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Sample records for flow cell studies

  1. Performance Mapping Studies in Redox Flow Cells

    NASA Technical Reports Server (NTRS)

    Hoberecht, M. A.; Thaller, L. H.

    1981-01-01

    Pumping power requirements in any flow battery system constitute a direct parasitic energy loss. It is therefore useful to determine the practical lower limit for reactant flow rates. Through the use of a theoretical framework based on electrochemical first principles, two different experimental flow mapping techniques were developed to evaluate and compare electrodes as a function of flow rate. For the carbon felt electrodes presently used in NASA-Lewis Redox cells, a flow rate 1.5 times greater than the stoichiometric rate seems to be the required minimum.

  2. Cell-cell interaction in blood flow in patients with coronary heart disease (in vitro study)

    NASA Astrophysics Data System (ADS)

    Malinova, Lidia I.; Simonenko, Georgy V.; Denisova, Tatyana P.; Tuchin, Valery V.

    2007-02-01

    Blood cell-cell and cell-vessel wall interactions are one of the key patterns in blood and vascular pathophysiology. We have chosen the method of reconstruction of pulsative blood flow in vitro in the experimental set. Blood flow structure was studied by PC integrated video camera with following slide by slide analysis. Studied flow was of constant volumetric blood flow velocity (1 ml/h). Diameter of tube in use was comparable with coronary arteries diameter. Glucose solution and unfractured heparin were used as the nonspecial irritants of studied flow. Erythrocytes space structure in flow differs in all groups of patients in our study (men with stable angina pectoris (SAP), myocardial infarction (MI) and practically healthy men (PHM). Intensity of erythrocytes aggregate formation was maximal in patients with SAP, but time of their "construction/deconstruction" at glucose injection was minimal. Phenomena of primary clotting formation in patients with SAP of high function class was reconstructed under experimental conditions. Heparin injection (10 000 ED) increased linear blood flow velocity both in patients with SAP, MI and PHP but modulated the cell profile in the flow. Received data correspond with results of animal model studies and noninvasive blood flow studies in human. Results of our study reveal differences in blood flow structure in patients with coronary heart disease and PHP under irritating conditions as the possible framework of metabolic model of coronary blood flow destabilization.

  3. A novel planar flow cell for studies of biofilm heterogeneity and flow-biofilm interactions

    PubMed Central

    Zhang, Wei; Sileika, Tadas S.; Chen, Cheng; Liu, Yang; Lee, Jisun; Packman, Aaron I.

    2012-01-01

    Biofilms are microbial communities growing on surfaces, and are ubiquitous in nature, in bioreactors, and in human infection. Coupling between physical, chemical, and biological processes is known to regulate the development of biofilms; however, current experimental systems do not provide sufficient control of environmental conditions to enable detailed investigations of these complex interactions. We developed a novel planar flow cell that supports biofilm growth under complex two-dimensional fluid flow conditions. This device provides precise control of flow conditions and can be used to create well-defined physical and chemical gradients that significantly affect biofilm heterogeneity. Moreover, the top and bottom of the flow chamber are transparent, so biofilm growth and flow conditions are fully observable using non-invasive confocal microscopy and high-resolution video imaging. To demonstrate the capability of the device, we observed the growth of Pseudomonas aeruginosa biofilms under imposed flow gradients. We found a positive relationship between patterns of fluid velocity and biofilm biomass because of faster microbial growth under conditions of greater local nutrient influx, but this relationship eventually reversed because high hydrodynamic shear leads to the detachment of cells from the surface. These results reveal that flow gradients play a critical role in the development of biofilm communities. By providing new capability for observing biofilm growth, solute and particle transport, and net chemical transformations under user-specified environmental gradients, this new planar flow cell system has broad utility for studies of environmental biotechnology and basic biofilm microbiology, as well as applications in bioreactor design, environmental engineering, biogeochemistry, geomicrobiology, and biomedical research. PMID:21656713

  4. A study of differential-flow-rate-cell corrosion in seawater

    SciTech Connect

    Miyasaka, M.; Kishimoto, K.; Aoki, S.

    1995-10-01

    Mechanisms of differential-flow-rate-cell corrosion (differential-aeration-cell corrosion caused by differential flow rates) of cast iron in seawater were studied. Potential and current density distributions produced by the differential-flow-rate-cell were on actual pumps and a model test cell. Boundary element analysis was also performed on differential-flow-rate-cell corrosion occurred in the model test cell. These studies demonstrate that differential-flow-rate-cell corrosion has characteristics similar to those of galvanic corrosion, and thus can be treated in the same manner as galvanic corrosion.

  5. Chemical responses of single yeast cells studied by fluorescence microspectroscopy under solution-flow conditions.

    PubMed

    Kogi, Osamu; Kim, Haeng-Boo; Kitamura, Noboru

    2002-07-01

    A microspectroscopy system combined with a fluid manifold was developed to manipulate and analyze "single" living cells. A sample buffer solution containing living cells was introduced into a flow cell set on a thermostated microscope stage and a few cells were allowed to attach to the bottom wall of the flow cell. With these living cells being attached to the wall, other floating cells were pumped out by flowing a buffer solution. These procedures made it possible to keep a few cells in the flow cell and to analyze single cells by fluorescence microspectroscopy. The technique was applied to study the time course of staining processes of single living yeast (Saccharomyces cerevisiae) cells by using two types of a fluorescent probe. The present methodology was shown to be of primary importance for obtaining biochemical/physiological information on single living cells and also for studying cell-to-cell variations in several characteristics.

  6. Imaging Flow Cytometry for the Study of Erythroid Cell Biology and Pathology

    PubMed Central

    Samsel, Leigh; McCoy, J Philip

    2015-01-01

    Erythroid cell maturation and diseases affecting erythrocytes are frequently accompanied by morphologic and immunophenotypic changes to these cells. In the past, these changes have been assessed primarily through the use of manual microscopy, which substantially limits the statistical rigor, throughput, and objectivity of these studies. Imaging flow cytometry provides a technology to examine both the morphology of cells as well as to quantify the staining intensity and signal distribution of numerous fluorescent markers on a cell-by-cell basis with high throughput in a statistically robust manner, and thus is ideally suited to studying erythroid cell biology. To date imaging flow cytometry has been used to study erythrocytes in three areas: 1) erythroid cell maturation, 2) sickle cell disease, and 3) infectious diseases such as malaria. In the maturation studies, imaging flow cytometry can closely recapitulate known stages of maturation and has led to the identification of a new population of erythroid cell precursors. In sickle cell disease, imaging flow cytometry provides a robust method to quantify sickled erythrocytes and to identify cellular aggregates linked to morbidities, and in malaria, imaging flow cytometry has been used to screen for new chemotherapeutic agents. These studies have demonstrated the value of imaging flow cytometry for investigations of erythrocyte biology and pathology. PMID:25858229

  7. Feasibility study of red blood cell debulking by magnetic field-flow fractionation with step-programmed flow.

    PubMed

    Moore, Lee R; Williams, P Stephen; Nehl, Franziska; Abe, Koji; Chalmers, Jeffrey J; Zborowski, Maciej

    2014-02-01

    Emerging applications of rare cell separation and analysis, such as separation of mature red blood cells from hematopoietic cell cultures, require efficient methods of red blood cell (RBC) debulking. We have tested the feasibility of magnetic RBC separation as an alternative to centrifugal separation using an approach based on the mechanism of magnetic field-flow fractionation (MgFFF). A specially designed permanent magnet assembly generated a quadrupole field having a maximum field of 1.68 T at the magnet pole tips, zero field at the aperture axis, and a nearly constant radial field gradient of 1.75 T/mm (with a negligible angular component) inside a cylindrical aperture of 1.9 mm (diameter) and 76 mm (length). The cell samples included high-spin hemoglobin RBCs obtained by chemical conversion of hemoglobin to methemoglobin (met RBC) or by exposure to anoxic conditions (deoxy RBC), low-spin hemoglobin obtained by exposure of RBC suspension to ambient air (oxy RBC), and mixtures of deoxy RBC and cells from a KG-1a white blood cell (WBC) line. The observation that met RBCs did not elute from the channel at the lower flow rate of 0.05 mL/min applied for 15 min but quickly eluted at the subsequent higher flow rate of 2.0 mL/min was in agreement with FFF theory. The well-defined experimental conditions (precise field and flow characteristics) and a well-established FFF theory verified by studies with model cell systems provided us with a strong basis for making predictions about potential practical applications of the magnetic RBC separation.

  8. An experimental study of mushroom shaped stall cells. [on finite wings with separated flow

    NASA Technical Reports Server (NTRS)

    Winkelmann, A. E.

    1982-01-01

    Surface patterns characterized by a pair of counter-rotating swirls have been observed in connection with the conduction of surface flow visualization experiments involving test geometries with separated flows. An example of this phenomenon occurring on a finite wing with trailing edge stall has been referred to by Winkelmann and Barlow (1980) as 'mushroom shaped'. A description is presented of a collection of experimental results which show or suggest the occurrence of mushroom shaped stall cells on a variety of test geometries. Investigations conducted with finite wings, airfoil models, and flat plates are considered, and attention is given to studies involving the use of bluff models, investigations of shock induced boundary layer separation, and mushroom shaped patterns observed in a number of miscellaneous cases. It is concluded that the mushroom shaped stall cell appears commonly in separated flow regions.

  9. An Analytical Study on Periodically Changing Flow Cells in Groundwater Basins

    NASA Astrophysics Data System (ADS)

    Jiang, X.; Zhao, K. Y.; Wang, J. Z.; Wan, L.; Wang, H.

    2014-12-01

    Current understanding on basin-scale groundwater flow is mainly based on Tóth's (1962, 1963) pioneering study, which assumed that water table is a subdued replica of topography and the annual mean water table controls the pattern of groundwater flow. In fact, water table is driven by dynamic forcing and changes with time. This study modifies the conceptual model of basin-scale groundwater flow by taking the fluctuating water-table into account. For both unit basin and complex basin, water table fluctuates throughout the basin cross-section except for at basin valleys. By the method of separation of variables, we derive the analytical solution of hydraulics head and stream function in the unit basin and the complex basin, and discuss the characteristics of the time-dependent flow cells. For the unit basin, the change in amplitude of hydraulic head fluctuation with depth is studied. For the complex basin, the time-dependent distribution of internal stagnation points is discussed. In addition, the relationship between flowing artesian zone and the fluctuating water-table is discussed. The results of the current study enhance our understanding on the transient nature of basin-scale groundwater flow.

  10. A histological and flow cytometric study of dog brain endothelial cell injuries in delayed radiation necrosis

    SciTech Connect

    Yamaguchi, N.; Yamashima, T.; Yamashita, J. )

    1991-04-01

    The pathogenesis of delayed cerebral radiation necrosis was studied histologically and biochemically in 25 dogs with special attention to vascular endothelial cell injuries. The dogs were sacrificed 3 to 30 months after irradiation with a single dose of 15 Gy to the head. Brain specimens were appropriately fixed for light and electron microscopic studies, and capillary endothelial cells were isolated for flow cytometric study. The endothelial cells were stained with acridine orange, then the cell ratios in the reproductive phase (S + G2 + M) were investigated with flow cytometry. Thereafter, Feulgen hydrolysis and computer analysis of the hydrolysis curves were performed to examine the qualitative changes in deoxyribonucleic acid (DNA) of endothelial cells after irradiation. Under light microscopy, spongy degeneration with small cell infiltration was observed, especially in the frontal white matter, at 6 months after irradiation. At 9 months, necrotic foci appeared and developed until 15 months after irradiation. Blood vessels around the necrotic area showed luminal narrowing with endothelial hyperplasia and proliferation. At 30 months, no fresh necrotic lesions were observed. Under electron microscopy, endothelial cells of capillaries and small vessels around the necrotic area showed an increase of pinocytosis, and in the nuclei there was an increase of infoldings and euchromatin. The cell ratios in the reproductive phase were 14.5% to 23.3% (maximum at 9 months) in the irradiated group compared to 6.4% in the control group. The rate constant of apurinic acid production, a parameter correlating with DNA transcriptional activity, was minimum at 3 months and maximum at 9 months after irradiation. The data suggest that impairment of the microcirculation plays an important role in the pathogenesis of delayed radiation necrosis.

  11. A microfluidic flow-cell for the study of the ultrafast dynamics of biological systems

    SciTech Connect

    Chauvet, Adrien Chergui, Majed; Tibiletti, Tania; Caffarri, Stefano

    2014-10-01

    The study of biochemical dynamics by ultrafast spectroscopic methods is often restricted by the limited amount of liquid sample available, while the high repetition rate of light sources can induce photodamage. In order to overcome these limitations, we designed a high flux, sub-ml, capillary flow-cell. While the 0.1 mm thin window of the 0.5 mm cross-section capillary ensures an optimal temporal resolution and a steady beam deviation, the cell-pump generates flows up to ~0.35 ml/s that are suitable to pump laser repetition rates up to ~14 kHz, assuming a focal spot-diameter of 100 μm. In addition, a decantation chamber efficiently removes bubbles and allows, via septum, for the addition of chemicals while preserving the closed atmosphere. The minimal useable amount of sample is ~250 μl.

  12. Influence of nanostructural environment and fluid flow on osteoblast-like cell behavior: a model for cell-mechanics studies.

    PubMed

    Prodanov, L; Semeins, C M; van Loon, J J W A; te Riet, J; Jansen, J A; Klein-Nulend, J; Walboomers, X F

    2013-05-01

    Introducing nanoroughness on various biomaterials has been shown to profoundly effect cell-material interactions. Similarly, physical forces act on a diverse array of cells and tissues. Particularly in bone, the tissue experiences compressive or tensile forces resulting in fluid shear stress. The current study aimed to develop an experimental setup for bone cell behavior, combining a nanometrically grooved substrate (200 nm wide, 50 nm deep) mimicking the collagen fibrils of the extracellular matrix, with mechanical stimulation by pulsatile fluid flow (PFF). MC3T3-E1 osteoblast-like cells were assessed for morphology, expression of genes involved in cell attachment and osteoblastogenesis and nitric oxide (NO) release. The results showed that both nanotexture and PFF did affect cellular morphology. Cells aligned on nanotexture substrate in a direction parallel to the groove orientation. PFF at a magnitude of 0.7 Pa was sufficient to induce alignment of cells on a smooth surface in a direction perpendicular to the applied flow. When environmental cues texture and flow were interacting, PFF of 1.4 Pa applied parallel to the nanogrooves initiated significant cellular realignment. PFF increased NO synthesis 15-fold in cells attached to both smooth and nanotextured substrates. Increased collagen and alkaline phosphatase mRNA expression was observed on the nanotextured substrate, but not on the smooth substrate. Furthermore, vinculin and bone sialoprotein were up-regulated after 1 h of PFF stimulation. In conclusion, the data show that interstitial fluid forces and structural cues mimicking extracellular matrix contribute to the final bone cell morphology and behavior, which might have potential application in tissue engineering.

  13. Myogenic progenitors and imaging single-cell flow analysis: a model to study commitment of adult muscle stem cells.

    PubMed

    Trapecar, Martin; Kelc, Robi; Gradisnik, Lidija; Vogrin, Matjaz; Rupnik, Marjan Slak

    2014-12-01

    Research on skeletal muscles suffers from a lack of appropriate human models to study muscle formation and regeneration on the regulatory level of single cells. This hampers both basic understanding and the development of new therapeutic approaches. The use of imaging multicolour flow cytometry and myogenic stem cells can help fill this void by allowing researchers to visualize and quantify the reaction of individual cultured cells to bioactives or other physiological impulses. As proof of concept, we subjected human CD56+ satellite cells to reference bioactives follistatin and Malva sylvestris extracts and then used imaging multicolor flow cytometry to visualize the stepwise activation of myogenic factors MyoD and myogenin in individual cells. This approach enabled us to evaluate the potency of these bioactives to stimulate muscle commitment. To validate this method, we used multi-photon confocal microscopy to confirm the potential of bioactives to stimulate muscle differentiation and expression of desmin. Imaging multicolor flow cytometry revealed statistically significant differences between treated and untreated groups of myogenic progenitors and we propose the utilization of this concept as an integral part of future muscle research strategies.

  14. Studying depletion kinetics of circulating prostate cancer cells by in vivo flow cytometer

    NASA Astrophysics Data System (ADS)

    Liu, Guangda; Gu, Zhengqin; Guo, Jin; Li, Yan; Chen, Yun; Chen, Tong; Wang, Cheng; Wei, Xunbin

    2011-03-01

    Prostate cancer is the most common malignancy in American men and the second leading cause of deaths from cancer, after lung cancer. The tumor usually grows slowly and remains confined to the gland for many years. During this time, the tumor produces little or no symptoms or outward signs. As the cancer advances, however, it can metastasize throughout other areas of the body, such as the bones, lungs, and liver. Surgical resection, hormonal therapy, chemotherapy and radiation therapy are the foundation of current prostate cancer therapies. Treatments for prostate cause both short- and long-term side effects that may be difficult to accept. Molecular mechanisms of prostate cancer metastasis need to be understood better and new therapies must be developed to selectively target to unique characteristics of cancer cell growth and metastasis. We have developed the "in vivo microscopy" to study the mechanisms that govern prostate cancer cell spread through the microenvironment in vivo in real-time confocal near-infrared fluorescence imaging. A recently developed "in vivo flow cytometer" and optical imaging are used to assess prostate cancer cell spreading and the circulation kinetics of prostate cancer cells. A real- time quantitative monitoring of circulating prostate cancer cells by the in vivo flow cytometer will be useful to assess the effectiveness of the potential therapeutic interventions.

  15. Depletion kinetics of circulating prostate cancer cells studied by in vivo flow cytometer

    NASA Astrophysics Data System (ADS)

    Liu, Guangda; Guo, Jin; Li, Yan; Chen, Yun; Gu, Zhengqin; Chen, Tong; Wang, Cheng; Wei, Xunbin

    2010-11-01

    Prostate cancer is the most common malignancy in American men and the second leading cause of deaths from cancer, after lung cancer. The tumor usually grows slowly and remains confined to the gland for many years. During this time, the tumor produces little or no symptoms or outward signs. As the cancer advances, however, it can metastasize throughout other areas of the body, such as the bones, lungs, and liver. Surgical resection, hormonal therapy, chemotherapy and radiation therapy are the foundation of current prostate cancer therapies. Treatments for prostate cause both short- and long-term side effects that may be difficult to accept. Molecular mechanisms of prostate cancer metastasis need to be understood better and new therapies must be developed to selectively target to unique characteristics of cancer cell growth and metastasis. We have developed the "in vivo microscopy" to study the mechanisms that govern prostate cancer cell spread through the microenvironment in vivo in real-time confocal nearinfrared fluorescence imaging. A recently developed "in vivo flow cytometer" and optical imaging are used to assess prostate cancer cell spreading and the circulation kinetics of prostate cancer cells. A real- time quantitative monitoring of circulating prostate cancer cells by the in vivo flow cytometer will be useful to assess the effectiveness of the potential therapeutic interventions.

