

In one embodiment, an apparatus includes a three dimensional structure comprising a semiconductor material, and at least one thin film in contact with at least one exterior surface of the three dimensional structure for inducing a strain in the structure, the thin film being characterized as providing at least one of: an induced strain of at least 0.05%, and an induced strain in at least 5% of a volume of the three dimensional structure. In another embodiment, a method includes forming a three dimensional structure comprising a semiconductor material, and depositing at least one thin film on at least one surface of the three dimensional structure for inducing a strain in the structure, the thin film being characterized as providing at least one of: an induced strain of at least 0.05%, and an induced strain in at least 5% of a volume of the structure.

Differentiation Potential of Human Chorion-Derived Mesenchymal Stem Cells into Motor Neuron-Like Cells in Two- and Three-Dimensional Culture Systems.

Science.gov (United States)

Faghihi, Faezeh; Mirzaei, Esmaeil; Ai, Jafar; Lotfi, Abolfazl; Sayahpour, Forough Azam; Barough, Somayeh Ebrahimi; Joghataei, Mohammad Taghi

2016-04-01

Many people worldwide suffer from motor neuron-related disorders such as amyotrophic lateral sclerosis and spinal cord injuries. Recently, several attempts have been made to recruit stem cells to modulate disease progression in ALS and also regenerate spinal cord injuries. Chorion-derived mesenchymal stem cells (C-MSCs), used to be discarded as postpartum medically waste product, currently represent a class of cells with self renewal property and immunomodulatory capacity. These cells are able to differentiate into mesodermal and nonmesodermal lineages such as neural cells. On the other hand, gelatin, as a simply denatured collagen, is a suitable substrate for cell adhesion and differentiation. It has been shown that electrospinning of scaffolds into fibrous structure better resembles the physiological microenvironment in comparison with two-dimensional (2D) culture system. Since there is no report on potential of human chorion-derived MSCs to differentiate into motor neuron cells in two- and three-dimensional (3D) culture systems, we set out to determine the effect of retinoic acid (RA) and sonic hedgehog (Shh) on differentiation of human C-MSCs into motor neuron-like cells cultured on tissue culture plates (2D) and electrospun nanofibrous gelatin scaffold (3D).

Three-Dimensional Organotypic Co-Culture Model of Intestinal Epithelial Cells and Macrophages to Study "Salmonella Enterica" Colonization Patterns

Science.gov (United States)

Ott, Mark; Yang, J; Barilla, J.; Crabbe, A.; Sarker, S. F.; Liu, Y.

2017-01-01

Three-dimensional/3-D organotypic models of human intestinal epithelium mimic the differentiated form and function of parental tissues often not exhibited by 2-D monolayers and respond to Salmonella in ways that reflect in vivo infections. To further enhance the physiological relevance of 3-D models to more closely approximate in vivo intestinal microenvironments during infection, we developed and validated a novel 3-D intestinal co-culture model containing multiple epithelial cell types and phagocytic macrophages, and applied to study enteric infection by different Salmonella pathovars.

Volume scanning three-dimensional display with an inclined two-dimensional display and a mirror scanner

Science.gov (United States)

Miyazaki, Daisuke; Kawanishi, Tsuyoshi; Nishimura, Yasuhiro; Matsushita, Kenji

2001-11-01

A new three-dimensional display system based on a volume-scanning method is demonstrated. To form a three-dimensional real image, an inclined two-dimensional image is rapidly moved with a mirror scanner while the cross-section patterns of a three-dimensional object are displayed sequentially. A vector-scan CRT display unit is used to obtain a high-resolution image. An optical scanning system is constructed with concave mirrors and a galvanometer mirror. It is confirmed that three-dimensional images, formed by the experimental system, satisfy all the criteria for human stereoscopic vision.

High Throughput Transcriptomics @ USEPA (Toxicology ...

Science.gov (United States)

The ideal chemical testing approach will provide complete coverage of all relevant toxicological responses. It should be sensitive and specific It should identify the mechanism/mode-of-action (with dose-dependence). It should identify responses relevant to the species of interest. Responses should ideally be translated into tissue-, organ-, and organism-level effects. It must be economical and scalable. Using a High Throughput Transcriptomics platform within US EPA provides broader coverage of biological activity space and toxicological MOAs and helps fill the toxicological data gap. Slide presentation at the 2016 ToxForum on using High Throughput Transcriptomics at US EPA for broader coverage biological activity space and toxicological MOAs.

Establishment of a long-term three-dimensional primary culture of mouse glandular stomach epithelial cells within the stem cell niche

International Nuclear Information System (INIS)

Katano, Takahito; Ootani, Akifumi; Mizoshita, Tsutomu; Tanida, Satoshi; Tsukamoto, Hironobu; Ozeki, Keiji; Ebi, Masahide; Mori, Yoshinori; Kataoka, Hiromi; Kamiya, Takeshi; Toda, Shuji; Joh, Takashi

2013-01-01

Highlights: ► We established a 3D culture system to allow long-term culture of stomach cells. ► In this culture system, gastric epithelial cells grew for about 3 months. ► The cultured cells differentiated into multi-units of the stomach. ► This culture method should be useful for elucidating the cause of gastric diseases. -- Abstract: Compared to the small intestine and colon, little is known about stem cells in the stomach because of a lack of specific stem cell markers and an in vitro system that allows long-term culture. Here we describe a long-term three-dimensional (3D) primary gastric culture system within the stem cell niche. Glandular stomach cells from neonatal mice cultured in collagen gel yielded expanding sphere-like structures for 3 months. The wall of the gastrospheres consisted of a highly polarized epithelial monolayer with an outer lining of myofibroblasts. The epithelial cells showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm as well as expression of MUC5AC, indicating differentiation into gastric surface mucous cells. These cells demonstrated the features of fully differentiated gastric surface mucous cells such as microvilli, junctional complexes, and glycogen and secretory granules. Fewer than 1% of cultured epithelial cells differentiated into enteroendocrine cells. Active proliferation of the epithelial cells and many apoptotic cells in the inner lumen revealed the rapid cell turnover in gastrospheres in vitro. This method enables us to investigate the role of signaling between cell–cell and epithelial–mesenchymal interactions in an environment that is extremely similar to the in vivo environment

Establishment of a long-term three-dimensional primary culture of mouse glandular stomach epithelial cells within the stem cell niche

Energy Technology Data Exchange (ETDEWEB)

Katano, Takahito [Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601 (Japan); Ootani, Akifumi [Department of Gastroenterology and GI Endoscopy Center, Shin-Kokura Hospital, Federation of National Public Service Personnel Mutual Aid Associations, 1-3-1 Kanada, Kokurakita-ku, Kitakyushu 803-0816 (Japan); Department of Internal Medicine, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga 849-8501 (Japan); Mizoshita, Tsutomu, E-mail: tmizoshi@med.nagoya-cu.ac.jp [Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601 (Japan); Tanida, Satoshi; Tsukamoto, Hironobu; Ozeki, Keiji; Ebi, Masahide; Mori, Yoshinori; Kataoka, Hiromi; Kamiya, Takeshi [Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601 (Japan); Toda, Shuji [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga 849-8501 (Japan); Joh, Takashi [Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601 (Japan)

2013-03-22

Highlights: ► We established a 3D culture system to allow long-term culture of stomach cells. ► In this culture system, gastric epithelial cells grew for about 3 months. ► The cultured cells differentiated into multi-units of the stomach. ► This culture method should be useful for elucidating the cause of gastric diseases. -- Abstract: Compared to the small intestine and colon, little is known about stem cells in the stomach because of a lack of specific stem cell markers and an in vitro system that allows long-term culture. Here we describe a long-term three-dimensional (3D) primary gastric culture system within the stem cell niche. Glandular stomach cells from neonatal mice cultured in collagen gel yielded expanding sphere-like structures for 3 months. The wall of the gastrospheres consisted of a highly polarized epithelial monolayer with an outer lining of myofibroblasts. The epithelial cells showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm as well as expression of MUC5AC, indicating differentiation into gastric surface mucous cells. These cells demonstrated the features of fully differentiated gastric surface mucous cells such as microvilli, junctional complexes, and glycogen and secretory granules. Fewer than 1% of cultured epithelial cells differentiated into enteroendocrine cells. Active proliferation of the epithelial cells and many apoptotic cells in the inner lumen revealed the rapid cell turnover in gastrospheres in vitro. This method enables us to investigate the role of signaling between cell–cell and epithelial–mesenchymal interactions in an environment that is extremely similar to the in vivo environment.

Application of ToxCast High-Throughput Screening and ...

Science.gov (United States)

Slide presentation at the SETAC annual meeting on High-Throughput Screening and Modeling Approaches to Identify Steroidogenesis Distruptors Slide presentation at the SETAC annual meeting on High-Throughput Screening and Modeling Approaches to Identify Steroidogenssis Distruptors

3D material cytometry (3DMaC): a very high-replicate, high-throughput analytical method using microfabricated, shape-specific, cell-material niches.

Science.gov (United States)

Parratt, Kirsten; Jeong, Jenny; Qiu, Peng; Roy, Krishnendu

2017-08-08

Studying cell behavior within 3D material niches is key to understanding cell biology in health and diseases, and developing biomaterials for regenerative medicine applications. Current approaches to studying these cell-material niches have low throughput and can only analyze a few replicates per experiment resulting in reduced measurement assurance and analytical power. Here, we report 3D material cytometry (3DMaC), a novel high-throughput method based on microfabricated, shape-specific 3D cell-material niches and imaging cytometry. 3DMaC achieves rapid and highly multiplexed analyses of very high replicate numbers ("n" of 10 4 -10 6 ) of 3D biomaterial constructs. 3DMaC overcomes current limitations of low "n", low-throughput, and "noisy" assays, to provide rapid and simultaneous analyses of potentially hundreds of parameters in 3D biomaterial cultures. The method is demonstrated here for a set of 85 000 events containing twelve distinct cell-biomaterial micro-niches along with robust, customized computational methods for high-throughput analytics with potentially unprecedented statistical power.

High-throughput bioinformatics with the Cyrille2 pipeline system

Directory of Open Access Journals (Sweden)

de Groot Joost CW

2008-02-01

Full Text Available Abstract Background Modern omics research involves the application of high-throughput technologies that generate vast volumes of data. These data need to be pre-processed, analyzed and integrated with existing knowledge through the use of diverse sets of software tools, models and databases. The analyses are often interdependent and chained together to form complex workflows or pipelines. Given the volume of the data used and the multitude of computational resources available, specialized pipeline software is required to make high-throughput analysis of large-scale omics datasets feasible. Results We have developed a generic pipeline system called Cyrille2. The system is modular in design and consists of three functionally distinct parts: 1 a web based, graphical user interface (GUI that enables a pipeline operator to manage the system; 2 the Scheduler, which forms the functional core of the system and which tracks what data enters the system and determines what jobs must be scheduled for execution, and; 3 the Executor, which searches for scheduled jobs and executes these on a compute cluster. Conclusion The Cyrille2 system is an extensible, modular system, implementing the stated requirements. Cyrille2 enables easy creation and execution of high throughput, flexible bioinformatics pipelines.

Simulating three-dimensional nonthermal high-energy photon emission in colliding-wind binaries

Energy Technology Data Exchange (ETDEWEB)

Reitberger, K.; Kissmann, R.; Reimer, A.; Reimer, O., E-mail: klaus.reitberger@uibk.ac.at [Institut für Astro- und Teilchenphysik and Institut für Theoretische Physik, Leopold-Franzens-Universität Innsbruck, A-6020 Innsbruck (Austria)

2014-07-01

Massive stars in binary systems have long been regarded as potential sources of high-energy γ rays. The emission is principally thought to arise in the region where the stellar winds collide and accelerate relativistic particles which subsequently emit γ rays. On the basis of a three-dimensional distribution function of high-energy particles in the wind collision region—as obtained by a numerical hydrodynamics and particle transport model—we present the computation of the three-dimensional nonthermal photon emission for a given line of sight. Anisotropic inverse Compton emission is modeled using the target radiation field of both stars. Photons from relativistic bremsstrahlung and neutral pion decay are computed on the basis of local wind plasma densities. We also consider photon-photon opacity effects due to the dense radiation fields of the stars. Results are shown for different stellar separations of a given binary system comprising of a B star and a Wolf-Rayet star. The influence of orbital orientation with respect to the line of sight is also studied by using different orbital viewing angles. For the chosen electron-proton injection ratio of 10{sup –2}, we present the ensuing photon emission in terms of two-dimensional projections maps, spectral energy distributions, and integrated photon flux values in various energy bands. Here, we find a transition from hadron-dominated to lepton-dominated high-energy emission with increasing stellar separations. In addition, we confirm findings from previous analytic modeling that the spectral energy distribution varies significantly with orbital orientation.

A long-term three dimensional liver co-culture system for improved prediction of clinically relevant drug-induced hepatotoxicity

International Nuclear Information System (INIS)

Kostadinova, Radina; Boess, Franziska; Applegate, Dawn; Suter, Laura; Weiser, Thomas; Singer, Thomas; Naughton, Brian; Roth, Adrian

2013-01-01

Drug-induced liver injury (DILI) is the major cause for liver failure and post-marketing drug withdrawals. Due to species-specific differences in hepatocellular function, animal experiments to assess potential liabilities of drug candidates can predict hepatotoxicity in humans only to a certain extent. In addition to animal experimentation, primary hepatocytes from rat or human are widely used for pre-clinical safety assessment. However, as many toxic responses in vivo are mediated by a complex interplay among different cell types and often require chronic drug exposures, the predictive performance of hepatocytes is very limited. Here, we established and characterized human and rat in vitro three-dimensional (3D) liver co-culture systems containing primary parenchymal and non-parenchymal hepatic cells. Our data demonstrate that cells cultured on a 3D scaffold have a preserved composition of hepatocytes, stellate, Kupffer and endothelial cells and maintain liver function for up to 3 months, as measured by the production of albumin, fibrinogen, transferrin and urea. Additionally, 3D liver co-cultures maintain cytochrome P450 inducibility, form bile canaliculi-like structures and respond to inflammatory stimuli. Upon incubation with selected hepatotoxicants including drugs which have been shown to induce idiosyncratic toxicity, we demonstrated that this model better detected in vivo drug-induced toxicity, including species-specific drug effects, when compared to monolayer hepatocyte cultures. In conclusion, our results underline the importance of more complex and long lasting in vitro cell culture models that contain all liver cell types and allow repeated drug-treatments for detection of in vivo-relevant adverse drug effects. - Highlights: ► 3D liver co-cultures maintain liver specific functions for up to three months. ► Activities of Cytochrome P450s remain drug- inducible accross three months. ► 3D liver co-cultures recapitulate drug-induced liver toxicity

A long-term three dimensional liver co-culture system for improved prediction of clinically relevant drug-induced hepatotoxicity

Energy Technology Data Exchange (ETDEWEB)

Kostadinova, Radina; Boess, Franziska [Non-Clinical Safety, Hoffmann-La Roche Ltd, Grenzacherstrasse 124, Building 73 / Room 117b, 4070 Basel (Switzerland); Applegate, Dawn [RegeneMed, 9855 Towne Centre Drive Suite 200, San Diego, CA 92121 (United States); Suter, Laura; Weiser, Thomas; Singer, Thomas [Non-Clinical Safety, Hoffmann-La Roche Ltd, Grenzacherstrasse 124, Building 73 / Room 117b, 4070 Basel (Switzerland); Naughton, Brian [RegeneMed, 9855 Towne Centre Drive Suite 200, San Diego, CA 92121 (United States); Roth, Adrian, E-mail: adrian_b.roth@roche.com [Non-Clinical Safety, Hoffmann-La Roche Ltd, Grenzacherstrasse 124, Building 73 / Room 117b, 4070 Basel (Switzerland)

2013-04-01

Drug-induced liver injury (DILI) is the major cause for liver failure and post-marketing drug withdrawals. Due to species-specific differences in hepatocellular function, animal experiments to assess potential liabilities of drug candidates can predict hepatotoxicity in humans only to a certain extent. In addition to animal experimentation, primary hepatocytes from rat or human are widely used for pre-clinical safety assessment. However, as many toxic responses in vivo are mediated by a complex interplay among different cell types and often require chronic drug exposures, the predictive performance of hepatocytes is very limited. Here, we established and characterized human and rat in vitro three-dimensional (3D) liver co-culture systems containing primary parenchymal and non-parenchymal hepatic cells. Our data demonstrate that cells cultured on a 3D scaffold have a preserved composition of hepatocytes, stellate, Kupffer and endothelial cells and maintain liver function for up to 3 months, as measured by the production of albumin, fibrinogen, transferrin and urea. Additionally, 3D liver co-cultures maintain cytochrome P450 inducibility, form bile canaliculi-like structures and respond to inflammatory stimuli. Upon incubation with selected hepatotoxicants including drugs which have been shown to induce idiosyncratic toxicity, we demonstrated that this model better detected in vivo drug-induced toxicity, including species-specific drug effects, when compared to monolayer hepatocyte cultures. In conclusion, our results underline the importance of more complex and long lasting in vitro cell culture models that contain all liver cell types and allow repeated drug-treatments for detection of in vivo-relevant adverse drug effects. - Highlights: ► 3D liver co-cultures maintain liver specific functions for up to three months. ► Activities of Cytochrome P450s remain drug- inducible accross three months. ► 3D liver co-cultures recapitulate drug-induced liver toxicity

A high throughput DNA extraction method with high yield and quality

Directory of Open Access Journals (Sweden)

Xin Zhanguo

2012-07-01

Full Text Available Abstract Background Preparation of large quantity and high quality genomic DNA from a large number of plant samples is a major bottleneck for most genetic and genomic analyses, such as, genetic mapping, TILLING (Targeting Induced Local Lesion IN Genome, and next-generation sequencing directly from sheared genomic DNA. A variety of DNA preparation methods and commercial kits are available. However, they are either low throughput, low yield, or costly. Here, we describe a method for high throughput genomic DNA isolation from sorghum [Sorghum bicolor (L. Moench] leaves and dry seeds with high yield, high quality, and affordable cost. Results We developed a high throughput DNA isolation method by combining a high yield CTAB extraction method with an improved cleanup procedure based on MagAttract kit. The method yielded large quantity and high quality DNA from both lyophilized sorghum leaves and dry seeds. The DNA yield was improved by nearly 30 fold with 4 times less consumption of MagAttract beads. The method can also be used in other plant species, including cotton leaves and pine needles. Conclusion A high throughput system for DNA extraction from sorghum leaves and seeds was developed and validated. The main advantages of the method are low cost, high yield, high quality, and high throughput. One person can process two 96-well plates in a working day at a cost of $0.10 per sample of magnetic beads plus other consumables that other methods will also need.

A versatile, high through-put, bead-based phagocytosis assay for Plasmodium falciparum

DEFF Research Database (Denmark)

Lloyd, Yukie M.; Ngati, Elise P.; Salanti, Ali

2017-01-01

Antibody-mediated phagocytosis is an important immune effector mechanism against Plasmodium falciparum-infected erythrocytes (IE); however, current phagocytosis assays use IE collected from infected individuals or from in vitro cultures of P. falciparum, making them prone to high variation....... A simple, high-throughput flow cytometric assay was developed that uses THP-1 cells and fluorescent beads covalently-coupled with the malarial antigen VAR2CSA. The assay is highly repeatable, provides both the overall percent phagocytosis and semi-quantitates the number of antigen-coupled beads...

The role of three-dimensional high-definition laparoscopic surgery for gynaecology.

Science.gov (United States)

Usta, Taner A; Gundogdu, Elif C

2015-08-01

This article reviews the potential benefits and disadvantages of new three-dimensional (3D) high-definition laparoscopic surgery for gynaecology. With the new-generation 3D high-definition laparoscopic vision systems (LVSs), operation time and learning period are reduced and procedural error margin is decreased. New-generation 3D high-definition LVSs enable to reduce operation time both for novice and experienced surgeons. Headache, eye fatigue or nausea reported with first-generation systems are not different than two-dimensional (2D) LVSs. The system's being more expensive, having the obligation to wear glasses, big and heavy camera probe in some of the devices are accounted for negative aspects of the system that need to be improved. Depth loss in tissues in 2D LVSs and associated adverse events can be eliminated with 3D high-definition LVSs. By virtue of faster learning curve, shorter operation time, reduced error margin and lack of side-effects reported by surgeons with first-generation systems, 3D LVSs seem to be a strong competition to classical laparoscopic imaging systems. Thanks to technological advancements, using lighter and smaller cameras and monitors without glasses is in the near future.

Computational tools for high-throughput discovery in biology

OpenAIRE

Jones, Neil Christopher

2007-01-01

High throughput data acquisition technology has inarguably transformed the landscape of the life sciences, in part by making possible---and necessary---the computational disciplines of bioinformatics and biomedical informatics. These fields focus primarily on developing tools for analyzing data and generating hypotheses about objects in nature, and it is in this context that we address three pressing problems in the fields of the computational life sciences which each require computing capaci...

High throughput generated micro-aggregates of chondrocytes stimulate cartilage formation in vitro and in vivo

Directory of Open Access Journals (Sweden)

LS Moreira Teixeira

2012-06-01

Full Text Available Cell-based cartilage repair strategies such as matrix-induced autologous chondrocyte implantation (MACI could be improved by enhancing cell performance. We hypothesised that micro-aggregates of chondrocytes generated in high-throughput prior to implantation in a defect could stimulate cartilaginous matrix deposition and remodelling. To address this issue, we designed a micro-mould to enable controlled high-throughput formation of micro-aggregates. Morphology, stability, gene expression profiles and chondrogenic potential of micro-aggregates of human and bovine chondrocytes were evaluated and compared to single-cells cultured in micro-wells and in 3D after encapsulation in Dextran-Tyramine (Dex-TA hydrogels in vitro and in vivo. We successfully formed micro-aggregates of human and bovine chondrocytes with highly controlled size, stability and viability within 24 hours. Micro-aggregates of 100 cells presented a superior balance in Collagen type I and Collagen type II gene expression over single cells and micro-aggregates of 50 and 200 cells. Matrix metalloproteinases 1, 9 and 13 mRNA levels were decreased in micro-aggregates compared to single-cells. Histological and biochemical analysis demonstrated enhanced matrix deposition in constructs seeded with micro-aggregates cultured in vitro and in vivo, compared to single-cell seeded constructs. Whole genome microarray analysis and single gene expression profiles using human chondrocytes confirmed increased expression of cartilage-related genes when chondrocytes were cultured in micro-aggregates. In conclusion, we succeeded in controlled high-throughput formation of micro-aggregates of chondrocytes. Compared to single cell-seeded constructs, seeding of constructs with micro-aggregates greatly improved neo-cartilage formation. Therefore, micro-aggregation prior to chondrocyte implantation in current MACI procedures, may effectively accelerate hyaline cartilage formation.

High throughput generated micro-aggregates of chondrocytes stimulate cartilage formation in vitro and in vivo.

Science.gov (United States)

Moreira Teixeira, L S; Leijten, J C H; Sobral, J; Jin, R; van Apeldoorn, A A; Feijen, J; van Blitterswijk, C; Dijkstra, P J; Karperien, M

2012-06-05

Cell-based cartilage repair strategies such as matrix-induced autologous chondrocyte implantation (MACI) could be improved by enhancing cell performance. We hypothesised that micro-aggregates of chondrocytes generated in high-throughput prior to implantation in a defect could stimulate cartilaginous matrix deposition and remodelling. To address this issue, we designed a micro-mould to enable controlled high-throughput formation of micro-aggregates. Morphology, stability, gene expression profiles and chondrogenic potential of micro-aggregates of human and bovine chondrocytes were evaluated and compared to single-cells cultured in micro-wells and in 3D after encapsulation in Dextran-Tyramine (Dex-TA) hydrogels in vitro and in vivo. We successfully formed micro-aggregates of human and bovine chondrocytes with highly controlled size, stability and viability within 24 hours. Micro-aggregates of 100 cells presented a superior balance in Collagen type I and Collagen type II gene expression over single cells and micro-aggregates of 50 and 200 cells. Matrix metalloproteinases 1, 9 and 13 mRNA levels were decreased in micro-aggregates compared to single-cells. Histological and biochemical analysis demonstrated enhanced matrix deposition in constructs seeded with micro-aggregates cultured in vitro and in vivo, compared to single-cell seeded constructs. Whole genome microarray analysis and single gene expression profiles using human chondrocytes confirmed increased expression of cartilage-related genes when chondrocytes were cultured in micro-aggregates. In conclusion, we succeeded in controlled high-throughput formation of micro-aggregates of chondrocytes. Compared to single cell-seeded constructs, seeding of constructs with micro-aggregates greatly improved neo-cartilage formation. Therefore, micro-aggregation prior to chondrocyte implantation in current MACI procedures, may effectively accelerate hyaline cartilage formation.

Preparation of three-dimensional graphene foam for high performance supercapacitors

Directory of Open Access Journals (Sweden)

Yunjie Ping

2017-04-01

Full Text Available Supercapacitor is a new type of energy-storage device, and has been attracted widely attentions. As a two dimensional (2D nanomaterials, graphene is considered to be a promising material of supercapacitor because of its excellent properties involving high electrical conductivity and large surface area. In this paper, the large-scale graphene is successfully fabricated via environmental-friendly electrochemical exfoliation of graphite, and then, the three dimensional (3D graphene foam is prepared by using nickel foam as template and FeCl3/HCl solution as etchant. Compared with the regular 2D graphene paper, the 3D graphene foam electrode shows better electrochemical performance, and exhibits the largest specific capacitance of approximately 128 F/g at the current density of 1 A/g in 6 M KOH electrolyte. It is expected that the 3D graphene foam will have a potential application in the supercapacitors.

Polycrystalline diamond detectors with three-dimensional electrodes

Energy Technology Data Exchange (ETDEWEB)

Lagomarsino, S., E-mail: lagomarsino@fi.infn.it [University of Florence, Department of Physics, Via Sansone 1, 50019 Sesto Fiorentino (Italy); INFN Firenze, Via B. Rossi 1, 50019 Sesto Fiorentino (Italy); Bellini, M. [INO-CNR Firenze, Largo E. Fermi 6, 50125 Firenze (Italy); Brianzi, M. [INFN Firenze, Via B. Rossi 1, 50019 Sesto Fiorentino (Italy); Carzino, R. [Smart Materials-Nanophysics, Istituto Italiano di Tecnologia, Genova, Via Morego 30, 16163 Genova (Italy); Cindro, V. [Joseph Stefan Institute, Jamova Cesta 39, 1000 Ljubljana (Slovenia); Corsi, C. [University of Florence, Department of Physics, Via Sansone 1, 50019 Sesto Fiorentino (Italy); LENS Firenze, Via N. Carrara 1, 50019 Sesto Fiorentino (Italy); Morozzi, A.; Passeri, D. [INFN Perugia, Perugia (Italy); Università degli Studi di Perugia, Dipartimento di Ingegneria, via G. Duranti 93, 06125 Perugia (Italy); Sciortino, S. [University of Florence, Department of Physics, Via Sansone 1, 50019 Sesto Fiorentino (Italy); INFN Firenze, Via B. Rossi 1, 50019 Sesto Fiorentino (Italy); Servoli, L. [INFN Perugia, Perugia (Italy)

2015-10-01

The three-dimensional concept in diamond detectors has been applied, so far, to high quality single-crystal material, in order to test this technology in the best available conditions. However, its application to polycrystalline chemical vapor deposited diamond could be desirable for two reasons: first, the short inter-electrode distance of three-dimensional detectors should improve the intrinsically lower collection efficiency of polycrystalline diamond, and second, at high levels of radiation damage the performances of the poly-crystal material are not expected to be much lower than those of the single crystal one. We report on the fabrication and test of three-dimensional polycrystalline diamond detectors with several inter-electrode distances, and we demonstrate that their collection efficiency is equal or higher than that obtained with conventional planar detectors fabricated with the same material. - Highlights: • Pulsed laser fabrication of polycristalline diamond detectors with 3D electrodes. • Measurement of the charge collection efficiency (CCE) under beta irradiation. • Comparation between the CCE of 3D and conventional planar diamond sensors. • A rationale for the behavior of three-dimensional and planar sensors is given.

Dielectric spectroscopy for non-invasive monitoring of epithelial cell differentiation within three-dimensional scaffolds

International Nuclear Information System (INIS)

Daoud, Jamal; Tabrizian, Maryam; Asami, Koji; Rosenberg, Lawrence

2012-01-01

In this study, we introduce a cellular differentiation cellular model based on dielectric spectroscopy that characterizes epithelial differentiation processes. Non-invasive cellular monitoring was achieved within a three-dimensional microenvironment consisting of a cell-containing collagen I gel seeded onto microfabricated scaffolds. In this proof-of-concept investigation, Madin–Darby canine kidney cells were cultured within microfabricated, geometrically controlled scaffolds and allowed us to differentiate to hollow cyst-like structures. This transformation within the three-dimensional environment is monitored and characterized through dielectric spectroscopy while maintaining cell culture in vitro. (paper)

Aluminum-based one- and two-dimensional micro fin array structures: high-throughput fabrication and heat transfer testing

International Nuclear Information System (INIS)

Primeaux, Philip A; Zhang, Bin; Zhang, Xiaoman; Miller, Jacob; Meng, W J; KC, Pratik; Moore, Arden L

2017-01-01

Microscale fin array structures were replicated onto surfaces of aluminum 1100 and aluminum 6061 alloy (Al1100/Al6061) sheet metals through room-temperature instrumented roll molding. Aluminum-based micro fin arrays were replicated at room temperature, and the fabrication process is one with high throughput and low cost. One-dimensional (1D) micro fin arrays were made through one-pass rolling, while two-dimensional (2D) micro fin arrays were made by sequential 90° cross rolling with the same roller sleeve. For roll molding of 1D micro fins, fin heights greater than 600 µ m were achieved and were shown to be proportional to the normal load force per feature width. At a given normal load force, the fin height was further shown to scale inversely with the hardness of the sheet metal. For sequential 90° cross rolling, morphologies of roll molded 2D micro fin arrays were examined, which provided clues to understand how plastic deformation occurred under cross rolling conditions. A series of pool boiling experiments on low profile Al micro fin array structures were performed within Novec 7100, a widely used commercial dielectric coolant. Results for both horizontal and vertical surface orientations show that roll molded Al micro fin arrays can increase heat flux at fixed surface temperature as compared to un-patterned Al sheet. The present results further suggest that many factors beyond just increased surface area can influence heat transfer performance, including surface finish and the important multiphase transport mechanisms in and around the fin geometry. These factors must also be considered when designing and optimizing micro fin array structures for heat transfer applications. (paper)

DockoMatic: automated peptide analog creation for high throughput virtual screening.

Science.gov (United States)

Jacob, Reed B; Bullock, Casey W; Andersen, Tim; McDougal, Owen M

2011-10-01

The purpose of this manuscript is threefold: (1) to describe an update to DockoMatic that allows the user to generate cyclic peptide analog structure files based on protein database (pdb) files, (2) to test the accuracy of the peptide analog structure generation utility, and (3) to evaluate the high throughput capacity of DockoMatic. The DockoMatic graphical user interface interfaces with the software program Treepack to create user defined peptide analogs. To validate this approach, DockoMatic produced cyclic peptide analogs were tested for three-dimensional structure consistency and binding affinity against four experimentally determined peptide structure files available in the Research Collaboratory for Structural Bioinformatics database. The peptides used to evaluate this new functionality were alpha-conotoxins ImI, PnIA, and their published analogs. Peptide analogs were generated by DockoMatic and tested for their ability to bind to X-ray crystal structure models of the acetylcholine binding protein originating from Aplysia californica. The results, consisting of more than 300 simulations, demonstrate that DockoMatic predicts the binding energy of peptide structures to within 3.5 kcal mol(-1), and the orientation of bound ligand compares to within 1.8 Å root mean square deviation for ligand structures as compared to experimental data. Evaluation of high throughput virtual screening capacity demonstrated that Dockomatic can collect, evaluate, and summarize the output of 10,000 AutoDock jobs in less than 2 hours of computational time, while 100,000 jobs requires approximately 15 hours and 1,000,000 jobs is estimated to take up to a week. Copyright © 2011 Wiley Periodicals, Inc.

Three-dimensional Magnetotelluric Characterization of the Xinzhou Geothermal Field, Southeastern China

Science.gov (United States)

Han, Q.; Hu, X.; Cai, J.; Wei, W.

2016-12-01

Xinzhou geothermal field is located in the Guangdong province and adjacent to the China South Sea, and its hot springs can reach up to 92 degree Celsius. Yanshanian granite expose widely in the south of this geothermal field and four faults cut across each other over it. A dense grid of 176 magnetotelluric (MT) sites with broadband has been acquired over the Xinzhou geothermal field and its surrounding area. Due to the related electromagnetic (EM) noise one permanent observatory was placed as a remote reference to suppress this cultural EM noise interference. The datasets are processed using the mutual reference technique, static shift correction, and structural strike and dimensionality analysis based on tensor decomposition. Data analysis reveals that the underground conductivity structure has obvious three-dimensional characterization. For the high resolution result ,two and three dimensional inversion are both applied in this area employing the non-linear conjugate gradient method (NLCG).These MT data sets are supposed to detect the deep subsurface resistivity structure correlated to the distribution of geothermal reservoir (such as faults and fractured granite) and investigate the channel of the upwelling magma. The whole and cold granite usually present high resistivity but once it functions as reservoir the resistivity will decrease, sometimes it is hard to separate the reservoir from the cap layer. The 3D inversion results delineate three high resistivity anomalies distributed in different locations. At last we put forward that the large areas of granite form the major thermal source for the study area and discuss whether any melt under these magma intrusions exists.

Three-dimensional, three-component wall-PIV

Science.gov (United States)

Berthe, André; Kondermann, Daniel; Christensen, Carolyn; Goubergrits, Leonid; Garbe, Christoph; Affeld, Klaus; Kertzscher, Ulrich

2010-06-01

This paper describes a new time-resolved three-dimensional, three-component (3D-3C) measurement technique called wall-PIV. It was developed to assess near wall flow fields and shear rates near non-planar surfaces. The method is based on light absorption according to Beer-Lambert’s law. The fluid containing a molecular dye and seeded with buoyant particles is illuminated by a monochromatic, diffuse light. Due to the dye, the depth of view is limited to the near wall layer. The three-dimensional particle positions can be reconstructed by the intensities of the particle’s projection on an image sensor. The flow estimation is performed by a new algorithm, based on learned particle trajectories. Possible sources of measurement errors related to the wall-PIV technique are analyzed. The accuracy analysis was based on single particle experiments and a three-dimensional artificial data set simulating a rotating sphere.