  16. Studying circulating prostate cancer cells by in-vivo flow cytometer

    NASA Astrophysics Data System (ADS)

    Guo, Jin; Gu, Zhengqin; Chen, Tong; Wang, Cheng; Wei, Xunbin

    2011-11-01

    Prostate cancer is the most common malignancy in American men and the second leading cause of deaths from cancer, after lung cancer. The tumor usually grows slowly and remains confined to the gland for many years. As the cancer advances, however, it can metastasize throughout other areas of the body, such as the bones, lungs, and liver. Surgical resection, hormonal therapy, chemotherapy and radiation therapy are the foundation of current prostate cancer therapies. Treatments for prostate cause both short- and long-term side effects that may be difficult to accept. Molecular mechanisms of prostate cancer metastasis need to be understood better and new therapies must be developed to selectively target to unique characteristics of cancer cell growth and metastasis. We have developed the "in vivo microscopy" to study the mechanisms that govern prostate cancer cell spread through the microenvironment in vivo in real-time confocal near-infrared fluorescence imaging. A recently developed "in vivo flow cytometer" and optical imaging are used to assess prostate cancer cell spreading and the circulation kinetics of prostate cancer cells. We have measured the depletion kinetics of cancer cells with different metastatic potential. Interestingly, more invasive PC-3 prostate cancer cells are depleted faster from the circulation than LNCaP cells.

  17. Studying circulating prostate cancer cells by in-vivo flow cytometer

    NASA Astrophysics Data System (ADS)

    Guo, Jin; Gu, Zhengqin; Chen, Tong; Wang, Cheng; Wei, Xunbin

    2012-03-01

    Prostate cancer is the most common malignancy in American men and the second leading cause of deaths from cancer, after lung cancer. The tumor usually grows slowly and remains confined to the gland for many years. As the cancer advances, however, it can metastasize throughout other areas of the body, such as the bones, lungs, and liver. Surgical resection, hormonal therapy, chemotherapy and radiation therapy are the foundation of current prostate cancer therapies. Treatments for prostate cause both short- and long-term side effects that may be difficult to accept. Molecular mechanisms of prostate cancer metastasis need to be understood better and new therapies must be developed to selectively target to unique characteristics of cancer cell growth and metastasis. We have developed the "in vivo microscopy" to study the mechanisms that govern prostate cancer cell spread through the microenvironment in vivo in real-time confocal near-infrared fluorescence imaging. A recently developed "in vivo flow cytometer" and optical imaging are used to assess prostate cancer cell spreading and the circulation kinetics of prostate cancer cells. We have measured the depletion kinetics of cancer cells with different metastatic potential. Interestingly, more invasive PC-3 prostate cancer cells are depleted faster from the circulation than LNCaP cells.

  18. Studying the role of macrophages in circulating prostate cancer cells by in vivo flow cytometry

    NASA Astrophysics Data System (ADS)

    Cui, Xiaojun; Guo, Jin; Gu, Zhengqin; Wei, Xunbin

    2012-12-01

    Metastasis is a very complicated multi-step process and accounts for the low survival rate of the cancerous patients. To metastasize, the malignant cells must detach from the primary tumor and migrate to secondary sites in the body through either blood or lymph circulation. Macrophages appear to be directly involved in tumor progression and metastasis. However, the role of macrophages in affecting cancer metastasis has not been fully elucidated. Here, we have utilized an emerging technique, namely in vivo flow cytometry (IVFC) to study the depletion kinetics of circulating prostate cancer cells in mice and how depletion of macrophages by the liposome-encapsulated clodronate affects the depletion kinetics. Our results show different depletion kinetics of PC-3 cells between macrophage-deficient group and the control group. The number of circulating tumor cells (CTCs) in macrophage-deficient group decreases in a slower manner compared to the control mice group. The differences in depletion kinetics indicate that the absence of macrophages facilitates the stay of prostate cancer cells in circulation. We speculate that macrophages might be able to arrest, phagocytose and digest PC-3 cells. Therefore, the phagocytosis may mainly contribute to the depletion kinetic differences. The developed methods here would be useful to study the relationship between macrophages and tumor metastasis in small animal cancer model.

  19. Miniaturized Electrochemical Flow Cells

    PubMed Central

    Sahlin, Eskil; Halle, Alexandra ter; Schaefer, Kathleen; Horn, Jeffery; Then, Matthew; Weber, Stephen G.

    2006-01-01

    Several novel types of miniaturized electrochemical flow cells are described. The flow cells are fabricated in fluorinated ethylene propylene using a novel technique where channels with inner diameters down to 13 μm are integrated with electrodes. The channel is formed by shrinking and simultaneous melting of a heat shrink/melt tubing around a channel template (a tungsten wire) and electrodes followed by removal of the channel template. The technique allows incorporation of different electrode materials of different sizes. The electrode configuration consists of one or two working electrodes inside the channel and a counter electrode located in the channel outlet reservoir. Electrode configurations with different channel and working electrode sizes, different electrode materials including carbon fibers, glassy carbon rods, poly(tetrafluoroethylene)/carbon composite material, and platinum wires, and different arrangements have been assembled. Hydrodynamic voltammograms in dual-electrode (generator–collector) experiments indicate good potential control for cells with 25-μm channels, while there is some iR drop in cells with 13-μm channels. Cells prepared with a cylindrical working electrode tangent and perpendicular to a flow channel show a flow rate dependence consistent with thin-layer cell behavior. Electrode areas can be made in the range of 10−10–10−8 m2. PMID:12622401

  20. Role of macrophages in circulating prostate cancer cells studied by in vivo flow cytometry

    NASA Astrophysics Data System (ADS)

    Liu, Rongrong; Guo, Jin; Gu, Zhengqin; Wei, Xunbin

    2013-02-01

    Macrophages appear to be directly involved in cancer progression and metastasis. However, the role of macrophages in influencing tumor metastasis has not been fully understood. Here, we have used an emerging technique, namely in vivo flow cytometry (IVFC) to study the depletion kinetics of circulating prostate cancer cells in mice and how depletion of macrophages by the liposome-encapsulated clodronate affects the depletion kinetics. Our results show different depletion kinetics of PC-3 prostate cancer cells between macrophage-deficient group and the control group. The number of circulating tumor cells (CTCs) in macrophage-deficient group decreases in a slower manner compared to the control mice group. The differences in depletion kinetics indicate that the absence of macrophages might facilitate the stay of prostate tumor cells in circulation. We speculate that macrophages might be able to arrest, phagocytose and digest PC-3 cancer cells. Therefore, the phagocytosis may mainly contribute to the differences in depletion kinetics. The developed methods here would be useful to study the relationship between macrophages and cancer metastasis in small animal tumor model.

  1. Growth of Myxococcus xanthus in continuous-flow-cell bioreactors as a method for studying development.

    PubMed

    Smaldone, Gregory T; Jin, Yujie; Whitfield, Damion L; Mu, Andrew Y; Wong, Edward C; Wuertz, Stefan; Singer, Mitchell

    2014-04-01

    Nutrient sensors and developmental timers are two classes of genes vital to the establishment of early development in the social soil bacterium Myxococcus xanthus. The products of these genes trigger and regulate the earliest events that drive the colony from a vegetative state to aggregates, which ultimately leads to the formation of fruiting bodies and the cellular differentiation of the individual cells. In order to more accurately identify the genes and pathways involved in the initiation of this multicellular developmental program in M. xanthus, we adapted a method of growing vegetative populations within a constant controllable environment by using flow cell bioreactors, or flow cells. By establishing an M. xanthus community within a flow cell, we are able to test developmental responses to changes in the environment with fewer concerns for effects due to nutrient depletion or bacterial waste production. This approach allows for greater sensitivity in investigating communal environmental responses, such as nutrient sensing. To demonstrate the versatility of our growth environment, we carried out time-lapse confocal laser scanning microscopy to visualize M. xanthus biofilm growth and fruiting body development, as well as fluorescence staining of exopolysaccharides deposited by biofilms. We also employed the flow cells in a nutrient titration to determine the minimum concentration required to sustain vegetative growth. Our data show that by using a flow cell, M. xanthus can be held in a vegetative growth state at low nutrient concentrations for long periods, and then, by slightly decreasing the nutrient concentration, cells can be allowed to initiate the developmental program.

  2. Ellipsoidal cell flow system

    DOEpatents

    Salzman, Gary C.; Mullaney, Paul F.

    1976-01-01

    The disclosure relates to a system incorporating an ellipsoidal flow chamber having light reflective walls for low level light detection in practicing cellular analysis. The system increases signal-to-noise ratio by a factor of ten over prior art systems. In operation, laser light passes through the primary focus of the ellipsoid. A controlled flow of cells simultaneously passes through this focus so that the laser light impinges on the cells and is modulated by the cells. The reflective walls of the ellipsoid reflect the cell-modulated light to the secondary focus of the ellipsoid. A tapered light guide at the secondary focus picks up a substantial portion of modulated reflective light and directs it onto a light detector to produce a signal. The signal is processed to obtain the intensity distribution of the modulated light and hence sought after characteristics of the cells. In addition, cells may be dyed so as to fluoresce in response to the laser light and their fluorescence may be processed as cell-modulated light above described. A light discriminating filter would be used to distinguish reflected modulated laser light from reflected fluorescent light.

  3. Poly(dimethylsiloxane) thin films as biocompatible coatings for microfluidic devices : cell culture and flow studies with glial cells.

    SciTech Connect

    Peterson, Sophie Louise; Sasaki, Darryl Yoshio; Gourley, Paul Lee; McDonald, Anthony Eugene

    2004-06-01

    Oxygen plasma treatment of poly(dimethylsiloxane) (PDMS) thin films produced a hydrophilic surface that was biocompatible and resistant to biofouling in microfluidic studies. Thin film coatings of PDMS were previously developed to provide protection for semiconductor-based microoptical devices from rapid degradation by biofluids. However, the hydrophobic surface of native PDMS induced rapid clogging of microfluidic channels with glial cells. To evaluate the various issues of surface hydrophobicity and chemistry on material biocompatibility, we tested both native and oxidized PDMS (ox-PDMS) coatings as well as bare silicon and hydrophobic alkane and hydrophilic oligoethylene glycol silane monolayer coated under both cell culture and microfluidic studies. For the culture studies, the observed trend was that the hydrophilic surfaces supported cell adhesion and growth, whereas the hydrophobic ones were inhibitive. However, for the fluidic studies, a glass-silicon microfluidic device coated with the hydrophilic ox-PDMS had an unperturbed flow rate over 14 min of operation, whereas the uncoated device suffered a loss in rate of 12%, and the native PDMS coating showed a loss of nearly 40%. Possible protein modification of the surfaces from the culture medium also were examined with adsorbed films of albumin, collagen, and fibrinogen to evaluate their effect on cell adhesion.

  4. Application Of Micro-Highspeed Flow Visualization In Study Of Blood Cells Rheology In Vivo

    NASA Astrophysics Data System (ADS)

    Gui-shah, Li; Ni, Liang; Yu-ju, Lin; Jian, Zhang; Qiang, Wang

    1990-01-01

    A new experimental method has been developed in study of rheological behaviour of single red blood cell (RBC) in passing through the capillaries in vivo, using the technique of micro-highspeed cinecamera and micro-highspeed video system. It is one of the most important topics in the study of microcirculatory theories that fur-ther understand the deformability of RBC, flow states, velocities and dynamic mechanimi. A micro-highspeed flow visualization system consisted of essential elements: a biological microscope, a highspeed cinecmera with 35 mm film, a highspeed motion analysis system SP2000 (Kodak U.S.A) and a cold-light source etc. We have investigated the rheological parameters of single RBC in vivo in single capillaries which are about 3.3 to 6.9 um in diameters. The RBCs velocities are 0.1 to 0.25 mm/sec, and maximum shear stress on the outside surface of RBC is 13.8 dyn/cml, and maximum extension of RBC is 10.3 um. In aforementioned experiment, the highspeed flow visualization system frequency at 530 frames/sec and 200 frames/sec were used respectively. In addition, the vasomotion of precapillary sphincters have been measured and a complicated coupling phenomena between the RBC and sphincter have also been recorded and analysed. The experiment were performed with intravital hamsters and frogs. The results obtained by this system shown that the method designed by us are an effective tool in the study of rheological behaviour of single RBC in passing through the blood capillaries in vivoz.

  5. Growth of Myxococcus xanthus in Continuous-Flow-Cell Bioreactors as a Method for Studying Development

    PubMed Central

    Smaldone, Gregory T.; Jin, Yujie; Whitfield, Damion L.; Mu, Andrew Y.; Wong, Edward C.; Wuertz, Stefan

    2014-01-01

    Nutrient sensors and developmental timers are two classes of genes vital to the establishment of early development in the social soil bacterium Myxococcus xanthus. The products of these genes trigger and regulate the earliest events that drive the colony from a vegetative state to aggregates, which ultimately leads to the formation of fruiting bodies and the cellular differentiation of the individual cells. In order to more accurately identify the genes and pathways involved in the initiation of this multicellular developmental program in M. xanthus, we adapted a method of growing vegetative populations within a constant controllable environment by using flow cell bioreactors, or flow cells. By establishing an M. xanthus community within a flow cell, we are able to test developmental responses to changes in the environment with fewer concerns for effects due to nutrient depletion or bacterial waste production. This approach allows for greater sensitivity in investigating communal environmental responses, such as nutrient sensing. To demonstrate the versatility of our growth environment, we carried out time-lapse confocal laser scanning microscopy to visualize M. xanthus biofilm growth and fruiting body development, as well as fluorescence staining of exopolysaccharides deposited by biofilms. We also employed the flow cells in a nutrient titration to determine the minimum concentration required to sustain vegetative growth. Our data show that by using a flow cell, M. xanthus can be held in a vegetative growth state at low nutrient concentrations for long periods, and then, by slightly decreasing the nutrient concentration, cells can be allowed to initiate the developmental program. PMID:24509931

  6. Numerical study of changing the geometry of the flow field of a PEM fuel cell

    NASA Astrophysics Data System (ADS)

    Khazaee, I.; Sabadbafan, H.

    2016-05-01

    The geometry of channels of a PEM fuel cell is an important parameter that affects the performance of it that the lower voltage loss in polarization curve can indicate the better performance. In this study a complete three-dimensional and single phase model is used to investigate the effect of increasing the number of serpentine channels in the bipolar plates and also increasing the area (depth) of channels of a PEM fuel cell with rectangular, triangular and elliptical cross-section geometry. A single set of conservation equations which are valid for the flow channels, gas-diffusion electrodes, catalyst layers, and the membrane region is developed and numerically solved using a finite volume based computational fluid dynamics technique. The results show that there are good agreement with the numerical results and experimental results of the previous work of authors. Also the results show that by increasing the number of channels from one to four and eight, the performance improved about 18 % and by decreasing the area of channels from 2 to 1 mm2 the performance improved about 13 %.

  7. Study of flow channel geometry using current distribution measurement in a high temperature polymer electrolyte membrane fuel cell

    NASA Astrophysics Data System (ADS)

    Lobato, Justo; Cañizares, Pablo; Rodrigo, Manuel A.; Pinar, F. Javier; Úbeda, Diego

    To improve fuel cell design and performance, research studies supported by a wide variety of physical and electrochemical methods have to be carried out. Among the different techniques, current distribution measurement owns the desired feature that can be performed during operation, revealing information about internal phenomena when the fuel cell is working. Moreover, short durability is one of the main problems that is hindering fuel cell wide implementation and it is known to be related to current density heterogeneities over the electrode surface. A good flow channel geometry design can favor a uniform current density profile, hence hypothetically extending fuel cell life. With this, it was thought that a study on the influence of flow channel geometry on the performance of a high temperature polymer electrolyte membrane (PEM) fuel cell using current distribution measurement should be a very solid work to optimize flow field design. Results demonstrate that the 4 step serpentine and pin-type geometries distribute the reactants more effectively, obtaining a relatively flat current density map at higher current densities than parallel or interdigitated ones and yielding maximum powers up to 25% higher when using oxygen as comburent. If air is the oxidant chosen, interdigitated flow channels perform almost as well as serpentine or pin-type due to that the flow conditions are very important for this geometry.

  8. Design of a Side-View Particle Imaging Velocimetry Flow System for Cell-Substrate Adhesion Studies

    PubMed Central

    Leyton-Mange, Jordan; Yang, Sung; Hoskins, Meghan H.; Kunz, Robert F.; Zahn, Jeffrey D.; Dong, Cheng

    2009-01-01

    Experimental models that mimic the flow conditions in microcapillaries have suggested that the local shear stresses and shear rates can mediate tumor cell and leukocyte arrest on the endothelium and subsequent sustained adhesion. However, further investigation has been limited by the lack of experimental models that allow quantitative measurement of the hydrodynamic environment over adherent cells. The purpose of this study was to develop a system capable of acquiring quantitative flow profiles over adherent cells. By combining the techniques of side-view imaging and particle image velocimetry (PIV), an in vitro model was constructed that is capable of obtaining quantitative flow data over cells adhering to the endothelium. The velocity over an adherent leukocyte was measured and the shear rate was calculated under low and high upstream wall shear. The microcapillary channel was modeled using computational fluid dynamics (CFD) and the calculated velocity profiles over cells under the low and high shear rates were compared to experimental results. The drag force applied to each cell by the fluid was then computed. This system provides a means for future study of the forces underlying adhesion by permitting characterization of the local hydrodynamic conditions over adherent cells. PMID:16524340

  9. Analytical and numerical study on cooling flow field designs performance of PEM fuel cell with variable heat flux

    NASA Astrophysics Data System (ADS)

    Afshari, Ebrahim; Ziaei-Rad, Masoud; Jahantigh, Nabi

    2016-06-01

    In PEM fuel cells, during electrochemical generation of electricity more than half of the chemical energy of hydrogen is converted to heat. This heat of reactions, if not exhausted properly, would impair the performance and durability of the cell. In general, large scale PEM fuel cells are cooled by liquid water that circulates through coolant flow channels formed in bipolar plates or in dedicated cooling plates. In this paper, a numerical method has been presented to study cooling and temperature distribution of a polymer membrane fuel cell stack. The heat flux on the cooling plate is variable. A three-dimensional model of fluid flow and heat transfer in cooling plates with 15 cm × 15 cm square area is considered and the performances of four different coolant flow field designs, parallel field and serpentine fields are compared in terms of maximum surface temperature, temperature uniformity and pressure drop characteristics. By comparing the results in two cases, the constant and variable heat flux, it is observed that applying constant heat flux instead of variable heat flux which is actually occurring in the fuel cells is not an accurate assumption. The numerical results indicated that the straight flow field model has temperature uniformity index and almost the same temperature difference with the serpentine models, while its pressure drop is less than all of the serpentine models. Another important advantage of this model is the much easier design and building than the spiral models.

  10. Minimal residual disease monitoring by 8-color flow cytometry in mantle cell lymphoma: an EU-MCL and LYSA study.

    PubMed

    Cheminant, Morgane; Derrieux, Coralie; Touzart, Aurore; Schmit, Stéphanie; Grenier, Adrien; Trinquand, Amélie; Delfau-Larue, Marie-Hélène; Lhermitte, Ludovic; Thieblemont, Catherine; Ribrag, Vincent; Cheze, Stéphane; Sanhes, Laurence; Jardin, Fabrice; Lefrère, François; Delarue, Richard; Hoster, Eva; Dreyling, Martin; Asnafi, Vahid; Hermine, Olivier; Macintyre, Elizabeth

    2016-03-01

    Quantification of minimal residual disease may guide therapeutic strategies in mantle cell lymphoma. While multiparameter flow cytometry is used for diagnosis, the gold standard method for minimal residual disease analysis is real-time quantitative polymerase chain reaction (RQ-PCR). In this European Mantle Cell Lymphoma network (EU-MCL) pilot study, we compared flow cytometry with RQ-PCR for minimal residual disease detection. Of 113 patients with at least one minimal residual disease sample, RQ-PCR was applicable in 97 (86%). A total of 284 minimal residual disease samples from 61 patients were analyzed in parallel by flow cytometry and RQ-PCR. A single, 8-color, 10-antibody flow cytometry tube allowed specific minimal residual disease assessment in all patients, with a robust sensitivity of 0.01%. Using this cut-off level, the true-positive-rate of flow cytometry with respect to RQ-PCR was 80%, whereas the true-negative-rate was 92%. As expected, RQ-PCR frequently detected positivity below this 0.01% threshold, which is insufficiently sensitive for prognostic evaluation and would ideally be replaced with robust quantification down to a 0.001% (10-5) threshold. In 10 relapsing patients, the transition from negative to positive by RQ-PCR (median 22.5 months before relapse) nearly always preceded transition by flow cytometry (4.5 months), but transition to RQ-PCR positivity above 0.01% (5 months) was simultaneous. Pre-emptive rituximab treatment of 2 patients at minimal residual disease relapse allowed re-establishment of molecular and phenotypic complete remission. Flow cytometry minimal residual disease is a complementary approach to RQ-PCR and a promising tool in individual mantle cell lymphoma therapeutic management. (clinicaltrials identifiers: 00209209 and 00209222).

  11. A flow cytometric study of chromosomes from rat kangaroo and Chinese hamster cells.

    PubMed

    Stöhr, M; Hutter, K J; Frank, M; Futterman, G; Goerttler, K

    1980-01-01

    Chromosomes from rat kangaroo (PTK) and chinese hamster (CHV 79) cells have been prepared for quantitative flow-cytometric analysis. The preparation time was otimized down to 30 (PTK) and 40 min (CHV 79). DAPI was used as a AT-sensitive fluorescent dye to stain for monoparameter DNA measurements. Simultaneous two-parameter DNA-protein analysis was carried out with DAPI and SR 101 (as a general protein fluorochrome) in combination. The karyotype of the PTK cells with 13 (14) chromosomes was separated into 10DNA peaks. The X-chromosome bearing the nucleolus organizer region generates a distinct peak. The karyotype of the CHV 79 cells with 22 chromosomes was separated inot 15 peaks. The DNA profile obtained indicates a geometric grading of the chromosomal amount of AT components in teh karyotype of this particular cell line. The simultaneous DNA-protein analysis performed show enough sensitivity of the instrument utilizing hihg power UV excitation illumination to discriminate the two color emission consisting of blue (DAPI) and red (SR 101) fluorescence. Color overlapping could be completely avoided. Additionally, the quality (number, location, and resolution of peaks) of the DNA distribution was not influences by the simultaneous application of a second fluorescent stain. Fluorescence activated electronic sorting applied on chromosomal fluorescence distributions providing purified fractions of chromosomes for subsequent biochemical and biological determinations is discussed.

  12. Preferential flow in heterogeneous forest-reclaimed lignitic mine soil I. Cell-lysimeter and multiple-tracer study

    NASA Astrophysics Data System (ADS)

    Hangen, E.; Gerke, H. H.; Schaaf, W.; Hüttl, R. F.

    2003-04-01

    Flow and transport processes in forest-reclaimed lignitic mine soils are required to quantify water and element budgets, which are important for long-term predictions of restored ecosystem stability and development of mining area water quality. Soil water pressure head and solute concentration measurements using tensiometers and suction cups showed strong spatial heterogeneity possibly indicating preferential flow effects. Properties and spatial structures of the mostly sandy mine soils and transport processes, however, have not sufficiently been known for detailed assessments. The objective of this study was to quantitatively analyse flow paths and measure amount and spatial distribtion of leaching. Water and element fluxes were studied at a reclaimed mine spoil site, which was afforested in 1982 with Pinus nigra. At a 3.3 m2 plot, the total percolating water was collected in 110 cm soil depth by 45 squared suction cells of 27 cm edge length each. A multi-tracer solution containing deuterium, bromide, and terbuthylazine was applied evenly at the plot surface and imposed to natural infiltration. Leaching was measured for a period of about 2 years. One third of the cells never delivered any drainage water while few cells had large drainage rates which in one case even exceeded local infiltration rates. About 71 % of the drainage was through 9 % of the area. The spatial distribution of the leached bromide tracer did not always correspond with that of drainage. Relative concentrations of bromide and deuterium were similar. Terbuthylazine was observed only sporadically during the first drainage period and at relatively small concentrations just above the analytical detection limit. Leaching patterns of the sorptive herbicide indicate only relatively small nonequilibrium-type preferential flow. Sediment structures, water repellent regions, and tree root distributions seem to be important for funneling and flow path formation.

  13. An ultra-high vacuum electrochemical flow cell for in situ/operando soft X-ray spectroscopy study

    SciTech Connect

    Bora, Debajeet K. E-mail: jguo@lbl.gov; Glans, Per-Anders; Pepper, John; Liu, Yi-Sheng; Guo, J.-H. E-mail: jguo@lbl.gov; Du, Chun; Wang, Dunwei

    2014-04-15

    An in situ flow electrochemical cell has been designed and fabricated to allow better seal under UHV chamber thus to achieve a good signal to noise ratio in fluorescence yield detection of X-ray absorption spectra for spectroelectrochemical study. The cell also stabilizes the thin silicon nitride membrane window in an effective manner so that the liquid cell remains intact during X-ray absorption experiments. With the improved design of the liquid cell, electrochemical experiments such as cyclic voltammetry have been performed for 10 cycles with a good stability of sample window. Also an operando electrochemical experiment during photoelectrochemistry has been performed on n-type hematite electrode deposited on silicon nitride window. The experiment allows us to observe the formation of two extra electronic transitions before pre edge of O K-edge spectra.

  14. MRI Study of Granular Flow in a Split-Bottomed Couette Cell

    NASA Astrophysics Data System (ADS)

    Cheng, Xiang

    2005-03-01

    Recent studies of dense granular flow in a split-bottomed Couette geometry have brought new insights into the concept of shear bands in granular systems [1]. However, to date experimental results have primarily focused on the flow at the top surface of the system. Here we present a study of the 3- dimensional structure of shear band formed in such a geometry using magnetic resonance imaging (MRI). We show that the angular velocity profiles in horizontal plane follow an error function as observed at the top surface. By measuring the center and the width of the shear band at the different heights in the bulk, we map out the 3-D shape of the shear band and investigate the behavior of the shear band as a function of the total filling height. We find that when the top of the shear band detaches from the surface of the bulk, its shape changes dramatically, similar to a first order transition as has been proposed by theory [2]. [1] D. Fenistein, J. W. van de Meent, and M. van Hecke, PRL 92, 094301 (2004). [2] T. Unger, J. Torok, J. Kertesz, D. E. Wolf, PRL 92, 214301 (2004).

  15. Bypass Flow Study

    SciTech Connect

    Richard Schultz

    2011-09-01

    The purpose of the fluid dynamics experiments in the MIR (Matched Index of-Refraction) flow system at Idaho National Laboratory (INL) is to develop benchmark databases for the assessment of Computational Fluid Dynamics (CFD) solutions of the momentum equations, scalar mixing, and turbulence models for the flow ratios between coolant channels and bypass gaps in the interstitial regions of typical prismatic standard fuel element (SFE) or upper reflector block geometries of typical Modular High-temperature Gas-cooled Reactors (MHTGR) in the limiting case of negligible buoyancy and constant fluid properties. The experiments use Particle Image Velocimetry (PIV) to measure the velocity fields that will populate the bypass flow study database.

  16. Fundamental studies of materials, designs, and models development for polymer electrolyte membrane fuel cell flow field distributors

    NASA Astrophysics Data System (ADS)

    Nikam, Vaibhav Vilas

    Fuel cells are becoming a popular source of energy due to their promising performance and availability. However, the high cost of fuel cell stack forbids its deployment to end user. Moreover, bipolar plate is one of the critical components in current polymer electrolyte membrane fuel cell (PEMFC) system, causing severe increase in manufacturing cost. The objective of this research work is to develop new materials, design and manufacturing process for bipolar plates. The materials proposed for use were tested for corrosion resistance in simulated fuel cell conditions. After corrosion studies copper alloy (C17200) and Low Temperature Carburized (LTC) SS 316 were selected as an alternative material for bipolar plate. It was observed that though the copper alloy offered good resistance in corrosive atmosphere, the major advantage of using the alloys was good conductivity even after formation of corrosion layer compared to SS 316. However, LTC SS 316 achieved the best corrosion resistance (ever reported in current open literature at relatively low cost) with decreased contact resistance, as compared to SS 316. Due to the expensive and tedious machining for bipolar plate manufacturing, the conventional machining process was not used. Bipolar plates were manufactured from thin corrugated sheets formed of the alloy. This research also proposed a novel single channel convoluted flow field design which was developed by increasing the tortuosity of conventional serpentine design. The CFD model for novel single channel convoluted design showed uniform distribution of velocity over the entire three dimensional domain. The novel design was further studied using pressure drop and permeability models. These modeling calculations showed substantial benefit in using corrugated sheet design and novel single channel convoluted flow field design. All the concepts of materials (except for LTC SS 316), manufacturing and design are validated using various tests like long term stability

  17. Transport studies in polymer electrolyte fuel cell with porous metallic flow field at ultra high current density

    NASA Astrophysics Data System (ADS)

    Srouji, Abdul-Kader

    Achieving cost reduction for polymer electrolyte fuel cells (PEFC) requires a simultaneous effort in increasing power density while reducing precious metal loading. In PEFCs, the cathode performance is often limiting due to both the slow oxygen reduction reaction (ORR), and mass transport limitation caused by limited oxygen diffusion and liquid water flooding at high current density. This study is motivated by the achievement of ultra-high current density through the elimination of the channel/land (C/L) paradigm in PEFC flow field design. An open metallic element (OME) flow field capable of operating at unprecedented ultra-high current density (3 A/cm2) introduces new advantages and limitations for PEFC operation. The first part of this study compares the OME with a conventional C/L flow field, through performance and electrochemical diagnostic tools such as electrochemical impedance spectroscopy (EIS). The results indicate the uniqueness of the OME's mass transport improvement. No sign of operation limitation due to flooding is noted. The second part specifically examines water management at high current density using the OME flow field. A unique experimental setup is developed to measure steady-state and transient net water drag across the membrane, in order to characterize the fundamental aspects of water transport at high current density with the OME. Instead of flooding, the new limitation is identified to be anode side dry-out of the membrane, caused by electroosmotic drag. The OME improves water removal from the cathode, which immediately improves oxygen transport and performance. However, the low water content in the cathode reduces back diffusion of water to the membrane, and electroosmotic drag dominates at high current density, leading to dry-out. The third part employs the OME flow field as a tool that avoids C/L effects endemic to a typical flow field, in order to study oxygen transport resistance at the catalyst layer of a PEFC. In open literature, a

  18. A Comparative Study of Electrolyte Flow and Slime Particle Transport in a Newly Designed Copper Electrolytic Cell and a Laboratory-Scale Conventional Electrolytic Cell

    NASA Astrophysics Data System (ADS)

    Zeng, Weizhi; Wang, Shijie; Free, Michael L.

    2016-08-01

    An innovative copper electrolytic cell was designed with its inlet at the cell top and its outlet near the cell bottom, in opposite to conventional electrolytic cells. It was modeled in COMSOL Multiphysics to simulate copper electrorefining process. Unlike conventional electrorefining cells, downward electrolyte flows are more dominant in the fluid flow field in this cell, which leads to faster settlement of slime particles and less contamination to the cathode. Copper concentration profiles, electrolyte flow velocity field, slime particle movements, and slime particle distributions were obtained as simulation results, which were compared with those in a laboratory-scale conventional electrolytic cell. Advantages of the newly designed electrolytic cell were found: copper ions are distributed more uniformly in the cell with a thinner diffusion layer near the cathode; stronger convection exists in the inter-electrode domain with dominant downward flows; and slime particles have larger possibilities to settle down and are less likely to reach the cathode.

  19. Biofilm responses to smooth flow fields and chemical gradients in novel microfluidic flow cells

    PubMed Central

    Song, Jisun L.; Au, Kelly H.; Huynh, Kimberly T.

    2013-01-01

    We present two novel microfluidic flow cells developed to provide reliable control of flow distributions and chemical gradients in biofilm studies. We developed a single-inlet microfluidic flow cell to support biofilm growth under a uniform velocity field, and a double-inlet flow cell to provide a very smooth transverse concentration gradient. Both flow cells consist of a layer of polydimethylsiloxane (PDMS) bonded to glass cover slips and were fabricated using the replica molding technique. We demonstrate the capabilities of the flow cells by quantifying flow patterns before and after growth of Pseudomonas aeruginosa biofilms through particle imaging velocimetry, and by evaluating concentration gradients within the double-inlet microfluidic flow cell. Biofilm growth substantially increased flow complexity by diverting flow around biomass, creating high- and low-velocity regions and surface friction. Under a glucose gradient in the double-inlet flow cell, P. aeruginosa biofilms grew in proportion to the local glucose concentration, producing distinct spatial patterns in biofilm biomass relative to the imposed glucose gradient. When biofilms were subjected to a ciprofloxacin gradient, spatial patterns of fractions of dead cells were also in proportion to the local antibiotic concentration. These results demonstrate that the microfluidic flow cells are suitable for quantifying flow complexities resulting from flow-biofilm interactions and investigating spatial patterns of biofilm growth under chemical gradients. These novel microfluidic flow cells will facilitate biofilm research that requires flow control and in situ imaging, particularly investigations of biofilm-environment interactions. PMID:24038055

  20. Biofilm responses to smooth flow fields and chemical gradients in novel microfluidic flow cells.

    PubMed

    Song, Jisun L; Au, Kelly H; Huynh, Kimberly T; Packman, Aaron I

    2014-03-01

    We present two novel microfluidic flow cells developed to provide reliable control of flow distributions and chemical gradients in biofilm studies. We developed a single-inlet microfluidic flow cell to support biofilm growth under a uniform velocity field, and a double-inlet flow cell to provide a very smooth transverse concentration gradient. Both flow cells consist of a layer of polydimethylsiloxane (PDMS) bonded to glass cover slips and were fabricated using the replica molding technique. We demonstrate the capabilities of the flow cells by quantifying flow patterns before and after growth of Pseudomonas aeruginosa biofilms through particle imaging velocimetry, and by evaluating concentration gradients within the double-inlet microfluidic flow cell. Biofilm growth substantially increased flow complexity by diverting flow around biomass, creating high- and low-velocity regions and surface friction. Under a glucose gradient in the double-inlet flow cell, P. aeruginosa biofilms grew in proportion to the local glucose concentration, producing distinct spatial patterns in biofilm biomass relative to the imposed glucose gradient. When biofilms were subjected to a ciprofloxacin gradient, spatial patterns of fractions of dead cells were also in proportion to the local antibiotic concentration. These results demonstrate that the microfluidic flow cells are suitable for quantifying flow complexities resulting from flow-biofilm interactions and investigating spatial patterns of biofilm growth under chemical gradients. These novel microfluidic flow cells will facilitate biofilm research that requires flow control and in situ imaging, particularly investigations of biofilm-environment interactions. PMID:24038055

  1. Studies on reactive flows

    SciTech Connect

    Sheu, Wenjenn.

    1989-01-01

    Theoretical, numerical and experimental studies are presented on the upstream interactions between premixed flamelets in turbulent combustions, the effect of centrifugal acceleration due to the rotation on Bunsen flames, extinction of counterflow diffusion flames in a counter-rotating finite jet, and nonplanar flame configurations in stagnation point flow. The processes during the upstream flame interactions are numerically investigated using two ozone decomposition planar flames propagating towards each other. The flame temperature and speed during the whole interaction process are also analyzed. One basic kind of rotational effects on the shape and stability of the premixed flames stabilized in Bunsen burner is theoretically and experimentally studied. The relation between the flame stabilization by the burner rim and rotation across the flame is examined. In addition the stable and unstable flame configuration are identified. In the study of the extinction characteristics of diffusion flames in opposed-jet flows with angular velocities which are equal in magnitude but opposite in directions, the extinction characteristics with vorticity in the flow were explored. The general configurations of premixed flame fronts in a stagnation point flow is pursued. Some flame shapes other than planar flames were verified. A simple mathematical model is given to provide the explicit expressions of flame fronts. The implications of these studies to the modeling of turbulent combustion are discussed.

  2. Micromodel foam flow study

    SciTech Connect

    Chambers, K.T.; Radke, C.J.

    1990-10-01

    Foams are often utilized as part of enhanced oil recovery techniques. This report presents the results of a micromodel foam flow study. Micromodels are valuable tools in uncovering capillary phenomena responsible for lamellae generation and coalescence during foam flow in porous media. Among the mechanisms observed are snap-off, weeping-flow breakup, and lamella division and leave behind. Coalescence mechanisms include dynamic capillary-pressure-induced lamella drainage and gas diffusion. These phenomena are sensitive to the mode of injection, the local capillary environment, and the geometry of the pore structure. An important consideration in presenting a tractable model of foam flow behavior is the ability to identify the pore-level mechanisms having the greatest impact on foam texture. The predominant mechanisms will vary depending upon the application for foam as an enhanced oil recovery (EOR) fluid. Both simultaneous gas and surfactant injection and surfactant alternating with gas injection (SAG) have been used to create foam for mobility control in EOR projects. The model developed is based on simultaneous gas and surfactant injection during steady-state conditions into a Berea sandstone core. The lamellae generation and coalescence mechanisms included in this model are snap-off, lamella division, and dynamic capillary-pressure-induced lamella drainage. This simplified steady-state model serves as a foundation for developing more complete rate expressions and for extending the population balance to handle transient foam flow behavior. 70 refs., 30 figs.

  3. Free flow cell electrophoresis using zwitterionic buffer

    NASA Technical Reports Server (NTRS)

    Rodkey, R. Scott

    1990-01-01

    Studies of a zwitterionic buffer formulated for cell electrophoresis were done using the McDonnell-Douglas Continuous Flow Electrophoresis System. Standard buffers were analyzed for their stability in the electrical field and the results showed that both buffers tested were inherently unstable. Further, titration studies showed that the standards buffers buffered poorly at the pH employed for electrophoresis. The zwitterionic buffer buffered well at its nominal pH and was shown to be stable in the electrical field. Comparative studies of the buffer with standard cell separation buffers using formalin fixed rabbit and goose red blood cells showed that the zwitterionic buffer gave better resolution of the fixed cells. Studies with viable hybridoma cells showed that buffer Q supported cell viability equal to Hank's Balanced Salt Solution and that hybridoma cells in different stages of the growth cycle demonstrated reproducible differences in electrophoretic mobility.

  4. Numerical study of a novel micro-diaphragm flow channel with piezoelectric device for proton exchange membrane fuel cells

    NASA Astrophysics Data System (ADS)

    Ma, H. K.; Huang, S. H.; Chen, B. R.; Cheng, L. W.

    Previous studies have shown that the amplitude of the vibration of a piezoelectric (PZT) device produces an oscillating flow that changes the chamber volume along with a curvature variation of the diaphragm. In this study, an actuating micro-diaphragm with piezoelectric effects is utilized as an air-flow channel in proton exchange membrane fuel cell (PEMFC) systems, called PZT-PEMFC. This newly designed gas pump, with a piezoelectric actuation structure, can feed air into the system of an air-breathing PEMFC. When the actuator moves outward to increase the cathode channel volume, the air is sucked into the chamber; moving inward decreases the channel's volume and thereby compresses air into the catalyst layer and enhancing the chemical reaction. The air-standard PZT-PEMFC cycle is proposed to describe an air-breathing PZT-PEMFC. A novel design for PZT-PEMFCs has been proposed and a three-dimensional, transitional model has been successfully built to account for its major phenomena and performance. Moreover, at high frequencies, PZT actuation leads to a more stable current output, more drained water, higher sucked air, higher hydrogen consumption, and also overcomes concentration losses.