A method for high throughput bioelectrochemical research based on small scale microbial electrolysis cells

KAUST Repository

Call, Douglas F.

2011-07-01

There is great interest in studying exoelectrogenic microorganisms, but existing methods can require expensive electrochemical equipment and specialized reactors. We developed a simple system for conducting high throughput bioelectrochemical research using multiple inexpensive microbial electrolysis cells (MECs) built with commercially available materials and operated using a single power source. MECs were small crimp top serum bottles (5mL) with a graphite plate anode (92m 2/m 3) and a cathode of stainless steel (SS) mesh (86m 2/m 3), graphite plate, SS wire, or platinum wire. The highest volumetric current density (240A/m 3, applied potential of 0.7V) was obtained using a SS mesh cathode and a wastewater inoculum (acetate electron donor). Parallel operated MECs (single power source) did not lead to differences in performance compared to non-parallel operated MECs, which can allow for high throughput reactor operation (>1000 reactors) using a single power supply. The utility of this method for cultivating exoelectrogenic microorganisms was demonstrated through comparison of buffer effects on pure (Geobacter sulfurreducens and Geobacter metallireducens) and mixed cultures. Mixed cultures produced current densities equal to or higher than pure cultures in the different media, and current densities for all cultures were higher using a 50mM phosphate buffer than a 30mM bicarbonate buffer. Only the mixed culture was capable of sustained current generation with a 200mM phosphate buffer. These results demonstrate the usefulness of this inexpensive method for conducting in-depth examinations of pure and mixed exoelectrogenic cultures. © 2011 Elsevier B.V.

Problems of high temperature superconductivity in three-dimensional systems

Energy Technology Data Exchange (ETDEWEB)

Geilikman, B T

1973-01-01

A review is given of more recent papers on this subject. These papers have dealt mainly with two-dimensional systems. The present paper extends the treatment to three-dimensional systems, under the following headings: systems with collective electrons of one group and localized electrons of another group (compounds of metals with non-metals-dielectrics, organic substances, undoped semiconductors, molecular crystals); experimental investigations of superconducting compounds of metals with organic compounds, dielectrics, semiconductors, and semi-metals; and systems with two or more groups of collective electrons. Mechanics are considered and models are derived. 86 references.

High-Throughput Process Development for Biopharmaceuticals.

Science.gov (United States)

Shukla, Abhinav A; Rameez, Shahid; Wolfe, Leslie S; Oien, Nathan

2017-11-14

The ability to conduct multiple experiments in parallel significantly reduces the time that it takes to develop a manufacturing process for a biopharmaceutical. This is particularly significant before clinical entry, because process development and manufacturing are on the "critical path" for a drug candidate to enter clinical development. High-throughput process development (HTPD) methodologies can be similarly impactful during late-stage development, both for developing the final commercial process as well as for process characterization and scale-down validation activities that form a key component of the licensure filing package. This review examines the current state of the art for HTPD methodologies as they apply to cell culture, downstream purification, and analytical techniques. In addition, we provide a vision of how HTPD activities across all of these spaces can integrate to create a rapid process development engine that can accelerate biopharmaceutical drug development. Graphical Abstract.

Statistical removal of background signals from high-throughput 1H NMR line-broadening ligand-affinity screens

International Nuclear Information System (INIS)

Worley, Bradley; Sisco, Nicholas J.; Powers, Robert

2015-01-01

NMR ligand-affinity screens are vital to drug discovery, are routinely used to screen fragment-based libraries, and used to verify chemical leads from high-throughput assays and virtual screens. NMR ligand-affinity screens are also a highly informative first step towards identifying functional epitopes of unknown proteins, as well as elucidating the biochemical functions of protein–ligand interaction at their binding interfaces. While simple one-dimensional 1 H NMR experiments are capable of indicating binding through a change in ligand line shape, they are plagued by broad, ill-defined background signals from protein 1 H resonances. We present an uncomplicated method for subtraction of protein background in high-throughput ligand-based affinity screens, and show that its performance is maximized when phase-scatter correction is applied prior to subtraction

Three-dimensional culture conditions differentially affect astrocyte modulation of brain endothelial barrier function in response to transforming growth factor β1.

Science.gov (United States)

Hawkins, Brian T; Grego, Sonia; Sellgren, Katelyn L

2015-05-22

Blood-brain barrier (BBB) function is regulated by dynamic interactions among cell types within the neurovascular unit, including astrocytes and endothelial cells. Co-culture models of the BBB typically involve astrocytes seeded on two-dimensional (2D) surfaces, which recent studies indicate cause astrocytes to express a phenotype similar to that of reactive astrocytes in situ. We hypothesized that the culture conditions of astrocytes would differentially affect their ability to modulate BBB function in vitro. Brain endothelial cells were grown alone or in co-culture with astrocytes. Astrocytes were grown either as conventional (2D) monolayers, or in a collagen-based gel which allows them to grow in a three-dimensional (3D) construct. Astrocytes were viable in 3D conditions, and displayed a marked reduction in their expression of glial fibrillary acidic protein (GFAP), suggesting reduced activation. Stimulation of astrocytes with transforming growth factor (TGF)β1 decreased transendothelial electrical resistance (TEER) and reduced expression of claudin-5 in co-cultures, whereas treatment of endothelial cells in the absence of astrocytes was without effect. The effect of TGFβ1 on TEER was significantly more pronounced in endothelial cells cultured with 3D astrocytes compared to 2D astrocytes. These results demonstrate that astrocyte culture conditions differentially affect their ability to modulate brain endothelial barrier function, and suggest a direct relationship between reactive gliosis and BBB permeability. Moreover, these studies demonstrate the potential importance of physiologically relevant culture conditions to in vitro modeling of disease processes that affect the neurovascular unit. Copyright © 2015 Elsevier B.V. All rights reserved.

Analysis of Operating Performance and Three Dimensional Magnetic Field of High Voltage Induction Motors with Stator Chute

Directory of Open Access Journals (Sweden)

WANG Qing-shan

2017-06-01

Full Text Available In view of the difficulties on technology of rotor chute in high voltage induction motor，the desig method adopted stator chute structure is put forward． The mathematical model of three dimensional nonlinear transient field for solving stator chute in high voltage induction motor is set up． Through the three dimensional entity model of motor，three dimensional finite element method based on T，ψ － ψ electromagnetic potential is adopted for the analysis and calculation of stator chute in high voltage induction motor under rated condition． The distributions long axial of fundamental wave magnetic field and tooth harmonic wave magnetic field are analyzed after stator chute，and the weakening effects on main tooth harmonic magnetic field are researched． Further more，the comparison analysis of main performance parameters of chute and straight slot is carried out under rated condition． The results show that the electrical performance of stator chute is better than that of straight slot in high voltage induction motor，and the tooth harmonic has been sharply decreased

Reduced, three-dimensional, nonlinear equations for high-β plasmas including toroidal effects

International Nuclear Information System (INIS)

Schmalz, R.

1980-11-01

The resistive MHD equations for toroidal plasma configurations are reduced by expanding to the second order in epsilon, the inverse aspect ratio, allowing for high β = μsub(o)p/B 2 of order epsilon. The result is a closed system of nonlinear, three-dimensional equations where the fast magnetohydrodynamic time scale is eliminated. In particular, the equation for the toroidal velocity remains decoupled. (orig.)

High-throughput and automated SAXS/USAXS experiment for industrial use at BL19B2 in SPring-8

Energy Technology Data Exchange (ETDEWEB)

Osaka, Keiichi, E-mail: k-osaka@spring8.or.jp; Inoue, Daisuke; Sato, Masugu; Sano, Norimichi [Industrial Application Division, Japan Synchrotron Radiation Research Institute (JASRI/SPring-8), 1-1-1, Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5198 (Japan); Matsumoto, Takuya; Taniguchi, Yosuke [SPring-8 Service Co., Ltd., 1-20-5, Kouto, Shingu, Tatsuno, Hyogo 679-5165 (Japan)

2016-07-27

A highly automated system combining a sample transfer robot with focused SR beam has been established for small-angle and ultra small-angle X-ray scattering (SAXS/USAXS) measurement at BL19B2 for industrial use of SPring-8. High-throughput data collection system can be realized by means of X-ray beam of high photon flux density concentrated by a cylindrical mirror, and a two-dimensional pixel detector PILATUS-2M. For SAXS measurement, we can obtain high-quality data within 1 minute for one exposure using this system. The sample transfer robot has a capacity of 90 samples with a large variety of shapes. The fusion of high-throughput and robotic system has enhanced the usability of SAXS/USAXS capability for industrial application.

Micropillar arrays as a high-throughput screening platform for therapeutics in multiple sclerosis.

Science.gov (United States)

Mei, Feng; Fancy, Stephen P J; Shen, Yun-An A; Niu, Jianqin; Zhao, Chao; Presley, Bryan; Miao, Edna; Lee, Seonok; Mayoral, Sonia R; Redmond, Stephanie A; Etxeberria, Ainhoa; Xiao, Lan; Franklin, Robin J M; Green, Ari; Hauser, Stephen L; Chan, Jonah R

2014-08-01

Functional screening for compounds that promote remyelination represents a major hurdle in the development of rational therapeutics for multiple sclerosis. Screening for remyelination is problematic, as myelination requires the presence of axons. Standard methods do not resolve cell-autonomous effects and are not suited for high-throughput formats. Here we describe a binary indicant for myelination using micropillar arrays (BIMA). Engineered with conical dimensions, micropillars permit resolution of the extent and length of membrane wrapping from a single two-dimensional image. Confocal imaging acquired from the base to the tip of the pillars allows for detection of concentric wrapping observed as 'rings' of myelin. The platform is formatted in 96-well plates, amenable to semiautomated random acquisition and automated detection and quantification. Upon screening 1,000 bioactive molecules, we identified a cluster of antimuscarinic compounds that enhance oligodendrocyte differentiation and remyelination. Our findings demonstrate a new high-throughput screening platform for potential regenerative therapeutics in multiple sclerosis.

Three-dimensional morphological imaging of human induced pluripotent stem cells by using low-coherence quantitative phase microscopy

Science.gov (United States)

Yamauchi, Toyohiko; Kakuno, Yumi; Goto, Kentaro; Fukami, Tadashi; Sugiyama, Norikazu; Iwai, Hidenao; Mizuguchi, Yoshinori; Yamashita, Yutaka

2014-03-01

There is an increasing need for non-invasive imaging techniques in the field of stem cell research. Label-free techniques are the best choice for assessment of stem cells because the cells remain intact after imaging and can be used for further studies such as differentiation induction. To develop a high-resolution label-free imaging system, we have been working on a low-coherence quantitative phase microscope (LC-QPM). LC-QPM is a Linnik-type interference microscope equipped with nanometer-resolution optical-path-length control and capable of obtaining three-dimensional volumetric images. The lateral and vertical resolutions of our system are respectively 0.5 and 0.93 μm and this performance allows capturing sub-cellular morphological features of live cells without labeling. Utilizing LC-QPM, we reported on three-dimensional imaging of membrane fluctuations, dynamics of filopodia, and motions of intracellular organelles. In this presentation, we report three-dimensional morphological imaging of human induced pluripotent stem cells (hiPS cells). Two groups of monolayer hiPS cell cultures were prepared so that one group was cultured in a suitable culture medium that kept the cells undifferentiated, and the other group was cultured in a medium supplemented with retinoic acid, which forces the stem cells to differentiate. The volumetric images of the 2 groups show distinctive differences, especially in surface roughness. We believe that our LC-QPM system will prove useful in assessing many other stem cell conditions.

Cytocompatibility of calcium silicate-based sealers in a three-dimensional cell culture model.

Science.gov (United States)

da Silva, Emmanuel João Nogueira Leal; Zaia, Alexandre A; Peters, Ove A

2017-06-01

The aim of the present study was to evaluate cytotoxic effects and cytokine production of calcium silicate-based sealers (EndoSeal, EndoSequence BC Sealer, and MTA Fillapex) using an in vitro root canal filling model and three-dimensional (3D) cell culture. AH Plus as a reference was compared to contemporary calcium silicate cements regarding cell viability and cytokine production. Root canals of 30 human maxillary incisors were prepared using a single-file reciprocating technique. The samples were randomly distributed and canals filled with either AH Plus, EndoSeal, EndoSequence BC Sealer, and MTA Fillapex (n = 6). In the negative control group, the root canal remained unfilled. Sealers were placed into the canals along with a gutta-percha cone placed to working length. Balb/c 3T3 fibroblasts, cultured in a type I collagen 3D scaffold, were exposed to filling material and the respective root apex for 24 h. Cytocompatibility of the materials was evaluated using the methyl-thiazoldiphenyl-tetrazolium (MTT) assay. The production of IL-1β, IL-6, and IL-8 was analyzed using enzyme-linked immunosorbent assay (ELISA). One-way analysis of variance was performed, and when the F-ratios were significant, data were compared by Duncan's multiple-range test. The alpha-type error was set at 0.05. EndoSeal, Endosequence BC Sealer and AH Plus showed cell viability that was similar to the negative control group (P > 0.05), while MTA Fillapex sealer was cytotoxic (P culture, AH Plus, EndoSeal, and EndoSequence BC Sealer were cytocompatible. These results may suggest that AH Plus, EndoSeal and EndoSequence BC Sealer may achieve better biological response when compared to MTA Fillapex.

Three-Dimensional, Transgenic Cell Models to Quantify Space Genotoxic Effects

Science.gov (United States)

Gonda, S. R.; Sognier, M. A.; Wu, H.; Pingerelli, P. L.; Glickman, B. W.; Dawson, David L. (Technical Monitor)

1999-01-01

The space environment contains radiation and chemical agents known to be mutagenic and carcinogenic to humans. Additionally, microgravity is a complicating factor that may modify or synergize induced genotoxic effects. Most in vitro models fail to use human cells (making risk extrapolation to humans more difficult), overlook the dynamic effect of tissue intercellular interactions on genotoxic damage, and lack the sensitivity required to measure low-dose effects. Currently a need exists for a model test system that simulates cellular interactions present in tissue, and can be used to quantify genotoxic damage induced by low levels of radiation and chemicals, and extrapolate assessed risk to humans. A state-of-the-art, three-dimensional, multicellular tissue equivalent cell culture model will be presented. It consists of mammalian cells genetically engineered to contain multiple copies of defined target genes for genotoxic assessment,. NASA-designed bioreactors were used to coculture mammalian cells into spheroids, The cells used were human mammary epithelial cells (H184135) and Stratagene's (Austin, Texas) Big Blue(TM) Rat 2 lambda fibroblasts. The fibroblasts were genetically engineered to contain -a high-density target gene for mutagenesis (60 copies of lacl/LacZ per cell). Tissue equivalent spheroids were routinely produced by inoculation of 2 to 7 X 10(exp 5) fibroblasts with Cytodex 3 beads (150 micrometers in diameter). at a 20:1 cell:bead ratio, into 50-ml HARV bioreactors (Synthecon, Inc.). Fibroblasts were cultured for 5 days, an equivalent number of epithelial cells added, and the fibroblast/epithelial cell coculture continued for 21 days. Three-dimensional spheroids with diameters ranging from 400 to 600 micrometers were obtained. Histological and immunohistochemical Characterization revealed i) both cell types present in the spheroids, with fibroblasts located primarily in the center, surrounded by epithelial cells; ii) synthesis of extracellular matrix

Accurate Classification of Protein Subcellular Localization from High-Throughput Microscopy Images Using Deep Learning

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Tanel Pärnamaa

2017-05-01

Full Text Available High-throughput microscopy of many single cells generates high-dimensional data that are far from straightforward to analyze. One important problem is automatically detecting the cellular compartment where a fluorescently-tagged protein resides, a task relatively simple for an experienced human, but difficult to automate on a computer. Here, we train an 11-layer neural network on data from mapping thousands of yeast proteins, achieving per cell localization classification accuracy of 91%, and per protein accuracy of 99% on held-out images. We confirm that low-level network features correspond to basic image characteristics, while deeper layers separate localization classes. Using this network as a feature calculator, we train standard classifiers that assign proteins to previously unseen compartments after observing only a small number of training examples. Our results are the most accurate subcellular localization classifications to date, and demonstrate the usefulness of deep learning for high-throughput microscopy.

Accurate Classification of Protein Subcellular Localization from High-Throughput Microscopy Images Using Deep Learning.

Science.gov (United States)

Pärnamaa, Tanel; Parts, Leopold

2017-05-05

High-throughput microscopy of many single cells generates high-dimensional data that are far from straightforward to analyze. One important problem is automatically detecting the cellular compartment where a fluorescently-tagged protein resides, a task relatively simple for an experienced human, but difficult to automate on a computer. Here, we train an 11-layer neural network on data from mapping thousands of yeast proteins, achieving per cell localization classification accuracy of 91%, and per protein accuracy of 99% on held-out images. We confirm that low-level network features correspond to basic image characteristics, while deeper layers separate localization classes. Using this network as a feature calculator, we train standard classifiers that assign proteins to previously unseen compartments after observing only a small number of training examples. Our results are the most accurate subcellular localization classifications to date, and demonstrate the usefulness of deep learning for high-throughput microscopy. Copyright © 2017 Parnamaa and Parts.

High Throughput PBTK: Open-Source Data and Tools for ...

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Presentation on High Throughput PBTK at the PBK Modelling in Risk Assessment meeting in Ispra, Italy Presentation on High Throughput PBTK at the PBK Modelling in Risk Assessment meeting in Ispra, Italy

Three Dimensional Dirac Semimetals

Science.gov (United States)

Zaheer, Saad

2014-03-01

Dirac points on the Fermi surface of two dimensional graphene are responsible for its unique electronic behavior. One can ask whether any three dimensional materials support similar pseudorelativistic physics in their bulk electronic spectra. This possibility has been investigated theoretically and is now supported by two successful experimental demonstrations reported during the last year. In this talk, I will summarize the various ways in which Dirac semimetals can be realized in three dimensions with primary focus on a specific theory developed on the basis of representations of crystal spacegroups. A three dimensional Dirac (Weyl) semimetal can appear in the presence (absence) of inversion symmetry by tuning parameters to the phase boundary separating a bulk insulating and a topological insulating phase. More generally, we find that specific rules governing crystal symmetry representations of electrons with spin lead to robust Dirac points at high symmetry points in the Brillouin zone. Combining these rules with microscopic considerations identifies six candidate Dirac semimetals. Another method towards engineering Dirac semimetals involves combining crystal symmetry and band inversion. Several candidate materials have been proposed utilizing this mechanism and one of the candidates has been successfully demonstrated as a Dirac semimetal in two independent experiments. Work carried out in collaboration with: Julia A. Steinberg, Steve M. Young, J.C.Y. Teo, C.L. Kane, E.J. Mele and Andrew M. Rappe.

Ultrastructural changes following electron irradiation in three-dimensional culture of normal human dermal fibroblasts

International Nuclear Information System (INIS)

Han, Chunmao; Ishikura, Naotaka; Tsukada, Sadao

1994-01-01

The present study was designed to examine the effect of electron irradiation on fibroblasts and extracellular matrices electron-microscopically. The three-dimensional dermal fibroblast culture was exposed to one, 4 or 10 Gy of electron beams. One day after irradiation, fibroblasts were vacuolated in all irradiated groups and intercellular spaces were increased in a dose-dependent manner. Seven days later, intercellular spaces became dense in both one and 4 Gy groups, although they were still extremely increased in the 10 Gy group. The remaining fibroblasts were still activated in all groups. Thirty days after irradiation, myofibroblastic cells were scarcely observed, but extracellular fine fibrils and collagen fibrils were observed in all irradiated groups. The other ultrastructural findings were similar to those in the control group. In conclusion, electron beams damaged not only cells but also extracellular matrix. The extracellular matrix may be repaired by activated residual fibroblasts, resulting in the mixture of new and old collagen fibrils having different diamters. (N.K.)

Three-dimensional effects in fracture mechanics

International Nuclear Information System (INIS)

Benitez, F.G.

1991-01-01

An overall view of the pioneering theories and works, which enlighten the three-dimensional nature of fracture mechanics during the last years is given. the main aim is not an exhaustive reviewing but the displaying of the last developments on this scientific field in a natural way. This work attempts to envisage the limits of disregarding the three-dimensional behaviour in theories, analyses and experiments. Moreover, it tries to draw attention on the scant fervour, although increasing, this three-dimensional nature of fracture has among the scientific community. Finally, a constructive discussion is presented on the use of two-dimensional solutions in the analysis of geometries which bear a three-dimensional configuration. the static two-dimensional solutions and its applications fields are reviewed. also, the static three-dimensional solutions, wherein a comparative analysis with elastoplastic and elastostatic solutions are presented. to end up, the dynamic three-dimensional solutions are compared to the asymptotic two-dimensional ones under the practical applications point of view. (author)

[Ideas and methods of two-dimensional zebrafish model combined with chromatographic techniques in high-throughput screening of active anti-osteoporosis components of traditional Chinese medicines].

Science.gov (United States)

Wei, Ying-Jie; Jing, Li-Jun; Zhan, Yang; Sun, E; Jia, Xiao-Bin

2014-05-01

To break through the restrictions of the evaluation model and the quantity of compounds by using the two-dimensional zebrafish model combined with chromatographic techniques, and establish a new method for the high-throughput screening of active anti-osteoporosis components. According to the research group-related studies and relevant foreign literatures, on the basis of the fact that the zebrafish osteoporosis model could efficiently evaluate the activity, the zebrafish metabolism model could efficiently enrich metabolites and the chromatographic techniques could efficiently separate and analyze components of traditional Chinese medicines, we proposed that the inherent combination of the three methods is expected to efficiently decode in vivo and in vitro efficacious anti-osteoporosis materials of traditional Chinese medicines. The method makes it simple and efficient in the enrichment, separation and analysis on components of traditional Chinese medicines, particularly micro-components and metabolites and the screening anti-osteoporosis activity, fully reflects that efficacious materials of traditional Chinese medicines contain original components and metabolites, with characteristic of "multi-components, multi-targets and integral effect", which provides new ideas and methods for the early and rapid discovery of active anti-osteoporosis components of traditional Chinese medicines.

Three-dimensional epithelial tissues generated from human embryonic stem cells.

Science.gov (United States)

Hewitt, Kyle J; Shamis, Yulia; Carlson, Mark W; Aberdam, Edith; Aberdam, Daniel; Garlick, Jonathan A

2009-11-01

The use of pluripotent human embryonic stem (hES) cells for tissue engineering may provide advantages over traditional sources of progenitor cells because of their ability to give rise to multiple cell types and their unlimited expansion potential. We derived cell populations with properties of ectodermal and mesenchymal cells in two-dimensional culture and incorporated these divergent cell populations into three-dimensional (3D) epithelial tissues. When grown in specific media and substrate conditions, two-dimensional cultures were enriched in cells (EDK1) with mesenchymal morphology and surface markers. Cells with a distinct epithelial morphology (HDE1) that expressed cytokeratin 12 and beta-catenin at cell junctions became the predominant cell type when EDK1 were grown on surfaces enriched in keratinocyte-derived extracellular matrix proteins. When these cells were incorporated into the stromal and epithelial tissue compartments of 3D tissues, they generated multilayer epithelia similar to those generated with foreskin-derived epithelium and fibroblasts. Three-dimensional tissues demonstrated stromal cells with morphologic features of mature fibroblasts, type IV collagen deposition in the basement membrane, and a stratified epithelium that expressed cytokeratin 12. By deriving two distinct cell lineages from a common hES cell source to fabricate complex tissues, it is possible to explore environmental cues that will direct hES-derived cells toward optimal tissue form and function.

Quantifying the correlation between spatially defined oxygen gradients and cell fate in an engineered three-dimensional culture model.

Science.gov (United States)

Ardakani, Amir G; Cheema, Umber; Brown, Robert A; Shipley, Rebecca J

2014-09-06

A challenge in three-dimensional tissue culture remains the lack of quantitative information linking nutrient delivery and cellular distribution. Both in vivo and in vitro, oxygen is delivered by diffusion from its source (blood vessel or the construct margins). The oxygen level at a defined distance from its source depends critically on the balance of diffusion and cellular metabolism. Cells may respond to this oxygen environment through proliferation, death and chemotaxis, resulting in spatially resolved gradients in cellular density. This study extracts novel spatially resolved and simultaneous data on tissue oxygenation, cellular proliferation, viability and chemotaxis in three-dimensional spiralled, cellular collagen constructs. Oxygen concentration gradients drove preferential cellular proliferation rates and viability in the higher oxygen zones and induced chemotaxis along the spiral of the collagen construct; an oxygen gradient of 1.03 mmHg mm(-1) in the spiral direction induced a mean migratory speed of 1015 μm day(-1). Although this movement was modest, it was effective in balancing the system to a stable cell density distribution, and provided insights into the natural cell mechanism for adapting cell number and activity to a prevailing oxygen regime.

Parameters in three-dimensional osteospheroids of telomerized human mesenchymal (stromal) stem cells grown on osteoconductive scaffolds that predict in vivo bone-forming potential

DEFF Research Database (Denmark)

Burns, Jorge S; Hansen, Pernille Lund; Larsen, Kenneth H

2010-01-01

Osteoblastic differentiation of human mesenchymal stem cells (hMSC) in monolayer culture is artefactual, lacking an organized bone-like matrix. We present a highly reproducible microwell protocol generating three-dimensional ex vivo multicellular aggregates of telomerized hMSC (hMSC-telomerase re......Osteoblastic differentiation of human mesenchymal stem cells (hMSC) in monolayer culture is artefactual, lacking an organized bone-like matrix. We present a highly reproducible microwell protocol generating three-dimensional ex vivo multicellular aggregates of telomerized hMSC (h......, was deposited in the scaffold concavities. Here, mature osteoblasts stained positively for differentiated osteoblast markers TAZ, biglycan, osteocalcin, and phospho-AKT. Quantification of collagen birefringence and relatively high expression of genes for matrix proteins, including type I collagen, biglycan...

Novel high-throughput cell-based hybridoma screening methodology using the Celigo Image Cytometer.

Science.gov (United States)

Zhang, Haohai; Chan, Leo Li-Ying; Rice, William; Kassam, Nasim; Longhi, Maria Serena; Zhao, Haitao; Robson, Simon C; Gao, Wenda; Wu, Yan

2017-08-01

Hybridoma screening is a critical step for antibody discovery, which necessitates prompt identification of potential clones from hundreds to thousands of hybridoma cultures against the desired immunogen. Technical issues associated with ELISA- and flow cytometry-based screening limit accuracy and diminish high-throughput capability, increasing time and cost. Conventional ELISA screening with coated antigen is also impractical for difficult-to-express hydrophobic membrane antigens or multi-chain protein complexes. Here, we demonstrate novel high-throughput screening methodology employing the Celigo Image Cytometer, which avoids nonspecific signals by contrasting antibody binding signals directly on living cells, with and without recombinant antigen expression. The image cytometry-based high-throughput screening method was optimized by detecting the binding of hybridoma supernatants to the recombinant antigen CD39 expressed on Chinese hamster ovary (CHO) cells. Next, the sensitivity of the image cytometer was demonstrated by serial dilution of purified CD39 antibody. Celigo was used to measure antibody affinities of commercial and in-house antibodies to membrane-bound CD39. This cell-based screening procedure can be completely accomplished within one day, significantly improving throughput and efficiency of hybridoma screening. Furthermore, measuring direct antibody binding to living cells eliminated both false positive and false negative hits. The image cytometry method was highly sensitive and versatile, and could detect positive antibody in supernatants at concentrations as low as ~5ng/mL, with concurrent K d binding affinity coefficient determination. We propose that this screening method will greatly facilitate antibody discovery and screening technologies. Copyright © 2017 Elsevier B.V. All rights reserved.

Poly(2-oxazoline) hydrogels as next generation three-dimensional cell supports

Science.gov (United States)

Dargaville, Tim R; Hollier, Brett G; Shokoohmand, Ali; Hoogenboom, Richard

2014-01-01

Synthetic hydrogels selectively decorated with cell adhesion motifs are rapidly emerging as promising substrates for 3D cell culture. When cells are grown in 3D they experience potentially more physiologically relevant cell–cell interactions and physical cues compared with traditional 2D cell culture on stiff surfaces. A newly developed polymer based on poly(2-oxazoline)s has been used for the first time to control attachment of fibroblast cells and is discussed here for its potential use in 3D cell culture with particular focus on cancer cells toward the ultimate aim of high-throughput screening of anticancer therapies. Advantages and limitations of using poly(2-oxazoline) hydrogels are discussed and compared with more established polymers, especially polyethylene glycol (PEG). PMID:24714592

High throughput experimentation for the discovery of new catalysts

International Nuclear Information System (INIS)

Thomson, S.; Hoffmann, C.; Johann, T.; Wolf, A.; Schmidt, H.-W.; Farrusseng, D.; Schueth, F.

2002-01-01

Full text: The use of combinatorial chemistry to obtain new materials has been developed extensively by the pharmaceutical and biochemical industries, but such approaches have been slow to impact on the field of heterogeneous catalysis. The reasons for this lie in with difficulties associated in the synthesis, characterisation and determination of catalytic properties of such materials. In many synthetic and catalytic reactions, the conditions used are difficult to emulate using High Throughput Experimentation (HTE). Furthermore, the ability to screen these catalysts simultaneously in real time, requires the development and/or modification of characterisation methods. Clearly, there is a need for both high throughput synthesis and screening of new and novel reactions, and we describe several new concepts that help to achieve these goals. Although such problems have impeded the development of combinatorial catalysis, the fact remains that many highly attractive processes still exist for which no suitable catalysts have been developed. The ability to decrease the tiFme needed to evaluate catalyst is therefore essential and this makes the use of high throughput techniques highly desirable. In this presentation we will describe the synthesis, catalytic testing, and novel screening methods developed at the Max Planck Institute. Automated synthesis procedures, performed by the use of a modified Gilson pipette robot, will be described, as will the development of two 16 and 49 sample fixed bed reactors and two 25 and 29 sample three phase reactors for catalytic testing. We will also present new techniques for the characterisation of catalysts and catalytic products using standard IR microscopy and infrared focal plane array detection, respectively

Three-dimensionally spiral structure of the water stream induced by a centrifugal stirrer in large aqua-cultural ponds

Science.gov (United States)

Itano, Tomoaki; Inagaki, Taishi; Nakamura, Choji; Sugihara-Seki, Masako; Hyodo, Jinsuke

2017-11-01

We have conducted measurements of the water stream produced by a mechanical stirrer (diameter 2.4[m], electric power 50[W]) located in shallow rectangular reservoirs (small 0.7[ha], large 3.7[ha]), which may be employed as a cost-efficient aerator for the aqua-cultural purpose, with the aid of both particle tracking velocimetry by passive tracers floating on the surface and direct measurement by electro-magnetic velocimeter under the surface. The present measurements indicate that the stirrer drives primarily the horizontally rotating water stream and secondarily the vertical convection between the surface and the bottom of the reservoir, which results in the three-dimensionally spiral-shaped water streams scaled vertically by just a meter but horizontally by more than ten meters. It is suggested that the spiral structure driven by the stirrer may activate the underwater vertical mixing and enhance dissolved oxygen at the bottom of aqua-cultural pond more effectively than the paddle-wheel aerators commonly used in aqua-cultural ponds. This research was financially supported in part by the Kansai University Fund for Supporting Young Scholars, 2016-2017.

Throughput Analysis on 3-Dimensional Underwater Acoustic Network with One-Hop Mobile Relay

Science.gov (United States)

Zhong, Xuefeng; Fan, Jiasheng; Guan, Quansheng; Ji, Fei; Yu, Hua

2018-01-01

Underwater acoustic communication network (UACN) has been considered as an essential infrastructure for ocean exploitation. Performance analysis of UACN is important in underwater acoustic network deployment and management. In this paper, we analyze the network throughput of three-dimensional randomly deployed transmitter–receiver pairs. Due to the long delay of acoustic channels, complicated networking protocols with heavy signaling overhead may not be appropriate. In this paper, we consider only one-hop or two-hop transmission, to save the signaling cost. That is, we assume the transmitter sends the data packet to the receiver by one-hop direct transmission, or by two-hop transmission via mobile relays. We derive the closed-form formulation of packet delivery rate with respect to the transmission delay and the number of transmitter–receiver pairs. The correctness of the derivation results are verified by computer simulations. Our analysis indicates how to obtain a precise tradeoff between the delay constraint and the network capacity. PMID:29337911

Throughput Analysis on 3-Dimensional Underwater Acoustic Network with One-Hop Mobile Relay.

Science.gov (United States)

Zhong, Xuefeng; Chen, Fangjiong; Fan, Jiasheng; Guan, Quansheng; Ji, Fei; Yu, Hua

2018-01-16

Underwater acoustic communication network (UACN) has been considered as an essential infrastructure for ocean exploitation. Performance analysis of UACN is important in underwater acoustic network deployment and management. In this paper, we analyze the network throughput of three-dimensional randomly deployed transmitter-receiver pairs. Due to the long delay of acoustic channels, complicated networking protocols with heavy signaling overhead may not be appropriate. In this paper, we consider only one-hop or two-hop transmission, to save the signaling cost. That is, we assume the transmitter sends the data packet to the receiver by one-hop direct transmission, or by two-hop transmission via mobile relays. We derive the closed-form formulation of packet delivery rate with respect to the transmission delay and the number of transmitter-receiver pairs. The correctness of the derivation results are verified by computer simulations. Our analysis indicates how to obtain a precise tradeoff between the delay constraint and the network capacity.

Three-dimensional iron sulfide-carbon interlocked graphene composites for high-performance sodium-ion storage

DEFF Research Database (Denmark)

Huang, Wei; Sun, Hongyu; Shangguan, Huihui

2018-01-01

Three-dimensional (3D) carbon-wrapped iron sulfide interlocked graphene (Fe7S8@C-G) composites for high-performance sodium-ion storage are designed and produced through electrostatic interactions and subsequent sulfurization. The iron-based metal–organic frameworks (MOFs, MIL-88-Fe) interact with...