  5. Metal-induced alteration of the cell membrane/cytoplasm complex studied by flow cytometry and detergent lysis

    SciTech Connect

    Zucker, R.M.; Elstein, K.H.; Easterling, R.E.; Massaro, E.J.

    1988-01-01

    Flow cytometric analysis of the cell cycle is most effectively accomplished with membrane-/cytoplasma-free (clean) nuclei. Non-ionic detergents (e.g. NP40 or Triton X-100) commonly are employed to solubilize cells membranes/cytoplasm to produce 'clean' nuclei. Treatment of murine erythroleukemic cells with tri-n-butylin methoxide, cadmium acetate, zinc sulfate, or lead acetate alters the properties of the cell membrane/cytoplasm complex making it resistant to NP40 dissolution. On a molar basis, the organotin compound was more effective in inducing resistance to detergent-mediated dissolution than the inorganic metal compounds. Resistance to NO40-mediated dissolution was manifested as an increase in the flow cytometric parameters 90 degree scatter and fluorescein isothiocyanate fluorescence and was confirmed by light microscopy.

  6. Flow over glycocalyx covered endothelial cells.

    NASA Astrophysics Data System (ADS)

    Waters, Sarah L.; Liu, Shu Q.; Grotberg, James B.

    1997-11-01

    Blood vessels are lined with a monolayer of endothelial cells that interact with the blood flow. Two structural features of the endothelial membrane may influence the fluid dynamics over the endothelium: 1) the endothelial membrane bumpiness; and 2) the porous glycocalyx layer covering the cell surface. The bumpiness of the endothelium may induce regions of recirculation, and the glycocalyx may effect the flow pattern at the cell surface. An analytical study is presented for pressure driven blood flow in the microcirculation. The vessel is modeled as a rigid, impermeable, symmetric two--dimensional channel, which has sinusoidally wavy walls. The vessel has two regions: 1) the glycocalyx layer which is modeled as a uniformly thick poroelastic deformable wall layer using biphasic mixture theory; and 2) the free lumen where the Navier--Stokes equations of motion apply. Analytical results are obtained by making the long wavelength approximation. The model predicts the fluid flow and hence the shear stress exerted by the flow on the individual endothelial cells and at the glycocalyx--lumen interface. Implications of the results for biological events such as molecular transport and signal transduction are considered.

  7. Flow cytometry total cell counts: a field study assessing microbiological water quality and growth in unchlorinated drinking water distribution systems.

    PubMed

    Liu, G; Van der Mark, E J; Verberk, J Q J C; Van Dijk, J C

    2013-01-01

    The objective of this study was to evaluate the application of flow cytometry total cell counts (TCCs) as a parameter to assess microbial growth in drinking water distribution systems and to determine the relationships between different parameters describing the biostability of treated water. A one-year sampling program was carried out in two distribution systems in The Netherlands. Results demonstrated that, in both systems, the biomass differences measured by ATP were not significant. TCC differences were also not significant in treatment plant 1, but decreased slightly in treatment plant 2. TCC values were found to be higher at temperatures above 15°C than at temperatures below 15°C. The correlation study of parameters describing biostability found no relationship among TCC, heterotrophic plate counts, and Aeromonas. Also no relationship was found between TCC and ATP. Some correlation was found between the subgroup of high nucleic acid content bacteria and ATP (R (2) = 0.63). Overall, the results demonstrated that TCC is a valuable parameter to assess the drinking water biological quality and regrowth; it can directly and sensitively quantify biomass, detect small changes, and can be used to determine the subgroup of active HNA bacteria that are related to ATP.

  8. Cell migration does not produce membrane flow

    PubMed Central

    1990-01-01

    We have previously reported that rearward migration of surface particles on slowly moving cells is not driven by membrane flow (Sheetz, M. P., S. Turney, H. Qian, and E. L. Elson. 1989. Nature (Lond.). 340:284-288) and recent photobleaching measurements have ruled out any rapid rearward lipid flow (Lee, J., M. Gustafsson, D. E. Magnussen, and K. Jacobson. 1990. Science (Wash. DC.) 247:1229-1233). It was not possible, however, to conclude from those studies that a slower or tank-tread membrane lipid flow does not occur. Therefore, we have used the technology of single particle tracking to examine the movements of diffusing particles on rapidly locomoting fish keratocytes where the membrane current is likely to be greatest. The keratocytes had a smooth lamellipodial surface on which bound Con A-coated gold particles were observed either to track toward the nuclear region (velocity of 0.35 +/- 0.15 micron/s) or to diffuse randomly (apparent diffusion coefficient of [3.5 +/- 2.0] x 10(-10) cm2/s). We detected no systematic drift relative to the cell edge of particles undergoing random diffusion even after the cell had moved many micrometers. The average net particle displacement was 0.01 +/- 2.7% of the cell displacement. These results strongly suggest that neither the motions of membrane proteins driven by the cytoskeleton nor other possible factors produce a bulk flow of membrane lipid. PMID:2211827

  9. Study of self-consistent particle flows in a plasma blob with particle-in-cell simulations

    SciTech Connect

    Hasegawa, Hiroki Ishiguro, Seiji

    2015-10-15

    The self-consistent particle flows in a filamentary coherent structure along the magnetic field line in scrape-off layer (SOL) plasma (plasma blob) have been investigated by means of a three-dimensional electrostatic particle-in-cell simulation code. The presence of the spiral current system composed of the diamagnetic and parallel currents in a blob is confirmed by the particle simulation without any assumed sheath boundary models. Furthermore, the observation of the electron and ion parallel velocity distributions in a blob shows that those distributions are far from Maxwellian due to modification with the sheath formation and that the electron temperature on the higher potential side in a blob is higher than that on the lower potential side. Also, it is found that the ions on the higher potential side are accelerated more intensively along the magnetic field line than those on the lower potential side near the edge. This study indicates that particle simulations are able to provide an exact current closure to analysis of blob dynamics and will bring more accurate prediction of plasma transport in the SOL without any empirical assumptions.

  10. Cancer cell glycocalyx mediates mechanotransduction and flow-regulated invasion.

    PubMed

    Qazi, Henry; Palomino, Rocio; Shi, Zhong-Dong; Munn, Lance L; Tarbell, John M

    2013-11-01

    Mammalian cells are covered by a surface proteoglycan (glycocalyx) layer, and it is known that blood vessel-lining endothelial cells use the glycocalyx to sense and transduce the shearing forces of blood flow into intracellular signals. Tumor cells in vivo are exposed to forces from interstitial fluid flow that may affect metastatic potential but are not reproduced by most in vitro cell motility assays. We hypothesized that glycocalyx-mediated mechanotransduction of interstitial flow shear stress is an un-recognized factor that can significantly enhance metastatic cell motility and play a role in augmentation of invasion. Involvement of MMP levels, cell adhesion molecules (CD44, α3 integrin), and glycocalyx components (heparan sulfate and hyaluronan) was investigated in a cell/collagen gel suspension model designed to mimic the interstitial flow microenvironment. Physiological levels of flow upregulated MMP levels and enhanced the motility of metastatic cells. Blocking the flow-enhanced expression of MMP activity or adhesion molecules (CD44 and integrins) resulted in blocking the flow-enhanced migratory activity. The presence of a glycocalyx-like layer was verified around tumor cells, and the degradation of this layer by hyaluronidase and heparinase blocked the flow-regulated invasion. This study shows for the first time that interstitial flow enhancement of metastatic cell motility can be mediated by the cell surface glycocalyx - a potential target for therapeutics.

  11. Cell-surface changes in cadmium-resistant Euglena: Studies using lectin-binding techniques and flow cytometry

    SciTech Connect

    Bonaly, J.; Brochiero, E.

    1994-01-01

    Most in vitro studies on contaminants focus on the short-term effects of pollutants on cells, without regard to long-term effects and the ability of cells or microorganisms to develop a specific resistance to a pollutant. Cadmium is ubiquitous environmental contaminant. This heavy metal enters the aquatic environment mainly through vapor emissions and fallout during smelting operations. Diverse mechanisms of algal resistance to toxic metals are known. Among these, the most general mechanism is the development of metal-binding proteins. In cadmium-resistant unicellular Euglena gracilis Z algae cells, the metal did not appear to be sequestered on soluble metal-binding ligands. Previous experiments have shown that resistance development is related to a diminution of cadmium penetration into cells, implicating cell surface or membrane alteration. This research investigates the mechanisms of development of cadmium resistance in Euglena cells at the cell-surface level. Sugar chains of glycoproteins and glycolipids are a predominant feature of the surface of cells. Moreover, the cell-response to environmental changes is often orchestrated through surface macromolecules such as glycoproteins. In this study, we applied this lectin method to investigate surface carbohydrate expression during and after resistance development. Our interest was twofold: (1) to learn more about the carbohydrate composition of the cell-surface of Euglena; and (2) to determine whether transition from wild cells to Cd-resistant cells changes the expression of cell-surface carbohydrates. 13 refs., 2 figs., 1 tab.

  12. Studies of shear flows

    NASA Astrophysics Data System (ADS)

    Kline, S. J.; Johnston, J. P.; Moffat, R. J.

    1986-03-01

    The objective of the work reported is construction of zonal models for accurate prediction of turbulent flows in rapid-running computer programs. The work follows the ideas set down in the discussion of zonal modeling by S. J. Kline in Vol. 2 of the Proceedings of the 1980-81 AFOSR-Stanford Conference on Complex Turbulent Flows. This discussion noted that the fast-running models available lack sufficient span to predict all classes of turbulent flows of engineering importance in a standard, invariant form. It therefore suggested that the models be treated as zonal. In the zonal approach, the constants in th models are adjusted for each important zone of the flow, where the word zone implies a region with a particular type of flow physics. In the proposal for the work, it was also noted that the parametrization of the flows and the selection of appropriate constants would ultimately need to be guided by the domain over which accurate results could be obtained, and that might well be different from pre-conceptions embodied in conventional taxonomies of the flows.

  13. Mirrored serpentine flow channels for fuel cell

    SciTech Connect

    Rock, Jeffrey Allan

    2000-08-08

    A PEM fuel cell having serpentine flow field channels wherein the input/inlet legs of each channel border the input/inlet legs of the next adjacent channels in the same flow field, and the output/exit legs of each channel border the output/exit legs of the next adjacent channels in the same flow field. The serpentine fuel flow channels may be longer, and may contain more medial legs, than the serpentine oxidant flow channels.

  14. Particle-in-cell simulation study of the scaling of asymmetric magnetic reconnection with in-plane flow shear

    NASA Astrophysics Data System (ADS)

    Doss, C. E.; Cassak, P. A.; Swisdak, M.

    2016-08-01

    We investigate magnetic reconnection in systems simultaneously containing asymmetric (anti-parallel) magnetic fields, asymmetric plasma densities and temperatures, and arbitrary in-plane bulk flow of plasma in the upstream regions. Such configurations are common in the high-latitudes of Earth's magnetopause and in tokamaks. We investigate the convection speed of the X-line, the scaling of the reconnection rate, and the condition for which the flow suppresses reconnection as a function of upstream flow speeds. We use two-dimensional particle-in-cell simulations to capture the mixing of plasma in the outflow regions better than is possible in fluid modeling. We perform simulations with asymmetric magnetic fields, simulations with asymmetric densities, and simulations with magnetopause-like parameters where both are asymmetric. For flow speeds below the predicted cutoff velocity, we find good scaling agreement with the theory presented in Doss et al. [J. Geophys. Res. 120, 7748 (2015)]. Applications to planetary magnetospheres, tokamaks, and the solar wind are discussed.

  15. Fuel cell with internal flow control

    SciTech Connect

    Haltiner, Jr., Karl J.; Venkiteswaran, Arun

    2012-06-12

    A fuel cell stack is provided with a plurality of fuel cell cassettes where each fuel cell cassette has a fuel cell with an anode and cathode. The fuel cell stack includes an anode supply chimney for supplying fuel to the anode of each fuel cell cassette, an anode return chimney for removing anode exhaust from the anode of each fuel cell cassette, a cathode supply chimney for supplying oxidant to the cathode of each fuel cell cassette, and a cathode return chimney for removing cathode exhaust from the cathode of each fuel cell cassette. A first fuel cell cassette includes a flow control member disposed between the anode supply chimney and the anode return chimney or between the cathode supply chimney and the cathode return chimney such that the flow control member provides a flow restriction different from at least one other fuel cell cassettes.

  16. High-speed flow cytometric analysis of nanoparticle targeting to rare leukemic stem cells in peripheral human blood: preliminary in-vitro studies

    NASA Astrophysics Data System (ADS)

    Cooper, Christy L.; Leary, James F.

    2014-03-01

    Leukemic cancer stem cells are both stem-like and leukemic-like. This complicates their detection as rare circulating tumor cells in peripheral blood of leukemia patients. The leukemic stem cells are also highly resistant to standard chemotherapeutic regimens so new therapeutic strategies need to be designed to kill the leukemic stem cells without killing normal stem cells. In these initial studies we have designed an antibody-targeted and fluorescent (Cy5.5) nanoparticle for targeting these leukemic stem cells and then introducing new strategies for killing them. Multicolor flow cytometric analyses were performed on a BD FACS Aria III. Human leukemic stem cell-like cell line RS4;11 (with putative immunophenotype CD123+/CD24+/CD38-/CD10-/Flt-3-) was used as a model human leukemic stem cell systems and were spiked into normal human peripheral blood cells containing normal blood stem-progenitor cells (immunophenotype CD123-/CD34+/CD38-) and Cy5.5-labeled nanoparticles with targeting molecule anti-CD123 antibody. An irrelevant antibody (CD71) which should not bind to any live leukemic stem cell or normal stem cell (binds erythrocytes) was used as a way of distinguishing between true-positive live and false-positive damaged/dead cells, the latter occurring at much higher frequencies than the very rare (e.g. 0.001 to 0.0001 percent frequency true leukemic stem cells). These studies are designed to measure the targeting sensitivity and specificity of the fluorescent nanoparticles to the putative rare leukemic stem cells with the eventual design to use the nanoparticles to direct killing therapeutic doses to the leukemic stem cells but not to the normal stem-progenitor cells.

  17. Lack of evidence of chalone activity in used medium and extract of JB-1 tumor cells in vitro. A flow cytometry study.

    PubMed

    Naeser, F K; Barfod, N M; Bichel, P

    1978-02-14

    In cell-free mouse ascites fluid from the JB-1 ascites tumor in the plateau phase of growth low-molecular chalone substances have been found which reversibly and specifically arrest JB-1 cells in the G1 and G2 phase of the cell cycle. The aim of this study was to investigate whether chalones were involved in the regulation of in vitro growth of JB-1 tumor cells. Used medium and cell extract from confluent, stationary JB-1 cell cultures were investigated for proliferation-inhibitory properties. JB-1 cells from stationary cultures were explanted in test cultures and the traverse of cells through the S phase was investigated by means of flow cytometry (FCM). Inhibition--expressed as a delay of the traverse of cells through the S phase--was not observed when a surplus of used medium, concentrated and fractionated used medium or concentrated and fractionated cell extract from JB-1 cells in vitro was added to test cultures. On the contrary, used medium and concentrated and fractionated used medium stimulated growth. Thus, no involvement of chalones in the growth regulation of JB-1 tumor cells in vitro was detected.

  18. Electrochemical study of multi-electrode microbial fuel cells under fed-batch and continuous flow conditions

    NASA Astrophysics Data System (ADS)

    Ren, Lijiao; Ahn, Yongtae; Hou, Huijie; Zhang, Fang; Logan, Bruce E.

    2014-07-01

    Power production of four hydraulically connected microbial fuel cells (MFCs) was compared with the reactors operated using individual electrical circuits (individual), and when four anodes were wired together and connected to four cathodes all wired together (combined), in fed-batch or continuous flow conditions. Power production under these different conditions could not be made based on a single resistance, but instead required polarization tests to assess individual performance relative to the combined MFCs. Based on the power curves, power produced by the combined MFCs (2.12 ± 0.03 mW, 200 Ω) was the same as the summed power (2.13 mW, 50 Ω) produced by the four individual reactors in fed-batch mode. With continuous flow through the four MFCs, the maximum power (0.59 ± 0.01 mW) produced by the combined MFCs was slightly lower than the summed maximum power of the four individual reactors (0.68 ± 0.02 mW). There was a small parasitic current flow from adjacent anodes and cathodes, but overall performance was relatively unaffected. These findings demonstrate that optimal power production by reactors hydraulically and electrically connected can be predicted from performance by individual reactors.

  19. Ultrasonic Enrichment of Flowing Blood Cells in Capillars: Influence of the Flow Rate

    NASA Astrophysics Data System (ADS)

    Carreras, Pilar; Gonzalez, Itziar; Ahumada, Oscar

    Red blood cells subjected to standing waves collect at the pressure nodes during their flow motion. Blood is a non-newtonian fluid whose density and other properties are defined by its flow velocity. Their drift motion is governed by the radiation force together with hydrodynamic conditions. This work presents a study of the blood cell enrichment performed in a rectangular capillar at f=1 MHz as a function of their flow motion. The cells collect along the central axis of the capillary in very few seconds, with a clearance in other lateral areas. Optimal flow rates below 100uL/min were found in the experiments.

  20. Recovery, Visualization, and Analysis of Actin and Tubulin Polymer Flow in Live Cells: A Fluorescent Speckle Microscopy Study

    PubMed Central

    Vallotton, P.; Ponti, A.; Waterman-Storer, C. M.; Salmon, E. D.; Danuser, G.

    2003-01-01

    Fluorescent speckle microscopy (FSM) is becoming the technique of choice for analyzing in vivo the dynamics of polymer assemblies, such as the cytoskeleton. The massive amount of data produced by this method calls for computational approaches to recover the quantities of interest; namely, the polymerization and depolymerization activities and the motions undergone by the cytoskeleton over time. Attempts toward this goal have been hampered by the limited signal-to-noise ratio of typical FSM data, by the constant appearance and disappearance of speckles due to polymer turnover, and by the presence of flow singularities characteristic of many cytoskeletal polymer assemblies. To deal with these problems, we present a particle-based method for tracking fluorescent speckles in time-lapse FSM image series, based on ideas from operational research and graph theory. Our software delivers the displacements of thousands of speckles between consecutive frames, taking into account that speckles may appear and disappear. In this article we exploit this information to recover the speckle flow field. First, the software is tested on synthetic data to validate our methods. We then apply it to mapping filamentous actin retrograde flow at the front edge of migrating newt lung epithelial cells. Our results confirm findings from previously published kymograph analyses and manual tracking of such FSM data and illustrate the power of automated tracking for generating complete and quantitative flow measurements. Third, we analyze microtubule poleward flux in mitotic metaphase spindles assembled in Xenopus egg extracts, bringing new insight into the dynamics of microtubule assemblies in this system. PMID:12885672

  1. Receptor-mediated cell attachment and detachment kinetics. II. Experimental model studies with the radial-flow detachment assay.