High throughput screening method for assessing heterogeneity of microorganisms

NARCIS (Netherlands)

Ingham, C.J.; Sprenkels, A.J.; van Hylckama Vlieg, J.E.T.; Bomer, Johan G.; de Vos, W.M.; van den Berg, Albert

2006-01-01

The invention relates to the field of microbiology. Provided is a method which is particularly powerful for High Throughput Screening (HTS) purposes. More specific a high throughput method for determining heterogeneity or interactions of microorganisms is provided.

Investigating Hydrocarbon Seep Environments with High-Resolution, Three-Dimensional Geographic Visualizations.

Science.gov (United States)

Doolittle, D. F.; Gharib, J. J.; Mitchell, G. A.

2015-12-01

Detailed photographic imagery and bathymetric maps of the seafloor acquired by deep submergence vehicles such as Autonomous Underwater Vehicles (AUV) and Remotely Operated Vehicles (ROV) are expanding how scientists and the public view and ultimately understand the seafloor and the processes that modify it. Several recently acquired optical and acoustic datasets, collected during ECOGIG (Ecosystem Impacts of Oil and Gas Inputs to the Gulf) and other Gulf of Mexico expeditions using the National Institute for Undersea Science Technology (NIUST) Eagle Ray, and Mola Mola AUVs, have been fused with lower resolution data to create unique three-dimensional geovisualizations. Included in these data are multi-scale and multi-resolution visualizations over hydrocarbon seeps and seep related features. Resolution of the data range from 10s of mm to 10s of m. When multi-resolution data is integrated into a single three-dimensional visual environment, new insights into seafloor and seep processes can be obtained from the intuitive nature of three-dimensional data exploration. We provide examples and demonstrate how integration of multibeam bathymetry, seafloor backscatter data, sub-bottom profiler data, textured photomosaics, and hull-mounted multibeam acoustic midwater imagery are made into a series a three-dimensional geovisualizations of actively seeping sites and associated chemosynthetic communities. From these combined and merged datasets, insights on seep community structure, morphology, ecology, fluid migration dynamics, and process geomorphology can be investigated from new spatial perspectives. Such datasets also promote valuable inter-comparisons of sensor resolution and performance.

High Dimensional Classification Using Features Annealed Independence Rules.

Science.gov (United States)

Fan, Jianqing; Fan, Yingying

2008-01-01

Classification using high-dimensional features arises frequently in many contemporary statistical studies such as tumor classification using microarray or other high-throughput data. The impact of dimensionality on classifications is largely poorly understood. In a seminal paper, Bickel and Levina (2004) show that the Fisher discriminant performs poorly due to diverging spectra and they propose to use the independence rule to overcome the problem. We first demonstrate that even for the independence classification rule, classification using all the features can be as bad as the random guessing due to noise accumulation in estimating population centroids in high-dimensional feature space. In fact, we demonstrate further that almost all linear discriminants can perform as bad as the random guessing. Thus, it is paramountly important to select a subset of important features for high-dimensional classification, resulting in Features Annealed Independence Rules (FAIR). The conditions under which all the important features can be selected by the two-sample t-statistic are established. The choice of the optimal number of features, or equivalently, the threshold value of the test statistics are proposed based on an upper bound of the classification error. Simulation studies and real data analysis support our theoretical results and demonstrate convincingly the advantage of our new classification procedure.

Three-Dimensional Digital Documentation of Heritage Sites Using Terrestrial Laser Scanning and Unmanned Aerial Vehicle Photogrammetry

Science.gov (United States)

Jo, Y. H.; Kim, J. Y.

2017-08-01

Three-dimensional digital documentation is an important technique for the maintenance and monitoring of cultural heritage sites. This study focuses on the three-dimensional digital documentation of the Magoksa Temple, Republic of Korea, using a combination of terrestrial laser scanning and unmanned aerial vehicle (UAV) photogrammetry. Terrestrial laser scanning mostly acquired the vertical geometry of the buildings. In addition, the digital orthoimage produced by UAV photogrammetry had higher horizontal data acquisition rate than that produced by terrestrial laser scanning. Thus, the scanning and UAV photogrammetry were merged by matching 20 corresponding points and an absolute coordinate system was established using seven ground control points. The final, complete threedimensional shape had perfect horizontal and vertical geometries. This study demonstrates the potential of integrating terrestrial laser scanning and UAV photogrammetry for three-dimensional digital documentation. This new technique is expected to contribute to the three-dimensional digital documentation and spatial analysis of cultural heritage sites.

Three-dimensional interconnected porous graphitic carbon derived from rice straw for high performance supercapacitors

Science.gov (United States)

Jin, Hong; Hu, Jingpeng; Wu, Shichao; Wang, Xiaolan; Zhang, Hui; Xu, Hui; Lian, Kun

2018-04-01

Three-dimensional interconnected porous graphitic carbon materials are synthesized via a combination of graphitization and activation process with rice straw as the carbon source. The physicochemical properties of the three-dimensional interconnected porous graphitic carbon materials are characterized by Nitrogen adsorption/desorption, Fourier-transform infrared spectroscopy, X-ray diffraction, Raman spectroscopy, Scanning electron microscopy and Transmission electron microscopy. The results demonstrate that the as-prepared carbon is a high surface area carbon material (a specific surface area of 3333 m2 g-1 with abundant mesoporous and microporous structures). And it exhibits superb performance in symmetric double layer capacitors with a high specific capacitance of 400 F g-1 at a current density of 0.1 A g-1, good rate performance with 312 F g-1 under a current density of 5 A g-1 and favorable cycle stability with 6.4% loss after 10000 cycles at a current density of 5 A g-1 in the aqueous electrolyte of 6M KOH. Thus, rice straw is a promising carbon source for fabricating inexpensive, sustainable and high performance supercapacitors' electrode materials.

Three-dimensional biomedical imaging

International Nuclear Information System (INIS)

Robb, R.A.

1985-01-01

Scientists in biomedical imaging provide researchers, physicians, and academicians with an understanding of the fundamental theories and practical applications of three-dimensional biomedical imaging methodologies. Succinct descriptions of each imaging modality are supported by numerous diagrams and illustrations which clarify important concepts and demonstrate system performance in a variety of applications. Comparison of the different functional attributes, relative advantages and limitations, complementary capabilities, and future directions of three-dimensional biomedical imaging modalities are given. Volume 1: Introductions to Three-Dimensional Biomedical Imaging Photoelectronic-Digital Imaging for Diagnostic Radiology. X-Ray Computed Tomography - Basic Principles. X-Ray Computed Tomography - Implementation and Applications. X-Ray Computed Tomography: Advanced Systems and Applications in Biomedical Research and Diagnosis. Volume II: Single Photon Emission Computed Tomography. Position Emission Tomography (PET). Computerized Ultrasound Tomography. Fundamentals of NMR Imaging. Display of Multi-Dimensional Biomedical Image Information. Summary and Prognostications

Three-dimensional bioprinting of embryonic stem cells directs highly uniform embryoid body formation

International Nuclear Information System (INIS)

Ouyang, Liliang; Yao, Rui; Mao, Shuangshuang; Sun, Wei; Chen, Xi; Na, Jie

2015-01-01

With the ability to manipulate cells temporarily and spatially into three-dimensional (3D) tissue-like construct, 3D bioprinting technology was used in many studies to facilitate the recreation of complex cell niche and/or to better understand the regulation of stem cell proliferation and differentiation by cellular microenvironment factors. Embryonic stem cells (ESCs) have the capacity to differentiate into any specialized cell type of the animal body, generally via the formation of embryoid body (EB), which mimics the early stages of embryogenesis. In this study, extrusion-based 3D bioprinting technology was utilized for biofabricating ESCs into 3D cell-laden construct. The influence of 3D printing parameters on ESC viability, proliferation, maintenance of pluripotency and the rule of EB formation was systematically studied in this work. Results demonstrated that ESCs were successfully printed with hydrogel into 3D macroporous construct. Upon process optimization, about 90% ESCs remained alive after the process of bioprinting and cell-laden construct formation. ESCs continued proliferating into spheroid EBs in the hydrogel construct, while retaining the protein expression and gene expression of pluripotent markers, like octamer binding transcription factor 4, stage specific embryonic antigen 1 and Nanog. In this novel technology, EBs were formed through cell proliferation instead of aggregation, and the quantity of EBs was tuned by the initial cell density in the 3D bioprinting process. This study introduces the 3D bioprinting of ESCs into a 3D cell-laden hydrogel construct for the first time and showed the production of uniform, pluripotent, high-throughput and size-controllable EBs, which indicated strong potential in ESC large scale expansion, stem cell regulation and fabrication of tissue-like structure and drug screening studies. (paper)

Three-dimensional bioprinting of embryonic stem cells directs highly uniform embryoid body formation.

Science.gov (United States)

Ouyang, Liliang; Yao, Rui; Mao, Shuangshuang; Chen, Xi; Na, Jie; Sun, Wei

2015-11-04

With the ability to manipulate cells temporarily and spatially into three-dimensional (3D) tissue-like construct, 3D bioprinting technology was used in many studies to facilitate the recreation of complex cell niche and/or to better understand the regulation of stem cell proliferation and differentiation by cellular microenvironment factors. Embryonic stem cells (ESCs) have the capacity to differentiate into any specialized cell type of the animal body, generally via the formation of embryoid body (EB), which mimics the early stages of embryogenesis. In this study, extrusion-based 3D bioprinting technology was utilized for biofabricating ESCs into 3D cell-laden construct. The influence of 3D printing parameters on ESC viability, proliferation, maintenance of pluripotency and the rule of EB formation was systematically studied in this work. Results demonstrated that ESCs were successfully printed with hydrogel into 3D macroporous construct. Upon process optimization, about 90% ESCs remained alive after the process of bioprinting and cell-laden construct formation. ESCs continued proliferating into spheroid EBs in the hydrogel construct, while retaining the protein expression and gene expression of pluripotent markers, like octamer binding transcription factor 4, stage specific embryonic antigen 1 and Nanog. In this novel technology, EBs were formed through cell proliferation instead of aggregation, and the quantity of EBs was tuned by the initial cell density in the 3D bioprinting process. This study introduces the 3D bioprinting of ESCs into a 3D cell-laden hydrogel construct for the first time and showed the production of uniform, pluripotent, high-throughput and size-controllable EBs, which indicated strong potential in ESC large scale expansion, stem cell regulation and fabrication of tissue-like structure and drug screening studies.

Three-dimensional neuroimaging

International Nuclear Information System (INIS)

Toga, A.W.

1990-01-01

This book reports on new neuroimaging technologies that are revolutionizing the study of the brain be enabling investigators to visualize its structure and entire pattern of functional activity in three dimensions. The book provides a theoretical and practical explanation of the new science of creating three-dimensional computer images of the brain. The coverage includes a review of the technology and methodology of neuroimaging, the instrumentation and procedures, issues of quantification, analytic protocols, and descriptions of neuroimaging systems. Examples are given to illustrate the use of three-dimensional enuroimaging to quantitate spatial measurements, perform analysis of autoradiographic and histological studies, and study the relationship between brain structure and function

Investigating the Role of Surface Materials and Three Dimensional Architecture on In Vitro Differentiation of Porcine Monocyte-Derived Dendritic Cells.

Science.gov (United States)

Hartmann, Sofie Bruun; Mohanty, Soumyaranjan; Skovgaard, Kerstin; Brogaard, Louise; Flagstad, Frederikke Bjergvang; Emnéus, Jenny; Wolff, Anders; Summerfield, Artur; Jungersen, Gregers

2016-01-01

In vitro generation of dendritic-like cells through differentiation of peripheral blood monocytes is typically done using two-dimensional polystyrene culture plates. In the process of optimising cell culture techniques, engineers have developed fluidic micro-devises usually manufactured in materials other than polystyrene and applying three-dimensional structures more similar to the in vivo environment. Polydimethylsiloxane (PDMS) is an often used polymer for lab-on-a-chip devices but not much is known about the effect of changing the culture surface material from polystyrene to PDMS. In the present study the differentiation of porcine monocytes to monocyte-derived dendritic cells (moDCs) was investigated using CD172apos pig blood monocytes stimulated with GM-CSF and IL-4. Monocytes were cultured on surfaces made of two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS and carbonised three-dimensional PDMS. Cells cultured conventionally (on two-dimensional polystyrene) differentiated into moDCs as expected. Interestingly, gene expression of a wide range of cytokines, chemokines, and pattern recognition receptors was influenced by culture surface material and architecture. Distinct clustering of cells, based on similar expression patterns of 46 genes of interest, was seen for cells isolated from two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS. Changing the material from polystyrene to PDMS resulted in cells with expression patterns usually associated with macrophage expression (upregulation of CD163 and downregulation of CD1a, FLT3, LAMP3 and BATF3). However, this was purely based on gene expression level, and no functional assays were included in this study which would be necessary in order to classify the cells as being macrophages. When changing to three-dimensional culture the cells became increasingly activated in terms of IL6, IL8, IL10 and CCR5 gene expression. Further stimulation with LPS resulted

Investigating the Role of Surface Materials and Three Dimensional Architecture on In Vitro Differentiation of Porcine Monocyte-Derived Dendritic Cells.

Directory of Open Access Journals (Sweden)

Sofie Bruun Hartmann

Full Text Available In vitro generation of dendritic-like cells through differentiation of peripheral blood monocytes is typically done using two-dimensional polystyrene culture plates. In the process of optimising cell culture techniques, engineers have developed fluidic micro-devises usually manufactured in materials other than polystyrene and applying three-dimensional structures more similar to the in vivo environment. Polydimethylsiloxane (PDMS is an often used polymer for lab-on-a-chip devices but not much is known about the effect of changing the culture surface material from polystyrene to PDMS. In the present study the differentiation of porcine monocytes to monocyte-derived dendritic cells (moDCs was investigated using CD172apos pig blood monocytes stimulated with GM-CSF and IL-4. Monocytes were cultured on surfaces made of two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS and carbonised three-dimensional PDMS. Cells cultured conventionally (on two-dimensional polystyrene differentiated into moDCs as expected. Interestingly, gene expression of a wide range of cytokines, chemokines, and pattern recognition receptors was influenced by culture surface material and architecture. Distinct clustering of cells, based on similar expression patterns of 46 genes of interest, was seen for cells isolated from two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS. Changing the material from polystyrene to PDMS resulted in cells with expression patterns usually associated with macrophage expression (upregulation of CD163 and downregulation of CD1a, FLT3, LAMP3 and BATF3. However, this was purely based on gene expression level, and no functional assays were included in this study which would be necessary in order to classify the cells as being macrophages. When changing to three-dimensional culture the cells became increasingly activated in terms of IL6, IL8, IL10 and CCR5 gene expression. Further stimulation

High-throughput sample adaptive offset hardware architecture for high-efficiency video coding

Science.gov (United States)

Zhou, Wei; Yan, Chang; Zhang, Jingzhi; Zhou, Xin

2018-03-01

A high-throughput hardware architecture for a sample adaptive offset (SAO) filter in the high-efficiency video coding video coding standard is presented. First, an implementation-friendly and simplified bitrate estimation method of rate-distortion cost calculation is proposed to reduce the computational complexity in the mode decision of SAO. Then, a high-throughput VLSI architecture for SAO is presented based on the proposed bitrate estimation method. Furthermore, multiparallel VLSI architecture for in-loop filters, which integrates both deblocking filter and SAO filter, is proposed. Six parallel strategies are applied in the proposed in-loop filters architecture to improve the system throughput and filtering speed. Experimental results show that the proposed in-loop filters architecture can achieve up to 48% higher throughput in comparison with prior work. The proposed architecture can reach a high-operating clock frequency of 297 MHz with TSMC 65-nm library and meet the real-time requirement of the in-loop filters for 8 K × 4 K video format at 132 fps.

High throughput nanostructure-initiator mass spectrometry screening of microbial growth conditions for maximal β-glucosidase production.

Science.gov (United States)

Cheng, Xiaoliang; Hiras, Jennifer; Deng, Kai; Bowen, Benjamin; Simmons, Blake A; Adams, Paul D; Singer, Steven W; Northen, Trent R

2013-01-01

Production of biofuels via enzymatic hydrolysis of complex plant polysaccharides is a subject of intense global interest. Microbial communities are known to express a wide range of enzymes necessary for the saccharification of lignocellulosic feedstocks and serve as a powerful reservoir for enzyme discovery. However, the growth temperature and conditions that yield high cellulase activity vary widely, and the throughput to identify optimal conditions has been limited by the slow handling and conventional analysis. A rapid method that uses small volumes of isolate culture to resolve specific enzyme activity is needed. In this work, a high throughput nanostructure-initiator mass spectrometry (NIMS)-based approach was developed for screening a thermophilic cellulolytic actinomycete, Thermobispora bispora, for β-glucosidase production under various growth conditions. Media that produced high β-glucosidase activity were found to be I/S + glucose or microcrystalline cellulose (MCC), Medium 84 + rolled oats, and M9TE + MCC at 45°C. Supernatants of cell cultures grown in M9TE + 1% MCC cleaved 2.5 times more substrate at 45°C than at all other temperatures. While T. bispora is reported to grow optimally at 60°C in Medium 84 + rolled oats and M9TE + 1% MCC, approximately 40% more conversion was observed at 45°C. This high throughput NIMS approach may provide an important tool in discovery and characterization of enzymes from environmental microbes for industrial and biofuel applications.

High throughput nanostructure-initiator mass spectrometry screening of microbial growth conditions for maximal β-glucosidase production

Directory of Open Access Journals (Sweden)

Xiaoliang eCheng

2013-12-01

Full Text Available Production of biofuels via enzymatic hydrolysis of complex plant polysaccharides is a subject of intense global interest. Microbial communities are known to express a wide range of enzymes necessary for the saccharification of lignocellulosic feedstocks and serve as a powerful reservoir for enzyme discovery. However, the growth temperature and conditions that yield high cellulase activity vary widely, and the throughput to identify optimal conditions has been limited by the slow handling and conventional analysis. A rapid method that uses small volumes of isolate culture to resolve specific enzyme activity is needed. In this work, a high throughput nanostructure-initiator mass spectrometry (NIMS based approach was developed for screening a thermophilic cellulolytic actinomycete, Thermobispora bispora, for β-glucosidase production under various growth conditions. Media that produced high β-glucosidase activity were found to be I/S + glucose or microcrystalline cellulose (MCC, Medium 84 + rolled oats, and M9TE + MCC at 45 °C. Supernatants of cell cultures grown in M9TE + 1% MCC cleaved 2.5 times more substrate at 45 °C than at all other temperatures. While T. bispora is reported to grow optimally at 60 °C in Medium 84 + rolled oats and M9TE + 1% MCC, approximately 40% more conversion was observed at 45 °C. This high throughput NIMS approach may provide an important tool in discovery and characterization of enzymes from environmental microbes for industrial and biofuel applications.

The Three-Dimensional Culture System with Matrigel and Neurotrophic Factors Preserves the Structure and Function of Spiral Ganglion Neuron In Vitro.

Science.gov (United States)

Sun, Gaoying; Liu, Wenwen; Fan, Zhaomin; Zhang, Daogong; Han, Yuechen; Xu, Lei; Qi, Jieyu; Zhang, Shasha; Gao, Bradley T; Bai, Xiaohui; Li, Jianfeng; Chai, Renjie; Wang, Haibo

2016-01-01

Whole organ culture of the spiral ganglion region is a resourceful model system facilitating manipulation and analysis of live sprial ganglion neurons (SGNs). Three-dimensional (3D) cultures have been demonstrated to have many biomedical applications, but the effect of 3D culture in maintaining the SGNs structure and function in explant culture remains uninvestigated. In this study, we used the matrigel to encapsulate the spiral ganglion region isolated from neonatal mice. First, we optimized the matrigel concentration for the 3D culture system and found the 3D culture system protected the SGNs against apoptosis, preserved the structure of spiral ganglion region, and promoted the sprouting and outgrowth of SGNs neurites. Next, we found the 3D culture system promoted growth cone growth as evidenced by a higher average number and a longer average length of filopodia and a larger growth cone area. 3D culture system also significantly elevated the synapse density of SGNs. Last, we found that the 3D culture system combined with neurotrophic factors had accumulated effects in promoting the neurites outgrowth compared with 3D culture or NFs treatment only groups. Together, we conclude that the 3D culture system preserves the structure and function of SGN in explant culture.

Ex vivo cultures of glioblastoma in three-dimensional hydrogel maintain the original tumor growth behavior and are suitable for preclinical drug and radiation sensitivity screening

Energy Technology Data Exchange (ETDEWEB)

Jiguet Jiglaire, Carine, E-mail: carine.jiguet-jiglaire@univ-amu.fr [Aix Marseille Université, Faculté de Médecine de la Timone, 27 boulevard Jean Moulin, 13284 Marseille (France); CRO2, UMR 911, Faculté de Médecine de la Timone, 27 boulevard Jean Moulin, 13284 Marseille Cedex (France); INSERM, U911, 13005 Marseille (France); Baeza-Kallee, Nathalie; Denicolaï, Emilie; Barets, Doriane [Aix Marseille Université, Faculté de Médecine de la Timone, 27 boulevard Jean Moulin, 13284 Marseille (France); CRO2, UMR 911, Faculté de Médecine de la Timone, 27 boulevard Jean Moulin, 13284 Marseille Cedex (France); INSERM, U911, 13005 Marseille (France); Metellus, Philippe [Aix Marseille Université, Faculté de Médecine de la Timone, 27 boulevard Jean Moulin, 13284 Marseille (France); CRO2, UMR 911, Faculté de Médecine de la Timone, 27 boulevard Jean Moulin, 13284 Marseille Cedex (France); INSERM, U911, 13005 Marseille (France); APHM, Timone Hospital, Department of Neurosurgery, 13005 Marseille (France); Timone Hospital, 264 Rue Saint Pierre, 13385 Marseille Cedex 5 (France); and others

2014-02-15

Identification of new drugs and predicting drug response are major challenges in oncology, especially for brain tumors, because total surgical resection is difficult and radiation therapy or chemotherapy is often ineffective. With the aim of developing a culture system close to in vivo conditions for testing new drugs, we characterized an ex vivo three-dimensional culture system based on a hyaluronic acid-rich hydrogel and compared it with classical two-dimensional culture conditions. U87-MG glioblastoma cells and seven primary cell cultures of human glioblastomas were subjected to radiation therapy and chemotherapy drugs. It appears that 3D hydrogel preserves the original cancer growth behavior and enables assessment of the sensitivity of malignant gliomas to radiation and drugs with regard to inter-tumoral heterogeneity of therapeutic response. It could be used for preclinical assessment of new therapies. - Highlights: • We have compared primary glioblastoma cell culture in a 2D versus 3D-matrix system. • In 3D morphology, organization and markers better recapitulate the original tumor. • 3D-matrix culture might represent a relevant system for more accurate drug screening.

Ex vivo cultures of glioblastoma in three-dimensional hydrogel maintain the original tumor growth behavior and are suitable for preclinical drug and radiation sensitivity screening

International Nuclear Information System (INIS)

Jiguet Jiglaire, Carine; Baeza-Kallee, Nathalie; Denicolaï, Emilie; Barets, Doriane; Metellus, Philippe

2014-01-01

Identification of new drugs and predicting drug response are major challenges in oncology, especially for brain tumors, because total surgical resection is difficult and radiation therapy or chemotherapy is often ineffective. With the aim of developing a culture system close to in vivo conditions for testing new drugs, we characterized an ex vivo three-dimensional culture system based on a hyaluronic acid-rich hydrogel and compared it with classical two-dimensional culture conditions. U87-MG glioblastoma cells and seven primary cell cultures of human glioblastomas were subjected to radiation therapy and chemotherapy drugs. It appears that 3D hydrogel preserves the original cancer growth behavior and enables assessment of the sensitivity of malignant gliomas to radiation and drugs with regard to inter-tumoral heterogeneity of therapeutic response. It could be used for preclinical assessment of new therapies. - Highlights: • We have compared primary glioblastoma cell culture in a 2D versus 3D-matrix system. • In 3D morphology, organization and markers better recapitulate the original tumor. • 3D-matrix culture might represent a relevant system for more accurate drug screening

Bioprinted three dimensional human tissues for toxicology and disease modeling.

Science.gov (United States)

Nguyen, Deborah G; Pentoney, Stephen L

2017-03-01

The high rate of attrition among clinical-stage therapies, due largely to an inability to predict human toxicity and/or efficacy, underscores the need for in vitro models that better recapitulate in vivo human biology. In much the same way that additive manufacturing has revolutionized the production of solid objects, three-dimensional (3D) bioprinting is enabling the automated production of more architecturally and functionally accurate in vitro tissue culture models. Here, we provide an overview of the most commonly used bioprinting approaches and how they are being used to generate complex in vitro tissues for use in toxicology and disease modeling research. Copyright © 2017 Elsevier Ltd. All rights reserved.

High Throughput Determinations of Critical Dosing Parameters (IVIVE workshop)

Science.gov (United States)

High throughput toxicokinetics (HTTK) is an approach that allows for rapid estimations of TK for hundreds of environmental chemicals. HTTK-based reverse dosimetry (i.e, reverse toxicokinetics or RTK) is used in order to convert high throughput in vitro toxicity screening (HTS) da...

Rotating three-dimensional dynamic culture of adult human bone marrow-derived cells for tissue engineering of hyaline cartilage.

Science.gov (United States)

Sakai, Shinsuke; Mishima, Hajime; Ishii, Tomoo; Akaogi, Hiroshi; Yoshioka, Tomokazu; Ohyabu, Yoshimi; Chang, Fei; Ochiai, Naoyuki; Uemura, Toshimasa

2009-04-01

The method of constructing cartilage tissue from bone marrow-derived cells in vitro is considered a valuable technique for hyaline cartilage regenerative medicine. Using a rotating wall vessel (RWV) bioreactor developed in a NASA space experiment, we attempted to efficiently construct hyaline cartilage tissue from human bone marrow-derived cells without using a scaffold. Bone marrow aspirates were obtained from the iliac crest of nine patients during orthopedic operation. After their proliferation in monolayer culture, the adherent cells were cultured in the RWV bioreactor with chondrogenic medium for 2 weeks. Cells from the same source were cultured in pellet culture as controls. Histological and immunohistological evaluations (collagen type I and II) and quantification of glycosaminoglycan were performed on formed tissues and compared. The engineered constructs obtained using the RWV bioreactor showed strong features of hyaline cartilage in terms of their morphology as determined by histological and immunohistological evaluations. The glycosaminoglycan contents per microg DNA of the tissues were 10.01 +/- 3.49 microg/microg DNA in the case of the RWV bioreactor and 6.27 +/- 3.41 microg/microg DNA in the case of the pellet culture, and their difference was significant. The RWV bioreactor could provide an excellent environment for three-dimensional cartilage tissue architecture that can promote the chondrogenic differentiation of adult human bone marrow-derived cells.

Applications of ambient mass spectrometry in high-throughput screening.

Science.gov (United States)

Li, Li-Ping; Feng, Bao-Sheng; Yang, Jian-Wang; Chang, Cui-Lan; Bai, Yu; Liu, Hu-Wei

2013-06-07

The development of rapid screening and identification techniques is of great importance for drug discovery, doping control, forensic identification, food safety and quality control. Ambient mass spectrometry (AMS) allows rapid and direct analysis of various samples in open air with little sample preparation. Recently, its applications in high-throughput screening have been in rapid progress. During the past decade, various ambient ionization techniques have been developed and applied in high-throughput screening. This review discusses typical applications of AMS, including DESI (desorption electrospray ionization), DART (direct analysis in real time), EESI (extractive electrospray ionization), etc., in high-throughput screening (HTS).

High-throughput sequencing of three Lemnoideae (duckweeds chloroplast genomes from total DNA.

Directory of Open Access Journals (Sweden)

Wenqin Wang

Full Text Available BACKGROUND: Chloroplast genomes provide a wealth of information for evolutionary and population genetic studies. Chloroplasts play a particularly important role in the adaption for aquatic plants because they float on water and their major surface is exposed continuously to sunlight. The subfamily of Lemnoideae represents such a collection of aquatic species that because of photosynthesis represents one of the fastest growing plant species on earth. METHODS: We sequenced the chloroplast genomes from three different genera of Lemnoideae, Spirodela polyrhiza, Wolffiella lingulata and Wolffia australiana by high-throughput DNA sequencing of genomic DNA using the SOLiD platform. Unfractionated total DNA contains high copies of plastid DNA so that sequences from the nucleus and mitochondria can easily be filtered computationally. Remaining sequence reads were assembled into contiguous sequences (contigs using SOLiD software tools. Contigs were mapped to a reference genome of Lemna minor and gaps, selected by PCR, were sequenced on the ABI3730xl platform. CONCLUSIONS: This combinatorial approach yielded whole genomic contiguous sequences in a cost-effective manner. Over 1,000-time coverage of chloroplast from total DNA were reached by the SOLiD platform in a single spot on a quadrant slide without purification. Comparative analysis indicated that the chloroplast genome was conserved in gene number and organization with respect to the reference genome of L. minor. However, higher nucleotide substitution, abundant deletions and insertions occurred in non-coding regions of these genomes, indicating a greater genomic dynamics than expected from the comparison of other related species in the Pooideae. Noticeably, there was no transition bias over transversion in Lemnoideae. The data should have immediate applications in evolutionary biology and plant taxonomy with increased resolution and statistical power.

High throughput generation and trapping of individual agarose microgel using microfluidic approach

KAUST Repository

Shi, Yang

2013-02-28

Microgel is a kind of biocompatible polymeric material, which has been widely used as micro-carriers in materials synthesis, drug delivery and cell biology applications. However, high-throughput generation of individual microgel for on-site analysis in a microdevice still remains a challenge. Here, we presented a simple and stable droplet microfluidic system to realize high-throughput generation and trapping of individual agarose microgels based on the synergetic effect of surface tension and hydrodynamic forces in microchannels and used it for 3-D cell culture in real-time. The established system was mainly composed of droplet generators with flow focusing T-junction and a series of array individual trap structures. The whole process including the independent agarose microgel formation, immobilization in trapping array and gelation in situ via temperature cooling could be realized on the integrated microdevice completely. The performance of this system was demonstrated by successfully encapsulating and culturing adenoid cystic carcinoma (ACCM) cells in the gelated agarose microgels. This established approach is simple, easy to operate, which can not only generate the micro-carriers with different components in parallel, but also monitor the cell behavior in 3D matrix in real-time. It can also be extended for applications in the area of material synthesis and tissue engineering. © 2013 Springer-Verlag Berlin Heidelberg.

High throughput generation and trapping of individual agarose microgel using microfluidic approach

KAUST Repository

Shi, Yang; Gao, Xinghua; Chen, Longqing; Zhang, Min; Ma, Jingyun; Zhang, Xixiang; Qin, Jianhua

2013-01-01

Microgel is a kind of biocompatible polymeric material, which has been widely used as micro-carriers in materials synthesis, drug delivery and cell biology applications. However, high-throughput generation of individual microgel for on-site analysis in a microdevice still remains a challenge. Here, we presented a simple and stable droplet microfluidic system to realize high-throughput generation and trapping of individual agarose microgels based on the synergetic effect of surface tension and hydrodynamic forces in microchannels and used it for 3-D cell culture in real-time. The established system was mainly composed of droplet generators with flow focusing T-junction and a series of array individual trap structures. The whole process including the independent agarose microgel formation, immobilization in trapping array and gelation in situ via temperature cooling could be realized on the integrated microdevice completely. The performance of this system was demonstrated by successfully encapsulating and culturing adenoid cystic carcinoma (ACCM) cells in the gelated agarose microgels. This established approach is simple, easy to operate, which can not only generate the micro-carriers with different components in parallel, but also monitor the cell behavior in 3D matrix in real-time. It can also be extended for applications in the area of material synthesis and tissue engineering. © 2013 Springer-Verlag Berlin Heidelberg.

High-speed three-dimensional plasma temperature determination of axially symmetric free-burning arcs

International Nuclear Information System (INIS)

Bachmann, B; Ekkert, K; Bachmann, J-P; Marques, J-L; Schein, J; Kozakov, R; Gött, G; Schöpp, H; Uhrlandt, D

2013-01-01

In this paper we introduce an experimental technique that allows for high-speed, three-dimensional determination of electron density and temperature in axially symmetric free-burning arcs. Optical filters with narrow spectral bands of 487.5–488.5 nm and 689–699 nm are utilized to gain two-dimensional spectral information of a free-burning argon tungsten inert gas arc. A setup of mirrors allows one to image identical arc sections of the two spectral bands onto a single camera chip. Two-different Abel inversion algorithms have been developed to reconstruct the original radial distribution of emission coefficients detected with each spectral window and to confirm the results. With the assumption of local thermodynamic equilibrium we calculate emission coefficients as a function of temperature by application of the Saha equation, the ideal gas law, the quasineutral gas condition and the NIST compilation of spectral lines. Ratios of calculated emission coefficients are compared with measured ones yielding local plasma temperatures. In the case of axial symmetry the three-dimensional plasma temperature distributions have been determined at dc currents of 100, 125, 150 and 200 A yielding temperatures up to 20000 K in the hot cathode region. These measurements have been validated by four different techniques utilizing a high-resolution spectrometer at different positions in the plasma. Plasma temperatures show good agreement throughout the different methods. Additionally spatially resolved transient plasma temperatures have been measured of a dc pulsed process employing a high-speed frame rate of 33000 frames per second showing the modulation of the arc isothermals with time and providing information about the sensitivity of the experimental approach. (paper)

Three-dimensional carbon nanotube networks with a supported nickel oxide nanonet for high-performance supercapacitors.

Science.gov (United States)

Wu, Mao-Sung; Zheng, Yo-Ru; Lin, Guan-Wei

2014-08-04

A three-dimensional porous carbon nanotube film with a supported NiO nanonet was prepared by simple electrophoretic deposition and hydrothermal synthesis, which could deliver a high specific capacitance of 1511 F g(-1) at a high discharge current of 50 A g(-1) due to the significantly improved transport of the electrolyte and electrons.

Detachably assembled microfluidic device for perfusion culture and post-culture analysis of a spheroid array.