    PubMed Central

    Cozens-Roberts, C; Quinn, J A; Lauffenburger, D A

    1990-01-01

    Quantitative information regarding the kinetics of receptor-mediated cell adhesion to a ligand-coated surface are crucial for understanding the role of certain key parameters in many physiological and biotechnology-related processes. Here, we use the probabilistic attachment and detachment models developed in the preceding paper to interpret transient data from well-defined experiments. These data are obtained with a simple model cell system that consists of receptor-coated latex beads (prototype cells) and a Radial-Flow Detachment Assay (RFDA) using a ligand-coated glass disc. The receptors and ligands used in this work are complementary antibodies. The beads enable us to examine transient behavior with particles that possess fairly uniform properties that can be varied systematically, and the RFDA is designed for direct observation of adhesion to the ligand-coated glass surface over a range of shear stresses. Our experiments focus on the effects of surface shear stress, receptor density, and ligand density. These data provide a crucial test of the probabilistic framework. We show that these data can be explained with the probabilistic analyses, whereas they cannot be readily interpreted on the basis of a deterministic analysis. In addition, we examine transient data on cell adhesion reported from other assays, demonstrating the consistency of these data with the predictions of the probabilistic models. Images FIGURE 2 PMID:2174272

  2. Blood Cell Interactions and Segregation in Flow

    PubMed Central

    Munn, Lance L.; Dupin, Michael M.

    2009-01-01

    For more than a century, pioneering researchers have been using novel experimental and computational approaches to probe the mysteries of blood flow. Thanks to their efforts, we know that blood cells generally prefer to migrate to the axis of flow, that red and white cells segregate in flow, and that cell deformability and their tendency to reversibly aggregate contribute to the non-Newtonian nature of this unique fluid. All of these properties have beneficial physiological consequences, allowing blood to perform a variety of critical functions. Our current understanding of these unusual flow properties of blood have been made possible by the ingenuity and diligence of a number of researchers, including Harry Goldsmith, who developed novel technologies to visualize and quantify the flow of blood at the level of individual cells. Here we summarize efforts in our lab to continue this tradition and to further our understanding of how blood cells interact with each other and with the blood vessel wall. PMID:18188702

  3. Study of gas pressure and flow rate influences on a 500 W PEM fuel cell, thanks to the experimental design methodology

    NASA Astrophysics Data System (ADS)

    Wahdame, B.; Candusso, D.; Kauffmann, Jean-Marie

    The behaviour of a 500 W PEM fuel cell stack, fed by pure hydrogen and humidified compressed air, is currently investigated on the fuel cell test platform of Belfort. In this paper, the influences on fuel cell performance of gas pressure and flow rate parameters are studied. The fuel cell is operated in the pressure regulation mode: the gas flow rates are regulated thanks to mass flow controllers placed upstream of the stack and the gas pressures at stack inlets are controlled by regulation valves located downstream of the stack. The choice of the various tests to perform is made thanks to experimental design methodology, which is a suitable technique to characterise, analyse and to improve a complex system such as a fuel cell generator. In this study, the four physical factors considered are both hydrogen/air pressures and anode/cathode flow rates. Each factor has two levels, leading to a full factorial design requiring 16 experiments (16 current-voltage curves). The test bench developed at the laboratory allows setting the other factors (for instance: stack temperature, relative humidity and dew point temperature of the air at stack inlet) at fixed values. The test responses are the maximal output power and the efficiency computed for this power. Statistical sensitivity analyses (ANOVA analyses) are used to compute the effects and the contributions of the various factors to the fuel cell maximal power. The use of fractional designs shows also how it is possible to reduce the number of experiments. Some graphic representations are employed in order to display the results of the statistical analyses made for different current values.

  4. Traction Forces of Endothelial Cells under Slow Shear Flow

    PubMed Central

    Perrault, Cecile M.; Brugues, Agusti; Bazellieres, Elsa; Ricco, Pierre; Lacroix, Damien; Trepat, Xavier

    2015-01-01

    Endothelial cells are constantly exposed to fluid shear stresses that regulate vascular morphogenesis, homeostasis, and disease. The mechanical responses of endothelial cells to relatively high shear flow such as that characteristic of arterial circulation has been extensively studied. Much less is known about the responses of endothelial cells to slow shear flow such as that characteristic of venous circulation, early angiogenesis, atherosclerosis, intracranial aneurysm, or interstitial flow. Here we used a novel, to our knowledge, microfluidic technique to measure traction forces exerted by confluent vascular endothelial cell monolayers under slow shear flow. We found that cells respond to flow with rapid and pronounced increases in traction forces and cell-cell stresses. These responses are reversible in time and do not involve reorientation of the cell body. Traction maps reveal that local cell responses to slow shear flow are highly heterogeneous in magnitude and sign. Our findings unveil a low-flow regime in which endothelial cell mechanics is acutely responsive to shear stress. PMID:26488643

  5. Modeling of flow field in polymer electrolyte membrane fuel cell

    NASA Astrophysics Data System (ADS)

    Karvonen, Suvi; Hottinen, Tero; Saarinen, Jaakko; Himanen, Olli

    Isothermal two- and three-dimensional polymer electrolyte membrane (PEM) fuel cell cathode flow field models were implemented to study the behavior of reactant and reaction product gas flow in a parallel channel flow field. The focus was on the flow distribution across the channels and the total pressure drop across the flow field. The effect of the density and viscosity variation in the gas resulting from the composition change due to cell reactions was studied and the models were solved with governing equations based on three different approximations. The focus was on showing how a uniform flow profile can be achieved by improving an existing channel design. The modeling results were verified experimentally. A close to uniform flow distribution was achieved in a parallel channel system.

  6. Platelet antibody screening by flow cytometry is more sensitive than solid phase red cell adherence assay and lymphocytotoxicity technique: a comparative study in Thai patients.

    PubMed

    Buakaew, Jarin; Promwong, Charuporn

    2010-01-01

    The objective of this study was to compare the sensitivity and specificity of lymphocytotoxicity test (LCT), solid phase red cell adherence assay (SPRCA) and flow cytometry in detecting platelet reactive antibodies against human leukocyte antigens (HLA) class I and human platelet antigens (HPA). Sera from 38 thrombocytopenic patients and 5 mothers of thrombocytopenic newborns were screened for platelet reactive antibodies by these three methods using screening platelets and/or lymphocytes panels derived from six subjects. The sensitivity and specificity of each method and levels of agreement were analysed. HLA antibodies were found in 18, 17 and 19 out of 43 patients' sera tested by LCT, SPRCA and flow cytometry, respectively. Four out of 43 patients' sera were reactive against HPA by flow cytometry, but were reactive to only 2 sera by SPRCA. Using flow cytometry as the reference method, the sensitivities/specificities of SPRCA and LCT in HLA antibody detection were 84.21/95.83% and 94.73/100%, respectively, with a good strength of agreement. SPRCA had 50% sensitivity and 100% specificity in HPA antibody detection compare to flow cytometry. Flow cytometry appeared to be the most sensitive technique compared with SPRCA and LCT for both HPA and HLA antibody screening. SPRCA sensitivity was too low for HPA antibody detection, but this might be because of the small number of samples. There was one serum from the mother of a baby suffering neonatal alloimmune thrombocytopenia (NAIT), in whom SPRCA could not detect HPA antibodies, while flow cytometry came out positive. Therefore, SPRCA should not be used in NAIT investigation and flow cytometry should be employed instead.

  7. Flow and Diffusion in Channel-Guided Cell Migration

    PubMed Central

    Marel, Anna-Kristina; Zorn, Matthias; Klingner, Christoph; Wedlich-Söldner, Roland; Frey, Erwin; Rädler, Joachim O.

    2014-01-01

    Collective migration of mechanically coupled cell layers is a notable feature of wound healing, embryonic development, and cancer progression. In confluent epithelial sheets, the dynamics have been found to be highly heterogeneous, exhibiting spontaneous formation of swirls, long-range correlations, and glass-like dynamic arrest as a function of cell density. In contrast, the flow-like properties of one-sided cell-sheet expansion in confining geometries are not well understood. Here, we studied the short- and long-term flow of Madin-Darby canine kidney (MDCK) cells as they moved through microchannels. Using single-cell tracking and particle image velocimetry (PIV), we found that a defined averaged stationary cell current emerged that exhibited a velocity gradient in the direction of migration and a plug-flow-like profile across the advancing sheet. The observed flow velocity can be decomposed into a constant term of directed cell migration and a diffusion-like contribution that increases with density gradient. The diffusive component is consistent with the cell-density profile and front propagation speed predicted by the Fisher-Kolmogorov equation. To connect diffusion-mediated transport to underlying cellular motility, we studied single-cell trajectories and occurrence of vorticity. We discovered that the directed large-scale cell flow altered fluctuations in cellular motion at short length scales: vorticity maps showed a reduced frequency of swirl formation in channel flow compared with resting sheets of equal cell density. Furthermore, under flow, single-cell trajectories showed persistent long-range, random-walk behavior superimposed on drift, whereas cells in resting tissue did not show significant displacements with respect to neighboring cells. Our work thus suggests that active cell migration manifests itself in an underlying, spatially uniform drift as well as in randomized bursts of short-range correlated motion that lead to a diffusion-mediated transport

  8. Overhead Projection Cell for Streamline Flow

    ERIC Educational Resources Information Center

    Waage, Harold M.

    1969-01-01

    Describes the construction and operation of an overhead projection apparatus designed to demonstrate streamline flow of a liquid. The apparatus consists of a Plexiglass tank containing water in which plates forming the cell are submerged, a constant level reservoir, an overflow device and a system for marking the flow lines with a dye. (LC)

  9. Flow regimes in a trapped vortex cell

    NASA Astrophysics Data System (ADS)

    Lasagna, D.; Iuso, G.

    2016-03-01

    This paper presents results of an experimental investigation on the flow in a trapped vortex cell, embedded into a flat plate, and interacting with a zero-pressure-gradient boundary layer. The objective of the work is to describe the flow features and elucidate some of the governing physical mechanisms, in the light of recent investigations on flow separation control using vortex cells. Hot-wire velocity measurements of the shear layer bounding the cell and of the boundary layers upstream and downstream are reported, together with spectral and correlation analyses of wall-pressure fluctuation measurements. Smoke flow visualisations provide qualitative insight into some relevant features of the internal flow, namely a large-scale flow unsteadiness and possible mechanisms driving the rotation of the vortex core. Results are presented for two very different regimes: a low-Reynolds-number case where the incoming boundary layer is laminar and its momentum thickness is small compared to the cell opening, and a moderately high-Reynolds-number case, where the incoming boundary layer is turbulent and the ratio between the momentum thickness and the opening length is significantly larger than in the first case. Implications of the present findings to flow control applications of trapped vortex cells are also discussed.

  10. Inverted annular flow experimental study

    SciTech Connect

    De Jarlais, G.; Ishii, M.

    1985-04-01

    Steady-state inverted annular flow of Freon 113 in up flow was established in a transparent test section. Using a special inlet configuration consisting of long aspect-ratio liquid nozzles coaxially centered within a heated quartz tube, idealized inverted annular flow initial geometry (cylindrical liquid core surrounded by coaxial annulus of gas) could be established. Inlet liquid and gas flowrates, liquid subcooling, and gas density (using various gas species) were measured and varied systematically. The hydrodynamic behavior of the liquid core, and the subsequent downstream break-up of this core into slugs, ligaments and/or droplets of various sizes, was observed. In general, for low inlet liquid velocities it was observed that after the initial formation of roll waves on the liquid core surface, an agitated region of high surface area, with attendant high momentum and energy transfers, occurs. This agitated region appears to propagate downsteam in a quasi-periodic pattern. Increased inlet liquid flow rates, and high gas annulus flow rates tend to diminish the significance of this agitated region. Observed inverted annular flow (and subsequent downstream flow pattern) hydrodynamic behavior is reported, and comparisons are drawn to data generated by previous experimenters studying post-CHF flow.

  11. Gas-Particle Interactions in a Microgravity Flow Cell

    NASA Technical Reports Server (NTRS)

    Louge, Michel; Jenkins, James

    1999-01-01

    We are developing a microgravity flow cell in which to study the interaction of a flowing gas with relatively massive particles that collide with each other and with the moving boundaries of the cell. The absence of gravity makes possible the independent control of the relative motion of the boundaries and the flow of the gas. The cell will permit gas-particle interactions to be studied over the entire range of flow conditions over which the mixture is not turbulent. Within this range, we shall characterize the viscous dissipation of the energy of the particle fluctuations, measure the influence of particle-phase viscosity on the pressure drop along the cell, and observe the development of localized inhomogeneities that are likely to be associated with the onset of clusters. These measurements and observations should contribute to an understanding of the essential physics of pneumatic transport.

  12. Chaotic dynamics of red blood cells in oscillating shear flow

    NASA Astrophysics Data System (ADS)

    Bagchi, Prosenjit; Cordasco, Daniel

    2015-11-01

    A 3D computational study of deformable red blood cells in dilute suspension and subject to sinusoidally oscillating shear flow is considered. It is observed that the cell exhibits either a periodic motion or a chaotic motion. In the periodic motion, the cell reverses its orientation either about the flow direction or about the flow gradient, depending on the initial conditions. In certain parameter range, the initial conditions are forgotten and the cells become entrained in the same sequence of horizontal reversals. The chaotic dynamics is characterized by a nonperiodic sequence of horizontal and vertical reversals, and swings. The study provides the first conclusive evidence of the chaotic dynamics of fully deformable cells in oscillating flow using a deterministic numerical model without the introduction of any stochastic noise. An analysis of the chaotic dynamics shows that chaos is only possible in certain frequency bands when the cell membrane can rotate by a certain amount allowing the cells to swing near the maximum shear rate. We make a novel observation that the occurrence of the vertical or horizontal reversal depends only on whether a critical angle, that is independent of the flow frequency, is exceeded at the instant of flow reversal.

  13. Ciliary Neurotrophic Factor Induces Genes Associated with Inflammation and Gliosis in the Retina: A Gene Profiling Study of Flow-Sorted, Müller Cells

    PubMed Central

    Dudley, V. Joseph; Brooks, Matthew; Swaroop, Anand; Sarthy, Vijay P.

    2011-01-01

    Background Ciliary neurotrophic factor (CNTF), a member of the interleukin-6 cytokine family, has been implicated in the development, differentiation and survival of retinal neurons. The mechanisms of CNTF action as well as its cellular targets in the retina are poorly understood. It has been postulated that some of the biological effects of CNTF are mediated through its action via retinal glial cells; however, molecular changes in retinal glia induced by CNTF have not been elucidated. We have, therefore, examined gene expression dynamics of purified Müller (glial) cells exposed to CNTF in vivo. Methodology/Principal Findings Müller cells were flow-sorted from mgfap-egfp transgenic mice one or three days after intravitreal injection of CNTF. Microarray analysis using RNA from purified Müller cells showed differential expression of almost 1,000 transcripts with two- to seventeen-fold change in response to CNTF. A comparison of transcriptional profiles from Müller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern. Ingenuity Pathway Analysis showed that the differentially regulated genes belong to distinct functional types such as cytokines, growth factors, G-protein coupled receptors, transporters and ion channels. Interestingly, many genes induced by CNTF were also highly expressed in reactive Müller cells from mice with inherited or experimentally induced retinal degeneration. Further analysis of gene profiles revealed 20–30% overlap in the transcription pattern among Müller cells, astrocytes and the RPE. Conclusions/Significance Our studies provide novel molecular insights into biological functions of Müller glial cells in mediating cytokine response. We suggest that CNTF remodels the gene expression profile of Müller cells leading to induction of networks associated with transcription, cell cycle regulation and inflammatory response. CNTF also appears to

  14. Electrically rechargeable REDOX flow cell

    NASA Technical Reports Server (NTRS)

    Thaller, L. H. (Inventor)

    1976-01-01

    A bulk energy storage system is designed with an electrically rechargeable reduction-oxidation (REDOX) cell divided into two compartments by a membrane, each compartment containing an electrode. An anode fluid is directed through the first compartment at the same time that a cathode fluid is directed through the second compartment. Means are provided for circulating the anode and cathode fluids, and the electrodes are connected to an intermittent or non-continuous electrical source, which when operating, supplies current to a load as well as to the cell to recharge it. Ancillary circuitry is provided for disconnecting the intermittent source from the cell at prescribed times and for circulating the anode and cathode fluids according to desired parameters and conditions.

  15. Flow cytometric immunofluorescence of rat anterior pituitary cells

    NASA Technical Reports Server (NTRS)

    Hatfield, J. Michael; Hymer, W. C.

    1985-01-01

    A flow cytometric immunofluorescence technique was developed for the quantification of growth hormone, prolactin, and luteinizing hormone producing cells. The procedure is based on indirect-immunofluorescence of intracellular hormone using an EPICS V cell sorter and can objectively count 50,000 cells in about 3 minutes. It can be used to study the dynamics of pituitary cell populations under various physiological and pharmacological conditions.

  16. Geophysical Fluid Flow Cell (GFFC) Simulation

    NASA Technical Reports Server (NTRS)

    1999-01-01

    These simulations of atmospheric flow use the same experimental parameters but started with slightly different initial conditions in the model. The simulations were part of data analysis for the Geophysical Fluid Flow Cell (GFFC), a planet in a test tube apparatus flown on Spacelab to mimic the atmospheres on gas giant planets and stars. (Credit: Dr. Tim Miller of Global Hydrology and Climate Center at the Marshall Space Flight Center)

  17. Theoretical and Experimental Flow Cell Studies of a Hydrogen-Bromine Fuel Cell, Part 1. M.S. Thesis. Final Report

    NASA Technical Reports Server (NTRS)

    Savinell, R. F.; Fritts, S. D.

    1986-01-01

    There is increasing interest in hydrogen-bromine fuel cells as both primary and regenerative energy storage systems. One promising design for a hydrogen-bromine fuel cell is a negative half cell having only a gas phase, which is separated by a cationic exchange membrane from a positive half cell having an aqueous electrolyte. The hydrogen gas and the aqueous bromide solution are stored external to the cell. In order to calculate the energy storage capacity and to predict and assess the performance of a single cell, the open circuit potential (OCV) must be estimated for different states of change, under various conditions. Theoretical expressions were derived to estimate the OCV of a hydrogen-bromine fuel cell. In these expressions temperature, hydrogen pressure, and bromine and hydrobromic acid concentrations were taken into consideration. Also included are the effects of the Nafion membrance separator and the various bromide complex species. Activity coefficients were taken into account in one of the expressions. The sensitivity of these parameters on the calculated OCV was studied.