Science.gov (United States)

Sakai, Yusuke; Hattori, Koji; Yanagawa, Fumiki; Sugiura, Shinji; Kanamori, Toshiyuki; Nakazawa, Kohji

2014-07-01

Microfluidic devices permit perfusion culture of three-dimensional (3D) tissue, mimicking the flow of blood in vascularized 3D tissue in our body. Here, we report a microfluidic device composed of a two-part microfluidic chamber chip and multi-microwell array chip able to be disassembled at the culture endpoint. Within the microfluidic chamber, an array of 3D tissue aggregates (spheroids) can be formed and cultured under perfusion. Subsequently, detailed post-culture analysis of the spheroids collected from the disassembled device can be performed. This device facilitates uniform spheroid formation, growth analysis in a high-throughput format, controlled proliferation via perfusion flow rate, and post-culture analysis of spheroids. We used the device to culture spheroids of human hepatocellular carcinoma (HepG2) cells under two controlled perfusion flow rates. HepG2 spheroids exhibited greater cell growth at higher perfusion flow rates than at lower perfusion flow rates, and exhibited different metabolic activity and mRNA and protein expression under the different flow rate conditions. These results show the potential of perfusion culture to precisely control the culture environment in microfluidic devices. The construction of spheroid array chambers allows multiple culture conditions to be tested simultaneously, with potential applications in toxicity and drug screening. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

High throughput second harmonic imaging for label-free biological applications

KAUST Repository

Macias Romero, Carlos; Didier, Marie E P; Jourdain, Pascal; Marquet, Pierre; Magistretti, Pierre J.; Tarun, Orly B.; Zubkovs, Vitalijs; Radenovic, Aleksandra; Roke, Sylvie

2014-01-01

Second harmonic generation (SHG) is inherently sensitive to the absence of spatial centrosymmetry, which can render it intrinsically sensitive to interfacial processes, chemical changes and electrochemical responses. Here, we seek to improve the imaging throughput of SHG microscopy by using a wide-field imaging scheme in combination with a medium-range repetition rate amplified near infrared femtosecond laser source and gated detection. The imaging throughput of this configuration is tested by measuring the optical image contrast for different image acquisition times of BaTiO3 nanoparticles in two different wide-field setups and one commercial point-scanning configuration. We find that the second harmonic imaging throughput is improved by 2-3 orders of magnitude compared to point-scan imaging. Capitalizing on this result, we perform low fluence imaging of (parts of) living mammalian neurons in culture.

Vibrational spectra and thermal rectification in three-dimensional anharmonic lattices

International Nuclear Information System (INIS)

Lan Jinghua; Li Baowen

2007-01-01

We study thermal rectification in a three-dimensional model consisting of two segments of anharmonic lattices. One segment consists of layers of harmonic oscillator arrays coupled to a substrate potential, which is a three-dimensional Frenkel-Kontorova model, and the other segment is a three-dimensional Fermi-Pasta-Ulam model. We study the vibrational bands of the two lattices analytically and numerically, and find that, by choosing the system parameters properly, the rectification can be as high as a few thousands, which is high enough to be observed in experiment. Possible experiments in nanostructures are discussed

High-throughput evaluation of interactions between biomaterials, proteins and cells using patterned superhydrophobic substrates

OpenAIRE

Neto, Ana I.; Custódio, Catarina A.; Wenlong Song; Mano, J. F.

2011-01-01

We propose a new low cost platform for high-throughput analysis that permits screening the biological performance of independent combinations of biomaterials, cells and culture media. Patterned superhydrophobic flat substrates with controlled wettable spots are used to produce microarray chips for accelerated multiplexing evaluation. This work was partially supported by Fundação para a Ciência e Tecnologia (FCT) under project PTDC/FIS/68517/2006.

Mosquitoes meet microfluidics: High-throughput microfluidic tools for insect-parasite ecology in field conditions

Science.gov (United States)

Prakash, Manu; Mukundarajan, Haripriya

2013-11-01

A simple bite from an insect is the transmission mechanism for many deadly diseases worldwide--including malaria, yellow fever, west nile and dengue. Very little is known about how populations of numerous insect species and disease-causing parasites interact in their natural habitats due to a lack of measurement techniques. At present, vector surveillance techniques involve manual capture by using humans as live bait, which is hard to justify on ethical grounds. Individual mosquitoes are manually dissected to isolate salivary glands to detect sporozites. With typical vector infection rates being very low even in endemic areas, it is almost impossible to get an accurate picture of disease distribution, in both space and time. Here we present novel high-throughput microfluidic tools for vector surveillance, specifically mosquitoes. A two-dimensional high density array with baits provide an integrated platform for multiplex PCR for detection of both vector and parasite species. Combining techniques from engineering and field ecology, methods and tools developed here will enable high-throughput measurement of infection rates for a number of diseases in mosquito populations in field conditions. Pew Foundation.

Three-dimensional visualization of cultural clusters in the 1878 yellow fever epidemic of New Orleans.

Science.gov (United States)

Curtis, Andrew J

2008-08-22

An epidemic may exhibit different spatial patterns with a change in geographic scale, with each scale having different conduits and impediments to disease spread. Mapping disease at each of these scales often reveals different cluster patterns. This paper will consider this change of geographic scale in an analysis of yellow fever deaths for New Orleans in 1878. Global clustering for the whole city, will be followed by a focus on the French Quarter, then clusters of that area, and finally street-level patterns of a single cluster. The three-dimensional visualization capabilities of a GIS will be used as part of a cluster creation process that incorporates physical buildings in calculating mortality-to-mortality distance. Including nativity of the deceased will also capture cultural connection. Twenty-two yellow fever clusters were identified for the French Quarter. These generally mirror the results of other global cluster and density surfaces created for the entire epidemic in New Orleans. However, the addition of building-distance, and disease specific time frame between deaths reveal that disease spread contains a cultural component. Same nativity mortality clusters emerge in a similar time frame irrespective of proximity. Italian nativity mortalities were far more densely grouped than any of the other cohorts. A final examination of mortalities for one of the nativity clusters reveals that further sub-division is present, and that this pattern would only be revealed at this scale (street level) of investigation. Disease spread in an epidemic is complex resulting from a combination of geographic distance, geographic distance with specific connection to the built environment, disease-specific time frame between deaths, impediments such as herd immunity, and social or cultural connection. This research has shown that the importance of cultural connection may be more important than simple proximity, which in turn might mean traditional quarantine measures should be

High-throughput screening (HTS) and modeling of the retinoid ...

Science.gov (United States)

Presentation at the Retinoids Review 2nd workshop in Brussels, Belgium on the application of high throughput screening and model to the retinoid system Presentation at the Retinoids Review 2nd workshop in Brussels, Belgium on the application of high throughput screening and model to the retinoid system

Three dimensional canonical transformations

International Nuclear Information System (INIS)

Tegmen, A.

2010-01-01

A generic construction of canonical transformations is given in three-dimensional phase spaces on which Nambu bracket is imposed. First, the canonical transformations are defined as based on cannonade transformations. Second, it is shown that determination of the generating functions and the transformation itself for given generating function is possible by solving correspondent Pfaffian differential equations. Generating functions of type are introduced and all of them are listed. Infinitesimal canonical transformations are also discussed as the complementary subject. Finally, it is shown that decomposition of canonical transformations is also possible in three-dimensional phase spaces as in the usual two-dimensional ones.

Progress of radiotherapy by three-dimensional treatment planning

International Nuclear Information System (INIS)

Imada, Hajime; Nomoto, Satoshi; Takahashi, Hiroyuki; Nakata, Hajime

1998-01-01

The recent progress of three-dimensional radiation treatment planning was reviewed. And clinical cases such as lung cancer and breast cancer are introduced. In the University of Occupational and Development Health, the treatment system FOCUS which is made up of CT simulator and linac was used mainly. Three-dimensional treatment planning was carried for about 90% of 330 patients who underwent radiotherapy for one year. The target becomes to be accurate and dose distribution with all CT slices in radiation field can be confirmed by using three-dimensional radiation treatment planning apparatus. High dose irradiation localized to tumor part is possible. Relations between total dose and volume of normal tissue and/or tumor can be estimated numerically and easily by DVH. A prediction of indication and affection became possible by this procedure. In conclusion, generalization of three-dimensional radiation treatment planning will bring progress of more effective radiotherapy with less adverse reaction. (K.H.). 21 refs

Three Dimensional Polarimetric Neutron Tomography of Magnetic Fields

DEFF Research Database (Denmark)

Sales, Morten; Strobl, Markus; Shinohara, Takenao

2018-01-01

Through the use of Time-of-Flight Three Dimensional Polarimetric Neutron Tomography (ToF 3DPNT) we have for the first time successfully demonstrated a technique capable of measuring and reconstructing three dimensional magnetic field strengths and directions unobtrusively and non-destructively wi......Through the use of Time-of-Flight Three Dimensional Polarimetric Neutron Tomography (ToF 3DPNT) we have for the first time successfully demonstrated a technique capable of measuring and reconstructing three dimensional magnetic field strengths and directions unobtrusively and non...... and reconstructed, thereby providing the proof-of-principle of a technique able to reveal hitherto unobtainable information on the magnetic fields in the bulk of materials and devices, due to a high degree of penetration into many materials, including metals, and the sensitivity of neutron polarisation to magnetic...... fields. The technique puts the potential of the ToF time structure of pulsed neutron sources to full use in order to optimise the recorded information quality and reduce measurement time....

Correction of Microplate Data from High-Throughput Screening.

Science.gov (United States)

Wang, Yuhong; Huang, Ruili

2016-01-01

High-throughput screening (HTS) makes it possible to collect cellular response data from a large number of cell lines and small molecules in a timely and cost-effective manner. The errors and noises in the microplate-formatted data from HTS have unique characteristics, and they can be generally grouped into three categories: run-wise (temporal, multiple plates), plate-wise (background pattern, single plate), and well-wise (single well). In this chapter, we describe a systematic solution for identifying and correcting such errors and noises, mainly basing on pattern recognition and digital signal processing technologies.

Three-dimensional reacting shock–bubble interaction

NARCIS (Netherlands)

Diegelmann, Felix; Hickel, S.; Adams, Nikolaus A.

2017-01-01

We investigate a reacting shock–bubble interaction through three-dimensional numerical simulations with detailed chemistry. The convex shape of the bubble focuses the shock and generates regions of high pressure and temperature, which are sufficient to ignite the diluted stoichiometric

Three-dimensional microbubble streaming flows

Science.gov (United States)

Rallabandi, Bhargav; Marin, Alvaro; Rossi, Massimiliano; Kaehler, Christian; Hilgenfeldt, Sascha

2014-11-01

Streaming due to acoustically excited bubbles has been used successfully for applications such as size-sorting, trapping and focusing of particles, as well as fluid mixing. Many of these applications involve the precise control of particle trajectories, typically achieved using cylindrical bubbles, which establish planar flows. Using astigmatic particle tracking velocimetry (APTV), we show that, while this two-dimensional picture is a useful description of the flow over short times, a systematic three-dimensional flow structure is evident over long time scales. We demonstrate that this long-time three-dimensional fluid motion can be understood through asymptotic theory, superimposing secondary axial flows (induced by boundary conditions at the device walls) onto the two-dimensional description. This leads to a general framework that describes three-dimensional flows in confined microstreaming systems, guiding the design of applications that profit from minimizing or maximizing these effects.

High-throughput scoring of seed germination

NARCIS (Netherlands)

Ligterink, Wilco; Hilhorst, Henk W.M.

2017-01-01

High-throughput analysis of seed germination for phenotyping large genetic populations or mutant collections is very labor intensive and would highly benefit from an automated setup. Although very often used, the total germination percentage after a nominated period of time is not very

Low-resistance gateless high electron mobility transistors using three-dimensional inverted pyramidal AlGaN/GaN surfaces

International Nuclear Information System (INIS)

So, Hongyun; Senesky, Debbie G.

2016-01-01

In this letter, three-dimensional gateless AlGaN/GaN high electron mobility transistors (HEMTs) were demonstrated with 54% reduction in electrical resistance and 73% increase in surface area compared with conventional gateless HEMTs on planar substrates. Inverted pyramidal AlGaN/GaN surfaces were microfabricated using potassium hydroxide etched silicon with exposed (111) surfaces and metal-organic chemical vapor deposition of coherent AlGaN/GaN thin films. In addition, electrical characterization of the devices showed that a combination of series and parallel connections of the highly conductive two-dimensional electron gas along the pyramidal geometry resulted in a significant reduction in electrical resistance at both room and high temperatures (up to 300 °C). This three-dimensional HEMT architecture can be leveraged to realize low-power and reliable power electronics, as well as harsh environment sensors with increased surface area

Low-resistance gateless high electron mobility transistors using three-dimensional inverted pyramidal AlGaN/GaN surfaces

Energy Technology Data Exchange (ETDEWEB)

So, Hongyun, E-mail: hyso@stanford.edu [Department of Aeronautics and Astronautics, Stanford University, Stanford, California 94305 (United States); Senesky, Debbie G. [Department of Aeronautics and Astronautics, Stanford University, Stanford, California 94305 (United States); Department of Electrical Engineering, Stanford University, Stanford, California 94305 (United States)

2016-01-04

In this letter, three-dimensional gateless AlGaN/GaN high electron mobility transistors (HEMTs) were demonstrated with 54% reduction in electrical resistance and 73% increase in surface area compared with conventional gateless HEMTs on planar substrates. Inverted pyramidal AlGaN/GaN surfaces were microfabricated using potassium hydroxide etched silicon with exposed (111) surfaces and metal-organic chemical vapor deposition of coherent AlGaN/GaN thin films. In addition, electrical characterization of the devices showed that a combination of series and parallel connections of the highly conductive two-dimensional electron gas along the pyramidal geometry resulted in a significant reduction in electrical resistance at both room and high temperatures (up to 300 °C). This three-dimensional HEMT architecture can be leveraged to realize low-power and reliable power electronics, as well as harsh environment sensors with increased surface area.

Cooperative simulation of lithography and topography for three-dimensional high-aspect-ratio etching

Science.gov (United States)

Ichikawa, Takashi; Yagisawa, Takashi; Furukawa, Shinichi; Taguchi, Takafumi; Nojima, Shigeki; Murakami, Sadatoshi; Tamaoki, Naoki

2018-06-01

A topography simulation of high-aspect-ratio etching considering transports of ions and neutrals is performed, and the mechanism of reactive ion etching (RIE) residues in three-dimensional corner patterns is revealed. Limited ion flux and CF2 diffusion from the wide space of the corner is found to have an effect on the RIE residues. Cooperative simulation of lithography and topography is used to solve the RIE residue problem.

A high-throughput method for quantifying metabolically active yeast cells

DEFF Research Database (Denmark)

Nandy, Subir Kumar; Knudsen, Peter Boldsen; Rosenkjær, Alexander

2015-01-01

By redesigning the established methylene blue reduction test for bacteria and yeast, we present a cheap and efficient methodology for quantitative physiology of eukaryotic cells applicable for high-throughput systems. Validation of themethod in fermenters and highthroughput systems proved....... The drop in metabolic activity associated with the diauxic shift in yeast proved more pronounced for the MBRT-derived curve compared with OD curves, consistent with a dramatic shift in the ratio between live and dead cells at this metabolic event. This method provides a tool with numerous applications, e.......g. characterizing the death phase of stationary phase cultures, or in drug screens with pathogenic yeasts....

[Application Progress of Three-dimensional Laser Scanning Technology in Medical Surface Mapping].

Science.gov (United States)

Zhang, Yonghong; Hou, He; Han, Yuchuan; Wang, Ning; Zhang, Ying; Zhu, Xianfeng; Wang, Mingshi

2016-04-01

The booming three-dimensional laser scanning technology can efficiently and effectively get spatial three-dimensional coordinates of the detected object surface and reconstruct the image at high speed,high precision and large capacity of information.Non-radiation,non-contact and the ability of visualization make it increasingly popular in three-dimensional surface medical mapping.This paper reviews the applications and developments of three-dimensional laser scanning technology in medical field,especially in stomatology,plastic surgery and orthopedics.Furthermore,the paper also discusses the application prospects in the future as well as the biomedical engineering problems it would encounter with.

Diffraction limited focusing with controllable arbitrary three-dimensional polarization

International Nuclear Information System (INIS)

Chen, Weibin; Zhan, Qiwen

2010-01-01

We propose a new approach that enables full control over the three-dimensional state of polarization and the field distribution near the focus of a high numerical aperture objective lens. By combining the electric dipole radiation and a vectorial diffraction method, the input field at the pupil plane for generating arbitrary three-dimensionally oriented linear polarization at the focal point with a diffraction limited spot size is found analytically by solving the inverse problem. Arbitrary three-dimensional elliptical polarization can be obtained by introducing a second electric dipole oriented in the orthogonal plane with appropriate amplitude and phase differences

Development of three-dimensional neoclassical transport simulation code with high performance Fortran on a vector-parallel computer

International Nuclear Information System (INIS)

Satake, Shinsuke; Okamoto, Masao; Nakajima, Noriyoshi; Takamaru, Hisanori

2005-11-01

A neoclassical transport simulation code (FORTEC-3D) applicable to three-dimensional configurations has been developed using High Performance Fortran (HPF). Adoption of computing techniques for parallelization and a hybrid simulation model to the δf Monte-Carlo method transport simulation, including non-local transport effects in three-dimensional configurations, makes it possible to simulate the dynamism of global, non-local transport phenomena with a self-consistent radial electric field within a reasonable computation time. In this paper, development of the transport code using HPF is reported. Optimization techniques in order to achieve both high vectorization and parallelization efficiency, adoption of a parallel random number generator, and also benchmark results, are shown. (author)

Optimization and high-throughput screening of antimicrobial peptides.

Science.gov (United States)

Blondelle, Sylvie E; Lohner, Karl

2010-01-01

While a well-established process for lead compound discovery in for-profit companies, high-throughput screening is becoming more popular in basic and applied research settings in academia. The development of combinatorial libraries combined with easy and less expensive access to new technologies have greatly contributed to the implementation of high-throughput screening in academic laboratories. While such techniques were earlier applied to simple assays involving single targets or based on binding affinity, they have now been extended to more complex systems such as whole cell-based assays. In particular, the urgent need for new antimicrobial compounds that would overcome the rapid rise of drug-resistant microorganisms, where multiple target assays or cell-based assays are often required, has forced scientists to focus onto high-throughput technologies. Based on their existence in natural host defense systems and their different mode of action relative to commercial antibiotics, antimicrobial peptides represent a new hope in discovering novel antibiotics against multi-resistant bacteria. The ease of generating peptide libraries in different formats has allowed a rapid adaptation of high-throughput assays to the search for novel antimicrobial peptides. Similarly, the availability nowadays of high-quantity and high-quality antimicrobial peptide data has permitted the development of predictive algorithms to facilitate the optimization process. This review summarizes the various library formats that lead to de novo antimicrobial peptide sequences as well as the latest structural knowledge and optimization processes aimed at improving the peptides selectivity.

Three-dimensional friction measurement during hip simulation.

Directory of Open Access Journals (Sweden)

Robert Sonntag

Full Text Available Wear of total hip replacements has been the focus of many studies. However, frictional effects, such as high loading on intramodular connections or the interface to the bone, as well as friction associated squeaking have recently increased interest about the amount of friction that is generated during daily activities. The aim of this study was thus to establish and validate a three-dimensional friction setup under standardized conditions.A standard hip simulator was modified to allow for high precision measurements of small frictional effects in the hip during three-dimensional hip articulation. The setup was verified by an ideal hydrostatic bearing and validated with a static-load physical pendulum and an extension-flexion rotation with a dynamic load profile. Additionally, a pendulum model was proposed for screening measurement of frictional effects based on the damping behavior of the angular oscillation without the need for any force/moment transducer. Finally, three-dimensional friction measurements have been realized for ceramic-on-polyethylene bearings of three different sizes (28, 36 and 40 mm.A precision of less than 0.2 Nm during three-dimensional friction measurements was reported, while increased frictional torque (resultant as well as taper torque was measured for larger head diameters. These effects have been confirmed by simple pendulum tests and the theoretical model. A comparison with current literature about friction measurements is presented.This investigation of friction is able to provide more information about a field that has been dominated by the reduction of wear. It should be considered in future pre-clinical testing protocols given by international organizations of standardization.

Three dimensional imaging in cardiac nuclear medicine

International Nuclear Information System (INIS)

Torizuka, Kanji; Ishii, Yasushi; Yonekura, Yoshiharu; Yamamoto, Kazutaka; Tamaki, Takeyoshi

1981-01-01

Methods to obtain three dimensional images of the heart were reviewed. Gated three dimensional display reconstructed from images using bidirectional collimator, was a useful method to detect akinesis of the heart wall. Tomographic observation of the heart can be carried out by a pinhole collimator to image ischemia with high sensitivity. However the focusing plane must be carefully selected to prevent false positives. In the case of emission CT (ECT), utilization of positron emitters gave a quantitative image without correction, whereas single photon ECT needed the correction due to the absorption of γ-ray. Though the reliability of the images by ECT was high, the time required for data acquisition was much longer than that by a 7 pinhole or bidirectional collimator. (Nakanishi, T.)

Chromatographic Monoliths for High-Throughput Immunoaffinity Isolation of Transferrin from Human Plasma

Directory of Open Access Journals (Sweden)

Irena Trbojević-Akmačić

2016-06-01

Full Text Available Changes in protein glycosylation are related to different diseases and have a potential as diagnostic and prognostic disease biomarkers. Transferrin (Tf glycosylation changes are common marker for congenital disorders of glycosylation. However, biological interindividual variability of Tf N-glycosylation and genes involved in glycosylation regulation are not known. Therefore, high-throughput Tf isolation method and large scale glycosylation studies are needed in order to address these questions. Due to their unique chromatographic properties, the use of chromatographic monoliths enables very fast analysis cycle, thus significantly increasing sample preparation throughput. Here, we are describing characterization of novel immunoaffinity-based monolithic columns in a 96-well plate format for specific high-throughput purification of human Tf from blood plasma. We optimized the isolation and glycan preparation procedure for subsequent ultra performance liquid chromatography (UPLC analysis of Tf N-glycosylation and managed to increase the sensitivity for approximately three times compared to initial experimental conditions, with very good reproducibility. This work is licensed under a Creative Commons Attribution 4.0 International License.

High-resolution liquid patterns via three-dimensional droplet shape control.

Science.gov (United States)

Raj, Rishi; Adera, Solomon; Enright, Ryan; Wang, Evelyn N

2014-09-25

Understanding liquid dynamics on surfaces can provide insight into nature's design and enable fine manipulation capability in biological, manufacturing, microfluidic and thermal management applications. Of particular interest is the ability to control the shape of the droplet contact area on the surface, which is typically circular on a smooth homogeneous surface. Here, we show the ability to tailor various droplet contact area shapes ranging from squares, rectangles, hexagons, octagons, to dodecagons via the design of the structure or chemical heterogeneity on the surface. We simultaneously obtain the necessary physical insights to develop a universal model for the three-dimensional droplet shape by characterizing the droplet side and top profiles. Furthermore, arrays of droplets with controlled shapes and high spatial resolution can be achieved using this approach. This liquid-based patterning strategy promises low-cost fabrication of integrated circuits, conductive patterns and bio-microarrays for high-density information storage and miniaturized biochips and biosensors, among others.

Graphene quantum dots-three-dimensional graphene composites for high-performance supercapacitors.

Science.gov (United States)

Chen, Qing; Hu, Yue; Hu, Chuangang; Cheng, Huhu; Zhang, Zhipan; Shao, Huibo; Qu, Liangti

2014-09-28

Graphene quantum dots (GQDs) have been successfully deposited onto the three-dimensional graphene (3DG) by a benign electrochemical method and the ordered 3DG structure remains intact after the uniform deposition of GQDs. In addition, the capacitive properties of the as-formed GQD-3DG composites are evaluated in symmetrical supercapacitors. It is found that the supercapacitor fabricated from the GQD-3DG composite is highly stable and exhibits a high specific capacitance of 268 F g(-1), representing a more than 90% improvement over that of the supercapacitor made from pure 3DG electrodes (136 F g(-1)). Owing to the convenience of the current method, it can be further used in other well-defined electrode materials, such as carbon nanotubes, carbon aerogels and conjugated polymers to improve the performance of the supercapacitors.

20180311 - High Throughput Transcriptomics: From screening to pathways (SOT 2018)

Science.gov (United States)

The EPA ToxCast effort has screened thousands of chemicals across hundreds of high-throughput in vitro screening assays. The project is now leveraging high-throughput transcriptomic (HTTr) technologies to substantially expand its coverage of biological pathways. The first HTTr sc...

High throughput label-free platform for statistical bio-molecular sensing

DEFF Research Database (Denmark)

Bosco, Filippo; Hwu, En-Te; Chen, Ching-Hsiu

2011-01-01

Sensors are crucial in many daily operations including security, environmental control, human diagnostics and patient monitoring. Screening and online monitoring require reliable and high-throughput sensing. We report on the demonstration of a high-throughput label-free sensor platform utilizing...

A comparison of three-dimensional culture systems to evaluate in vitro chondrogenesis of equine bone marrow-derived mesenchymal stem cells.

Science.gov (United States)

Watts, Ashlee E; Ackerman-Yost, Jeremy C; Nixon, Alan J

2013-10-01

To compare in vitro three-dimensional (3D) culture systems that model chondrogenesis of bone marrow-derived mesenchymal stem cells (MSCs). MSCs from five horses 2-3 years of age were consolidated in fibrin 0.3% alginate, 1.2% alginate, 2.5×10(5) cell pellets, 5×10(5) cell pellets, and 2% agarose, and maintained in chondrogenic medium with supplemental TGF-β1 for 4 weeks. Pellets and media were tested at days 1, 14, and 28 for gene expression of markers of chondrogenic maturation and hypertrophy (ACAN, COL2B, COL10, SOX9, 18S), and evaluated by histology (hematoxylin and eosin, Toluidine Blue) and immunohistochemistry (collagen type II and X). alginate, fibrin alginate (FA), and both pellet culture systems resulted in chondrogenic transformation. Adequate RNA was not obtained from agarose cultures at any time point. There was increased COL2B, ACAN, and SOX9 expression on day 14 from both pellet culture systems. On day 28, increased expression of COL2B was maintained in 5×10(5) cell pellets and there was no difference in ACAN and SOX9 between FA and both pellet cultures. COL10 expression was significantly lower in FA cultures on day 28. Collagen type II was abundantly formed in all culture systems except alginate and collagen type X was least in FA hydrogels. equine MSCs respond to 3D culture in FA blended hydrogel and both pellet culture systems with chondrogenic induction. For prevention of terminal differentiation and hypertrophy, FA culture may be superior to pellet culture systems.

Thermal Investigation of Three-Dimensional GaN-on-SiC High Electron Mobility Transistors

Science.gov (United States)

2017-07-01

University of L’Aquila, (2011). 23 Rao, H. & Bosman, G. Hot-electron induced defect generation in AlGaN/GaN high electron mobility transistors. Solid...AFRL-RY-WP-TR-2017-0143 THERMAL INVESTIGATION OF THREE- DIMENSIONAL GaN-on-SiC HIGH ELECTRON MOBILITY TRANSISTORS Qing Hao The University of Arizona...clarification memorandum dated 16 Jan 09. This report is available to the general public, including foreign nationals. Copies may be obtained from the

3D Cell Culture in a Self-Assembled Nanofiber Environment.

Directory of Open Access Journals (Sweden)

Yi Wen Chai

Full Text Available The development and utilization of three-dimensional cell culture platforms has been gaining more traction. Three-dimensional culture platforms are capable of mimicking in vivo microenvironments, which provide greater physiological relevance in comparison to conventional two-dimensional cultures. The majority of three-dimensional culture platforms are challenged by the lack of cell attachment, long polymerization times, and inclusion of undefined xenobiotics, and cytotoxic cross-linkers. In this study, we review the use of a highly defined material composed of naturally occurring compounds, hyaluronic acid and chitosan, known as Cell-Mate3DTM. Moreover, we provide an original measurement of Young's modulus using a uniaxial unconfined compression method to elucidate the difference in microenvironment rigidity for acellular and cellular conditions. When hydrated into a tissue-like hybrid hydrocolloid/hydrogel, Cell-Mate3DTM is a highly versatile three-dimensional culture platform that enables downstream applications such as flow cytometry, immunostaining, histological staining, and functional studies to be applied with relative ease.

Development of three-dimensional lung multicellular spheroids in air- and liquid-interface culture for the evaluation of anticancer therapeutics.

Science.gov (United States)

Meenach, Samantha A; Tsoras, Alexandra N; McGarry, Ronald C; Mansour, Heidi M; Hilt, J Zach; Anderson, Kimberly W

2016-04-01

Three-dimensional (3D) lung multicellular spheroids (MCS) in liquid-covered culture (LCC) and air-interface culture (AIC) conditions have both been developed for the evaluation of aerosol anticancer therapeutics in solution and aerosols, respectively. The MCS were formed by seeding lung cancer cells on top of collagen where they formed spheroids due to the prevalence of cell-to-cell interactions. LCC MCS were exposed to paclitaxel (PTX) in media whereas AIC MCS were exposed to dry powder PEGylated phospholipid aerosol microparticles containing paclitaxel. The difference in viability for 2D versus 3D culture for both LCC and AIC was evaluated along with the effects of the particles on lung epithelium via transepithelial electrical resistance (TEER) measurements. For LCC and AIC conditions, the 3D spheroids were more resistant to treatment with higher IC50 values for A549 and H358 cell lines. TEER results initially indicated a decrease in resistance upon drug or particle exposure, however, these values increased over the course of several days indicating the ability of the cells to recover. Overall, these studies offer a comprehensive in vitro evaluation of aerosol particles used in the treatment of lung cancer while introducing a new method for culturing lung cancer MCS in both LCC and AIC conditions.

Characterization of A Three-Dimensional Organotypic Co-Culture Skin Model for Epidermal Differentiation of Rat Adipose-Derived Stem Cells.

Science.gov (United States)

Ghanavati, Zeinab; Orazizadeh, Mahmoud; Bayati, Vahid; Abbaspour, Mohammad Reza; Khorsandi, Layasadat; Mansouri, Esrafil; Neisi, Niloofar

2016-01-01

The organotypic co-culture is a well-known technique to examine cellular interactions and their roles in stem cell proliferation and differentiation. This study aims to evaluate the effects of dermal fibroblasts (DFs) on epidermal differentiation of adipose-derived stem cells (ASCs) using a three-dimensional (3D) organotypic co- culture technique. In this experimental research study, rat DFs and ASCs were isolated and cultured separately on electrospun polycaprolactone (PCL) matrices. The PCL matrices seeded by ASCs were superimposed on to the matrices seeded by DFs in order to create a 3D organotypic co-culture. In the control groups, PCL matrices seeded by ASCs were placed on matrices devoid of DFs. After 10 days, we assessed the expressions of keratinocyte-related genes by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and expression of pan-cytokeratin protein by immunofluorescence in the differentiated keratinocyte-like cells from co- culture and control groups. Keratinocyte-like cell morphologies were also observed by scanning electron microscopy (SEM). The early, intermediate, and terminal differentiation keratinocyte markers-Cytokeratin14, Filaggrin, and Involucrin significantly expressed in the co-culture groups com- pared to the control ones (P<0.05). We observed pan-cytokeratin in keratinocyte-like cells of both groups by immunofluorescence. SEM observation of the co-culture groups showed that the differentiated keratinocyte-like cells developed a polygonal cobblestone shape, considered characteristic of keratinocytes. The 3D organotypic co-culture bilayered construct that consisted of DFs and ASCs was an effective technique for epidermal differentiation of ASCs. This co-culture might be useful for epidermal differentiation of stem cells for future applications in skin regeneration.

COSMO-PAFOG: Three-dimensional fog forecasting with the high-resolution COSMO-model

Science.gov (United States)

Hacker, Maike; Bott, Andreas

2017-04-01

The presence of fog can have critical impact on shipping, aviation and road traffic increasing the risk of serious accidents. Besides these negative impacts of fog, in arid regions fog is explored as a supplementary source of water for human settlements. Thus the improvement of fog forecasts holds immense operational value. The aim of this study is the development of an efficient three-dimensional numerical fog forecast model based on a mesoscale weather prediction model for the application in the Namib region. The microphysical parametrization of the one-dimensional fog forecast model PAFOG (PArameterized FOG) is implemented in the three-dimensional nonhydrostatic mesoscale weather prediction model COSMO (COnsortium for Small-scale MOdeling) developed and maintained by the German Meteorological Service. Cloud water droplets are introduced in COSMO as prognostic variables, thus allowing a detailed description of droplet sedimentation. Furthermore, a visibility parametrization depending on the liquid water content and the droplet number concentration is implemented. The resulting fog forecast model COSMO-PAFOG is run with kilometer-scale horizontal resolution. In vertical direction, we use logarithmically equidistant layers with 45 of 80 layers in total located below 2000 m. Model results are compared to satellite observations and synoptic observations of the German Meteorological Service for a domain in the west of Germany, before the model is adapted to the geographical and climatological conditions in the Namib desert. COSMO-PAFOG is able to represent the horizontal structure of fog patches reasonably well. Especially small fog patches typical of radiation fog can be simulated in agreement with observations. Ground observations of temperature are also reproduced. Simulations without the PAFOG microphysics yield unrealistically high liquid water contents. This in turn reduces the radiative cooling of the ground, thus inhibiting nocturnal temperature decrease. The

High-Throughput Analysis of Enzyme Activities

Energy Technology Data Exchange (ETDEWEB)

Lu, Guoxin [Iowa State Univ., Ames, IA (United States)

2007-01-01

High-throughput screening (HTS) techniques have been applied to many research fields nowadays. Robot microarray printing technique and automation microtiter handling technique allows HTS performing in both heterogeneous and homogeneous formats, with minimal sample required for each assay element. In this dissertation, new HTS techniques for enzyme activity analysis were developed. First, patterns of immobilized enzyme on nylon screen were detected by multiplexed capillary system. The imaging resolution is limited by the outer diameter of the capillaries. In order to get finer images, capillaries with smaller outer diameters can be used to form the imaging probe. Application of capillary electrophoresis allows separation of the product from the substrate in the reaction mixture, so that the product doesn't have to have different optical properties with the substrate. UV absorption detection allows almost universal detection for organic molecules. Thus, no modifications of either the substrate or the product molecules are necessary. This technique has the potential to be used in screening of local distribution variations of specific bio-molecules in a tissue or in screening of multiple immobilized catalysts. Another high-throughput screening technique is developed by directly monitoring the light intensity of the immobilized-catalyst surface using a scientific charge-coupled device (CCD). Briefly, the surface of enzyme microarray is focused onto a scientific CCD using an objective lens. By carefully choosing the detection wavelength, generation of product on an enzyme spot can be seen by the CCD. Analyzing the light intensity change over time on an enzyme spot can give information of reaction rate. The same microarray can be used for many times. Thus, high-throughput kinetic studies of hundreds of catalytic reactions are made possible. At last, we studied the fluorescence emission spectra of ADP and obtained the detection limits for ADP under three different

Three-dimensional CT imaging of soft-tissue anatomy

International Nuclear Information System (INIS)

Fishman, E.K.; Ney, D.R.; Magid, D.; Kuhlman, J.E.

1988-01-01

Three-dimensional display of computed tomographic data has been limited to skeletal structures. This was in part related to the reconstruction algorithm used, which relied on a binary classification scheme. A new algorithm, volumetric rendering with percentage classification, provides the ability to display three-dimensional images of muscle and soft tissue. A review was conducted of images in 35 cases in which muscle and/or soft tissue were part of the clinical problem. In all cases, individual muscle groups could be clearly identified and discriminated. Branching vessels in the range of 2.3 mm could be identified. Similarly, lymph nodes could be clearly defined. High-resolution three-dimensional images were found to be useful both in providing an increased understanding of complex muscle and soft tissue anatomy and in surgical planning

Combinatorial chemoenzymatic synthesis and high-throughput screening of sialosides.

Science.gov (United States)

Chokhawala, Harshal A; Huang, Shengshu; Lau, Kam; Yu, Hai; Cheng, Jiansong; Thon, Vireak; Hurtado-Ziola, Nancy; Guerrero, Juan A; Varki, Ajit; Chen, Xi

2008-09-19

Although the vital roles of structures containing sialic acid in biomolecular recognition are well documented, limited information is available on how sialic acid structural modifications, sialyl linkages, and the underlying glycan structures affect the binding or the activity of sialic acid-recognizing proteins and related downstream biological processes. A novel combinatorial chemoenzymatic method has been developed for the highly efficient synthesis of biotinylated sialosides containing different sialic acid structures and different underlying glycans in 96-well plates from biotinylated sialyltransferase acceptors and sialic acid precursors. By transferring the reaction mixtures to NeutrAvidin-coated plates and assaying for the yields of enzymatic reactions using lectins recognizing sialyltransferase acceptors but not the sialylated products, the biotinylated sialoside products can be directly used, without purification, for high-throughput screening to quickly identify the ligand specificity of sialic acid-binding proteins. For a proof-of-principle experiment, 72 biotinylated alpha2,6-linked sialosides were synthesized in 96-well plates from 4 biotinylated sialyltransferase acceptors and 18 sialic acid precursors using a one-pot three-enzyme system. High-throughput screening assays performed in NeutrAvidin-coated microtiter plates show that whereas Sambucus nigra Lectin binds to alpha2,6-linked sialosides with high promiscuity, human Siglec-2 (CD22) is highly selective for a number of sialic acid structures and the underlying glycans in its sialoside ligands.

Yeast diversity during the fermentation of Andean chicha: A comparison of high-throughput sequencing and culture-dependent approaches.

Science.gov (United States)

Mendoza, Lucía M; Neef, Alexander; Vignolo, Graciela; Belloch, Carmela

2017-10-01

Diversity and dynamics of yeasts associated with the fermentation of Argentinian maize-based beverage chicha was investigated. Samples taken at different stages from two chicha productions were analyzed by culture-dependent and culture-independent methods. Five hundred and ninety six yeasts were isolated by classical microbiological methods and 16 species identified by RFLPs and sequencing of D1/D2 26S rRNA gene. Genetic typing of isolates from the dominant species, Saccharomyces cerevisiae, by PCR of delta elements revealed up to 42 different patterns. High-throughput sequencing (HTS) of D1/D2 26S rRNA gene amplicons from chicha samples detected more than one hundred yeast species and almost fifty filamentous fungi taxa. Analysis of the data revealed that yeasts dominated the fermentation, although, a significant percentage of filamentous fungi appeared in the first step of the process. Statistical analysis of results showed that very few taxa were represented by more than 1% of the reads per sample at any step of the process. S. cerevisiae represented more than 90% of the reads in the fermentative samples. Other yeast species dominated the pre-fermentative steps and abounded in fermented samples when S. cerevisiae was in percentages below 90%. Most yeasts species detected by pyrosequencing were not recovered by cultivation. In contrast, the cultivation-based methodology detected very few yeast taxa, and most of them corresponded with very few reads in the pyrosequencing analysis. Copyright © 2017 Elsevier Ltd. All rights reserved.

High-throughput sequence alignment using Graphics Processing Units

Directory of Open Access Journals (Sweden)

Trapnell Cole

2007-12-01

Full Text Available Abstract Background The recent availability of new, less expensive high-throughput DNA sequencing technologies has yielded a dramatic increase in the volume of sequence data that must be analyzed. These data are being generated for several purposes, including genotyping, genome resequencing, metagenomics, and de novo genome assembly projects. Sequence alignment programs such as MUMmer have proven essential for analysis of these data, but researchers will need ever faster, high-throughput alignment tools running on inexpensive hardware to keep up with new sequence technologies. Results This paper describes MUMmerGPU, an open-source high-throughput parallel pairwise local sequence alignment program that runs on commodity Graphics Processing Units (GPUs in common workstations. MUMmerGPU uses the new Compute Unified Device Architecture (CUDA from nVidia to align multiple query sequences against a single reference sequence stored as a suffix tree. By processing the queries in parallel on the highly parallel graphics card, MUMmerGPU achieves more than a 10-fold speedup over a serial CPU version of the sequence alignment kernel, and outperforms the exact alignment component of MUMmer on a high end CPU by 3.5-fold in total application time when aligning reads from recent sequencing projects using Solexa/Illumina, 454, and Sanger sequencing technologies. Conclusion MUMmerGPU is a low cost, ultra-fast sequence alignment program designed to handle the increasing volume of data produced by new, high-throughput sequencing technologies. MUMmerGPU demonstrates that even memory-intensive applications can run significantly faster on the relatively low-cost GPU than on the CPU.

Miniaturizing 3D assay for high-throughput drug and genetic screens for small patient-derived tumor samples (Conference Presentation)

Science.gov (United States)

Rotem, Asaf; Garraway, Levi; Su, Mei-Ju; Basu, Anindita; Regev, Aviv; Struhl, Kevin

2017-02-01

Three-dimensional growth conditions reflect the natural environment of cancer cells and are crucial to be performed at drug screens. We developed a 3D assay for cellular transformation that involves growth in low attachment (GILA) conditions and is strongly correlated with the 50-year old benchmark assay-soft agar. Using GILA, we performed high-throughput screens for drugs and genes that selectively inhibit or increase transformation, but not proliferation. This phenotypic approach is complementary to our genetic approach that utilizes single-cell RNA-sequencing of a patient sample to identify putative oncogenes that confer sensitivity to drugs designed to specifically inhibit the identified oncoprotein. Currently, we are dealing with a big challenge in our field- the limited number of cells that might be extracted from a biopsy. Small patient-derived samples are hard to test in the traditional multiwell plate and it will be helpful to minimize the culture area and the experimental system. We managed to design a suitable microfluidic device for limited number of cells and perform the assay using image analysis. We aim to test drugs on tumor cells, outside of the patient body- and recommend on the ideal treatment that is tailored to the individual. This device will help to minimize biopsy-sampling volumes and minimize interventions in the patient's tumor.

High throughput imaging cytometer with acoustic focussing.

Science.gov (United States)

Zmijan, Robert; Jonnalagadda, Umesh S; Carugo, Dario; Kochi, Yu; Lemm, Elizabeth; Packham, Graham; Hill, Martyn; Glynne-Jones, Peter

2015-10-31

We demonstrate an imaging flow cytometer that uses acoustic levitation to assemble cells and other particles into a sheet structure. This technique enables a high resolution, low noise CMOS camera to capture images of thousands of cells with each frame. While ultrasonic focussing has previously been demonstrated for 1D cytometry systems, extending the technology to a planar, much higher throughput format and integrating imaging is non-trivial, and represents a significant jump forward in capability, leading to diagnostic possibilities not achievable with current systems. A galvo mirror is used to track the images of the moving cells permitting exposure times of 10 ms at frame rates of 50 fps with motion blur of only a few pixels. At 80 fps, we demonstrate a throughput of 208 000 beads per second. We investigate the factors affecting motion blur and throughput, and demonstrate the system with fluorescent beads, leukaemia cells and a chondrocyte cell line. Cells require more time to reach the acoustic focus than beads, resulting in lower throughputs; however a longer device would remove this constraint.

High-throughput GPU-based LDPC decoding

Science.gov (United States)

Chang, Yang-Lang; Chang, Cheng-Chun; Huang, Min-Yu; Huang, Bormin

2010-08-01

Low-density parity-check (LDPC) code is a linear block code known to approach the Shannon limit via the iterative sum-product algorithm. LDPC codes have been adopted in most current communication systems such as DVB-S2, WiMAX, WI-FI and 10GBASE-T. LDPC for the needs of reliable and flexible communication links for a wide variety of communication standards and configurations have inspired the demand for high-performance and flexibility computing. Accordingly, finding a fast and reconfigurable developing platform for designing the high-throughput LDPC decoder has become important especially for rapidly changing communication standards and configurations. In this paper, a new graphic-processing-unit (GPU) LDPC decoding platform with the asynchronous data transfer is proposed to realize this practical implementation. Experimental results showed that the proposed GPU-based decoder achieved 271x speedup compared to its CPU-based counterpart. It can serve as a high-throughput LDPC decoder.

Evaluating High Throughput Toxicokinetics and Toxicodynamics for IVIVE (WC10)

Science.gov (United States)

High-throughput screening (HTS) generates in vitro data for characterizing potential chemical hazard. TK models are needed to allow in vitro to in vivo extrapolation (IVIVE) to real world situations. The U.S. EPA has created a public tool (R package “httk” for high throughput tox...

Investigating the Role of Surface Materials and Three Dimensional Architecture on In Vitro Differentiation of Porcine Monocyte-Derived Dendritic Cells

DEFF Research Database (Denmark)

Hartmann, Sofie Bruun; Mohanty, Soumyaranjan; Skovgaard, Kerstin

2016-01-01

In vitro generation of dendritic-like cells through differentiation of peripheral blood monocytes is typically done using two-dimensional polystyrene culture plates. In the process of optimising cell culture techniques, engineers have developed fluidic micro-devises usually manufactured in materi......In vitro generation of dendritic-like cells through differentiation of peripheral blood monocytes is typically done using two-dimensional polystyrene culture plates. In the process of optimising cell culture techniques, engineers have developed fluidic micro-devises usually manufactured......-dimensional PDMS and carbonised three-dimensional PDMS. Cells cultured conventionally (on two-dimensional polystyrene) differentiated into moDCs as expected. Interestingly, gene expression of a wide range of cytokines, chemokines, and pattern recognition receptors was influenced by culture surface material...... and IL23A) but the influence of the surfaces was unchanged. These findings highlights future challenges of combining and comparing data generated from microfluidic cell culture-devices made using alternative materials to data generated using conventional polystyrene plates used by most laboratories today....

Three-dimensional magnetospheric equilibrium with isotropic pressure

International Nuclear Information System (INIS)

Cheng, C.Z.

1995-05-01

In the absence of the toroidal flux, two coupled quasi two-dimensional elliptic equilibrium equations have been derived to describe self-consistent three-dimensional static magnetospheric equilibria with isotropic pressure in an optimal (Ψ,α,χ) flux coordinate system, where Ψ is the magnetic flux function, χ is a generalized poloidal angle, α is the toroidal angle, α = φ - δ(Ψ,φ,χ) is the toroidal angle, δ(Ψ,φ,χ) is periodic in φ, and the magnetic field is represented as rvec B = ∇Ψ x ∇α. A three-dimensional magnetospheric equilibrium code, the MAG-3D code, has been developed by employing an iterative metric method. The main difference between the three-dimensional and the two-dimensional axisymmetric solutions is that the field-aligned current and the toroidal magnetic field are finite for the three-dimensional case, but vanish for the two-dimensional axisymmetric case. With the same boundary flux surface shape, the two-dimensional axisymmetric results are similar to the three-dimensional magnetosphere at each local time cross section

Three dimensional visualization of medical images

International Nuclear Information System (INIS)

Suto, Yasuzo

1992-01-01

Three dimensional visualization is a stereoscopic technique that allows the diagnosis and treatment of complicated anatomy site of the bone and organ. In this article, the current status and technical application of three dimensional visualization are introduced with special reference to X-ray CT and MRI. The surface display technique is the most common for three dimensional visualization, consisting of geometric model, voxel element, and stereographic composition techniques. Recent attention has been paid to display method of the content of the subject called as volume rendering, whereby information on the living body is provided accurately. The application of three dimensional visualization is described in terms of diagnostic imaging and surgical simulation. (N.K.)

(Weakly) three-dimensional caseology

International Nuclear Information System (INIS)

Pomraning, G.C.

1996-01-01

The singular eigenfunction technique of Case for solving one-dimensional planar symmetry linear transport problems is extended to a restricted class of three-dimensional problems. This class involves planar geometry, but with forcing terms (either boundary conditions or internal sources) which are weakly dependent upon the transverse spatial variables. Our analysis involves a singular perturbation about the classic planar analysis, and leads to the usual Case discrete and continuum modes, but modulated by weakly dependent three-dimensional spatial functions. These functions satisfy parabolic differential equations, with a different diffusion coefficient for each mode. Representative one-speed time-independent transport problems are solved in terms of these generalised Case eigenfunctions. Our treatment is very heuristic, but may provide an impetus for more rigorous analysis. (author)

Programmable full-adder computations in communicating three-dimensional cell cultures.

Science.gov (United States)

Ausländer, David; Ausländer, Simon; Pierrat, Xavier; Hellmann, Leon; Rachid, Leila; Fussenegger, Martin

2018-01-01

Synthetic biologists have advanced the design of trigger-inducible gene switches and their assembly into input-programmable circuits that enable engineered human cells to perform arithmetic calculations reminiscent of electronic circuits. By designing a versatile plug-and-play molecular-computation platform, we have engineered nine different cell populations with genetic programs, each of which encodes a defined computational instruction. When assembled into 3D cultures, these engineered cell consortia execute programmable multicellular full-adder logics in response to three trigger compounds.

Three-dimensional visualization of cultural clusters in the 1878 yellow fever epidemic of New Orleans

Directory of Open Access Journals (Sweden)

Curtis Andrew J

2008-08-01

Full Text Available Abstract Background An epidemic may exhibit different spatial patterns with a change in geographic scale, with each scale having different conduits and impediments to disease spread. Mapping disease at each of these scales often reveals different cluster patterns. This paper will consider this change of geographic scale in an analysis of yellow fever deaths for New Orleans in 1878. Global clustering for the whole city, will be followed by a focus on the French Quarter, then clusters of that area, and finally street-level patterns of a single cluster. The three-dimensional visualization capabilities of a GIS will be used as part of a cluster creation process that incorporates physical buildings in calculating mortality-to-mortality distance. Including nativity of the deceased will also capture cultural connection. Results Twenty-two yellow fever clusters were identified for the French Quarter. These generally mirror the results of other global cluster and density surfaces created for the entire epidemic in New Orleans. However, the addition of building-distance, and disease specific time frame between deaths reveal that disease spread contains a cultural component. Same nativity mortality clusters emerge in a similar time frame irrespective of proximity. Italian nativity mortalities were far more densely grouped than any of the other cohorts. A final examination of mortalities for one of the nativity clusters reveals that further sub-division is present, and that this pattern would only be revealed at this scale (street level of investigation. Conclusion Disease spread in an epidemic is complex resulting from a combination of geographic distance, geographic distance with specific connection to the built environment, disease-specific time frame between deaths, impediments such as herd immunity, and social or cultural connection. This research has shown that the importance of cultural connection may be more important than simple proximity, which in

Three-dimensional porous hollow fibre copper electrodes for efficient and high-rate electrochemical carbon dioxide reduction

NARCIS (Netherlands)

Kas, Recep; Hummadi, Khalid Khazzal; Kortlever, Ruud; de Wit, Patrick; Milbrat, Alexander; Luiten-Olieman, Maria W.J.; Benes, Nieck Edwin; Koper, Marc T.M.; Mul, Guido

2016-01-01

Aqueous-phase electrochemical reduction of carbon dioxide requires an active, earth-abundant electrocatalyst, as well as highly efficient mass transport. Here we report the design of a porous hollow fibre copper electrode with a compact three-dimensional geometry, which provides a large area,

Three-dimensional imaging of flat natural and cultural heritage objects by a Compton scattering modality

Science.gov (United States)

Guerrero Prado, Patricio; Nguyen, Mai K.; Dumas, Laurent; Cohen, Serge X.

2017-01-01

Characterization and interpretation of flat ancient material objects, such as those found in archaeology, paleoenvironments, paleontology, and cultural heritage, have remained a challenging task to perform by means of conventional x-ray tomography methods due to their anisotropic morphology and flattened geometry. To overcome the limitations of the mentioned methodologies for such samples, an imaging modality based on Compton scattering is proposed in this work. Classical x-ray tomography treats Compton scattering data as noise in the image formation process, while in Compton scattering tomography the conditions are set such that Compton data become the principal image contrasting agent. Under these conditions, we are able, first, to avoid relative rotations between the sample and the imaging setup, and second, to obtain three-dimensional data even when the object is supported by a dense material by exploiting backscattered photons. Mathematically this problem is addressed by means of a conical Radon transform and its inversion. The image formation process and object reconstruction model are presented. The feasibility of this methodology is supported by numerical simulations.

Two-Dimensional High Definition Versus Three-Dimensional Endoscopy in Endonasal Skull Base Surgery: A Comparative Preclinical Study.

Science.gov (United States)

Rampinelli, Vittorio; Doglietto, Francesco; Mattavelli, Davide; Qiu, Jimmy; Raffetti, Elena; Schreiber, Alberto; Villaret, Andrea Bolzoni; Kucharczyk, Walter; Donato, Francesco; Fontanella, Marco Maria; Nicolai, Piero

2017-09-01

Three-dimensional (3D) endoscopy has been recently introduced in endonasal skull base surgery. Only a relatively limited number of studies have compared it to 2-dimensional, high definition technology. The objective was to compare, in a preclinical setting for endonasal endoscopic surgery, the surgical maneuverability of 2-dimensional, high definition and 3D endoscopy. A group of 68 volunteers, novice and experienced surgeons, were asked to perform 2 tasks, namely simulating grasping and dissection surgical maneuvers, in a model of the nasal cavities. Time to complete the tasks was recorded. A questionnaire to investigate subjective feelings during tasks was filled by each participant. In 25 subjects, the surgeons' movements were continuously tracked by a magnetic-based neuronavigator coupled with dedicated software (ApproachViewer, part of GTx-UHN) and the recorded trajectories were analyzed by comparing jitter, sum of square differences, and funnel index. Total execution time was significantly lower with 3D technology (P < 0.05) in beginners and experts. Questionnaires showed that beginners preferred 3D endoscopy more frequently than experts. A minority (14%) of beginners experienced discomfort with 3D endoscopy. Analysis of jitter showed a trend toward increased effectiveness of surgical maneuvers with 3D endoscopy. Sum of square differences and funnel index analyses documented better values with 3D endoscopy in experts. In a preclinical setting for endonasal skull base surgery, 3D technology appears to confer an advantage in terms of time of execution and precision of surgical maneuvers. Copyright © 2017 Elsevier Inc. All rights reserved.

Variations in gene and protein expression in canine chondrodystrophic nucleus pulposus cells following long-term three-dimensional culture.

Directory of Open Access Journals (Sweden)

Munetaka Iwata

Full Text Available Intervertebral disc (IVD degeneration greatly affects quality of life. The nucleus pulposus (NP of chondrodystrophic dog breeds (CDBs is similar to the human NP, because the cells disappear with age and are replaced by fibrochondrocyte-like cells. However, because IVD develops as early as within the first year of life, we used canines as a model to investigate in vitro the mechanisms underlying IVD degeneration. Specifically, we evaluated the potential of a three-dimensional (3D culture of healthy NP as an in vitro model system to investigate the mechanisms of IVD degeneration. Agarose hydrogels were populated with healthy NP cells from beagles after performing magnetic resonance imaging, and mRNA expression profiles and pericellular extracellular matrix (ECM protein distribution were determined. After 25 days of 3D culture, there was a tendency for redifferentiation into the native NP phenotype, and mRNA levels of Col2A1, COMP, and CK18 were not significantly different from those of freshly isolated cells. Our findings suggest that long-term 3D culture promoted chondrodystrophic NP redifferentiation through reconstruction of the pericellular microenvironment. Further, lipopolysaccharide (LPS induced expression of TNF-α, MMP3, MMP13, VEGF, and PGES mRNA in the 3D cultures, creating a molecular milieu that mimics that of degenerated NP. These results suggest that this in vitro model represents a reliable and cost-effective tool for evaluating new therapies for disc degeneration.

Three-dimensional quick response code based on inkjet printing of upconversion fluorescent nanoparticles for drug anti-counterfeiting

Science.gov (United States)

You, Minli; Lin, Min; Wang, Shurui; Wang, Xuemin; Zhang, Ge; Hong, Yuan; Dong, Yuqing; Jin, Guorui; Xu, Feng

2016-05-01

Medicine counterfeiting is a serious issue worldwide, involving potentially devastating health repercussions. Advanced anti-counterfeit technology for drugs has therefore aroused intensive interest. However, existing anti-counterfeit technologies are associated with drawbacks such as the high cost, complex fabrication process, sophisticated operation and incapability in authenticating drug ingredients. In this contribution, we developed a smart phone recognition based upconversion fluorescent three-dimensional (3D) quick response (QR) code for tracking and anti-counterfeiting of drugs. We firstly formulated three colored inks incorporating upconversion nanoparticles with RGB (i.e., red, green and blue) emission colors. Using a modified inkjet printer, we printed a series of colors by precisely regulating the overlap of these three inks. Meanwhile, we developed a multilayer printing and splitting technology, which significantly increases the information storage capacity per unit area. As an example, we directly printed the upconversion fluorescent 3D QR code on the surface of drug capsules. The 3D QR code consisted of three different color layers with each layer encoded by information of different aspects of the drug. A smart phone APP was designed to decode the multicolor 3D QR code, providing the authenticity and related information of drugs. The developed technology possesses merits in terms of low cost, ease of operation, high throughput and high information capacity, thus holds great potential for drug anti-counterfeiting.Medicine counterfeiting is a serious issue worldwide, involving potentially devastating health repercussions. Advanced anti-counterfeit technology for drugs has therefore aroused intensive interest. However, existing anti-counterfeit technologies are associated with drawbacks such as the high cost, complex fabrication process, sophisticated operation and incapability in authenticating drug ingredients. In this contribution, we developed a

Three-dimensional optical reconstruction of vocal fold kinematics using high-speed video with a laser projection system

Science.gov (United States)

Luegmair, Georg; Mehta, Daryush D.; Kobler, James B.; Döllinger, Michael

2015-01-01

Vocal fold kinematics and its interaction with aerodynamic characteristics play a primary role in acoustic sound production of the human voice. Investigating the temporal details of these kinematics using high-speed videoendoscopic imaging techniques has proven challenging in part due to the limitations of quantifying complex vocal fold vibratory behavior using only two spatial dimensions. Thus, we propose an optical method of reconstructing the superior vocal fold surface in three spatial dimensions using a high-speed video camera and laser projection system. Using stereo-triangulation principles, we extend the camera-laser projector method and present an efficient image processing workflow to generate the three-dimensional vocal fold surfaces during phonation captured at 4000 frames per second. Initial results are provided for airflow-driven vibration of an ex vivo vocal fold model in which at least 75% of visible laser points contributed to the reconstructed surface. The method captures the vertical motion of the vocal folds at a high accuracy to allow for the computation of three-dimensional mucosal wave features such as vibratory amplitude, velocity, and asymmetry. PMID:26087485

Three-dimensional metamaterials fabricated using Proton Beam Writing

Energy Technology Data Exchange (ETDEWEB)

Bettiol, A.A., E-mail: a.bettiol@nus.edu.sg [Centre for Ion Beam Applications, Department of Physics, National University of Singapore, 2 Science Dr. 3, Singapore 117542 (Singapore); Turaga, S.P.; Yan, Y.; Vanga, S.K. [Centre for Ion Beam Applications, Department of Physics, National University of Singapore, 2 Science Dr. 3, Singapore 117542 (Singapore); Chiam, S.Y. [NUS High School for Maths and Science, 20 Clementi Avenue 1, Singapore 129957 (Singapore)

2013-07-01

Proton Beam Writing (PBW) is a direct write lithographic technique that has recently been applied to the fabrication of three dimensional metamaterials. In this work, we show that the unique capabilities of PBW, namely the ability to fabricate arrays of high resolution, high aspect ratio microstructures in polymer or replicated into metal, is well suited to metamaterials research. We have also developed a novel method for selectively electroless plating silver directly onto polymer structures that were fabricated using PBW. This method opens up new avenues for utilizing PBW for making metamaterials and other sub-wavelength metallic structures. Several potential applications of three dimensional metamaterials fabricated using PBW are discussed, including sensing and negative refractive index materials.

Multiplex enrichment quantitative PCR (ME-qPCR): a high-throughput, highly sensitive detection method for GMO identification.

Science.gov (United States)

Fu, Wei; Zhu, Pengyu; Wei, Shuang; Zhixin, Du; Wang, Chenguang; Wu, Xiyang; Li, Feiwu; Zhu, Shuifang

2017-04-01

Among all of the high-throughput detection methods, PCR-based methodologies are regarded as the most cost-efficient and feasible methodologies compared with the next-generation sequencing or ChIP-based methods. However, the PCR-based methods can only achieve multiplex detection up to 15-plex due to limitations imposed by the multiplex primer interactions. The detection throughput cannot meet the demands of high-throughput detection, such as SNP or gene expression analysis. Therefore, in our study, we have developed a new high-throughput PCR-based detection method, multiplex enrichment quantitative PCR (ME-qPCR), which is a combination of qPCR and nested PCR. The GMO content detection results in our study showed that ME-qPCR could achieve high-throughput detection up to 26-plex. Compared to the original qPCR, the Ct values of ME-qPCR were lower for the same group, which showed that ME-qPCR sensitivity is higher than the original qPCR. The absolute limit of detection for ME-qPCR could achieve levels as low as a single copy of the plant genome. Moreover, the specificity results showed that no cross-amplification occurred for irrelevant GMO events. After evaluation of all of the parameters, a practical evaluation was performed with different foods. The more stable amplification results, compared to qPCR, showed that ME-qPCR was suitable for GMO detection in foods. In conclusion, ME-qPCR achieved sensitive, high-throughput GMO detection in complex substrates, such as crops or food samples. In the future, ME-qPCR-based GMO content identification may positively impact SNP analysis or multiplex gene expression of food or agricultural samples. Graphical abstract For the first-step amplification, four primers (A, B, C, and D) have been added into the reaction volume. In this manner, four kinds of amplicons have been generated. All of these four amplicons could be regarded as the target of second-step PCR. For the second-step amplification, three parallels have been taken for

Three dimensional reconstruction of tomographic images of the retina

International Nuclear Information System (INIS)

Glittenberg, C.; Zeiler, F.; Falkner, C.; Binder, S.; Povazay, B.; Hermann, B.; Drexler, W.

2007-01-01

The development of a new display system for the three-dimensional visualization of tomographic images in ophthalmology. Specifically, a system that can use stacks of B-mode scans from an ultrahigh resolution optical tomography examination to vividly display retinal specimens as three-dimensional objects. Several subroutines were programmed in the rendering and raytracing program Cinema 4D XL 9.102 Studio Bundle (Maxon Computer Inc., Friedrichsburg, Germany), which could process stacks of tomographic scans into three-dimensional objects. Ultrahigh resolution optical coherence tomography examinations were performed on patients with various retinal pathologies and post processed with the subroutines that had been designed. All ultrahigh resolution optical coherence tomographies were performed with a titanium: sapphire based ultra broad bandwidth (160 nm) femtosecond laser system (INTEGRAL, Femtolasers Productions GmbH. Vienna Austria) with an axial resolution of 3 μm. A new three dimensional display system for tomographic images in ophthalmology was developed, which allows a highly vivid display of physiological and pathological structures of the retina. The system also distinguishes itself through its high interactivity and adaptability. This new display system allows the visualization of physiological and pathological structures of the retina in a new way, which will give us new insight into their morphology and development. (author) [de

High-throughput optical system for HDES hyperspectral imager

Science.gov (United States)

Václavík, Jan; Melich, Radek; Pintr, Pavel; Pleštil, Jan

2015-01-01

Affordable, long-wave infrared hyperspectral imaging calls for use of an uncooled FPA with high-throughput optics. This paper describes the design of the optical part of a stationary hyperspectral imager in a spectral range of 7-14 um with a field of view of 20°×10°. The imager employs a push-broom method made by a scanning mirror. High throughput and a demand for simplicity and rigidity led to a fully refractive design with highly aspheric surfaces and off-axis positioning of the detector array. The design was optimized to exploit the machinability of infrared materials by the SPDT method and a simple assemblage.

High-throughput isolation of giant viruses in liquid medium using automated flow cytometry and fluorescence staining.

Directory of Open Access Journals (Sweden)

Jacques Yaacoub Bou Khalil

2016-01-01

Full Text Available The isolation of giant viruses using amoeba co-culture is tedious and fastidious. Recently, the procedure was successfully associated with a method that detects amoebal lysis on agar plates. However, the procedure remains time-consuming and is limited to protozoa growing on agar. We present here advances for the isolation of giant viruses. A high-throughput automated method based on flow cytometry and fluorescent staining was used to detect the presence of giant viruses in liquid medium. Development was carried out with the Acanthamoeba polyphaga strain widely used in past and current co-culture experiments. The proof of concept was validated with virus suspensions: artificially contaminated samples but also environmental samples from which viruses were previously isolated. After validating the technique, and fortuitously isolating a new Mimivirus, we automated the technique on 96-well plates and tested it on clinical and environmental samples using other protozoa. This allowed us to detect more than ten strains of previously known species of giant viruses and 7 new strains of a new virus lineage. This automated high-throughput method demonstrated significant time saving, and higher sensitivity than older techniques. It thus creates the means to isolate giant viruses at high speed.

RSCA genotyping of MHC for high-throughput evolutionary studies in the model organism three-spined stickleback Gasterosteus aculeatus

Science.gov (United States)

Lenz, Tobias L; Eizaguirre, Christophe; Becker, Sven; Reusch, Thorsten BH

2009-01-01

Background In all jawed vertebrates, highly polymorphic genes of the major histocompatibility complex (MHC) encode antigen presenting molecules that play a key role in the adaptive immune response. Their polymorphism is composed of multiple copies of recently duplicated genes, each possessing many alleles within populations, as well as high nucleotide divergence between alleles of the same species. Experimental evidence is accumulating that MHC polymorphism is a result of balancing selection by parasites and pathogens. In order to describe MHC diversity and analyse the underlying mechanisms that maintain it, a reliable genotyping technique is required that is suitable for such highly variable genes. Results We present a genotyping protocol that uses Reference Strand-mediated Conformation Analysis (RSCA), optimised for recently duplicated MHC class IIB genes that are typical for many fish and bird species, including the three-spined stickleback, Gasterosteus aculeatus. In addition we use a comprehensive plasmid library of MHC class IIB alleles to determine the nucleotide sequence of alleles represented by RSCA allele peaks. Verification of the RSCA typing by cloning and sequencing demonstrates high congruency between both methods and provides new insight into the polymorphism of classical stickleback MHC genes. Analysis of the plasmid library additionally reveals the high resolution and reproducibility of the RSCA technique. Conclusion This new RSCA genotyping protocol offers a fast, but sensitive and reliable way to determine the MHC allele repertoire of three-spined sticklebacks. It therefore provides a valuable tool to employ this highly polymorphic and adaptive marker in future high-throughput studies of host-parasite co-evolution and ecological speciation in this emerging model organism. PMID:19291291

RSCA genotyping of MHC for high-throughput evolutionary studies in the model organism three-spined stickleback Gasterosteus aculeatus

Directory of Open Access Journals (Sweden)

Becker Sven

2009-03-01

Full Text Available Abstract Background In all jawed vertebrates, highly polymorphic genes of the major histocompatibility complex (MHC encode antigen presenting molecules that play a key role in the adaptive immune response. Their polymorphism is composed of multiple copies of recently duplicated genes, each possessing many alleles within populations, as well as high nucleotide divergence between alleles of the same species. Experimental evidence is accumulating that MHC polymorphism is a result of balancing selection by parasites and pathogens. In order to describe MHC diversity and analyse the underlying mechanisms that maintain it, a reliable genotyping technique is required that is suitable for such highly variable genes. Results We present a genotyping protocol that uses Reference Strand-mediated Conformation Analysis (RSCA, optimised for recently duplicated MHC class IIB genes that are typical for many fish and bird species, including the three-spined stickleback, Gasterosteus aculeatus. In addition we use a comprehensive plasmid library of MHC class IIB alleles to determine the nucleotide sequence of alleles represented by RSCA allele peaks. Verification of the RSCA typing by cloning and sequencing demonstrates high congruency between both methods and provides new insight into the polymorphism of classical stickleback MHC genes. Analysis of the plasmid library additionally reveals the high resolution and reproducibility of the RSCA technique. Conclusion This new RSCA genotyping protocol offers a fast, but sensitive and reliable way to determine the MHC allele repertoire of three-spined sticklebacks. It therefore provides a valuable tool to employ this highly polymorphic and adaptive marker in future high-throughput studies of host-parasite co-evolution and ecological speciation in this emerging model organism.

Three-dimensional parametrization of photon-initiated high energy showers

International Nuclear Information System (INIS)

De Angelis, A.

1988-01-01

A three-dimensional parametrization of photon-initiated showers in a homogeneous absorber is presented. The form, suggested by a model assimilating the transverse shower development to a random walk process, displays a simple scaling with the primary energy, and is very suitable for numerical integration. The parameters are explicitly calculated for the case of showers in SF5 lead glass, and the results are compared with the explicit simulation by GEANT3.11. Fields of application are investigated. (orig.)