  18. Flow of Red Blood Cells in Stenosed Microvessels

    PubMed Central

    Vahidkhah, Koohyar; Balogh, Peter; Bagchi, Prosenjit

    2016-01-01

    A computational study is presented on the flow of deformable red blood cells in stenosed microvessels. It is observed that the Fahraeus-Lindqvist effect is significantly enhanced due to the presence of a stenosis. The apparent viscosity of blood is observed to increase by several folds when compared to non-stenosed vessels. An asymmetric distribution of the red blood cells, caused by geometric focusing in stenosed vessels, is observed to play a major role in the enhancement. The asymmetry in cell distribution also results in an asymmetry in average velocity and wall shear stress along the length of the stenosis. The discrete motion of the cells causes large time-dependent fluctuations in flow properties. The root-mean-square of flow rate fluctuations could be an order of magnitude higher than that in non-stenosed vessels. Several folds increase in Eulerian velocity fluctuation is also observed in the vicinity of the stenosis. Surprisingly, a transient flow reversal is observed upstream a stenosis but not downstream. The asymmetry and fluctuations in flow quantities and the flow reversal would not occur in absence of the cells. It is concluded that the flow physics and its physiological consequences are significantly different in micro- versus macrovascular stenosis. PMID:27319318

  19. Flow of Red Blood Cells in Stenosed Microvessels

    NASA Astrophysics Data System (ADS)

    Vahidkhah, Koohyar; Balogh, Peter; Bagchi, Prosenjit

    2016-06-01

    A computational study is presented on the flow of deformable red blood cells in stenosed microvessels. It is observed that the Fahraeus-Lindqvist effect is significantly enhanced due to the presence of a stenosis. The apparent viscosity of blood is observed to increase by several folds when compared to non-stenosed vessels. An asymmetric distribution of the red blood cells, caused by geometric focusing in stenosed vessels, is observed to play a major role in the enhancement. The asymmetry in cell distribution also results in an asymmetry in average velocity and wall shear stress along the length of the stenosis. The discrete motion of the cells causes large time-dependent fluctuations in flow properties. The root-mean-square of flow rate fluctuations could be an order of magnitude higher than that in non-stenosed vessels. Several folds increase in Eulerian velocity fluctuation is also observed in the vicinity of the stenosis. Surprisingly, a transient flow reversal is observed upstream a stenosis but not downstream. The asymmetry and fluctuations in flow quantities and the flow reversal would not occur in absence of the cells. It is concluded that the flow physics and its physiological consequences are significantly different in micro- versus macrovascular stenosis.

  20. Flow distribution in the manifold of PEM fuel cell stack

    NASA Astrophysics Data System (ADS)

    Chen, Chung-Hsien; Jung, Shiauh-Ping; Yen, Shi-Chern

    In this study, the pressure variation and the flow distribution in the manifold of a fuel-cell stack are simulated by a computational fluid dynamics (CFD) approach. Two dimensional stack model composed of 72 cells filled with porous media is constructed to evaluate pressure drop caused by channel flow resistance. In order to simplify this model, electrochemical reactions, heat and mass transport phenomena are ignored and air is treated as working fluid to investigate flow distribution in stacks. Design parameters such as the permeability of the porous media, the manifold width and the air feeding rate were changed to estimate uniformity of the flow distribution in the manifold. A momentum-balance theory and a pressure-drop model are presented to explain the physical mechanism of flow distribution. Modeling results indicate that both the channel resistance and the manifold width can enhance the uniformity of the flow distribution. In addition, a lower air feeding rate can also enhance the uniformity of flow distribution. However, excessive pressure drop is not beneficial for realistic applications of a fuel-cell stack and hence enhanced manifold width is a better solution for flow distribution.

  1. Debris flow study in Malaysia

    NASA Astrophysics Data System (ADS)

    Bahrin Jaafar, Kamal

    2016-04-01

    The phenomenon of debris flow occurs in Malaysia occasionally. The topography of Peningsular Malysia is characterized by the central mountain ranges running from south to north. Several parts of hilly areas with steep slopes, combined with high saturation of soil strata that deliberately increase the pore water pressure underneath the hill slope. As a tropical country Malaysia has very high intensity rainfall which is triggered the landslide. In the study area where the debris flow are bound to occur, there are a few factors that contribute to this phenomenon such as high rainfall intensity, very steep slope which an inclination more than 35 degree and sandy clay soil type which is easily change to liquidity soil. This paper will discuss the study of rainfall, mechanism, modeling and design of mitigation measure to avoid repeated failure in future in same area.

  2. Swan falls instream flow study

    SciTech Connect

    Anglin, D.R.; Cummings, T.R.; Ecklund, A.E.

    1992-10-01

    The purpose of the Swan Falls Instream Flow Study was to define the relationship between streamflows and instream habitat for resident fish species and to assess the relative impact of several different hydrographs on resident fish habitat. Specific objectives included the following: (1) Conduct a literature search to compile life history, distribution, and habitat requirements for species of interest. Physical and hydrologic characteristics of the Snake River were also compiled. (2) Determine physical habitat versus discharge relationships and conduct habitat time series analysis for each species/lifestage using the Instream Flow Incremental Methodology (IFIM) developed by the U.S. Fish and Wildlife Service. (3) Examine the impacts on resident fish habitat of proposed hydrographs, including Swan Falls Agreement flows, relative to current conditions. (4) Characterize water quality conditions, including water temperature and dissolved oxygen, in the vicinity of the study area and determine the implications of those conditions for the resident species of interest. (5) Determine streamflows necessary to protect and maintain resident fish habitat in the study area.

  3. Segmentation of touching cell nuclei using gradient flow tracking.

    PubMed

    Li, G; Liu, T; Nie, J; Guo, L; Chen, J; Zhu, J; Xia, W; Mara, A; Holley, S; Wong, S T C

    2008-07-01

    Reliable cell nuclei segmentation is an important yet unresolved problem in biological imaging studies. This paper presents a novel computerized method for robust cell nuclei segmentation based on gradient flow tracking. This method is composed of three key steps: (1) generate a diffused gradient vector flow field; (2) perform a gradient flow tracking procedure to attract points to the basin of a sink; and (3) separate the image into small regions, each containing one nucleus and nearby peripheral background, and perform local adaptive thresholding in each small region to extract the cell nucleus from the background. To show the generality of the proposed method, we report the validation and experimental results using microscopic image data sets from three research labs, with both over-segmentation and under-segmentation rates below 3%. In particular, this method is able to segment closely juxtaposed or clustered cell nuclei, with high sensitivity and specificity in different situations.

  4. Artificial Hair Cells for Sensing Flows

    NASA Technical Reports Server (NTRS)

    Chen, Jack

    2007-01-01

    The purpose of this article is to present additional information about the flow-velocity sensors described briefly in the immediately preceding article. As noted therein, these sensors can be characterized as artificial hair cells that implement an approximation of the sensory principle of flow-sensing cilia of fish: A cilium is bent by an amount proportional to the flow to which it is exposed. A nerve cell at the base of the cilium senses the flow by sensing the bending of the cilium. In an artificial hair cell, the artificial cilium is a microscopic cantilever beam, and the bending of an artificial cilium is measured by means of a strain gauge at its base (see Figure 1). Figure 2 presents cross sections of a representative sensor of this type at two different stages of its fabrication process. The process consists of relatively- low-temperature metallization, polymer-deposition, microfabrication, and surface-micromachining subprocesses, including plastic-deformation magnetic assembly (PDMA), which is described below. These subprocesses are suitable for a variety of substrate materials, including silicon, some glasses, and some polymers. Moreover, because it incorporates a polymeric supporting structure, this sensor is more robust, relative to its silicon-based counterparts.

  5. High speed flow cytometric separation of viable cells

    DOEpatents

    Sasaki, D.T.; Van den Engh, G.J.; Buckie, A.M.

    1995-11-14

    Hematopoietic cell populations are separated to provide cell sets and subsets as viable cells with high purity and high yields, based on the number of original cells present in the mixture. High-speed flow cytometry is employed using light characteristics of the cells to separate the cells, where high flow speeds are used to reduce the sorting time.

  6. High speed flow cytometric separation of viable cells

    DOEpatents

    Sasaki, Dennis T.; Van den Engh, Gerrit J.; Buckie, Anne-Marie

    1995-01-01

    Hematopoietic cell populations are separated to provide cell sets and subsets as viable cells with high purity and high yields, based on the number of original cells present in the mixture. High-speed flow cytometry is employed using light characteristics of the cells to separate the cells, where high flow speeds are used to reduce the sorting time.

  7. Cell stretching in extensional flows for assaying cell mechanics

    NASA Astrophysics Data System (ADS)

    Gossett, Daniel; Tse, Henry; Adeyiga, Oladunni; Yang, Otto; Rao, Jianyu; di Carlo, Dino

    2013-03-01

    There is growing evidence that cell deformability is a useful indicator of cell state and may be a label-free biomarker of metastatic potential, degree of differentiation, and leukocyte activation. In order for deformability measurements to be clinically valuable given the heterogeneity of biological samples, there exists a need for a high-throughput assay of this biophysical property. We developed a robust method for obtaining high-throughput (>1,000 cells/sec) single-cell mechanical measurements which employs coupled hydrodynamic lift forces and curvature-induced secondary flows to uniformly position cells in flow, extensional flow stretching, high-speed imaging, and automated image analysis to extract diameter and deformability parameters. Using this method we have assayed numerous in vitro models of cellular transformations and clinical fluids where malignant cells manifest. We found transformations associated with increased motility or invasiveness increased deformability and the presence of large and deformable cells within clinical pleural fluids correlated well with cytological diagnoses of malignancy. This agrees with the hypothesis that cancerous cells are deformable by necessity-to be able to transverse tight endothelial gaps and invade tissues.

  8. Determination of natural killer cell function by flow cytometry.

    PubMed Central

    Kane, K L; Ashton, F A; Schmitz, J L; Folds, J D

    1996-01-01

    Natural killer cells (NK cells) are a subset of peripheral blood lymphocytes that mediate non-major histocompatibility complex-restricted cytotoxicity of foreign target cells. The "gold standard" assay for NK cell activity has been the chromium release assay. This method is not easily performed in the clinical laboratory because of difficulties with disposal of radioactive and hazardous materials, short reagent half-lives, expense, and difficulties with assay standardization. We describe a flow cytometric assay for the clinical measurement of NK cell activity. This study compared the chromium release assay and the flow cytometric assay by using clinically relevant specimens. There were no significant differences between the two assays in the measurement of lytic activity for 17 peripheral blood specimens or in reproducibility in repeated samplings of healthy individuals. We also established a normal range of values for NK activity in healthy adults and identified a small cluster of individuals who have exceptionally high or low levels of NK activity. The flow cytometric assay was validated by testing specimens from subjects expected to have abnormally low levels of NK activity (pregnant women) and specimens from healthy individuals in whom the activity of NK cells was enhanced by exposure to interleukin-2 or alpha interferon. Treatment with these agents was associated with a significant increase in NK activity. These results confirm and extend those of others, showing that the flow cytometric assay is a viable alternative to the chromium release assay for measuring NK cell activity. PMID:8705672

  9. Full cell study of Diels Alder poly(phenylene) anion and cation exchange membranes in vanadium redox flow batteries

    DOE PAGES

    Pezeshki, Alan M.; Fujimoto, Cy; Sun, Che -Nan; Mench, Matthew M.; Zawodzinski, Thomas A.; Tang, Z. J.

    2015-11-14

    In this paper, we report on the performance of Diels Alder poly(phenylene) membranes in vanadium redox flow batteries. The membranes were functionalized with quaternary ammonium groups to form an anion exchange membrane (QDAPP) and with sulfonic acid groups to form a cation exchange membrane (SDAPP). Both membrane classes showed similar conductivities in the battery environment, suggesting that the ion conduction mechanism in the material is not strongly affected by the moieties along the polymer backbone. The resistance to vanadium permeation in QDAPP was not improved relative to SDAPP, further suggesting that the polarity of the functional groups do not playmore » a significant role in the membrane materials tested. Both QDAPP and SDAPP outperformed Nafion membranes in cycling tests, with both achieving voltage efficiencies above 85% while maintaining 95% coulombic efficiency while at a current density of 200 mA/cm2.« less

  10. Full cell study of Diels Alder poly(phenylene) anion and cation exchange membranes in vanadium redox flow batteries

    SciTech Connect

    Pezeshki, Alan M.; Fujimoto, Cy; Sun, Che -Nan; Mench, Matthew M.; Zawodzinski, Thomas A.; Tang, Z. J.

    2015-11-14

    In this paper, we report on the performance of Diels Alder poly(phenylene) membranes in vanadium redox flow batteries. The membranes were functionalized with quaternary ammonium groups to form an anion exchange membrane (QDAPP) and with sulfonic acid groups to form a cation exchange membrane (SDAPP). Both membrane classes showed similar conductivities in the battery environment, suggesting that the ion conduction mechanism in the material is not strongly affected by the moieties along the polymer backbone. The resistance to vanadium permeation in QDAPP was not improved relative to SDAPP, further suggesting that the polarity of the functional groups do not play a significant role in the membrane materials tested. Both QDAPP and SDAPP outperformed Nafion membranes in cycling tests, with both achieving voltage efficiencies above 85% while maintaining 95% coulombic efficiency while at a current density of 200 mA/cm2.

  11. Studies of planetary scale waves and instabilities in support of the geophysical fluid flow cell experiment on USML-2

    NASA Technical Reports Server (NTRS)

    Hart, J. E.

    1995-01-01

    High resolution numerical simulations of thermal convection in a rapidly rotating channel with gravity perpendicular to the rotation vector are described. The convecting columns are subject to a beta-effect resulting from cross-channel topographic vortex stretching. The symmetries of the problem allow many invariant wavenumber sets, and this property is associated with the existence of stable multiple-equilibria at modest supercriticality. The transition to chaotic behavior involves the production of intermittent unstable orbits off a two-torus in energy space. At very high Rayleigh number (of order 10(exp 6) to 10(exp 7)) the motion can be turbulent, depending on the size of beta. However, the turbulence is usually characterized by an almost-periodic formation of patches of small scale convection that cause regular pulsations in the accompanying strong zonal jets. The processes maintaining these flows may be related to those responsible for the zonal currents on Jupiter and for cyclic variability on the Sun.

  12. Deterministic sequential isolation of floating cancer cells under continuous flow.

    PubMed

    Tran, Quang D; Kong, Tian Fook; Hu, Dinglong; Marcos; Lam, Raymond H W

    2016-08-01

    Isolation of rare cells, such as circulating tumor cells, has been challenging because of their low abundance and limited timeframes of expressions of relevant cell characteristics. In this work, we devise a novel hydrodynamic mechanism to sequentially trap and isolate floating cells in biosamples. We develop a microfluidic device for the sequential isolation of floating cancer cells through a series of microsieves to obtain up to 100% trapping yield and >95% sequential isolation efficiency. We optimize the trappers' dimensions and locations through both computational and experimental analyses using microbeads and cells. Furthermore, we investigated the functional range of flow rates for effective sequential cell isolation by taking the cell deformability into account. We verify the cell isolation ability using the human breast cancer cell line MDA-MB-231 with perfect agreement with the microbead results. The viability of the isolated cells can be maintained for direct identification of any cell characteristics within the device. We further demonstrate that this device can be applied to isolate the largest particles from a sample containing multiple sizes of particles, revealing its possible applicability in isolation of circulating tumor cells in cancer patients' blood. Our study provides a promising sequential cell isolation strategy with high potential for rapid detection and analysis of general floating cells, including circulating tumor cells and other rare cell types. PMID:27387093

  13. Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study.

    PubMed

    Mandruzzato, Susanna; Brandau, Sven; Britten, Cedrik M; Bronte, Vincenzo; Damuzzo, Vera; Gouttefangeas, Cécile; Maurer, Dominik; Ottensmeier, Christian; van der Burg, Sjoerd H; Welters, Marij J P; Walter, Steffen

    2016-02-01

    There is an increasing interest for monitoring circulating myeloid-derived suppressor cells (MDSCs) in cancer patients, but there are also divergences in their phenotypic definition. To overcome this obstacle, the Cancer Immunoguiding Program under the umbrella of the Association of Cancer Immunotherapy is coordinating a proficiency panel program that aims at harmonizing MDSC phenotyping. After a consultation period, a two-stage approach was designed to harmonize MDSC phenotype. In the first step, an international consortium of 23 laboratories immunophenotyped 10 putative MDSC subsets on pretested, peripheral blood mononuclear cells of healthy donors to assess the level of concordance and define robust marker combinations for the identification of circulating MDSCs. At this stage, no mandatory requirements to standardize reagents or protocols were introduced. Data analysis revealed a small intra-laboratory, but very high inter-laboratory variance for all MDSC subsets, especially for the granulocytic subsets. In particular, the use of a dead-cell marker altered significantly the reported percentage of granulocytic MDSCs, confirming that these cells are especially sensitive to cryopreservation and/or thawing. Importantly, the gating strategy was heterogeneous and associated with high inter-center variance. Overall, our results document the high variability in MDSC phenotyping in the multicenter setting if no harmonization/standardization measures are applied. Although the observed variability depended on a number of identified parameters, the main parameter associated with variation was the gating strategy. Based on these findings, we propose further efforts to harmonize marker combinations and gating parameters to identify strategies for a robust enumeration of MDSC subsets.

  14. Deterministic Aperiodic Sickle Cell Blood Flows

    NASA Astrophysics Data System (ADS)

    Atsaves, Louis; Harris, Wesley

    2013-11-01

    In this paper sickle cell blood flow in the capillaries is modeled as a hydrodynamical system. The hydrodynamical system consists of the axisymmetric unsteady, incompressible Navier-Stokes equations and a set of constitutive equations for oxygen transport. Blood cell deformation is not considered in this paper. The hydrodynamical system is reduced to a system of non-linear partial differential equations that are then transformed into a system of three autonomous non-linear ordinary differential equations and a set of algebraic equations. We examine the hydrodynamical system to discern stable/unstable, periodic/nonperiodic, reversible/irreversible properties of the system. The properties of the solutions are driven in large part by the coefficients of the governing system of equations. These coefficients depend on the physiological properties of the sickle cell blood. The chaotic nature of the onset of crisis in sickle cell patients is identified. Research Assistant.

  15. Blood Flow through an Open-Celled Foam

    NASA Astrophysics Data System (ADS)

    Ortega, Jason; Maitland, Duncan

    2011-11-01

    The Hazen-Dupuit-Darcy (HDD) equation is commonly used in engineering applications to model the pressure gradient of flow through a porous media. One major advantage of this equation is that it simplifies the complex geometric details of the porous media into two coefficients: the permeability, K, and form factor, C. However through this simplification, the flow details within the porous media are no longer accessible, making it difficult to study the phenomena that contribute to changes in K and C due to clotting of blood flow. To obtain a more detailed understanding of blood flow through a porous media, a direct assessment of the complex interstitial geometry and flow is required. In this study, we solve the Navier-Stokes equations for Newtonian and non-Newtonian blood flow through an open-celled foam geometry obtained from a micro-CT scan. The nominal strut size of the foam sample is of O(10e-5) m and the corresponding Reynolds number based upon this length ranges up to O(10). Fitting the pressure gradient vs. Darcy velocity data with the HDD equation demonstrates that both viscous and inertial forces play an important role in the flow through the foam at these Reynolds numbers. Recirculation zones are observed to form in the wake of the pore struts, producing regions of flow characterized by both low shear rates and long fluid residence times, factors of which have been shown in previous studies to promote blood clotting.

  16. A dynamic plug flow reactor model for a vanadium redox flow battery cell

    NASA Astrophysics Data System (ADS)

    Li, Yifeng; Skyllas-Kazacos, Maria; Bao, Jie

    2016-04-01

    A dynamic plug flow reactor model for a single cell VRB system is developed based on material balance, and the Nernst equation is employed to calculate cell voltage with consideration of activation and concentration overpotentials. Simulation studies were conducted under various conditions to investigate the effects of several key operation variables including electrolyte flow rate, upper SOC limit and input current magnitude on the cell charging performance. The results show that all three variables have a great impact on performance, particularly on the possibility of gassing during charging at high SOCs or inadequate flow rates. Simulations were also carried out to study the effects of electrolyte imbalance during long term charging and discharging cycling. The results show the minimum electrolyte flow rate needed for operation within a particular SOC range in order to avoid gassing side reactions during charging. The model also allows scheduling of partial electrolyte remixing operations to restore capacity and also avoid possible gassing side reactions during charging. Simulation results also suggest the proper placement for cell voltage monitoring and highlight potential problems associated with setting the upper charging cut-off limit based on the inlet SOC calculated from the open-circuit cell voltage measurement.

  17. Porcine skin flow-through diffusion cell system.

    PubMed

    Baynes, R E

    2001-11-01

    Porcine Skin Flow-Through Diffusion Cell System (Ronald E. Baynes, North Carolina State University, Raleigh, North Carolina). Porcine skin can be used in a diffusion cell apparatus to study the rate and extent of absorption of topically applied chemicals through the skin. Although the skin of a number of animals can be used in this system, that of the pig most closely approximates human skin anatomically and physiologically.

  18. Red blood cell in simple shear flow

    NASA Astrophysics Data System (ADS)

    Chien, Wei; Hew, Yayu; Chen, Yeng-Long

    2013-03-01

    The dynamics of red blood cells (RBC) in blood flow is critical for oxygen transport, and it also influences inflammation (white blood cells), thrombosis (platelets), and circulatory tumor migration. The physical properties of a RBC can be captured by modeling RBC as lipid membrane linked to a cytoskeletal spectrin network that encapsulates cytoplasm rich in hemoglobin, with bi-concave equilibrium shape. Depending on the shear force, RBC elasticity, membrane viscosity, and cytoplasm viscosity, RBC can undergo tumbling, tank-treading, or oscillatory motion. We investigate the dynamic state diagram of RBC in shear and pressure-driven flow using a combined immersed boundary-lattice Boltzmann method with a multi-scale RBC model that accurately captures the experimentally established RBC force-deformation relation. It is found that the tumbling (TU) to tank-treading (TT) transition occurs as shear rate increases for cytoplasm/outer fluid viscosity ratio smaller than 0.67. The TU frequency is found to be half of the TT frequency, in agreement with experiment observations. Larger viscosity ratios lead to the disappearance of stable TT phase and unstable complex dynamics, including the oscillation of the symmetry axis of the bi-concave shape perpendicular to the flow direction. The dependence on RBC bending rigidity, shear modulus, the order of membrane spectrin network and fluid field in the unstable region will also be discussed.

  19. On-line preferential solvation studies of polymers by coupled chromatographic-Fourier transform infrared spectroscopic flow-cell technique.

    PubMed

    Malanin, M; Eichhorn, K-J; Lederer, A; Treppe, P; Adam, G; Fischer, D; Voigt, D

    2009-12-18

    Qualitative and quantitative comparison between liquid chromatography (LC) and LC coupled with Fourier transform infrared spectroscopy (LC-FTIR) to evaluate preferential solvation phenomenon of polymers in a mixed solvent has been performed. These studies show that LC-FTIR technique leads to detailed structural information without the requirement for determination of additional parameters for quantitative analysis except calibration. Appropriate experimental conditions for preferential solvation study have been established by variation of polymer concentration, molar mass and eluent content.

  20. The Geophysical Fluid Flow Cell Experiment

    NASA Technical Reports Server (NTRS)

    Hart, J. E.; Ohlsen, D.; Kittleman, S.; Borhani, N.; Leslie, F.; Miller, T.

    1999-01-01

    The Geophysical Fluid Flow Cell (GFFC) experiment performed visualizations of thermal convection in a rotating differentially heated spherical shell of fluid. In these experiments dielectric polarization forces are used to generate a radially directed buoyancy force. This enables the laboratory simulation of a number of geophysically and astrophysically important situations in which sphericity and rotation both impose strong constraints on global scale fluid motions. During USML-2 a large set of experiments with spherically symmetric heating were carried out. These enabled the determination of critical points for the transition to various forms of nonaxisymmetric convection and, for highly turbulent flows, the transition latitudes separating the different modes of motion. This paper presents a first analysis of these experiments as well as data on the general performance of the instrument during the USML-2 flight.

  1. Estimating the efficiency of cell capture and arrest in flow chambers: study of neutrophil binding via E-selectin and ICAM-1.

    PubMed Central

    Zhang, Yi; Neelamegham, Sriram

    2002-01-01

    A mathematical model was developed to quantify the efficiency of cell-substrate attachment in the parallel-plate flow chamber. The model decouples the physical features of the system that affect cell-substrate collision rates from the biological features that influence cellular adhesivity. Thus, experimental data on cell rolling and adhesion density are converted into "frequency" parameters that quantify the "efficiency" with which cells in the flow chamber progress from the free stream to rolling, and transition from rolling to firm arrest. The model was partially validated by comparing simulation results with experiments where neutrophils rolled and adhered onto substrates composed of cotransfected cells bearing E-selectin and intercellular adhesion molecule-1 (ICAM-1). Results suggest that: 1) Neutrophils contact the E-selectin substrate on average for 4-8.5s before tethering. This contact duration is insensitive to applied shear stress. 2) At 2 dyn/cm(2), approximately 28% of the collisions between the cells and substrate result in primary capture. Also, approximately 5-7% of collisions between neutrophils in the free stream and previously recruited neutrophils bound on the substrate result in secondary capture. These percentages were higher at lower shears. 3) An adherent cell may influence the flow streams in its vicinity up to a distance of 2.5 cell diameters away. 4) Our estimates of selectin on-rate in cellular systems compare favorably with data from reconstituted systems with immobilized soluble E-selectin. In magnitude, the observed on-rates occur in the order, L-selectin > P-selectin > E-selectin. PMID:12324413

  2. Predicting dynamics and rheology of blood flow: A comparative study of multiscale and low-dimensional models of red blood cells

    SciTech Connect

    Pan, Wenxiao; Fedosov, Dmitry A.; Caswell, Bruce; Karniadakis, George E.

    2011-05-27

    In this work we compare the predictive capability of two mathematical models for red blood cells (RBCs) focusing on blood flow in capillaries and arterioles. Both RBC models as well as their corresponding blood flows are based on the dissipative particle dynamics (DPD) method, a coarse-grained molecular dynamics approach. The first model employs a multiscale description of the RBC (MS-RBC), with its membrane represented by hundreds or even thousands of DPD-particles connected by springs into a triangular network in combination with out-of-plane elastic bending resistance. Extra dissipation within the network accounts for membrane viscosity, while the characteristic biconcave RBC shape is achieved by imposition of constraints for constant membrane area and constant cell volume. The second model is based on a low-dimensional description (LD-RBC) constructed as a closed torus-like ring of only 10 large DPD colloidal particles. They are connected into a ring by worm-like chain (WLC) springs combined with bending resistance. The LD-RBC model can be fitted to represent the entire range of nonlinear elastic deformations as measured by optical-tweezers for healthy and for infected RBCs in malaria. MS-RBCs suspensions model the dynamics and rheology of blood flow accurately for any size vessel but this approach is computationally expensive above 100 microns. Surprisingly, the much more economical suspensions of LD-RBCs also capture the blood flow dynamics and rheology accurately except for vessels with sizes comparable to RBC diameter. In particular, the LD-RBC suspensions are shown to properly capture the experimental data for the apparent viscosity of blood and its cell-free layer (CFL) in tube flow. Taken together, these findings suggest a hierarchical approach in modeling blood flow in the arterial tree, whereby the MS-RBC model should be employed for capillaries and arterioles below 100 microns, the LD-RBC model for arterioles, and the continuum description for

  3. Predicting dynamics and rheology of blood flow: A comparative study of multiscale and low-dimensional models of red blood cells

    PubMed Central

    Pan, Wenxiao; Fedosov, Dmitry A.; Caswell, Bruce; Karniadakis, George Em

    2011-01-01

    We compare the predictive capability of two mathematical models for red blood cells (RBCs) focusing on blood flow in capillaries and arterioles. Both RBC models as well as their corresponding blood flows are based on the dissipative particle dynamics (DPD) method, a coarse-grained molecular dynamics approach. The first model employs a multiscale description of the RBC (MS-RBC), with its membrane represented by hundreds or even thousands of DPD-particles connected by springs into a triangular network in combination with out-of-plane elastic bending resistance. Extra dissipation within the network accounts for membrane viscosity, while the characteristic biconcave RBC shape is achieved by imposition of constraints for constant membrane area and constant cell volume. The second model is based on a low-dimensional description (LD-RBC) constructed as a closed torus-like ring of only 10 large DPD colloidal particles. They are connected into a ring by worm-like chain (WLC) springs combined with bending resistance. The LD-RBC model can be fitted to represent the entire range of nonlinear elastic deformations as measured by optical-tweezers for healthy and for infected RBCs in malaria. MS-RBCs suspensions model the dynamics and rheology of blood flow accurately for any vessel size but this approach is computationally expensive for vessel diameters above 100 microns. Surprisingly, the much more economical suspensions of LD-RBCs also capture the blood flow dynamics and rheology accurately except for small-size vessels comparable to RBC diameter. In particular, the LD-RBC suspensions are shown to properly capture the experimental data for the apparent viscosity of blood and its cell-free layer (CFL) in tube flow. Taken together, these findings suggest a hierarchical approach in modeling blood flow in the arterial tree, whereby the MS-RBC model should be employed for capillaries and arterioles below 100 microns, the LD-RBC model for arterioles, and the continuum description for

  4. Parallel-plate fluid flow systems for bone cell stimulation.

    PubMed

    Huesa, Carmen; Helfrich, Miep H; Aspden, Richard M

    2010-04-19

    Bone responds to changes in its mechanical environment, but the mechanisms by which it does so are poorly understood. One hypothesis of mechanosensing in bone states that osteocytes can sense the flow of fluid through the canalicular system. To study this in vitro a number of fluid flow devices have been designed in which cells are placed between parallel plates in sealed chambers. Fluid flows through the chambers at controlled rates, most commonly driven by a peristaltic pump. In addition to fluid flow, high pressures have been observed in these chambers, but the effect of this on the cellular responses has generally been ignored or considered irrelevant, something challenged by recent cellular experiments using pressure only. We have, therefore, devised a system in which we can considerably reduce the pressure while maintaining the flow rate to enable study of their effects individually and in combination. As reducing pressure also reduces the risk of leaks in flow chambers, our system is suitable for real-time microscopical experiments. We present details of the new systems and of experiments with osteoblasts to illustrate the effects of fluid flow with and without additional pressure on the translocation of beta-catenin to the nucleus.

  5. Schlieren System For Flow Studies In Round Glass Pipes

    NASA Technical Reports Server (NTRS)

    Costen, Robert C.; Rhodes, David B.; Jones, Stephen B.

    1990-01-01

    In schlieren system for studying flow of gas in transparent pipe of circular cross section, cylindrical lenses placed on opposite sides of pipe compensate for refraction caused by wall of pipe. Enables direct visualization of such phenomena as laminar or turbulent flow, shock waves, vortexes, and flow separations in systems having inherently cylindrical geometry; potentially unreliable extrapolations from results in flat-sided test cells no longer necessary.

  6. Feasibility study of laminar flow bodies in fully turbulent flow

    SciTech Connect

    Sarkar, T.; Sayer, P.G.; Fraser, S.M.

    1994-12-31

    One of the most important design requirements of long range autonomous underwater vehicles (AUVs) is to minimize propulsive power. An important and relatively easy way of achieving this is by careful selection of hull shape. Two main schools of thought in this respect are: if laminar flow can be maintained for a long length of the body, the effective drag can be reduced; it is not possible to maintain laminar flow for a significant length of the body and hull design should be based on turbulent flow conditions. In this paper, a feasibility study of laminar flow designs is undertaken under the assumption that flow will be turbulent over the entire length. For comparison two laminar flow designs X-35 and F-57 are selected and results are compared with those of two typical torpedo shaped bodies, namely AFTERBODY1 and AFTERBODY2 of DTNSRDC. It has been shown that laminar flow bodies have 10--15% higher drag when flow is turbulent over the entire length. Hence there is some hydrodynamic risk involved in adopting such laminar bodies without further consideration.

  7. Flow Cytometric Analysis of Immune Cells Within Murine Aorta.

    PubMed

    Gjurich, Breanne N; Taghavie-Moghadam, Parésa L; Galkina, Elena V

    2015-01-01

    The immune system plays a critical role in the modulation of atherogenesis at all stages of the disease. However, there are many technical difficulties when studying the immune system within murine aortas. Common techniques such as PCR and immunohistochemistry have answered many questions about the presence of immune cells and mediators of inflammation within the aorta yet many questions remain unanswered due to the limitations of these techniques. On the other hand, cumulatively the flow cytometry approach has propelled the immunology field forward but it has been challenging to apply this technique to aortic tissues. Here, we describe the methodology to isolate and characterize the immune cells within the murine aorta and provide examples of functional assays for aortic leukocytes using flow cytometry. The method involves the harvesting and enzymatic digestion of the aorta, extracellular and intracellular protein staining, and a subsequent flow cytometric analysis. PMID:26445788

  8. Multiscale modeling of mechanosensing channels on vesicles and cell membranes in 3D constricted flows and shear flows

    NASA Astrophysics Data System (ADS)

    Peng, Zhangli; Pak, On Shun; Young, Yuan-Nan; Liu, Allen; Stone, Howard

    2015-11-01

    We investigate the gating of mechanosensing channels (Mscls) on vesicles and cell membranes under different flow conditions using a multiscale approach. At the cell level (microns), the membrane tension is calculated using a 3D two-component whole-cell membrane model based on dissipative particle dynamics (DPD), including the cortex cytoskeleton and its interactions with the lipid bilayer. At the Mscl level (nanometers), we predict the relation between channel gating and the membrane tension obtained from a cell-level model using a semi-analytical model based on the bilayer hydrophobic mismatch energy. We systematically study the gating of Mscls of vesicles and cell membranes in constricted channel flows and shear flows, and explore the dependence of the gating on flow rate, cell shape and size. The results provide guidance for future experiments in inducing Mscl opening for various purposes such as drug delivery.

  9. Spatial response variations within biosensor flow cells

    NASA Astrophysics Data System (ADS)

    Cant, Nicola; Harrison, Sarah

    2012-02-01

    Biosensors are currently being developed for the detection of a wide range of analytes in a variety of scenarios. One such area is that of environmental monitoring for the presence of biological threats, from toxins through to viruses and bacteria. The varying nature, and in particular disparate size, of such a variety of analytes poses a significant challenge in the development of effective high confidence instruments. Many existing biosensors employ functionalised flow cells in which spatially defined arrays of surface immobilised recognition elements are present to specifically capture their analyte of interest. Experimental data obtained using a grating coupled SPR instrument, the BIAcore Flexchip, has revealed spatial dependency differences in response behaviours between proteinaceous and particulate analytes. In particular, the magnitude of responses seen with Bacillus anthracis spores across the instruments flow cell appear to be influenced by shear and gravitational effects whilst those from soluble proteins are more uniform. We have explored this dependence to understand its fundamental impact on the successful implementation of multi-analyte environmental biological detection systems.

  10. Co-flow planar SOFC fuel cell stack

    DOEpatents

    Chung, Brandon W.; Pham, Ai Quoc; Glass, Robert S.

    2004-11-30

    A co-flow planar solid oxide fuel cell stack with an integral, internal manifold and a casing/holder to separately seal the cell. This construction improves sealing and gas flow, and provides for easy manifolding of cell stacks. In addition, the stack construction has the potential for an improved durability and operation with an additional increase in cell efficiency. The co-flow arrangement can be effectively utilized in other electrochemical systems requiring gas-proof separation of gases.

  11. Studies of two phase flow

    NASA Technical Reports Server (NTRS)

    Witte, Larry C.

    1994-01-01

    The development of instrumentation for the support of research in two-phase flow in simulated microgravity conditions was performed. The funds were expended in the development of a technique for characterizing the motion and size distribution of small liquid droplets dispersed in a flowing gas. Phenomena like this occur in both microgravity and normal earth gravity situations inside of conduits that are carrying liquid-vapor mixtures at high flow rates. Some effort to develop a conductance probe for the measurement of liquid film thickness was also expended.

  12. Morphological analysis of tumor cell/endothelial cell interactions under shear flow.

    PubMed

    Chotard-Ghodsnia, Roxana; Haddad, Oualid; Leyrat, Anne; Drochon, Agnès; Verdier, Claude; Duperray, Alain

    2007-01-01

    In the process of hematogenous cancer metastasis, tumor cells (TCs) must shed into the blood stream, survive in the blood circulation, migrate through the vascular endothelium (extravasation) and proliferate in the target organs. However, the precise mechanisms by which TCs penetrate the endothelial cell (EC) junctions remain one of the least understood aspects of TC extravasation. This question has generally been addressed under static conditions, despite the important role of flow induced mechanical stress on the circulating cell-endothelium interactions. Moreover, flow studies were generally focused on transient or firm adhesion steps of TC-EC interactions and did not consider TCs spreading or extravasation. In this paper, we used a parallel-plate flow chamber to investigate TC-EC interactions under flow conditions. An EC monolayer was cultured on the lower plate of the flow chamber to model the endothelial barrier. Circulating TCs were introduced into the flow channel under a well-defined flow field and TC cell shape changes on the EC monolayer were followed in vitro with live phase contrast and fluorescence microscopy. Two spreading patterns were observed: radial spreading which corresponds to TC extravasation, and axial spreading where TCs formed a mosaic TC-EC monolayer. By investigating the changes in area and minor/major aspect ratio, we have established a simple quantitative basis for comparing spreading modes under various shear stresses. Contrary to radial spreading, the extent of axial spreading was increased by shear stress.

  13. Studies on best dose of X-ray for Hep-2 cells by using FTIR, UV-vis absorption spectroscopy and flow cytometry

    NASA Astrophysics Data System (ADS)

    Liu, Renming; Tang, Weiyue; Kang, Yipu; Si, Minzhen

    2009-08-01

    We report here the use of Fourier transform infrared spectroscopy (FTIR), ultraviolet-visible (UV-vis) absorption spectroscopy, and flow cytometry (FCM) to analysis the best dose of X-ray for human laryngeal squamous cell carcinoma cell lines (Hep-2). Our analysis indicates specific FTIR and UV-vis spectral differences between X-irradiated and normal Hep-2 cells. In addition, striking spectral differences are seen in FTIR spectra in the ratios at 2925/2958 and 1654/1542 cm -1. These two ratios of the X-irradiated cells for 8 Gy dose group with value of 1.07 ± 0.025 and 1.184 ± 0.013, respectively, were more notable (mean ± S.D., n = 5, P < 0.05) compared with that of the cells for the controls. UV-vis absorption spectra analysis shows X-ray irradiation disturbed the metabolism of phenylalanine and tyrosine intracellular, maybe, which was caused by cell cycle arrest. Spectroscopy analysis suggests 8 Gy is a better dose of X-ray for lowering the canceration degree of Hep-2 cells. Moreover, FCM analysis shows the apoptosis of X-irradiated cells depended on the radiation dose to some extent, but it was not linear. The total apoptosis ratio with value of (20.793 ± 1.133)% ( P < 0.01, n = 5) for the 12 Gy dose group was the maximum, however, the maximum apoptosis ratio per Gray (total apoptosis ratio/radiation dose) was the cells of the 2 Gy dose group with value of (4.887 ± 0.211)% ( P < 0.05, n = 5). Our data suggest that Hep-2 cells are given 2 Gy radiation of X-ray once a time, 8 Gy per week (accumulatively), the effect for lowering the canceration degree and restraining the proliferation of Hep-2 cells will be better.