High Performance Computing Modernization Program Kerberos Throughput Test Report

Science.gov (United States)

2017-10-26

Naval Research Laboratory Washington, DC 20375-5320 NRL/MR/5524--17-9751 High Performance Computing Modernization Program Kerberos Throughput Test ...NUMBER 5d. PROJECT NUMBER 5e. TASK NUMBER 5f. WORK UNIT NUMBER 2. REPORT TYPE1. REPORT DATE (DD-MM-YYYY) 4. TITLE AND SUBTITLE 6. AUTHOR(S) 8. PERFORMING...PAGE 18. NUMBER OF PAGES 17. LIMITATION OF ABSTRACT High Performance Computing Modernization Program Kerberos Throughput Test Report Daniel G. Gdula* and

Development of three-dimensional cellular culture system for testing of biological effects of radiations in tumoral and non-tumoral models

International Nuclear Information System (INIS)

Bonfim, Leticia; Vieira, Daniel Perez; Oliveira, Karina

2017-01-01

Pre-clinical drug testing is currently based on based on monolayer or 2D (2D) cell cultures and, despite the large-scale use of this form of culture, there is already scientific evidence that the cellular disposition in monolayers does not adequately simulate tissue physiology, as it prevents cells from expressing their characteristics in a manner analogous to that found in the organism. For this purpose, the work aimed to produce three-dimensional structures, referred as spheroids, using magnetic levitation by adding iron nanoparticles to the cultures and with the aid of magnets. Electron microscopy showed particles with about 20nm in diameter. FTIR (Fourier-transform infrared spectroscopy) analysis showed stretches compatible with iron and amino acid (Lysine) binding. The images showed the formation of spherical bodies until the ninth day. LnCap spheroid diameter varied from (mean ± error) 434.407 ± 50.018 μm (5 th day) to 264.574 ± 13.184 μm (9 t 'h day). Cultures of CHO ranged from 229.237 ± 5.278 μm to 236.719 ± 12.910 μm in the same period. Spheres generated by magnetic levitation could be measured by digital means and compared throughout the experiment. The tool can be used to test the biological effects of radiation and / or radiopharmaceuticals in culture. (author)

Development of three-dimensional cellular culture system for testing of biological effects of radiations in tumoral and non-tumoral models

Energy Technology Data Exchange (ETDEWEB)

Bonfim, Leticia; Vieira, Daniel Perez, E-mail: leticia.bonfim@ipen.br [Instituto de Pesquisas Energéticas e Nucleares (IPEN/CNEN-SP), São Paulo, SP (Brazil). Lab. de Radiobiologia; Oliveira, Karina [Universidade Federal de Sao Paulo (UNIFESP), Diadema, SP (Brazil). Departamento de Fisica

2017-07-01

Pre-clinical drug testing is currently based on based on monolayer or 2D (2D) cell cultures and, despite the large-scale use of this form of culture, there is already scientific evidence that the cellular disposition in monolayers does not adequately simulate tissue physiology, as it prevents cells from expressing their characteristics in a manner analogous to that found in the organism. For this purpose, the work aimed to produce three-dimensional structures, referred as spheroids, using magnetic levitation by adding iron nanoparticles to the cultures and with the aid of magnets. Electron microscopy showed particles with about 20nm in diameter. FTIR (Fourier-transform infrared spectroscopy) analysis showed stretches compatible with iron and amino acid (Lysine) binding. The images showed the formation of spherical bodies until the ninth day. LnCap spheroid diameter varied from (mean ± error) 434.407 ± 50.018 μm (5{sup th} day) to 264.574 ± 13.184 μm (9{sup t}'h day). Cultures of CHO ranged from 229.237 ± 5.278 μm to 236.719 ± 12.910 μm in the same period. Spheres generated by magnetic levitation could be measured by digital means and compared throughout the experiment. The tool can be used to test the biological effects of radiation and / or radiopharmaceuticals in culture. (author)

Effects of ethanol and acetaldehyde on tight junction integrity: in vitro study in a three dimensional intestinal epithelial cell culture model.

Directory of Open Access Journals (Sweden)

Elhaseen Elamin

Full Text Available BACKGROUND: Intestinal barrier dysfunction and translocation of endotoxins are involved in the pathogenesis of alcoholic liver disease. Exposure to ethanol and its metabolite, acetaldehyde at relatively high concentrations have been shown to disrupt intestinal epithelial tight junctions in the conventional two dimensional cell culture models. The present study investigated quantitatively and qualitatively the effects of ethanol at concentrations detected in the blood after moderate ethanol consumption, of its metabolite acetaldehyde and of the combination of both compounds on intestinal barrier function in a three-dimensional cell culture model. METHODS AND FINDINGS: Caco-2 cells were grown in a basement membrane matrix (Matrigel™ to induce spheroid formation and were then exposed to the compounds at the basolateral side. Morphological differentiation of the spheroids was assessed by immunocytochemistry and transmission electron microscopy. The barrier function was assessed by the flux of FITC-labeled dextran from the basal side into the spheroids' luminal compartment using confocal microscopy. Caco-2 cells grown on Matrigel assembled into fully differentiated and polarized spheroids with a central lumen, closely resembling enterocytes in vivo and provide an excellent model to study epithelial barrier functionality. Exposure to ethanol (10-40 mM or acetaldehyde (25-200 µM for 3 h, dose-dependently and additively increased the paracellular permeability and induced redistribution of ZO-1 and occludin without affecting cell viability or tight junction-encoding gene expression. Furthermore, ethanol and acetaldehyde induced lysine residue and microtubules hyperacetylation. CONCLUSIONS: These results indicate that ethanol at concentrations found in the blood after moderate drinking and acetaldehyde, alone and in combination, can increase the intestinal epithelial permeability. The data also point to the involvement of protein hyperacetylation in

Use of three-dimensional, high-resolution seismic technology to optimize the location of remedial systems

International Nuclear Information System (INIS)

Bainer, R.W.; Adams, M.L.

1993-02-01

Two three-dimensional (3-D), high-resolution seismic reflection pilot studies were conducted in California at two sites, where the primary contaminants of concern are solvents. Identify pathways of contaminant migration. Determine the subsurface stratigraphy and structure to optimize the location for placement of remedial systems. The geology at the first site, located at the Lawrence Livermore National Laboratory in Livermore, California, is characterized by unconsolidated alluvium. Ground water varies in depth from about 30 to 100 ft. The site typically is subjected to extensive cultural noise. The second site, in Southern California, is located in a broad, synclinal depression in the Transverse Range. Shallow alluvium overlies a marine turbidite sequence that crops out as massive sandstone beds. Field work for both surveys took place in August 1992. A Bison Model 90120-A, 120-channel (DIFP) seismograph was used to record the data. Thirty-hertz, natural-frequency geophones were used to receive the data, and an Elastic Wave Generator (EWG) was used as the seismic source. The use of a signal-stacking, noninvasive source was found to be an effective method of overriding background noise at the sites. Prior to the commencement of the 3-D pilot studies, a two-dimensional (2-D) profile was recorded to test the acquisition parameters, which included the geometry of the survey, digital sample rate, and analog filter settings. The data were monitored in the field with a Bison 486 Explorer outdoor computer. The 2-D data were processed and displayed in the field. Both sites displayed coherent seismic reflections from the depths of interest on the field-stacked sections

Reassembly of anterior pituitary organization by hanging drop three-dimensional cell culture.

Science.gov (United States)

Tsukada, Takehiro; Kouki, Tom; Fujiwara, Ken; Ramadhani, Dini; Horiguchi, Kotaro; Kikuchi, Motoshi; Yashiro, Takashi

2013-08-29

The anterior pituitary gland comprises 5 types of hormone-producing cells and non-endocrine cells, such as folliculostellate (FS) cells. The cells form a lobular structure surrounded by extracellular matrix (ECM) but are not randomly distributed in each lobule; hormone-producing cells have affinities for specific cell types (topographic affinity), and FS cells form a homotypic meshwork. To determine whether this cell and ECM organization can be reproduced in vitro, we developed a 3-dimensional (3D) model that utilizes hanging drop cell culture. We found that the topographic affinities of hormone-producing cells were indeed maintained (ie, GH to ACTH cells, GH to TSH cells, PRL to LH/FSH cells). Fine structures in hormone-producing cells retained their normal appearance. In addition, FS cells displayed well-developed cytoplasmic protrusions, which interconnected with adjacent FS cells to form a 3D meshwork. In addition, reassembly of gap junctions and pseudofollicles among FS cells was observed in cell aggregates. Major ECM components-collagens and laminin-were deposited and distributed around the cells. In sum, the dissociated anterior pituitary cells largely maintained their in vivo anterior pituitary architectures. This culture system appears to be a powerful experimental tool for detailed analysis of anterior pituitary cell organization.

High-resolution computer-generated reflection holograms with three-dimensional effects written directly on a silicon surface by a femtosecond laser.

Science.gov (United States)

Wædegaard, Kristian J; Balling, Peter

2011-02-14

An infrared femtosecond laser has been used to write computer-generated holograms directly on a silicon surface. The high resolution offered by short-pulse laser ablation is employed to write highly detailed holograms with resolution up to 111 kpixels/mm2. It is demonstrated how three-dimensional effects can be realized in computer-generated holograms. Three-dimensional effects are visualized as a relative motion between different parts of the holographic reconstruction, when the hologram is moved relative to the reconstructing laser beam. Potential security applications are briefly discussed.

Printing Proteins as Microarrays for High-Throughput Function Determination

Science.gov (United States)

MacBeath, Gavin; Schreiber, Stuart L.

2000-09-01

Systematic efforts are currently under way to construct defined sets of cloned genes for high-throughput expression and purification of recombinant proteins. To facilitate subsequent studies of protein function, we have developed miniaturized assays that accommodate extremely low sample volumes and enable the rapid, simultaneous processing of thousands of proteins. A high-precision robot designed to manufacture complementary DNA microarrays was used to spot proteins onto chemically derivatized glass slides at extremely high spatial densities. The proteins attached covalently to the slide surface yet retained their ability to interact specifically with other proteins, or with small molecules, in solution. Three applications for protein microarrays were demonstrated: screening for protein-protein interactions, identifying the substrates of protein kinases, and identifying the protein targets of small molecules.

Exploring the sources of bacterial spoilers in beefsteaks by culture-independent high-throughput sequencing.

Directory of Open Access Journals (Sweden)

Francesca De Filippis

Full Text Available Microbial growth on meat to unacceptable levels contributes significantly to change meat structure, color and flavor and to cause meat spoilage. The types of microorganisms initially present in meat depend on several factors and multiple sources of contamination can be identified. The aims of this study were to evaluate the microbial diversity in beefsteaks before and after aerobic storage at 4°C and to investigate the sources of microbial contamination by examining the microbiota of carcasses wherefrom the steaks originated and of the processing environment where the beef was handled. Carcass, environmental (processing plant and meat samples were analyzed by culture-independent high-throughput sequencing of 16S rRNA gene amplicons. The microbiota of carcass swabs was very complex, including more than 600 operational taxonomic units (OTUs belonging to 15 different phyla. A significant association was found between beef microbiota and specific beef cuts (P<0.01 indicating that different cuts of the same carcass can influence the microbial contamination of beef. Despite the initially high complexity of the carcass microbiota, the steaks after aerobic storage at 4°C showed a dramatic decrease in microbial complexity. Pseudomonas sp. and Brochothrix thermosphacta were the main contaminants, and Acinetobacter, Psychrobacter and Enterobacteriaceae were also found. Comparing the relative abundance of OTUs in the different samples it was shown that abundant OTUs in beefsteaks after storage occurred in the corresponding carcass. However, the abundance of these same OTUs clearly increased in environmental samples taken in the processing plant suggesting that spoilage-associated microbial species originate from carcasses, they are carried to the processing environment where the meat is handled and there they become a resident microbiota. Such microbiota is then further spread on meat when it is handled and it represents the starting microbial association

Production and release of infectious hepatitis C virus from human liver cell cultures in the three-dimensional radial-flow bioreactor

International Nuclear Information System (INIS)

Aizaki, Hideki; Nagamori, Seishi; Matsuda, Mami; Kawakami, Hayato; Hashimoto, Osamu; Ishiko, Hiroaki; Kawada, Masaaki; Matsuura, Tomokazu; Hasumura, Satoshi; Matsuura, Yoshiharu; Suzuki, Tetsuro; Miyamura, Tatsuo

2003-01-01

Lack of efficient culture systems for hepatitis C virus (HCV) has been a major obstacle in HCV research. Human liver cells grown in a three-dimensional radial-flow bioreactor were successfully infected following inoculation with plasma from an HCV carrier. Subsequent detection of increased HCV RNA suggested viral replication. Furthermore, transfection of HCV RNA transcribed from full-length cDNA also resulted in the production and release of HCV virions into supernatant. Infectivity was shown by successful secondary passage to a new culture. Introduction of mutations in RNA helicase and polymerase regions of HCV cDNA abolished virus replication, indicating that reverse genetics of this system is possible. The ability to replicate and detect the extracellular release of HCV might provide clues with regard to the persistent nature of HCV infection. It will also accelerate research into the pathogenicity of HCV, as well as the development of prophylactic agents and new therapy

High-throughput theoretical design of lithium battery materials

International Nuclear Information System (INIS)

Ling Shi-Gang; Gao Jian; Xiao Rui-Juan; Chen Li-Quan

2016-01-01

The rapid evolution of high-throughput theoretical design schemes to discover new lithium battery materials is reviewed, including high-capacity cathodes, low-strain cathodes, anodes, solid state electrolytes, and electrolyte additives. With the development of efficient theoretical methods and inexpensive computers, high-throughput theoretical calculations have played an increasingly important role in the discovery of new materials. With the help of automatic simulation flow, many types of materials can be screened, optimized and designed from a structural database according to specific search criteria. In advanced cell technology, new materials for next generation lithium batteries are of great significance to achieve performance, and some representative criteria are: higher energy density, better safety, and faster charge/discharge speed. (topical review)

Three-dimensional bicontinuous nanoporous Au/polyaniline hybrid films for high-performance electrochemical supercapacitors

Science.gov (United States)

Lang, Xingyou; Zhang, Ling; Fujita, Takeshi; Ding, Yi; Chen, Mingwei

2012-01-01

We report three-dimensional bicontinuous nanoporous Au/polyaniline (PANI) composite films made by one-step electrochemical polymerization of PANI shell onto dealloyed nanoporous gold (NPG) skeletons for the applications in electrochemical supercapacitors. The NPG/PANI based supercapacitors exhibit ultrahigh volumetric capacitance (∼1500 F cm-3) and energy density (∼0.078 Wh cm-3), which are seven and four orders of magnitude higher than these of electrolytic capacitors, with the same power density up to ∼190 W cm-3. The outstanding capacitive performances result from a novel nanoarchitecture in which pseudocapacitive PANI shells are incorporated into pore channels of highly conductive NPG, making them promising candidates as electrode materials in supercapacitor devices combing high-energy storage densities with high-power delivery.

An Ultra-High Fluorescence Enhancement and High Throughput Assay for Revealing Expression and Internalization of Chemokine Receptor CXCR4.

Science.gov (United States)

He, Hua; Wang, Xiaojuan; Cheng, Tiantian; Xia, Yongqing; Lao, Jun; Ge, Baosheng; Ren, Hao; Khan, Naseer Ullah; Huang, Fang

2016-04-18

Revealing chemokine receptor CXCR4 expression, distribution, and internalization levels in different cancers helps to evaluate cancer progression or prognosis and to set personalized treatment strategy. We here describe a sensitive and high-throughput immunoassay for determining CXCR4 expression and distribution in cancer cells. The assay is accessible to a wide range of users in an ordinary lab only by dip-coating poly(styrene-co-N-isopropylacrylamide) spheres on the glass substrate. The self- assembled spheres form three-dimensional photonic colloidal crystals which enhance the fluorescence of CF647 and Alexa Fluor 647 by a factor of up to 1000. CXCR4 in cells is detected by using the sandwich immunoassay, where the primary antibody recognizes CXCR4 and the secondary antibody is labeled with CF647. With the newly established assay, we quantified the total expression of CXCR4, its distribution on the cell membrane and cytoplasm, and revealed their internalization level upon SDF-1α activation in various cancer cells, even for those with extremely low expression level. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Mechanotransductive Regulation of Gap-Junction Activity Between MLO-Y4 Osteocyte-Like and MC3T3-E1 Osteoblast-Like Cells in Three-Dimensional Co-Culture.

Science.gov (United States)

Juran, C. M.; Blaber, E. A.; Almeida, E. A. C.

2016-01-01

Cell and animal studies conducted onboard the International Space Station and formerly on Shuttle flights have provided groundbreaking data illuminating the deleterious biological response of bone to mechanical unloading. However the intercellular communicative mechanisms associated with the regulation of bone synthesis and bone resorption cells are still largely unknown. Connexin-43 (CX43), a gap junction protein, is hypothesized to play a significant role in osteoblast and osteocyte signaling. The purpose of this investigation was to evaluate within a novel three-dimensional microenvironment how the osteocyte-osteoblast gap-junction expression changes when cultures are exposed to exaggerated mechanical load. MLO-Y4 osteocyte-like cells were cultured on a 3D-Biotek polystyrene insert and placed in direct contact with an MC3T3-E1 pre-osteoblast co-cultured monolayer and exposed to 48 h of mechanical stimulation (pulsatile fluid flow (PFF) or monolayer cyclic stretch (MCS)) then evaluated for viability, proliferation, metabolism, and CX43 expression. Mono-cultured MLO-Y4 and MC3T3-E1 control experiments were conducted under PFF and MCS stimulation to observe how strain application stimuli (PFF cell membrane shear or MCS cell focal adhesion/attachment loading) initiates different signaling pathways or downstream regulatory controls. TotalLive cell count, viability and metabolic reduction (Trypan Blue, LIVEDead and Alamar Blue analysis respectively) indicate that mechanical activation of MC3T3-E1 cells inhibits proliferation while maintaining an average 1.04E4 reductioncell metabolic rate, *p0.05 n4. MLO-Y4s in monolayer culture increase in number when exposed to MCS loading but the percent of live cells within the population is low (46.3 total count, *p0.05 n4), these results may indicate an apoptotic signaling cascade. PFF stimulation of the three-dimensional co-cultures elicits a universal increase in CX43 in MLO-Y4 and MC3T3-E1 cells, illustrated by

Rapid Screening of Acetylcholinesterase Inhibitors by Effect-Directed Analysis Using LC × LC Fractionation, a High Throughput in Vitro Assay, and Parallel Identification by Time of Flight Mass Spectrometry.

Science.gov (United States)

Ouyang, Xiyu; Leonards, Pim E G; Tousova, Zuzana; Slobodnik, Jaroslav; de Boer, Jacob; Lamoree, Marja H

2016-02-16

Effect-directed analysis (EDA) is a useful tool to identify bioactive compounds in complex samples. However, identification in EDA is usually challenging, mainly due to limited separation power of the liquid chromatography based fractionation. In this study, comprehensive two-dimensional liquid chromatography (LC × LC) based microfractionation combined with parallel high resolution time of flight (HR-ToF) mass spectrometric detection and a high throughput acetylcholinesterase (AChE) assay was developed. The LC × LC fractionation method was validated using analytical standards and a C18 and pentafluorophenyl (PFP) stationary phase combination was selected for the two-dimensional separation and fractionation in four 96-well plates. The method was successfully applied to identify AChE inhibitors in a wastewater treatment plant (WWTP) effluent. Good orthogonality (>0.9) separation was achieved and three AChE inhibitors (tiapride, amisulpride, and lamotrigine), used as antipsychotic medicines, were identified and confirmed by two-dimensional retention alignment as well as their AChE inhibition activity.

High-Throughput Block Optical DNA Sequence Identification.

Science.gov (United States)

Sagar, Dodderi Manjunatha; Korshoj, Lee Erik; Hanson, Katrina Bethany; Chowdhury, Partha Pratim; Otoupal, Peter Britton; Chatterjee, Anushree; Nagpal, Prashant

2018-01-01

Optical techniques for molecular diagnostics or DNA sequencing generally rely on small molecule fluorescent labels, which utilize light with a wavelength of several hundred nanometers for detection. Developing a label-free optical DNA sequencing technique will require nanoscale focusing of light, a high-throughput and multiplexed identification method, and a data compression technique to rapidly identify sequences and analyze genomic heterogeneity for big datasets. Such a method should identify characteristic molecular vibrations using optical spectroscopy, especially in the "fingerprinting region" from ≈400-1400 cm -1 . Here, surface-enhanced Raman spectroscopy is used to demonstrate label-free identification of DNA nucleobases with multiplexed 3D plasmonic nanofocusing. While nanometer-scale mode volumes prevent identification of single nucleobases within a DNA sequence, the block optical technique can identify A, T, G, and C content in DNA k-mers. The content of each nucleotide in a DNA block can be a unique and high-throughput method for identifying sequences, genes, and other biomarkers as an alternative to single-letter sequencing. Additionally, coupling two complementary vibrational spectroscopy techniques (infrared and Raman) can improve block characterization. These results pave the way for developing a novel, high-throughput block optical sequencing method with lossy genomic data compression using k-mer identification from multiplexed optical data acquisition. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Three-Dimensional Porous Iron Vanadate Nanowire Arrays as a High-Performance Lithium-Ion Battery.

Science.gov (United States)

Cao, Yunhe; Fang, Dong; Liu, Ruina; Jiang, Ming; Zhang, Hang; Li, Guangzhong; Luo, Zhiping; Liu, Xiaoqing; Xu, Jie; Xu, Weilin; Xiong, Chuanxi

2015-12-23

Development of three-dimensional nanoarchitectures on current collectors has emerged as an effective strategy for enhancing rate capability and cycling stability of the electrodes. Herein, a new type of three-dimensional porous iron vanadate (Fe0.12V2O5) nanowire arrays on a Ti foil has been synthesized by a hydrothermal method. The as-prepared Fe0.12V2O5 nanowires are about 30 nm in diameter and several micrometers in length. The effect of reaction time on the resulting morphology is investigated and the mechanism for the nanowire formation is proposed. As an electrode material used in lithium-ion batteries, the unique configuration of the Fe0.12V2O5 nanowire arrays presents enhanced capacitance, satisfying rate capability and good cycling stability, as evaluated by cyclic voltammetry and galvanostatic discharge-charge cycling. It delivers a high discharge capacity of 293 mAh·g(-1) at 2.0-3.6 V or 382.2 mAh·g(-1) at 1.0-4.0 V after 50 cycles at 30 mA·g(-1).

Three-dimensional particle image velocimetry measurement technique

International Nuclear Information System (INIS)

Hassan, Y.A.; Seeley, C.H.; Henderson, J.A.; Schmidl, W.D.

2004-01-01

The experimental flow visualization tool, Particle Image Velocimetry (PIV), is being used to determine the velocity field in two-dimensional fluid flows. In the past few years, the technique has been improved to allow the capture of flow fields in three dimensions. This paper describes changes which were made to two existing two-dimensional tracking algorithms to enable them to track three-dimensional PIV data. Results of the tests performed on these three-dimensional routines with synthetic data are presented. Experimental data was also used to test the tracking algorithms. The test setup which was used to acquire the three-dimensional experimental data is described, along with the results from both of the tracking routines which were used to analyze the experimental data. (author)

Three-Dimensional Triplet Tracking for LHC and Future High Rate Experiments

CERN Document Server

Schöning, Andre

2014-10-20

The hit combinatorial problem is a main challenge for track reconstruction and triggering at high rate experiments. At hadron colliders the dominant fraction of hits is due to low momentum tracks for which multiple scattering (MS) effects dominate the hit resolution. MS is also the dominating source for hit confusion and track uncertainties in low energy precision experiments. In all such environments, where MS dominates, track reconstruction and fitting can be largely simplified by using three-dimensional (3D) hit-triplets as provided by pixel detectors. This simplification is possible since track uncertainties are solely determined by MS if high precision spatial information is provided. Fitting of hit-triplets is especially simple for tracking detectors in solenoidal magnetic fields. The over-constrained 3D-triplet method provides a complete set of track parameters and is robust against fake hit combinations. The triplet method is ideally suited for pixel detectors where hits can be treated as 3D-space poi...

High-throughput image analysis of tumor spheroids: a user-friendly software application to measure the size of spheroids automatically and accurately.

Science.gov (United States)

Chen, Wenjin; Wong, Chung; Vosburgh, Evan; Levine, Arnold J; Foran, David J; Xu, Eugenia Y

2014-07-08

The increasing number of applications of three-dimensional (3D) tumor spheroids as an in vitro model for drug discovery requires their adaptation to large-scale screening formats in every step of a drug screen, including large-scale image analysis. Currently there is no ready-to-use and free image analysis software to meet this large-scale format. Most existing methods involve manually drawing the length and width of the imaged 3D spheroids, which is a tedious and time-consuming process. This study presents a high-throughput image analysis software application - SpheroidSizer, which measures the major and minor axial length of the imaged 3D tumor spheroids automatically and accurately; calculates the volume of each individual 3D tumor spheroid; then outputs the results in two different forms in spreadsheets for easy manipulations in the subsequent data analysis. The main advantage of this software is its powerful image analysis application that is adapted for large numbers of images. It provides high-throughput computation and quality-control workflow. The estimated time to process 1,000 images is about 15 min on a minimally configured laptop, or around 1 min on a multi-core performance workstation. The graphical user interface (GUI) is also designed for easy quality control, and users can manually override the computer results. The key method used in this software is adapted from the active contour algorithm, also known as Snakes, which is especially suitable for images with uneven illumination and noisy background that often plagues automated imaging processing in high-throughput screens. The complimentary "Manual Initialize" and "Hand Draw" tools provide the flexibility to SpheroidSizer in dealing with various types of spheroids and diverse quality images. This high-throughput image analysis software remarkably reduces labor and speeds up the analysis process. Implementing this software is beneficial for 3D tumor spheroids to become a routine in vitro model

High-dimensional data: p >> n in mathematical statistics and bio-medical applications

OpenAIRE

Van De Geer, Sara A.; Van Houwelingen, Hans C.

2004-01-01

The workshop 'High-dimensional data: p >> n in mathematical statistics and bio-medical applications' was held at the Lorentz Center in Leiden from 9 to 20 September 2002. This special issue of Bernoulli contains a selection of papers presented at that workshop. ¶ The introduction of high-throughput micro-array technology to measure gene-expression levels and the publication of the pioneering paper by Golub et al. (1999) has brought to life a whole new branch of data analysis under the name of...

Three-dimensional growth of human endothelial cells in an automated cell culture experiment container during the SpaceX CRS-8 ISS space mission - The SPHEROIDS project.

Science.gov (United States)

Pietsch, Jessica; Gass, Samuel; Nebuloni, Stefano; Echegoyen, David; Riwaldt, Stefan; Baake, Christin; Bauer, Johann; Corydon, Thomas J; Egli, Marcel; Infanger, Manfred; Grimm, Daniela

2017-04-01

Human endothelial cells (ECs) were sent to the International Space Station (ISS) to determine the impact of microgravity on the formation of three-dimensional structures. For this project, an automatic experiment unit (EU) was designed allowing cell culture in space. In order to enable a safe cell culture, cell nourishment and fixation after a pre-programmed timeframe, the materials used for construction of the EUs were tested in regard to their biocompatibility. These tests revealed a high biocompatibility for all parts of the EUs, which were in contact with the cells or the medium used. Most importantly, we found polyether ether ketones for surrounding the incubation chamber, which kept cellular viability above 80% and allowed the cells to adhere as long as they were exposed to normal gravity. After assembling the EU the ECs were cultured therein, where they showed good cell viability at least for 14 days. In addition, the functionality of the automatic medium exchange, and fixation procedures were confirmed. Two days before launch, the ECs were cultured in the EUs, which were afterwards mounted on the SpaceX CRS-8 rocket. 5 and 12 days after launch the cells were fixed. Subsequent analyses revealed a scaffold-free formation of spheroids in space. Copyright © 2017 Elsevier Ltd. All rights reserved.

Three-Dimensional Numerical Analysis of an Operating Helical Rotor Pump at High Speeds and High Pressures including Cavitation

Directory of Open Access Journals (Sweden)

Zhou Yang

2017-01-01

Full Text Available High pressures, high speeds, low noise and miniaturization is the direction of development in hydraulic pump. According to the development trend, an operating helical rotor pump (HRP at high speeds and high pressures has been designed and produced, which rotational speed can reach 12000r/min and outlet pressure is as high as 25MPa. Three-dimensional simulation with and without cavitation inside the HRP is completed by the means of the computational fluid dynamics (CFD in this paper, which contributes to understand the complex fluid flow inside it. Moreover, the influences of the rotational speeds of the HRP with and without cavitation has been simulated at 25MPa.

Structures of two-dimensional three-body systems

International Nuclear Information System (INIS)

Ruan, W.Y.; Liu, Y.Y.; Bao, C.G.

1996-01-01

Features of the structure of L = 0 states of a two-dimensional three-body model system have been investigated. Three types of permutation symmetry of the spatial part, namely symmetric, antisymmetric, and mixed, have been considered. A comparison has been made between the two-dimensional system and the corresponding three-dimensional one. The effect of symmetry on microscopic structures is emphasized. (author)

High throughput salt separation from uranium deposits

Energy Technology Data Exchange (ETDEWEB)

Kwon, S.W.; Park, K.M.; Kim, J.G.; Kim, I.T.; Park, S.B., E-mail: swkwon@kaeri.re.kr [Korea Atomic Energy Research Inst. (Korea, Republic of)

2014-07-01

It is very important to increase the throughput of the salt separation system owing to the high uranium content of spent nuclear fuel and high salt fraction of uranium dendrites in pyroprocessing. Multilayer porous crucible system was proposed to increase a throughput of the salt distiller in this study. An integrated sieve-crucible assembly was also investigated for the practical use of the porous crucible system. The salt evaporation behaviors were compared between the conventional nonporous crucible and the porous crucible. Two step weight reductions took place in the porous crucible, whereas the salt weight reduced only at high temperature by distillation in a nonporous crucible. The first weight reduction in the porous crucible was caused by the liquid salt penetrated out through the perforated crucible during the temperature elevation until the distillation temperature. Multilayer porous crucibles have a benefit to expand the evaporation surface area. (author)

A DETERMINISTIC METHOD FOR TRANSIENT, THREE-DIMENSIONAL NUETRON TRANSPORT

International Nuclear Information System (INIS)

S. GOLUOGLU, C. BENTLEY, R. DEMEGLIO, M. DUNN, K. NORTON, R. PEVEY I.SUSLOV AND H.L. DODDS

1998-01-01

A deterministic method for solving the time-dependent, three-dimensional Boltzmam transport equation with explicit representation of delayed neutrons has been developed and evaluated. The methodology used in this study for the time variable of the neutron flux is known as the improved quasi-static (IQS) method. The position, energy, and angle-dependent neutron flux is computed deterministically by using the three-dimensional discrete ordinates code TORT. This paper briefly describes the methodology and selected results. The code developed at the University of Tennessee based on this methodology is called TDTORT. TDTORT can be used to model transients involving voided and/or strongly absorbing regions that require transport theory for accuracy. This code can also be used to model either small high-leakage systems, such as space reactors, or asymmetric control rod movements. TDTORT can model step, ramp, step followed by another step, and step followed by ramp type perturbations. It can also model columnwise rod movement can also be modeled. A special case of columnwise rod movement in a three-dimensional model of a boiling water reactor (BWR) with simple adiabatic feedback is also included. TDTORT is verified through several transient one-dimensional, two-dimensional, and three-dimensional benchmark problems. The results show that the transport methodology and corresponding code developed in this work have sufficient accuracy and speed for computing the dynamic behavior of complex multidimensional neutronic systems

Three dimensional CT imaging of ossicular chain: a preliminary study

International Nuclear Information System (INIS)

Hu Chunhong; Zhong Shenbin; Fu Yindi; Zhu Wei; Wang Xueyuan; Chen Jianhua; Ding Yi

2001-01-01

Objective: To analysis the features of normal and abnormal ossicular chain in three dimensional images and asses the best parameters and its usefulness in diagnosis and treatment of chronic otitis media (COM). Methods: All patients, including 43 patients with normal ears and 24 ears with COM, were examined using spiral CT with inner ear software, 1-mm slice width and 1 pitch. SSD method was used in three dimensional reconstruction and the threshold was 100-300 Hu. Results: In normal cases, Malleus, incus, stapes crura, incudomalleal joints and incudostapedial joints were displayed well, but stapes footplate unsatisfactorily. The disruption of the ossicular chain showed in three-dimensional images in cases of chronic otitis media was in accord with that seen in the operation. Conclusion: It is very important for imaging with high quality through selecting proper parameters, and three-dimensional image can provide valuable information for surgery

A novel three-dimensional and high-definition flexible scope.

Science.gov (United States)

Nishiyama, Kenichi; Natori, Yoshihiro; Oka, Kazunari

2014-06-01

Recent high-tech innovations in digital surgical technology have led to advances in three-dimensional (3D) and high-definition (HD) operating scopes. We introduce a novel 3D-HD flexible surgical scope called "3D-Eye-Flex" and evaluate its utility as an alternative to the operating microscope. The 3D-Eye-Flex has a 15 mm long 3D-HD scope-head with a 15 mm outer diameter, a focus distance of 18-100 mm and 80° angle of view. Attached to a 615-mm-long flexible bellows, 3D-Eye-Flex can be easily fixed to the operating table. Microsurgical dissection of wet brain tissue and drilling a skull base model were performed under the scope while using the 3D-HD video monitor. This scope system provided excellent illumination and image quality during the procedures. A large depth of field with stereoscopic vision had a greater advantage over using an operating microscope. 3D-Eye-Flex was easy to manipulate and provided an abundance of space above the operative field. Surgeons felt comfortable while working and could easily shift the position of the scope. This novel 3D-HD flexible scope is an effective alternative to the operating microscope as a new surgeon's eye and will be suitable for digital image-based surgery with further refinement.

High throughput, low set-up time reconfigurable linear feedback shift registers

NARCIS (Netherlands)

Nas, R.J.M.; Berkel, van C.H.

2010-01-01

This paper presents a hardware design for a scalable, high throughput, configurable LFSR. High throughput is achieved by producing L consecutive outputs per clock cycle with a clock cycle period that, for practical cases, increases only logarithmically with the block size L and the length of the

High-throughput epitope identification for snakebite antivenom

DEFF Research Database (Denmark)

Engmark, Mikael; De Masi, Federico; Laustsen, Andreas Hougaard

Insight into the epitopic recognition pattern for polyclonal antivenoms is a strong tool for accurate prediction of antivenom cross-reactivity and provides a basis for design of novel antivenoms. In this work, a high-throughput approach was applied to characterize linear epitopes in 966 individua...... toxins from pit vipers (Crotalidae) using the ICP Crotalidae antivenom. Due to an abundance of snake venom metalloproteinases and phospholipase A2s in the venoms used for production of the investigated antivenom, this study focuses on these toxin families.......Insight into the epitopic recognition pattern for polyclonal antivenoms is a strong tool for accurate prediction of antivenom cross-reactivity and provides a basis for design of novel antivenoms. In this work, a high-throughput approach was applied to characterize linear epitopes in 966 individual...

Elastocapillary fabrication of three-dimensional microstructures

NARCIS (Netherlands)

van Honschoten, J.W.; Berenschot, Johan W.; Ondarcuhu, T.; Sanders, Remco G.P.; Sundaram, J.; Elwenspoek, Michael Curt; Tas, Niels Roelof

2010-01-01

We describe the fabrication of three-dimensional microstructures by means of capillary forces. Using an origami-like technique, planar silicon nitride structures of various geometries are folded to produce three-dimensional objects of 50–100 m. Capillarity is a particularly effective mechanism since

Three dimensional transport model for toroidal plasmas

International Nuclear Information System (INIS)

Copenhauer, C.

1980-12-01

A nonlinear MHD model, developed for three-dimensional toroidal geometries (asymmetric) and for high β (β approximately epsilon), is used as a basis for a three-dimensional transport model. Since inertia terms are needed in describing evolving magnetic islands, the model can calculate transport, both in the transient phase before nonlinear saturation of magnetic islands and afterwards on the resistive time scale. In the β approximately epsilon ordering, the plasma does not have sufficient energy to compress the parallel magnetic field, which allows the Alfven wave to be eliminated in the reduced nonlinear equations, and the model then follows the slower time scales. The resulting perpendicular and parallel plasma drift velocities can be identified with those of guiding center theory

High performance supercapacitors based on three-dimensional ultralight flexible manganese oxide nanosheets/carbon foam composites

Science.gov (United States)

He, Shuijian; Chen, Wei

2014-09-01

The syntheses and capacitance performances of ultralight and flexible MnO2/carbon foam (MnO2/CF) hybrids are systematically studied. Flexible carbon foam with a low mass density of 6.2 mg cm-3 and high porosity of 99.66% is simply obtained by carbonization of commercially available and low-cost melamine resin foam. With the high porous carbon foam as framework, ultrathin MnO2 nanosheets are grown through in situ redox reaction between KMnO4 and carbon foam. The three-dimensional (3D) MnO2/CF networks exhibit highly ordered hierarchical pore structure. Attributed to the good flexibility and ultralight weight, the MnO2/CF nanomaterials can be directly fabricated into supercapacitor electrodes without any binder and conductive agents. Moreover, the pseudocapacitance of the MnO2 nanosheets is enhanced by the fast ion diffusion in the three-dimensional porous architecture and by the conductive carbon foam skeleton as well as good contact of carbon/oxide interfaces. Supercapacitor based on the MnO2/CF composite with 3.4% weight percent of MnO2 shows a high specific capacitance of 1270.5 F g-1 (92.7% of the theoretical specific capacitance of MnO2) and high energy density of 86.2 Wh kg-1. The excellent capacitance performance of the present 3D ultralight and flexible nanomaterials make them promising candidates as electrode materials for supercapacitors.

In vitro three-dimensional coculturing poly3-hydroxybutyrate-co-3-hydroxyhexanoate with mouse-induced pluripotent stem cells for myocardial patch application.

Science.gov (United States)

Shijun, Xu; Junsheng, Mu; Jianqun, Zhang; Ping, Bo

2016-03-01

Identifying a suitable polymeric biomaterial for myocardial patch repair following myocardial infarction, cerebral infarction, and cartilage injury is essential. This study aimed to investigate the effect of the novel polymer material, poly3-hydroxybutyrate-co-3-hydroxyhexanoate, on the adhesion, proliferation, and differentiation of mouse-induced pluripotent stem cells in vitro. Mouse-induced pluripotent stem cells were isolated, expanded, and cultured on either two-dimensional or three-dimensional poly3-hydroxybutyrate-co-3-hydroxyhexanoate films (membranes were perforated to imitate three-dimensional space). Following attachment onto the films, mouse-induced pluripotent stem cell morphology was visualized using scanning electron microscopy. Cell vitality was detected using the Cell Counting Kit-8 assay and cell proliferation was observed using fluorescent 4',6-diamidino-2-phenylindole (DAPI) staining. Mouse-induced pluripotent stem cells were induced into cardiomyocytes by differentiation medium containing vitamin C. A control group in the absence of an inducer was included. Mouse-induced pluripotent stem cell survival and differentiation were observed using immunofluorescence and flow cytometry, respectively. Mouse-induced pluripotent stem cells growth, proliferation, and differentiation were observed on both two-dimensional and three-dimensional poly3-hydroxybutyrate-co-3-hydroxyhexanoate films. Vitamin C markedly improved the efficiency of mouse-induced pluripotent stem cells differentiation into cardiomyocytes on poly3-hydroxybutyrate-co-3-hydroxyhexanoate films. Three-dimensional culture was better at promoting mouse-induced pluripotent stem cell proliferation and differentiation compared with two-dimensional culture. © The Author(s) 2016.

High-throughput microfluidic mixing and multiparametric cell sorting for bioactive compound screening.

Science.gov (United States)

Young, Susan M; Curry, Mark S; Ransom, John T; Ballesteros, Juan A; Prossnitz, Eric R; Sklar, Larry A; Edwards, Bruce S

2004-03-01

HyperCyt, an automated sample handling system for flow cytometry that uses air bubbles to separate samples sequentially introduced from multiwell plates by an autosampler. In a previously documented HyperCyt configuration, air bubble separated compounds in one sample line and a continuous stream of cells in another are mixed in-line for serial flow cytometric cell response analysis. To expand capabilities for high-throughput bioactive compound screening, the authors investigated using this system configuration in combination with automated cell sorting. Peptide ligands were sampled from a 96-well plate, mixed in-line with fluo-4-loaded, formyl peptide receptor-transfected U937 cells, and screened at a rate of 3 peptide reactions per minute with approximately 10,000 cells analyzed per reaction. Cell Ca(2+) responses were detected to as little as 10(-11) M peptide with no detectable carryover between samples at up to 10(-7) M peptide. After expansion in culture, cells sort-purified from the 10% highest responders exhibited enhanced sensitivity and more sustained responses to peptide. Thus, a highly responsive cell subset was isolated under high-throughput mixing and sorting conditions in which response detection capability spanned a 1000-fold range of peptide concentration. With single-cell readout systems for protein expression libraries, this technology offers the promise of screening millions of discrete compound interactions per day.

Novel Acoustic Loading of a Mass Spectrometer: Toward Next-Generation High-Throughput MS Screening.

Science.gov (United States)

Sinclair, Ian; Stearns, Rick; Pringle, Steven; Wingfield, Jonathan; Datwani, Sammy; Hall, Eric; Ghislain, Luke; Majlof, Lars; Bachman, Martin

2016-02-01

High-throughput, direct measurement of substrate-to-product conversion by label-free detection, without the need for engineered substrates or secondary assays, could be considered the "holy grail" of drug discovery screening. Mass spectrometry (MS) has the potential to be part of this ultimate screening solution, but is constrained by the limitations of existing MS sample introduction modes that cannot meet the throughput requirements of high-throughput screening (HTS). Here we report data from a prototype system (Echo-MS) that uses acoustic droplet ejection (ADE) to transfer femtoliter-scale droplets in a rapid, precise, and accurate fashion directly into the MS. The acoustic source can load samples into the MS from a microtiter plate at a rate of up to three samples per second. The resulting MS signal displays a very sharp attack profile and ions are detected within 50 ms of activation of the acoustic transducer. Additionally, we show that the system is capable of generating multiply charged ion species from simple peptides and large proteins. The combination of high speed and low sample volume has significant potential within not only drug discovery, but also other areas of the industry. © 2015 Society for Laboratory Automation and Screening.

A high-throughput and quantitative method to assess the mutagenic potential of translesion DNA synthesis

Science.gov (United States)

Taggart, David J.; Camerlengo, Terry L.; Harrison, Jason K.; Sherrer, Shanen M.; Kshetry, Ajay K.; Taylor, John-Stephen; Huang, Kun; Suo, Zucai

2013-01-01

Cellular genomes are constantly damaged by endogenous and exogenous agents that covalently and structurally modify DNA to produce DNA lesions. Although most lesions are mended by various DNA repair pathways in vivo, a significant number of damage sites persist during genomic replication. Our understanding of the mutagenic outcomes derived from these unrepaired DNA lesions has been hindered by the low throughput of existing sequencing methods. Therefore, we have developed a cost-effective high-throughput short oligonucleotide sequencing assay that uses next-generation DNA sequencing technology for the assessment of the mutagenic profiles of translesion DNA synthesis catalyzed by any error-prone DNA polymerase. The vast amount of sequencing data produced were aligned and quantified by using our novel software. As an example, the high-throughput short oligonucleotide sequencing assay was used to analyze the types and frequencies of mutations upstream, downstream and at a site-specifically placed cis–syn thymidine–thymidine dimer generated individually by three lesion-bypass human Y-family DNA polymerases. PMID:23470999

High-throughput screening of small molecule libraries using SAMDI mass spectrometry.

Science.gov (United States)

Gurard-Levin, Zachary A; Scholle, Michael D; Eisenberg, Adam H; Mrksich, Milan

2011-07-11

High-throughput screening is a common strategy used to identify compounds that modulate biochemical activities, but many approaches depend on cumbersome fluorescent reporters or antibodies and often produce false-positive hits. The development of "label-free" assays addresses many of these limitations, but current approaches still lack the throughput needed for applications in drug discovery. This paper describes a high-throughput, label-free assay that combines self-assembled monolayers with mass spectrometry, in a technique called SAMDI, as a tool for screening libraries of 100,000 compounds in one day. This method is fast, has high discrimination, and is amenable to a broad range of chemical and biological applications.

Three-dimensional printing and pediatric liver disease.

Science.gov (United States)

Alkhouri, Naim; Zein, Nizar N

2016-10-01

Enthusiastic physicians and medical researchers are investigating the role of three-dimensional printing in medicine. The purpose of the current review is to provide a concise summary of the role of three-dimensional printing technology as it relates to the field of pediatric hepatology and liver transplantation. Our group and others have recently demonstrated the feasibility of printing three-dimensional livers with identical anatomical and geometrical landmarks to the native liver to facilitate presurgical planning of complex liver surgeries. Medical educators are exploring the use of three-dimensional printed organs in anatomy classes and surgical residencies. Moreover, mini-livers are being developed by regenerative medicine scientist as a way to test new drugs and, eventually, whole livers will be grown in the laboratory to replace organs with end-stage disease solving the organ shortage problem. From presurgical planning to medical education to ultimately the bioprinting of whole organs for transplantation, three-dimensional printing will change medicine as we know in the next few years.

A Novel Multiparametric Drug-Scoring Method for High-Throughput Screening of 3D Multicellular Tumor Spheroids Using the Celigo Image Cytometer.

Science.gov (United States)

Cribbes, Scott; Kessel, Sarah; McMenemy, Scott; Qiu, Jean; Chan, Leo Li-Ying

2017-06-01

Three-dimensional (3D) tumor models have been increasingly used to investigate and characterize cancer drug compounds. The ability to perform high-throughput screening of 3D multicellular tumor spheroids (MCTS) can highly improve the efficiency and cost-effectiveness of discovering potential cancer drug candidates. Previously, the Celigo Image Cytometer has demonstrated a novel method for high-throughput screening of 3D multicellular tumor spheroids. In this work, we employed the Celigo Image Cytometer to examine the effects of 14 cancer drug compounds on 3D MCTS of the glioblastoma cell line U87MG in 384-well plates. Using parameters such as MCTS diameter and invasion area, growth and invasion were monitored for 9 and 3 d, respectively. Furthermore, fluorescent staining with calcein AM, propidium iodide, Hoechst 33342, and caspase 3/7 was performed at day 9 posttreatment to measure viability and apoptosis. Using the kinetic and endpoint data generated, we created a novel multiparametric drug-scoring system for 3D MCTS that can be used to identify and classify potential drug candidates earlier in the drug discovery process. Furthermore, the combination of quantitative and qualitative image data can be used to delineate differences between drugs that induce cytotoxic and cytostatic effects. The 3D MCTS-based multiparametric scoring method described here can provide an alternative screening method to better qualify tested drug compounds.

Preliminary High-Throughput Metagenome Assembly

Energy Technology Data Exchange (ETDEWEB)

Dusheyko, Serge; Furman, Craig; Pangilinan, Jasmyn; Shapiro, Harris; Tu, Hank

2007-03-26

Metagenome data sets present a qualitatively different assembly problem than traditional single-organism whole-genome shotgun (WGS) assembly. The unique aspects of such projects include the presence of a potentially large number of distinct organisms and their representation in the data set at widely different fractions. In addition, multiple closely related strains could be present, which would be difficult to assemble separately. Failure to take these issues into account can result in poor assemblies that either jumble together different strains or which fail to yield useful results. The DOE Joint Genome Institute has sequenced a number of metagenomic projects and plans to considerably increase this number in the coming year. As a result, the JGI has a need for high-throughput tools and techniques for handling metagenome projects. We present the techniques developed to handle metagenome assemblies in a high-throughput environment. This includes a streamlined assembly wrapper, based on the JGI?s in-house WGS assembler, Jazz. It also includes the selection of sensible defaults targeted for metagenome data sets, as well as quality control automation for cleaning up the raw results. While analysis is ongoing, we will discuss preliminary assessments of the quality of the assembly results (http://fames.jgi-psf.org).

Three dimensional investigation of oceanic active faults. A demonstration survey

Energy Technology Data Exchange (ETDEWEB)

Nakao, Seizo; Kishimoto, Kiyoyuki; Kuramoto, Shinichi; Sato, Mikio [Geological Survey of Japan, Tsukuba, Ibaraki (Japan)

1998-02-01

In order to upgrade probability of activity and action potential evaluation of oceanic active faults which have some important effects on nuclear facilities, trench type oceanic active fault was investigated three dimensionally. Contents of the investigation were high precision sea bottom topographic survey and sea bottom back scattering wave image data observation by using a sea bottom topography acoustic imaginator. And, by high resolution earthquake wave survey, high precision survey of an active fault under sea bottom was conducted to detect oceanic active faults three-dimensionally. Furthermore, the generally issued data were summarized to promote to construct a data base for evaluating the active faults. (G.K.)

Three dimensional investigation of oceanic active faults. A demonstration survey

International Nuclear Information System (INIS)

Nakao, Seizo; Kishimoto, Kiyoyuki; Kuramoto, Shinichi; Sato, Mikio

1998-01-01

In order to upgrade probability of activity and action potential evaluation of oceanic active faults which have some important effects on nuclear facilities, trench type oceanic active fault was investigated three dimensionally. Contents of the investigation were high precision sea bottom topographic survey and sea bottom back scattering wave image data observation by using a sea bottom topography acoustic imaginator. And, by high resolution earthquake wave survey, high precision survey of an active fault under sea bottom was conducted to detect oceanic active faults three-dimensionally. Furthermore, the generally issued data were summarized to promote to construct a data base for evaluating the active faults. (G.K.)

Visual Interpretation with Three-Dimensional Annotations (VITA): Three-Dimensional Image Interpretation Tool for Radiological Reporting

OpenAIRE

Roy, Sharmili; Brown, Michael S.; Shih, George L.

2013-01-01

This paper introduces a software framework called Visual Interpretation with Three-Dimensional Annotations (VITA) that is able to automatically generate three-dimensional (3D) visual summaries based on radiological annotations made during routine exam reporting. VITA summaries are in the form of rotating 3D volumes where radiological annotations are highlighted to place important clinical observations into a 3D context. The rendered volume is produced as a Digital Imaging and Communications i...

Three-dimensional reconstruction of functional brain images

International Nuclear Information System (INIS)

Inoue, Masato; Shoji, Kazuhiko; Kojima, Hisayoshi; Hirano, Shigeru; Naito, Yasushi; Honjo, Iwao

1999-01-01

We consider PET (positron emission tomography) measurement with SPM (Statistical Parametric Mapping) analysis to be one of the most useful methods to identify activated areas of the brain involved in language processing. SPM is an effective analytical method that detects markedly activated areas over the whole brain. However, with the conventional presentations of these functional brain images, such as horizontal slices, three directional projection, or brain surface coloring, makes understanding and interpreting the positional relationships among various brain areas difficult. Therefore, we developed three-dimensionally reconstructed images from these functional brain images to improve the interpretation. The subjects were 12 normal volunteers. The following three types of images were constructed: routine images by SPM, three-dimensional static images, and three-dimensional dynamic images, after PET images were analyzed by SPM during daily dialog listening. The creation of images of both the three-dimensional static and dynamic types employed the volume rendering method by VTK (The Visualization Toolkit). Since the functional brain images did not include original brain images, we synthesized SPM and MRI brain images by self-made C++ programs. The three-dimensional dynamic images were made by sequencing static images with available software. Images of both the three-dimensional static and dynamic types were processed by a personal computer system. Our newly created images showed clearer positional relationships among activated brain areas compared to the conventional method. To date, functional brain images have been employed in fields such as neurology or neurosurgery, however, these images may be useful even in the field of otorhinolaryngology, to assess hearing and speech. Exact three-dimensional images based on functional brain images are important for exact and intuitive interpretation, and may lead to new developments in brain science. Currently, the surface

Three-dimensional reconstruction of functional brain images

Energy Technology Data Exchange (ETDEWEB)

Inoue, Masato; Shoji, Kazuhiko; Kojima, Hisayoshi; Hirano, Shigeru; Naito, Yasushi; Honjo, Iwao [Kyoto Univ. (Japan)

1999-08-01

We consider PET (positron emission tomography) measurement with SPM (Statistical Parametric Mapping) analysis to be one of the most useful methods to identify activated areas of the brain involved in language processing. SPM is an effective analytical method that detects markedly activated areas over the whole brain. However, with the conventional presentations of these functional brain images, such as horizontal slices, three directional projection, or brain surface coloring, makes understanding and interpreting the positional relationships among various brain areas difficult. Therefore, we developed three-dimensionally reconstructed images from these functional brain images to improve the interpretation. The subjects were 12 normal volunteers. The following three types of images were constructed: routine images by SPM, three-dimensional static images, and three-dimensional dynamic images, after PET images were analyzed by SPM during daily dialog listening. The creation of images of both the three-dimensional static and dynamic types employed the volume rendering method by VTK (The Visualization Toolkit). Since the functional brain images did not include original brain images, we synthesized SPM and MRI brain images by self-made C++ programs. The three-dimensional dynamic images were made by sequencing static images with available software. Images of both the three-dimensional static and dynamic types were processed by a personal computer system. Our newly created images showed clearer positional relationships among activated brain areas compared to the conventional method. To date, functional brain images have been employed in fields such as neurology or neurosurgery, however, these images may be useful even in the field of otorhinolaryngology, to assess hearing and speech. Exact three-dimensional images based on functional brain images are important for exact and intuitive interpretation, and may lead to new developments in brain science. Currently, the surface

Prognostic value of three-dimensional ultrasound for fetal hydronephrosis

Science.gov (United States)

WANG, JUNMEI; YING, WEIWEN; TANG, DAXING; YANG, LIMING; LIU, DONGSHENG; LIU, YUANHUI; PAN, JIAOE; XIE, XING

2015-01-01

The present study evaluated the prognostic value of three-dimensional ultrasound for fetal hydronephrosis. Pregnant females with fetal hydronephrosis were enrolled and a novel three-dimensional ultrasound indicator, renal parenchymal volume/kidney volume, was introduced to predict the postnatal prognosis of fetal hydronephrosis in comparison with commonly used ultrasound indicators. All ultrasound indicators of fetal hydronephrosis could predict whether postnatal surgery was required for fetal hydronephrosis; however, the predictive performance of renal parenchymal volume/kidney volume measurements as an individual indicator was the highest. In conclusion, ultrasound is important in predicting whether postnatal surgery is required for fetal hydronephrosis, and the three-dimensional ultrasound indicator renal parenchymal volume/kidney volume has a high predictive performance. Furthermore, the majority of cases of fetal hydronephrosis spontaneously regress subsequent to birth, and the regression time is closely associated with ultrasound indicators. PMID:25667626

Three-dimensional metallic opals fabricated by double templating

International Nuclear Information System (INIS)

Yan Qingfeng; Nukala, Pavan; Chiang, Yet-Ming; Wong, C.C.

2009-01-01

We report a simple and cost-effective double templating method for fabricating large-area three-dimensional metallic photonic crystals of controlled thickness. A self-assembled polystyrene opal was used as the first template to fabricate a silica inverse opal on a gold-coated glass substrate via sol-gel processing. Gold was subsequently infiltrated to the pores of the silica inverse opal using electrochemical deposition. A high-quality three-dimensional gold photonic crystal was obtained after removal of the secondary template (silica inverse opal). The effects of template sphere size and deposition current density on the gold growth rate, and the resulting morphology and growth mechanism of the gold opal, were investigated.

Three-dimensional growth patterns of various human tumor cell lines in simulated microgravity of a NASA bioreactor.

Science.gov (United States)

Ingram, M; Techy, G B; Saroufeem, R; Yazan, O; Narayan, K S; Goodwin, T J; Spaulding, G F

1997-06-01

Growth patterns of a number of human tumor cell lines that from three-dimensional structures of various architectures when cultured without carrier beads in a NASA rotary cell culture system are described and illustrated. The culture system, which was designed to mimic microgravity, maintained cells in suspension under very low-shear stress throughout culture. Spheroid (particulate) production occurred within a few hours after culture was started, and spheroids increased in size by cell division and fusion of small spheroids, usually stabilizing at a spheroid diameter of about 0.5 mm. Architecture of spheroids varied with cell type. Cellular interactions that occurred in spheroids resulted in conformation and shape changes of cells, and some cell lines produced complex, epithelial-like architectures. Expression of the cell adhesion molecules, CD44 and E cadherin, was upregulated in the three-dimensional constructs. Coculture of fibroblast spheroids with PC3 prostate cancer cells induced tenascin expression by the fibroblasts underlying the adherent prostate epithelial cells. Invasion of the fibroblast spheroids by the malignant epithelium was also demonstrated.

Application of Simulated Three Dimensional CT Image in Orthognathic Surgery

Energy Technology Data Exchange (ETDEWEB)

Kim, Hyun Don; Park, Chang Seo [Dept. of Dental Radiology, College of Dentistry, Yensei University, Seoul (Korea, Republic of); Yoo, Sun Kook; Lee, Kyoung Sang [Dept. of Medical Engineering, College of Medicine, Yensei University, Seoul (Korea, Republic of)

1998-08-15

In orthodontics and orthognathic surgery, cephalogram has been routine practice in diagnosis and treatment evaluation of craniofacial deformity. But its inherent distortion of actual length and angles during projecting three dimensional object to two dimensional plane might cause errors in quantitative analysis of shape and size. Therefore, it is desirable that three dimensional object is diagnosed and evaluated three dimensionally and three dimensional CT image is best for three dimensional analysis. Development of clinic necessitates evaluation of result of treatment and comparison before and after surgery. It is desirable that patient that was diagnosed and planned by three dimensional computed tomography before surgery is evaluated by three dimensional computed tomography after surgery, too. But Because there is no standardized normal values in three dimension now and three dimensional Computed Tomography needs expensive equipment and because of its expenses and amount of exposure to radiation, limitations still remain to be solved in its application to routine practice. If postoperative three dimensional image is constructed by pre and postoperative lateral and postero-anterior cephalograms and preoperative three dimensional computed tomogram, pre and postoperative image will be compared and evaluated three dimensionally without three dimensional computed tomography after surgery and that will contribute to standardize normal values in three dimension. This study introduced new method that computer-simulated three dimensional image was constructed by preoperative three dimensional computed tomogram and pre and postoperative lateral and postero-anterior cephalograms, and for validation of new method, in four cases of dry skull that position of mandible was displaced and four patients of orthognathic surgery, computer-simulated three dimensional image and actual postoperative three dimensional image were compared. The results were as follows. 1. In four cases of

Application of Simulated Three Dimensional CT Image in Orthognathic Surgery

International Nuclear Information System (INIS)

Kim, Hyun Don; Park, Chang Seo; Yoo, Sun Kook; Lee, Kyoung Sang

1998-01-01

In orthodontics and orthognathic surgery, cephalogram has been routine practice in diagnosis and treatment evaluation of craniofacial deformity. But its inherent distortion of actual length and angles during projecting three dimensional object to two dimensional plane might cause errors in quantitative analysis of shape and size. Therefore, it is desirable that three dimensional object is diagnosed and evaluated three dimensionally and three dimensional CT image is best for three dimensional analysis. Development of clinic necessitates evaluation of result of treatment and comparison before and after surgery. It is desirable that patient that was diagnosed and planned by three dimensional computed tomography before surgery is evaluated by three dimensional computed tomography after surgery, too. But Because there is no standardized normal values in three dimension now and three dimensional Computed Tomography needs expensive equipment and because of its expenses and amount of exposure to radiation, limitations still remain to be solved in its application to routine practice. If postoperative three dimensional image is constructed by pre and postoperative lateral and postero-anterior cephalograms and preoperative three dimensional computed tomogram, pre and postoperative image will be compared and evaluated three dimensionally without three dimensional computed tomography after surgery and that will contribute to standardize normal values in three dimension. This study introduced new method that computer-simulated three dimensional image was constructed by preoperative three dimensional computed tomogram and pre and postoperative lateral and postero-anterior cephalograms, and for validation of new method, in four cases of dry skull that position of mandible was displaced and four patients of orthognathic surgery, computer-simulated three dimensional image and actual postoperative three dimensional image were compared. The results were as follows. 1. In four cases of

Three-dimensional polypyrrole-derived carbon nanotube framework for dye adsorption and electrochemical supercapacitor

Energy Technology Data Exchange (ETDEWEB)

Xin, Shengchang; Yang, Na; Gao, Fei [School of Life Sciences, State Key Laboratory of Coordination Chemistry and Collaborative Innovation Center of Chemistry for Life Sciences, Institute of Chemistry and BioMedical Sciences, Nanjing University, Nanjing 210093 (China); Zhao, Jing, E-mail: jingzhao@nju.edu.cn [School of Life Sciences, State Key Laboratory of Coordination Chemistry and Collaborative Innovation Center of Chemistry for Life Sciences, Institute of Chemistry and BioMedical Sciences, Nanjing University, Nanjing 210093 (China); Li, Liang, E-mail: msell08@163.com [School of Materials Science and Engineering, Wuhan Institute of Technology, Wuhan 430073 (China); Teng, Chao, E-mail: tengc@pkusz.edu.cn [Guangdong Provincial Key Laboratory of Nano-Micro Materials Research, School of Chemical Biology & Biotechnology, Peking University Shenzhen Graduate School, Shenzhen 518055 (China)

2017-08-31

Highlights: • Three-dimensional polypyrrole-derived carbon nanotube frameworks are prepared. • They display outstanding absorption capacity (609 mg g{sup −1}) towards methylene blue. • They possess high specific capacitance (167 F g{sup −1}) and good rate capability (64%). • They have excellent cycling performance with no capacitance loss over 1000 cycles. - Abstract: Three-dimensional carbon nanotube frameworks have been prepared via pyrolysis of polypyrrole nanotube aerogels that are synthesized by the simultaneous self-degraded template synthesis and hydrogel assembly followed by freeze-drying. The microstructure and composition of the materials are investigated by thermal gravimetric analysis, Raman spectrum, X-ray photoelectron spectroscopy, transmission electron microscopy, and specific surface analyzer. The results confirm the formation of three-dimensional carbon nanotube frameworks with low density, high mechanical properties, and high specific surface area. Compared with PPy aerogel precursor, the as-prepared three-dimensional carbon nanotube frameworks exhibit outstanding adsorption capacity towards organic dyes. Moreover, electrochemical tests show that the products possess high specific capacitance, good rate capability and excellent cycling performance with no capacitance loss over 1000 cycles. These characteristics collectively indicate the potential of three-dimensional polypyrrole-derived carbon nanotube framework as a promising macroscopic device for the applications in environmental and energy storages.

Nano-engineering of three-dimensional core/shell nanotube arrays for high performance supercapacitors

Science.gov (United States)

Grote, Fabian; Wen, Liaoyong; Lei, Yong

2014-06-01

Large-scale arrays of core/shell nanostructures are highly desirable to enhance the performance of supercapacitors. Here we demonstrate an innovative template-based fabrication technique with high structural controllability, which is capable of synthesizing well-ordered three-dimensional arrays of SnO2/MnO2 core/shell nanotubes for electrochemical energy storage in supercapacitor applications. The SnO2 core is fabricated by atomic layer deposition and provides a highly electrical conductive matrix. Subsequently a thin MnO2 shell is coated by electrochemical deposition onto the SnO2 core, which guarantees a short ion diffusion length within the shell. The core/shell structure shows an excellent electrochemical performance with a high specific capacitance of 910 F g-1 at 1 A g-1 and a good rate capability of remaining 217 F g-1 at 50 A g-1. These results shall pave the way to realize aqueous based asymmetric supercapacitors with high specific power and high specific energy.

Large three-dimensional photonic crystals based on monocrystalline liquid crystal blue phases.

Science.gov (United States)

Chen, Chun-Wei; Hou, Chien-Tsung; Li, Cheng-Chang; Jau, Hung-Chang; Wang, Chun-Ta; Hong, Ching-Lang; Guo, Duan-Yi; Wang, Cheng-Yu; Chiang, Sheng-Ping; Bunning, Timothy J; Khoo, Iam-Choon; Lin, Tsung-Hsien

2017-09-28

Although there have been intense efforts to fabricate large three-dimensional photonic crystals in order to realize their full potential, the technologies developed so far are still beset with various material processing and cost issues. Conventional top-down fabrications are costly and time-consuming, whereas natural self-assembly and bottom-up fabrications often result in high defect density and limited dimensions. Here we report the fabrication of extraordinarily large monocrystalline photonic crystals by controlling the self-assembly processes which occur in unique phases of liquid crystals that exhibit three-dimensional photonic-crystalline properties called liquid-crystal blue phases. In particular, we have developed a gradient-temperature technique that enables three-dimensional photonic crystals to grow to lateral dimensions of ~1 cm (~30,000 of unit cells) and thickness of ~100 μm (~ 300 unit cells). These giant single crystals exhibit extraordinarily sharp photonic bandgaps with high reflectivity, long-range periodicity in all dimensions and well-defined lattice orientation.Conventional fabrication approaches for large-size three-dimensional photonic crystals are problematic. By properly controlling the self-assembly processes, the authors report the fabrication of monocrystalline blue phase liquid crystals that exhibit three-dimensional photonic-crystalline properties.

Three-dimensional propagation and absorption of high frequency Gaussian beams in magnetoactive plasmas

International Nuclear Information System (INIS)

Nowak, S.; Orefice, A.

1994-01-01

In today's high frequency systems employed for plasma diagnostics, power heating, and current drive the behavior of the wave beams is appreciably affected by the self-diffraction phenomena due to their narrow collimation. In the present article the three-dimensional propagation of Gaussian beams in inhomogeneous and anisotropic media is analyzed, starting from a properly formulated dispersion relation. Particular attention is paid, in the case of electromagnetic electron cyclotron (EC) waves, to the toroidal geometry characterizing tokamak plasmas, to the power density evolution on the advancing wave fronts, and to the absorption features occurring when a beam crosses an EC resonant layer

Poly(N-isopropylacrylamide) hydrogel/chitosan scaffold hybrid for three-dimensional stem cell culture and cartilage tissue engineering.

Science.gov (United States)

Mellati, Amir; Kiamahalleh, Meisam Valizadeh; Madani, S Hadi; Dai, Sheng; Bi, Jingxiu; Jin, Bo; Zhang, Hu

2016-11-01

Providing a controllable and definable three-dimensional (3D) microenvironment for chondrogenic differentiation of mesenchymal stem cells (MSCs) remains a great challenge for cartilage tissue engineering. In this work, poly(N-isopropylacrylamide) (PNIPAAm) polymers with the degrees of polymerization of 100 and 400 (NI100 and NI400) were prepared and the polymer solutions were introduced into the preprepared chitosan porous scaffolds (CS) to form hybrids (CSNI100 and CSNI400, respectively). SEM images indicated that the PNIPAAm gel partially occupied chitosan pores while the interconnected porous structure of chitosan was preserved. MSCs were incorporated within the hybrid and cell proliferation and chondrogenic differentiation were monitored. After 7-day incubation of the cell-laden constructs in a growth medium, the cell viability in CSNI100 and CSNI400 were 54 and 108% higher than that in CS alone, respectively. Glycosaminoglycan and total collagen contents increased 2.6- and 2.5-fold after 28-day culture of cell-laden CSNI400 in the chondrogenic medium. These results suggest that the hybrid structure composed of the chitosan porous scaffold and the well-defined PNIPAAm hydrogel, in particular CSNI400, is suitable for 3D stem cell culture and cartilage tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2764-2774, 2016. © 2016 Wiley Periodicals, Inc.

High-throughput transformation of Saccharomyces cerevisiae using liquid handling robots.

Directory of Open Access Journals (Sweden)

Guangbo Liu

Full Text Available Saccharomyces cerevisiae (budding yeast is a powerful eukaryotic model organism ideally suited to high-throughput genetic analyses, which time and again has yielded insights that further our understanding of cell biology processes conserved in humans. Lithium Acetate (LiAc transformation of yeast with DNA for the purposes of exogenous protein expression (e.g., plasmids or genome mutation (e.g., gene mutation, deletion, epitope tagging is a useful and long established method. However, a reliable and optimized high throughput transformation protocol that runs almost no risk of human error has not been described in the literature. Here, we describe such a method that is broadly transferable to most liquid handling high-throughput robotic platforms, which are now commonplace in academic and industry settings. Using our optimized method, we are able to comfortably transform approximately 1200 individual strains per day, allowing complete transformation of typical genomic yeast libraries within 6 days. In addition, use of our protocol for gene knockout purposes also provides a potentially quicker, easier and more cost-effective approach to generating collections of double mutants than the popular and elegant synthetic genetic array methodology. In summary, our methodology will be of significant use to anyone interested in high throughput molecular and/or genetic analysis of yeast.

A high-throughput multiplex method adapted for GMO detection.

Science.gov (United States)

Chaouachi, Maher; Chupeau, Gaëlle; Berard, Aurélie; McKhann, Heather; Romaniuk, Marcel; Giancola, Sandra; Laval, Valérie; Bertheau, Yves; Brunel, Dominique

2008-12-24

A high-throughput multiplex assay for the detection of genetically modified organisms (GMO) was developed on the basis of the existing SNPlex method designed for SNP genotyping. This SNPlex assay allows the simultaneous detection of up to 48 short DNA sequences (approximately 70 bp; "signature sequences") from taxa endogenous reference genes, from GMO constructions, screening targets, construct-specific, and event-specific targets, and finally from donor organisms. This assay avoids certain shortcomings of multiplex PCR-based methods already in widespread use for GMO detection. The assay demonstrated high specificity and sensitivity. The results suggest that this assay is reliable, flexible, and cost- and time-effective for high-throughput GMO detection.

Continuation of full-scale three-dimensional numerical experiments on high-intensity particle and laser beam-matter interactions

Energy Technology Data Exchange (ETDEWEB)

Mori, Warren, B.

2012-12-01

We present results from the grant entitled, Continuation of full-scale three-dimensional numerical experiments on high-intensity particle and laser beam-matter interactions. The research significantly advanced the understanding of basic high-energy density science (HEDS) on ultra intense laser and particle beam plasma interactions. This advancement in understanding was then used to to aid in the quest to make 1 GeV to 500 GeV plasma based accelerator stages. The work blended basic research with three-dimensions fully nonlinear and fully kinetic simulations including full-scale modeling of ongoing or planned experiments. The primary tool was three-dimensional particle-in-cell simulations. The simulations provided a test bed for theoretical ideas and models as well as a method to guide experiments. The research also included careful benchmarking of codes against experiment. High-fidelity full-scale modeling provided a means to extrapolate parameters into regimes that were not accessible to current or near term experiments, thereby allowing concepts to be tested with confidence before tens to hundreds of millions of dollars were spent building facilities. The research allowed the development of a hierarchy of PIC codes and diagnostics that is one of the most advanced in the world.

A phasor approach analysis of multiphoton FLIM measurements of three-dimensional cell culture models

Science.gov (United States)

Lakner, P. H.; Möller, Y.; Olayioye, M. A.; Brucker, S. Y.; Schenke-Layland, K.; Monaghan, M. G.

2016-03-01

Fluorescence lifetime imaging microscopy (FLIM) is a useful approach to obtain information regarding the endogenous fluorophores present in biological samples. The concise evaluation of FLIM data requires the use of robust mathematical algorithms. In this study, we developed a user-friendly phasor approach for analyzing FLIM data and applied this method on three-dimensional (3D) Caco-2 models of polarized epithelial luminal cysts in a supporting extracellular matrix environment. These Caco-2 based models were treated with epidermal growth factor (EGF), to stimulate proliferation in order to determine if FLIM could detect such a change in cell behavior. Autofluorescence from nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) in luminal Caco-2 cysts was stimulated by 2-photon laser excitation. Using a phasor approach, the lifetimes of involved fluorophores and their contribution were calculated with fewer initial assumptions when compared to multiexponential decay fitting. The phasor approach simplified FLIM data analysis, making it an interesting tool for non-experts in numerical data analysis. We observed that an increased proliferation stimulated by EGF led to a significant shift in fluorescence lifetime and a significant alteration of the phasor data shape. Our data demonstrates that multiphoton FLIM analysis with the phasor approach is a suitable method for the non-invasive analysis of 3D in vitro cell culture models qualifying this method for monitoring basic cellular features and the effect of external factors.

Three-dimensional low-energy topological invariants

International Nuclear Information System (INIS)

Bakalarska, M.; Broda, B.

2000-01-01

A description of the one-loop approximation formula for the partition function of a three-dimensional abelian version of the Donaldson-Witten theory is proposed. The one-loop expression is shown to contain such topological invariants of a three-dimensional manifold M like the Reidemeister-Ray-Singer torsion τ R and Betti numbers. (orig.)

Instrumented cardiac microphysiological devices via multimaterial three-dimensional printing

Science.gov (United States)

Lind, Johan U.; Busbee, Travis A.; Valentine, Alexander D.; Pasqualini, Francesco S.; Yuan, Hongyan; Yadid, Moran; Park, Sung-Jin; Kotikian, Arda; Nesmith, Alexander P.; Campbell, Patrick H.; Vlassak, Joost J.; Lewis, Jennifer A.; Parker, Kevin K.

2017-03-01

Biomedical research has relied on animal studies and conventional cell cultures for decades. Recently, microphysiological systems (MPS), also known as organs-on-chips, that recapitulate the structure and function of native tissues in vitro, have emerged as a promising alternative. However, current MPS typically lack integrated sensors and their fabrication requires multi-step lithographic processes. Here, we introduce a facile route for fabricating a new class of instrumented cardiac microphysiological devices via multimaterial three-dimensional (3D) printing. Specifically, we designed six functional inks, based on piezo-resistive, high-conductance, and biocompatible soft materials that enable integration of soft strain gauge sensors within micro-architectures that guide the self-assembly of physio-mimetic laminar cardiac tissues. We validated that these embedded sensors provide non-invasive, electronic readouts of tissue contractile stresses inside cell incubator environments. We further applied these devices to study drug responses, as well as the contractile development of human stem cell-derived laminar cardiac tissues over four weeks.

High-throughput tri-colour flow cytometry technique to assess Plasmodium falciparum parasitaemia in bioassays

DEFF Research Database (Denmark)

Tiendrebeogo, Regis W; Adu, Bright; Singh, Susheel K

2014-01-01

BACKGROUND: Unbiased flow cytometry-based methods have become the technique of choice in many laboratories for high-throughput, accurate assessments of malaria parasites in bioassays. A method to quantify live parasites based on mitotracker red CMXRos was recently described but consistent...... distinction of early ring stages of Plasmodium falciparum from uninfected red blood cells (uRBC) remains a challenge. METHODS: Here, a high-throughput, three-parameter (tri-colour) flow cytometry technique based on mitotracker red dye, the nucleic acid dye coriphosphine O (CPO) and the leucocyte marker CD45...... for enumerating live parasites in bioassays was developed. The technique was applied to estimate the specific growth inhibition index (SGI) in the antibody-dependent cellular inhibition (ADCI) assay and compared to parasite quantification by microscopy and mitotracker red staining. The Bland-Altman analysis...

Reversed thermo-switchable molecular sieving membranes composed of two-dimensional metal-organic nanosheets for gas separation

Science.gov (United States)

Wang, Xuerui; Chi, Chenglong; Zhang, Kang; Qian, Yuhong; Gupta, Krishna M.; Kang, Zixi; Jiang, Jianwen; Zhao, Dan

2017-02-01

It is highly desirable to reduce the membrane thickness in order to maximize the throughput and break the trade-off limitation for membrane-based gas separation. Two-dimensional membranes composed of atomic-thick graphene or graphene oxide nanosheets have gas transport pathways that are at least three orders of magnitude higher than the membrane thickness, leading to reduced gas permeation flux and impaired separation throughput. Here we present nm-thick molecular sieving membranes composed of porous two-dimensional metal-organic nanosheets. These membranes possess pore openings parallel to gas concentration gradient allowing high gas permeation flux and high selectivity, which are proven by both experiment and molecular dynamics simulation. Furthermore, the gas transport pathways of these membranes exhibit a reversed thermo-switchable feature, which is attributed to the molecular flexibility of the building metal-organic nanosheets.

Three-dimensional analysis of harmonic generation in high-gain free-electron lasers

International Nuclear Information System (INIS)

Huang, Zhirong; Kim, Kwang-Je

2000-01-01

In a high-gain free-electron laser (FEL) employing a planar undulator, strong bunching at the fundamental wavelength can drive substantial bunching and power levels at the harmonic frequencies. In this paper we investigate the three-dimensional evolution of harmonic radiation based on the coupled Maxwell-Klimontovich equations that take into account nonlinear harmonic interactions. Each harmonic field is a sum of a linear amplification term and a term driven by nonlinear harmonic interactions. After a certain stage of exponential growth, the dominant nonlinear term is determined by interactions of the lower nonlinear harmonics and the fundamental radiation. As a result, the gain length, transverse profile, and temporal structure of the first few harmonics are eventually governed by those of the fundamental. Transversely coherent third-harmonic radiation power is found to approach 1% of the fundamental power level for current high-gain FEL projects

Characterization of highly anisotropic three-dimensionally nanostructured surfaces

International Nuclear Information System (INIS)

Schmidt, Daniel

2014-01-01

Generalized ellipsometry, a non-destructive optical characterization technique, is employed to determine geometrical structure parameters and anisotropic dielectric properties of highly spatially coherent three-dimensionally nanostructured thin films grown by glancing angle deposition. The (piecewise) homogeneous biaxial layer model approach is discussed, which can be universally applied to model the optical response of sculptured thin films with different geometries and from diverse materials, and structural parameters as well as effective optical properties of the nanostructured thin films are obtained. Alternative model approaches for slanted columnar thin films, anisotropic effective medium approximations based on the Bruggeman formalism, are presented, which deliver results comparable to the homogeneous biaxial layer approach and in addition provide film constituent volume fraction parameters as well as depolarization or shape factors. Advantages of these ellipsometry models are discussed on the example of metal slanted columnar thin films, which have been conformally coated with a thin passivating oxide layer by atomic layer deposition. Furthermore, the application of an effective medium approximation approach to in-situ growth monitoring of this anisotropic thin film functionalization process is presented. It was found that structural parameters determined with the presented optical model equivalents for slanted columnar thin films agree very well with scanning electron microscope image estimates. - Highlights: • Summary of optical model strategies for sculptured thin films with arbitrary geometries • Application of the rigorous anisotropic Bruggeman effective medium applications • In-situ growth monitoring of atomic layer deposition on biaxial metal slanted columnar thin film

Noninvasive Quantitative Imaging of Collagen Microstructure in Three-Dimensional Hydrogels Using High-Frequency Ultrasound.

Science.gov (United States)

Mercado, Karla P; Helguera, María; Hocking, Denise C; Dalecki, Diane

2015-07-01

Collagen I is widely used as a natural component of biomaterials for both tissue engineering and regenerative medicine applications. The physical and biological properties of fibrillar collagens are strongly tied to variations in collagen fiber microstructure. The goal of this study was to develop the use of high-frequency quantitative ultrasound to assess collagen microstructure within three-dimensional (3D) hydrogels noninvasively and nondestructively. The integrated backscatter coefficient (IBC) was employed as a quantitative ultrasound parameter to detect, image, and quantify spatial variations in collagen fiber density and diameter. Collagen fiber microstructure was varied by fabricating hydrogels with different collagen concentrations or polymerization temperatures. IBC values were computed from measurements of the backscattered radio-frequency ultrasound signals collected using a single-element transducer (38-MHz center frequency, 13-47 MHz bandwidth). The IBC increased linearly with increasing collagen concentration and decreasing polymerization temperature. Parametric 3D images of the IBC were generated to visualize and quantify regional variations in collagen microstructure throughout the volume of hydrogels fabricated in standard tissue culture plates. IBC parametric images of corresponding cell-embedded collagen gels showed cell accumulation within regions having elevated collagen IBC values. The capability of this ultrasound technique to noninvasively detect and quantify spatial differences in collagen microstructure offers a valuable tool to monitor the structural properties of collagen scaffolds during fabrication, to detect functional differences in collagen microstructure, and to guide fundamental research on the interactions of cells and collagen matrices.

Three-dimensional MR imaging of the cerebrospinal system with the RARE technique

International Nuclear Information System (INIS)

Hennig, J.; Ott, D.; Ylayasski, J.

1987-01-01

Three-dimensional RARE myelography is a fast technique for high-resolution imaging of the cerebrospinal fluid. A data set with 1 x 1 x 1-mm resolution can be generated with a 12-minute acquisition time. Sophisticated three-dimensional display algorithms allow reconstruction of planes at arbitrary angles and full three-dimensional displays, which yield extremely useful information for neurosurgical planning. Additionally, the injection of contrast agent can be simulated on the computer and communication pathways between structures of interest can be found noninvasively

High Throughput Analysis of Photocatalytic Water Purification

NARCIS (Netherlands)

Sobral Romao, J.I.; Baiao Barata, David; Habibovic, Pamela; Mul, Guido; Baltrusaitis, Jonas

2014-01-01

We present a novel high throughput photocatalyst efficiency assessment method based on 96-well microplates and UV-Vis spectroscopy. We demonstrate the reproducibility of the method using methyl orange (MO) decomposition, and compare kinetic data obtained with those provided in the literature for

Growing three-dimensional biomorphic graphene powders using naturally abundant diatomite templates towards high solution processability

Science.gov (United States)

Chen, Ke; Li, Cong; Shi, Liurong; Gao, Teng; Song, Xiuju; Bachmatiuk, Alicja; Zou, Zhiyu; Deng, Bing; Ji, Qingqing; Ma, Donglin; Peng, Hailin; Du, Zuliang; Rümmeli, Mark Hermann; Zhang, Yanfeng; Liu, Zhongfan

2016-11-01

Mass production of high-quality graphene with low cost is the footstone for its widespread practical applications. We present herein a self-limited growth approach for producing graphene powders by a small-methane-flow chemical vapour deposition process on naturally abundant and industrially widely used diatomite (biosilica) substrates. Distinct from the chemically exfoliated graphene, thus-produced biomorphic graphene is highly crystallized with atomic layer-thickness controllability, structural designability and less noncarbon impurities. In particular, the individual graphene microarchitectures preserve a three-dimensional naturally curved surface morphology of original diatom frustules, effectively overcoming the interlayer stacking and hence giving excellent dispersion performance in fabricating solution-processible electrodes. The graphene films derived from as-made graphene powders, compatible with either rod-coating, or inkjet and roll-to-roll printing techniques, exhibit much higher electrical conductivity (~110,700 S m-1 at 80% transmittance) than previously reported solution-based counterparts. This work thus puts forward a practical route for low-cost mass production of various powdery two-dimensional materials.

High Throughput Synthesis and Screening for Agents Inhibiting Androgen Receptor Mediated Gene Transcription

National Research Council Canada - National Science Library

Boger, Dale L

2005-01-01

.... This entails the high throughput synthesis of DNA binding agents related to distamycin, their screening for binding to androgen response elements using a new high throughput DNA binding screen...

High Throughput Synthesis and Screening for Agents Inhibiting Androgen Receptor Mediated Gene Transcription

National Research Council Canada - National Science Library

Boger, Dale

2004-01-01

.... This entails the high throughput synthesis of DNA binding agents related to distamycin, their screening for binding to androgen response elements using a new high throughput DNA binding screen...

A CRISPR CASe for High-Throughput Silencing

Directory of Open Access Journals (Sweden)

Jacob eHeintze

2013-10-01

Full Text Available Manipulation of gene expression on a genome-wide level is one of the most important systematic tools in the post-genome era. Such manipulations have largely been enabled by expression cloning approaches using sequence-verified cDNA libraries, large-scale RNA interference libraries (shRNA or siRNA and zinc finger nuclease technologies. More recently, the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated (Cas9-mediated gene editing technology has been described that holds great promise for future use of this technology in genomic manipulation. It was suggested that the CRISPR system has the potential to be used in high-throughput, large-scale loss of function screening. Here we discuss some of the challenges in engineering of CRISPR/Cas genomic libraries and some of the aspects that need to be addressed in order to use this technology on a high-throughput scale.

[Bone drilling simulation by three-dimensional imaging].

Science.gov (United States)

Suto, Y; Furuhata, K; Kojima, T; Kurokawa, T; Kobayashi, M

1989-06-01

The three-dimensional display technique has a wide range of medical applications. Pre-operative planning is one typical application: in orthopedic surgery, three-dimensional image processing has been used very successfully. We have employed this technique in pre-operative planning for orthopedic surgery, and have developed a simulation system for bone-drilling. Positive results were obtained by pre-operative rehearsal; when a region of interest is indicated by means of a mouse on the three-dimensional image displayed on the CRT, the corresponding region appears on the slice image which is displayed simultaneously. Consequently, the status of the bone-drilling is constantly monitored. In developing this system, we have placed emphasis on the quality of the reconstructed three-dimensional images, on fast processing, and on the easy operation of the surgical planning simulation.

Three-Dimensional Printing Surgical Applications.

Science.gov (United States)

AlAli, Ahmad B; Griffin, Michelle F; Butler, Peter E

2015-01-01

Three-dimensional printing, a technology used for decades in the industrial field, gains a lot of attention in the medical field for its potential benefits. With advancement of desktop printers, this technology is accessible and a lot of research is going on in the medical field. To evaluate its application in surgical field, which may include but not limited to surgical planning, surgical education, implants, and prosthesis, which are the focus of this review. Research was conducted by searching PubMed, Web of science, and other reliable sources. We included original articles and excluded articles based on animals, those more than 10 years old, and those not in English. These articles were evaluated, and relevant studies were included in this review. Three-dimensional printing shows a potential benefit in surgical application. Printed implants were used in patient in a few cases and show successful results; however, longer follow-up and more trials are needed. Surgical and medical education is believed to be more efficient with this technology than the current practice. Printed surgical instrument and surgical planning are also believed to improve with three-dimensional printing. Three-dimensional printing can be a very powerful tool in the near future, which can aid the medical field that is facing a lot of challenges and obstacles. However, despite the reported results, further research on larger samples and analytical measurements should be conducted to ensure this technology's impact on the practice.

Quantitative volumetric Raman imaging of three dimensional cell cultures

KAUST Repository

Kallepitis, Charalambos; Bergholt, Mads S.; Mazo, Manuel M.; Leonardo, Vincent; Skaalure, Stacey C.; Maynard, Stephanie A.; Stevens, Molly M.

2017-01-01

in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies

High throughput route selection in multi-rate wireless mesh networks

Institute of Scientific and Technical Information of China (English)

WEI Yi-fei; GUO Xiang-li; SONG Mei; SONG Jun-de

2008-01-01

Most existing Ad-hoc routing protocols use the shortest path algorithm with a hop count metric to select paths. It is appropriate in single-rate wireless networks, but has a tendency to select paths containing long-distance links that have low data rates and reduced reliability in multi-rate networks. This article introduces a high throughput routing algorithm utilizing the multi-rate capability and some mesh characteristics in wireless fidelity (WiFi) mesh networks. It uses the medium access control (MAC) transmission time as the routing metric, which is estimated by the information passed up from the physical layer. When the proposed algorithm is adopted, the Ad-hoc on-demand distance vector (AODV) routing can be improved as high throughput AODV (HT-AODV). Simulation results show that HT-AODV is capable of establishing a route that has high data-rate, short end-to-end delay and great network throughput.

The Three-dimensional Digital Factory for Shipbuilding Technology Research

Directory of Open Access Journals (Sweden)

Xu Wei

2016-01-01

Full Text Available The three-dimensional digital factory technology research is the hotspot in shipbuilding recently. The three-dimensional digital factory technology not only focus on design the components of the product, but also discuss on the simulation and analyses of the production process.Based on the three-dimensional model, the basic data layer, application control layer and the presentation layer of hierarchical structure are established in the three-dimensional digital factory of shipbuilding in this paper. And the key technologies of three-dimensional digital factory of shipbuilding are analysed. Finally, a case study is applied and the results show that the three-dimensional digital factory will play an important role in the future.

Three-dimensional time-lapse velocity tomography of an underground longwall panel

Energy Technology Data Exchange (ETDEWEB)

Luxbacher, K.; Westman, E.; Swanson, P.; Karfakis, M. [Virginia Tech., Blacksburg, VA (United States). Dept. of Mining & Minerals Engineering

2008-06-15

Three-dimensional velocity tomograms were generated to image the stress redistribution around an underground coal longwall panel to produce a better understanding of the mechanisms that lead to ground failure, especially rockbursts. Mining-induced microseismic events provided passive sources for the three-dimensional velocity tomography. Surface-mounted geophones monitored microseismic activity for 18 days. Eighteen tomograms were generated and high-velocity regions correlated with high abutment stresses predicted by numerical modeling. Additionally, the high-velocity regions were observed to redistribute as the longwall face retreated, indicating that velocity tomography may be an appropriate technology for monitoring stress redistribution in underground mines.

Alginate Immobilization of Metabolic Enzymes (AIME) for High-Throughput Screening Assays (SOT)

Science.gov (United States)

Alginate Immobilization of Metabolic Enzymes (AIME) for High-Throughput Screening Assays DE DeGroot, RS Thomas, and SO SimmonsNational Center for Computational Toxicology, US EPA, Research Triangle Park, NC USAThe EPA’s ToxCast program utilizes a wide variety of high-throughput s...

Development of a high-throughput microscale cell disruption platform for Pichia pastoris in rapid bioprocess design.

Science.gov (United States)

Bláha, Benjamin A F; Morris, Stephen A; Ogonah, Olotu W; Maucourant, Sophie; Crescente, Vincenzo; Rosenberg, William; Mukhopadhyay, Tarit K

2018-01-01

The time and cost benefits of miniaturized fermentation platforms can only be gained by employing complementary techniques facilitating high-throughput at small sample volumes. Microbial cell disruption is a major bottleneck in experimental throughput and is often restricted to large processing volumes. Moreover, for rigid yeast species, such as Pichia pastoris, no effective high-throughput disruption methods exist. The development of an automated, miniaturized, high-throughput, noncontact, scalable platform based on adaptive focused acoustics (AFA) to disrupt P. pastoris and recover intracellular heterologous protein is described. Augmented modes of AFA were established by investigating vessel designs and a novel enzymatic pretreatment step. Three different modes of AFA were studied and compared to the performance high-pressure homogenization. For each of these modes of cell disruption, response models were developed to account for five different performance criteria. Using multiple responses not only demonstrated that different operating parameters are required for different response optima, with highest product purity requiring suboptimal values for other criteria, but also allowed for AFA-based methods to mimic large-scale homogenization processes. These results demonstrate that AFA-mediated cell disruption can be used for a wide range of applications including buffer development, strain selection, fermentation process development, and whole bioprocess integration. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:130-140, 2018. © 2017 American Institute of Chemical Engineers.

Measurement invariance of the Mental Health Continuum-Short Form (MHC-SF) across three cultural groups

NARCIS (Netherlands)

Joshanloo, Mohsen; Wissing, Marie P.; Khumalo, Itumeleng P.; Lamers, S.M.A.

2013-01-01

This study investigated the factorial structure and invariance of the Mental Health Continuum-Short Form (MHC-SF) across cultural groups from three nations, namely, the Netherlands, South Africa, and Iran (N = 1120). The three-dimensional structure of mental well-being was supported in all the

High-throughput phenotyping and genomic selection: the frontiers of crop breeding converge.

Science.gov (United States)

Cabrera-Bosquet, Llorenç; Crossa, José; von Zitzewitz, Jarislav; Serret, María Dolors; Araus, José Luis

2012-05-01

Genomic selection (GS) and high-throughput phenotyping have recently been captivating the interest of the crop breeding community from both the public and private sectors world-wide. Both approaches promise to revolutionize the prediction of complex traits, including growth, yield and adaptation to stress. Whereas high-throughput phenotyping may help to improve understanding of crop physiology, most powerful techniques for high-throughput field phenotyping are empirical rather than analytical and comparable to genomic selection. Despite the fact that the two methodological approaches represent the extremes of what is understood as the breeding process (phenotype versus genome), they both consider the targeted traits (e.g. grain yield, growth, phenology, plant adaptation to stress) as a black box instead of dissecting them as a set of secondary traits (i.e. physiological) putatively related to the target trait. Both GS and high-throughput phenotyping have in common their empirical approach enabling breeders to use genome profile or phenotype without understanding the underlying biology. This short review discusses the main aspects of both approaches and focuses on the case of genomic selection of maize flowering traits and near-infrared spectroscopy (NIRS) and plant spectral reflectance as high-throughput field phenotyping methods for complex traits such as crop growth and yield. © 2012 Institute of Botany, Chinese Academy of Sciences.

Towards three-dimensional optical metamaterials

Science.gov (United States)

Tanaka, Takuo; Ishikawa, Atsushi

2017-12-01

Metamaterials have opened up the possibility of unprecedented and fascinating concepts and applications in optics and photonics. Examples include negative refraction, perfect lenses, cloaking, perfect absorbers, and so on. Since these metamaterials are man-made materials composed of sub-wavelength structures, their development strongly depends on the advancement of micro- and nano-fabrication technologies. In particular, the realization of three-dimensional metamaterials is one of the big challenges in this research field. In this review, we describe recent progress in the fabrication technologies for three-dimensional metamaterials, as well as proposed applications.

Secondary instability and transition in three-dimensional boundary layers

Energy Technology Data Exchange (ETDEWEB)

Stolte, A.; Bertolotti, F.P.; Koch, W. (Deutsche Forschungsanstalt fuer Luft- und Raumfahrt e.V. (DLR), Goettingen (Germany). Inst. fuer Stroemungsmechanik)

1999-01-01

Stationary and traveling crossflow modes are the most prominent disturbances in the highly accelerated three-dimensional flow near the leading edge of a swept wing. Near transition onset, secondary three-dimensional instabilities of high frequency can be observed in such flows. A model flow on the basis of a DLR swept plate experiment allows a detailed study of transition scenarios triggered by crossflow instabilities, since the favorable pressure gradient over the whole plate inhibits instabilities of Tollmien-Schlichting type. In order to shed some light upon the role of the high-frequency secondary instabilities, the saturation characteristics of crossflow vortices in this model flow are investigated using the parabolized stability equations. In contrast to nonlinear equilibrium solutions of steady crossflow vortices, the nonlinear Polarized Stability Equations (PSE) results yield different maximal disturbance amplitudes for different initial amplitudes. (orig./AKF)

Secondary instability and transition in three-dimensional boundary layers

Energy Technology Data Exchange (ETDEWEB)

Stolte, A.; Bertolotti, F.P.; Koch, W. [Deutsche Forschungsanstalt fuer Luft- und Raumfahrt e.V. (DLR), Goettingen (Germany). Inst. fuer Stroemungsmechanik

1999-12-01

Stationary and traveling crossflow modes are the most prominent disturbances in the highly accelerated three-dimensional flow near the leading edge of a swept wing. Near transition onset, secondary three-dimensional instabilities of high frequency can be observed in such flows. A model flow on the basis of a DLR swept plate experiment allows a detailed study of transition scenarios triggered by crossflow instabilities, since the favorable pressure gradient over the whole plate inhibits instabilities of Tollmien-Schlichting type. In order to shed some light upon the role of the high-frequency secondary instabilities, the saturation characteristics of crossflow vortices in this model flow are investigated using the parabolized stability equations. In contrast to nonlinear equilibrium solutions of steady crossflow vortices, the nonlinear Polarized Stability Equations (PSE) results yield different maximal disturbance amplitudes for different initial amplitudes. (orig./AKF)

High-throughput single nucleotide polymorphism genotyping using nanofluidic Dynamic Arrays

Directory of Open Access Journals (Sweden)

Crenshaw Andrew

2009-01-01

Full Text Available Abstract Background Single nucleotide polymorphisms (SNPs have emerged as the genetic marker of choice for mapping disease loci and candidate gene association studies, because of their high density and relatively even distribution in the human genomes. There is a need for systems allowing medium multiplexing (ten to hundreds of SNPs with high throughput, which can efficiently and cost-effectively generate genotypes for a very large sample set (thousands of individuals. Methods that are flexible, fast, accurate and cost-effective are urgently needed. This is also important for those who work on high throughput genotyping in non-model systems where off-the-shelf assays are not available and a flexible platform is needed. Results We demonstrate the use of a nanofluidic Integrated Fluidic Circuit (IFC - based genotyping system for medium-throughput multiplexing known as the Dynamic Array, by genotyping 994 individual human DNA samples on 47 different SNP assays, using nanoliter volumes of reagents. Call rates of greater than 99.5% and call accuracies of greater than 99.8% were achieved from our study, which demonstrates that this is a formidable genotyping platform. The experimental set up is very simple, with a time-to-result for each sample of about 3 hours. Conclusion Our results demonstrate that the Dynamic Array is an excellent genotyping system for medium-throughput multiplexing (30-300 SNPs, which is simple to use and combines rapid throughput with excellent call rates, high concordance and low cost. The exceptional call rates and call accuracy obtained may be of particular interest to those working on validation and replication of genome- wide- association (GWA studies.

Three-dimensional imaging modalities in endodontics

Science.gov (United States)

Mao, Teresa

2014-01-01

Recent research in endodontics has highlighted the need for three-dimensional imaging in the clinical arena as well as in research. Three-dimensional imaging using computed tomography (CT) has been used in endodontics over the past decade. Three types of CT scans have been studied in endodontics, namely cone-beam CT, spiral CT, and peripheral quantitative CT. Contemporary endodontics places an emphasis on the use of cone-beam CT for an accurate diagnosis of parameters that cannot be visualized on a two-dimensional image. This review discusses the role of CT in endodontics, pertaining to its importance in the diagnosis of root canal anatomy, detection of peri-radicular lesions, diagnosis of trauma and resorption, presurgical assessment, and evaluation of the treatment outcome. PMID:25279337

Three-dimensional imaging modalities in endodontics

Energy Technology Data Exchange (ETDEWEB)

Mao, Teresa; Neelakantan, Prasanna [Dept. of Conservative Dentistry and Endodontics, Saveetha Dental College and Hospitals, Saveetha University, Chennai (India)

2014-09-15

Recent research in endodontics has highlighted the need for three-dimensional imaging in the clinical arena as well as in research. Three-dimensional imaging using computed tomography (CT) has been used in endodontics over the past decade. Three types of CT scans have been studied in endodontics, namely cone-beam CT, spiral CT, and peripheral quantitative CT. Contemporary endodontics places an emphasis on the use of cone-beam CT for an accurate diagnosis of parameters that cannot be visualized on a two-dimensional image. This review discusses the role of CT in endodontics, pertaining to its importance in the diagnosis of root canal anatomy, detection of peri-radicular lesions, diagnosis of trauma and resorption, presurgical assessment, and evaluation of the treatment outcome.

Three-dimensional imaging modalities in endodontics

International Nuclear Information System (INIS)

Mao, Teresa; Neelakantan, Prasanna

2014-01-01

Recent research in endodontics has highlighted the need for three-dimensional imaging in the clinical arena as well as in research. Three-dimensional imaging using computed tomography (CT) has been used in endodontics over the past decade. Three types of CT scans have been studied in endodontics, namely cone-beam CT, spiral CT, and peripheral quantitative CT. Contemporary endodontics places an emphasis on the use of cone-beam CT for an accurate diagnosis of parameters that cannot be visualized on a two-dimensional image. This review discusses the role of CT in endodontics, pertaining to its importance in the diagnosis of root canal anatomy, detection of peri-radicular lesions, diagnosis of trauma and resorption, presurgical assessment, and evaluation of the treatment outcome

HTTK: R Package for High-Throughput Toxicokinetics

Science.gov (United States)

Thousands of chemicals have been profiled by high-throughput screening programs such as ToxCast and Tox21; these chemicals are tested in part because most of them have limited or no data on hazard, exposure, or toxicokinetics. Toxicokinetic models aid in predicting tissue concent...

A three-dimensional field solutions of Halbach

International Nuclear Information System (INIS)

Chen Jizhong; Xiao Jijun; Zhang Yiming; Xu Chunyan

2008-01-01

A three-dimensional field solutions are presented for Halback cylinder magnet. Based on Ampere equivalent current methods, the permanent magnets are taken as distributing of current density. For getting the three-dimensional field solution of ideal polarized permanent magnets, the solution method entails the use of the vector potential and involves the closed-form integration of the free-space Green's function. The programmed field solution are ideal for performing rapid parametric studies of the dipole Halback cylinder magnets made from rare earth materials. The field solutions are verified by both an analytical two-dimensional algorithm and three-dimensional finite element software. A rapid method is presented for extensive analyzing and optimizing Halbach cylinder magnet. (authors)

A novel hanging spherical drop system for the generation of cellular spheroids and high throughput combinatorial drug screening.

Science.gov (United States)

Neto, A I; Correia, C R; Oliveira, M B; Rial-Hermida, M I; Alvarez-Lorenzo, C; Reis, R L; Mano, J F

2015-04-01

We propose a novel hanging spherical drop system for anchoring arrays of droplets of cell suspension based on the use of biomimetic superhydrophobic flat substrates, with controlled positional adhesion and minimum contact with a solid substrate. By facing down the platform, it was possible to generate independent spheroid bodies in a high throughput manner, in order to mimic in vivo tumour models on the lab-on-chip scale. To validate this system for drug screening purposes, the toxicity of the anti-cancer drug doxorubicin in cell spheroids was tested and compared to cells in 2D culture. The advantages presented by this platform, such as feasibility of the system and the ability to control the size uniformity of the spheroid, emphasize its potential to be used as a new low cost toolbox for high-throughput drug screening and in cell or tissue engineering.

Three-dimensional image signals: processing methods

Science.gov (United States)

Schiopu, Paul; Manea, Adrian; Craciun, Anca-Ileana; Craciun, Alexandru

2010-11-01

Over the years extensive studies have been carried out to apply coherent optics methods in real-time processing, communications and transmission image. This is especially true when a large amount of information needs to be processed, e.g., in high-resolution imaging. The recent progress in data-processing networks and communication systems has considerably increased the capacity of information exchange. We describe the results of literature investigation research of processing methods for the signals of the three-dimensional images. All commercially available 3D technologies today are based on stereoscopic viewing. 3D technology was once the exclusive domain of skilled computer-graphics developers with high-end machines and software. The images capture from the advanced 3D digital camera can be displayed onto screen of the 3D digital viewer with/ without special glasses. For this is needed considerable processing power and memory to create and render the complex mix of colors, textures, and virtual lighting and perspective necessary to make figures appear three-dimensional. Also, using a standard digital camera and a technique called phase-shift interferometry we can capture "digital holograms." These are holograms that can be stored on computer and transmitted over conventional networks. We present some research methods to process "digital holograms" for the Internet transmission and results.

In-Field High-Throughput Phenotyping of Cotton Plant Height Using LiDAR

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