  14. SAXS/WAXS studies of flow-induced crystallization of poly(1-butene) in shear flow

    NASA Astrophysics Data System (ADS)

    Luo, Binbin; Burghardt, Wesley

    2013-03-01

    Flow-induced crystallization of poly(1-butene) was studied in shear flow. Flow was produced using a Linkam shear cell that has been modified to allow x-ray access for in situ studies of polymer structure using synchrotron x-ray scattering techniques. After loading in the the shear cell, samples were first heated well into the melt, and then cooled to a crystallization temperature selected such that negligible quiescent crystallization would occur on reasonable time scales. A short burst of shear flow was then applied at various rates, after which simultaneous wide- and small-angle x-ray scattering (SAXS and WAXS) data were collected to study the resulting accelerated crystallization kinetics, as well as the morphology of the resulting crystallites (e.g. degree of crystallite orientation). The impact of both deformation rate and total applied strain on the crystallization process were examined.

  15. A flow cytometric approach for studying alterations in the cytoplasmic concentration of calcium ions in immune cells following stimulation with thymic peptides.

    PubMed

    Papaioannou, Nikos E; Voutsas, Ioannis F; Samara, Pinelopi; Tsitsilonis, Ourania E

    2016-04-01

    [Ca(2+)]i alterations are vital in signaling pathways of cell activation. We tried to detect such changes, in intracellular signaling pathways downstream TLR4 in immune cells, following stimulation with prothymosin alpha (proTα) and its decapeptide proTα(100-109). Human leukocytes were activated with LPS, proTα or proTα(100-109), directly or after 24h stimulation, while neutrophils were directly challenged. Cells were loaded with Fluo-4 and cytoplasmic Ca(2+) alterations were recorded by flow cytometry. Direct challenge with 20 μg/mL LPS induced a measurable [Ca(2+)]i increase in macrophages and neutrophils. Monocytes and macrophages incubated for 24h with LPS, proTα or proTα(100-109) and challenged with LPS, displayed a robust response. Lymphocytes and iDCs exhibited no alterations. Conclusively, we assessed a flow cytometry-based method for monitoring Ca(2+) ion influx changes in immune cells. Their stimulation with proTα or proTα(100-109) generates an activating background, similar to LPS, allowing for the detection of [Ca(2+)]i alterations induced upon subsequent challenge.

  16. Predicting dynamics and rheology of blood flow: A comparative study of multiscale and low-dimensional models of red blood cells.

    PubMed

    Pan, Wenxiao; Fedosov, Dmitry A; Caswell, Bruce; Karniadakis, George Em

    2011-09-01

    We compare the predictive capability of two mathematical models for red blood cells (RBCs) focusing on blood flow in capillaries and arterioles. Both RBC models as well as their corresponding blood flows are based on the dissipative particle dynamics (DPD) method, a coarse-grained molecular dynamics approach. The first model employs a multiscale description of the RBC (MS-RBC), with its membrane represented by hundreds or even thousands of DPD-particles connected by springs into a triangular network in combination with out-of-plane elastic bending resistance. Extra dissipation within the network accounts for membrane viscosity, while the characteristic biconcave RBC shape is achieved by imposition of constraints for constant membrane area and constant cell volume. The second model is based on a low-dimensional description (LD-RBC) constructed as a closed torus-like ring of only 10 large DPD colloidal particles. They are connected into a ring by worm-like chain (WLC) springs combined with bending resistance. The LD-RBC model can be fitted to represent the entire range of nonlinear elastic deformations as measured by optical-tweezers for healthy and for infected RBCs in malaria. MS-RBCs suspensions model the dynamics and rheology of blood flow accurately for any vessel size but this approach is computationally expensive for vessel diameters above 100μm. Surprisingly, the much more economical suspensions of LD-RBCs also capture the blood flow dynamics and rheology accurately except for small-size vessels comparable to RBC diameter. In particular, the LD-RBC suspensions are shown to properly capture the experimental data for the apparent viscosity of blood and its cell-free layer (CFL) in tube flow. Taken together, these findings suggest a hierarchical approach in modeling blood flow in the arterial tree, whereby the MS-RBC model should be employed for capillaries and arterioles below 100μm, the LD-RBC model for arterioles, and the continuum description for arteries.

  17. Hybrid laminar flow control study

    NASA Technical Reports Server (NTRS)

    1982-01-01

    Hybrid laminar flow control (HLFC) in which leading edge suction is used in conjunction with wing pressure distribution tailoring to postpone boundary layer transition and reduce friction drag was examined. Airfoil design characteristics required for laminar flow control (LFC) were determined. The aerodynamic design of the HLFC wing for a 178 passenger commercial turbofan transport was developed, and a drag was estimated. Systems changes required to install HLFC were defined, and weights and fuel economy were estimated. The potential for 9% fuel reduction for a 3926-km (2120-nmi) mission is identified.

  18. Alternative experiments using the geophysical fluid flow cell

    NASA Technical Reports Server (NTRS)

    Hart, J. E.

    1984-01-01

    This study addresses the possibility of doing large scale dynamics experiments using the Geophysical Fluid Flow Cell. In particular, cases where the forcing generates a statically stable stratification almost everywhere in the spherical shell are evaluated. This situation is typical of the Earth's atmosphere and oceans. By calculating the strongest meridional circulation expected in the spacelab experiments, and testing its stability using quasi-geostrophic stability theory, it is shown that strongly nonlinear baroclinic waves on a zonally symmetric modified thermal wind will not occur. The Geophysical Fluid Flow Cell does not have a deep enough fluid layer to permit useful studies of large scale planetary wave processes arising from instability. It is argued, however, that by introducing suitable meridional barriers, a significant contribution to the understanding of the oceanic thermocline problem could be made.

  19. Breakthrough Flow Battery Cell Stack: Transformative Electrochemical Flow Storage System (TEFSS)

    SciTech Connect

    2010-09-09

    GRIDS Project: UTRC is developing a flow battery with a unique design that provides significantly more power than today's flow battery systems. A flow battery is a cross between a traditional battery and a fuel cell. Flow batteries store their energy in external tanks instead of inside the cell itself. Flow batteries have traditionally been expensive because the battery cell stack, where the chemical reaction takes place, is costly. In this project, UTRC is developing a new stack design that achieves 10 times higher power than today’s flow batteries. This high power output means the size of the cell stack can be smaller, reducing the amount of expensive materials that are needed. UTRC’s flow battery will reduce the cost of storing electricity for the electric grid, making widespread use feasible.

  20. Ex-situ experimental studies on serpentine flow field design for redox flow battery systems

    NASA Astrophysics Data System (ADS)

    Jyothi Latha, T.; Jayanti, S.

    2014-02-01

    Electrolyte distribution using parallel flow field for redox flow battery (RFB) applications shows severe non-uniformity, while the conventional design of using the carbon felt itself as the flow distributor gives too high pressure drop. An optimized flow field design for uniform flow distribution at a minimal parasitic power loss is therefore needed for RFB systems. Since the materials and geometrical dimensions in RFBs are very different from those used in fuel cells, the hydrodynamics of the flow fields in RFBs is likely to be very different. In the present paper, we report on a fundamental study of the hydrodynamics of a serpentine flow field relevant to RFB applications. The permeability of the porous medium has been measured under different compression ratios and this is found to be in the range of 5-8 × 10-11 m2. The pressure drop in two serpentine flow fields of different geometric characteristics has been measured over a range of Reynolds numbers. Further analysis using computational fluid dynamics simulations brings out the importance of the compression of the porous medium as an additional parameter in determining the flow distribution and pressure drop in these flow fields.

  1. Counter-Flow Cooling Tower Test Cell

    NASA Astrophysics Data System (ADS)

    Dvořák, Lukáš; Nožička, Jiří

    2014-03-01

    The article contains a design of a functional experimental model of a cross-flow mechanical draft cooling tower and the results and outcomes of measurements. This device is primarily used for measuring performance characteristics of cooling fills, but with a simple rebuild, it can be used for measuring other thermodynamic processes that take part in so-called wet cooling. The main advantages of the particular test cell lie in the accuracy, size, and the possibility of changing the water distribution level. This feature is very useful for measurements of fills of different heights without the influence of the spray and rain zone. The functionality of this test cell has been verified experimentally during assembly, and data from the measurement of common film cooling fills have been compared against the results taken from another experimental line. For the purpose of evaluating the data gathered, computational scripts were created in the MATLAB numerical computing environment. The first script is for exact calculation of the thermal balance of the model, and the second is for determining Merkel's number via Chebyshev's method.

  2. Flow-cell fibre-optic enzyme sensor for phenols

    SciTech Connect

    Papkovsky, D.B.; Ghindilis, A.L.; Kurochkin, I.N. )

    1993-07-01

    A solid-state fibre-optic luminescent oxygen sensor was used for flow-through measurements. It acts as a transducer in a new flow-cell enzyme sensor arrangement. This arrangement comprises a flow path, sample injector, microcolumn with the immobilized enzyme, oxygen membrane and fibre-optic connector joined together to form an integral unit. Laccase enzyme was used as a recognition system which provided specific oxidation of the substrates with the dissolved oxygen being monitored. The assay procedure was optimized and performance of the new system studied. The sensor was applied to the determination polyphenol content in tea, brandy, etc. (quality control test). The sensitivity to some important phenolic compounds was tested with the view of industrial wastewater control applications. 5 refs., 6 figs., 1 tab.

  3. Comparative in vitro dissolution study of carbamazepine immediate-release products using the USP paddles method and the flow-through cell system

    PubMed Central

    Medina, José Raúl; Salazar, Dulce Karina; Hurtado, Marcela; Cortés, Alma Rosa; Domínguez-Ramírez, Adriana Miriam

    2013-01-01

    Dissolution profiles of four carbamazepine immediate-release generic products (200 mg tablets) and the reference product Tegretol® were evaluated using the USP paddles method and an alternative method with the flow-through cell system, USP Apparatus 4. Under official conditions all products met the Q specification, dissolution profiles of generic products were similar to the dissolution profile of the reference product (f2 > 50) and model-independent parameters showed non significant differences to the reference product except mean dissolution time for product A (p < 0.05). On the other hand, when the flow-through cell system was used, none of the products met the pharmacopeial specification at 15 min and product A did not reach dissolution criteria at 60 min, dissolution profiles of all generic products were not similar to the reference product profile (f2 < 50) and all model-independent parameters showed significant differences compared to the reference product (p < 0.05). Weibull’s model was more useful for adjusting the dissolution data of all products in both USP apparatuses and Td values showed significant differences compared to the reference product (p < 0.05) when USP Apparatus 4 was used. These results indicate that the proposed method, using the flow-through cell system, is more discriminative in evaluating both, rate and extent of carbamazepine dissolution process from immediate-release generic products. PMID:24648826

  4. Aqueous semi-solid flow cell: demonstration and analysis.

    PubMed

    Li, Zheng; Smith, Kyle C; Dong, Yajie; Baram, Nir; Fan, Frank Y; Xie, Jing; Limthongkul, Pimpa; Carter, W Craig; Chiang, Yet-Ming

    2013-10-14

    An aqueous Li-ion flow cell using suspension-based flow electrodes based on the LiTi2(PO4)3-LiFePO4 couple is demonstrated. Unlike conventional flow batteries, the semi-solid approach utilizes fluid electrodes that are electronically conductive. A model of simultaneous advection and electrochemical transport is developed and used to separate flow-induced losses from those due to underlying side reactions. The importance of plug flow to achieving high energy efficiency in flow batteries utilizing highly non-Newtonian flow electrodes is emphasized.

  5. Aqueous semi-solid flow cell: demonstration and analysis

    SciTech Connect

    Li, Z; Smith, KC; Dong, YJ; Baram, N; Fan, FY; Xie, J; Limthongkul, P; Carter, WC; Chiang, YM

    2013-01-01

    An aqueous Li-ion flow cell using suspension-based flow electrodes based on the LiTi2(PO4)(3)-LiFePO4 couple is demonstrated. Unlike conventional flow batteries, the semi-solid approach utilizes fluid electrodes that are electronically conductive. A model of simultaneous advection and electrochemical transport is developed and used to separate flow-induced losses from those due to underlying side reactions. The importance of plug flow to achieving high energy efficiency in flow batteries utilizing highly non-Newtonian flow electrodes is emphasized.

  6. Micropatterned surfaces for controlling cell adhesion and rolling under flow.

    PubMed

    Nalayanda, Divya D; Kalukanimuttam, Mahendran; Schmidtke, David W

    2007-04-01

    Cell adhesion and rolling on the vascular wall is critical to both inflammation and thrombosis. In this study we demonstrate the feasibility of using microfluidic patterning for controlling cell adhesion and rolling under physiological flow conditions. By controlling the width of the lines (50-1000 microm) and the spacing between them (50-100 microm) we were able to fabricate surfaces with well-defined patterns of adhesion molecules. We demonstrate the versatility of this technique by patterning surfaces with 3 different adhesion molecules (P-selectin, E-selectin, and von Willebrand Factor) and controlling the adhesion and rolling of three different cell types (neutrophils, Chinese Hamster Ovary cells, and platelets). By varying the concentration of the incubating solution we could control the surface ligand density and hence the cell rolling velocity. Finally by patterning surfaces with both P-selectin and von Willebrand Factor we could control the rolling of both leukocytes and platelets simultaneously. The technique described in this paper provides and effective and inexpensive way to fabricate patterned surfaces for use in cell rolling assays under physiologic flow conditions. PMID:17160704

  7. Redox flow cell development and demonstration project, calendar year 1977

    NASA Technical Reports Server (NTRS)

    1979-01-01

    Research and development on the redox flow cell conducted from January 1, 1977, to December 31, 1977, are described in this report. The major focus of the effort during 1977 was the key technology issues that directly influence the fundamental feasibility of the overall redox concept. These issues were the development of a suitable ion exchange membrane for the system, the screening and study of candidate redox couples to achieve optimum cell performance, and the carrying out of systems analysis and modeling to develop system performance goals and cost estimates.

  8. Rapid flow-induced responses in endothelial cells

    NASA Technical Reports Server (NTRS)

    Stamatas, G. N.; McIntire, L. V.

    2001-01-01

    Endothelial cells alter their morphology, growth rate, and metabolism in response to fluid shear stress. To study rapid flow-induced responses in the 3D endothelial cell morphology and calcium distribution, coupled fluorescence microscopy with optical sectioning, digital imaging, and numerical deconvolution techniques have been utilized. Results demonstrate that within the first minutes of flow application nuclear calcium is increasing. In the same time frame whole cell height and nuclear height are reduced by about 1 microm. Whole cell height changes may facilitate reduction of shear stress gradients on the luminal surface, whereas nuclear structural changes may be important for modulating endothelial growth rate and metabolism. To study the role of the cytoskeleton in these responses, endothelial cells have been treated with specific disrupters (acrylamide, cytochalasin D, and colchicine) of each of the cytoskeleton elements (intermediate filaments, microfilaments, and microtubules, respectively). None of these compounds had any effect on the shear-induced calcium response. Cytochalasin D and acrylamide did not affect the shear-induced nuclear morphology changes. Colchicine, however, completely abrogated the response, indicating that microtubules may be implicated in force transmission from the plasma membrane to the nucleus. A pedagogical model based on tensegrity theory principles is presented that is consistent with the results on the 3D endothelial morphology.

  9. Oxygen transport and cell viability in an annular flow bioreactor: comparison of laminar Couette and Taylor-vortex flow regimes.

    PubMed

    Curran, Stephen J; Black, Richard A

    2005-03-30

    Rotating wall vessel bioreactors have been proposed as a means of controlling the fluid dynamic environment during long-term culture of mammalian cells and engineered tissues. In this study, we show how the delivery of oxygen to cells in an annular flow bioreactor is enhanced by the forced convective transport afforded by Taylor vortex flows. A fiberoptic oxygen probe with negligible lag time was used to measure the dissolved oxygen concentration in real time and under carefully controlled aeration conditions. From these data, the overall mass transfer coefficients were calculated and mass transport correlations determined under laminar Couette flow conditions and discrete Taylor vortex flow regimes, including laminar, wavy, and turbulent flows. While oxygen transport in Taylor vortex flows was significantly greater, and the available oxygen exceeded that consumed by murine fibroblasts in free suspension, the proportion of cells that remained viable decreased with increasing Reynolds number (101.8 < Rei < 1018), which we attribute to the action of fluid shear stresses on the cells as opposed to any limitation in mass transport. Nevertheless, the results of this study suggest that laminar Taylor-vortex flow regimes provide an effective means of maintaining the levels of oxygen transport required for long-term cell culture. PMID:15696514

  10. Evaluation of cytotoxicity of aqueous extract of Graviola leaves on squamous cell carcinoma cell-25 cell lines by 3-(4,5-dimethylthiazol-2-Yl) -2,5-diphenyltetrazolium bromide assay and determination of percentage of cell inhibition at G2M phase of cell cycle by flow cytometry: An in vitro study

    PubMed Central

    Magadi, Visveswaraiah Paranjyothi; Ravi, Venkatadasappa; Arpitha, Anantharaju; Litha; Kumaraswamy, Kikkerilakshminarayana; Manjunath, Krishnappa

    2015-01-01

    Introduction: Malignancies constitute a wide variety of disorders having high mortality and morbidity rates. Current protocols for management include surgical intervention, chemotherapy, and radiation which possess numerous adverse effects. Many phytochemicals are available with anticancer properties similar to anticancer drugs. Major benefit of these compounds is apparent lack of toxicity to normal tissues. Graviola (botanical name: Annona Muricata) contain bioactive compound “annonaceous acetogenins” known for anticancer activity on cancer cell lines. Aims: To determine cytotoxicity of Graviola and percentage cell inhibition at G2M phase of cell cycle. Settings and Design: The cytotoxicity of aqueous extract of Graviola leaves on squamous cell carcinoma (SCC-25) cell lines at various concentrations evaluated using 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The percentage of SCC-25 cell inhibition at G2M phase of cell cycle determined using flow cytometry. Methods: Graviola Leaves, American Type Culture Collection SCC-25 cell lines were procured from Skanda Laboratories, Bengaluru. The cytotoxicity of aqueous extract of Graviola on SCC-25 cells at various concentrations evaluated using MTT assay. The percentage of SCC-25 cell inhibition at G2M phase of cell cycle determined using flow cytometry. Statistical Analysis: Statistical analysis was done using one-way ANOVA. Results: MTT assay showed statistically significant (P < 0.001) dose-dependent inhibition of SCC-25 cell lines by Graviola with IC50 value of 12.42 μg/ml. Flow cytometry revealed that Graviola at 25 and 50 g/ml arrested 53.39% and 52.09% cells in G2M phase of cell cycle respectively, which was statistically significant. Conclusion: Graviola showed significant cytotoxic activity and percentage of cell inhibition at G2M phase cell cycle against SCC-25 cell lines. PMID:26681860

  11. Parallel-plate flow chamber and continuous flow circuit to evaluate endothelial progenitor cells under laminar flow shear stress.

    PubMed

    Lane, Whitney O; Jantzen, Alexandra E; Carlon, Tim A; Jamiolkowski, Ryan M; Grenet, Justin E; Ley, Melissa M; Haseltine, Justin M; Galinat, Lauren J; Lin, Fu-Hsiung; Allen, Jason D; Truskey, George A; Achneck, Hardean E

    2012-01-01

    The overall goal of this method is to describe a technique to subject adherent cells to laminar flow conditions and evaluate their response to well quantifiable fluid shear stresses. Our flow chamber design and flow circuit (Fig. 1) contains a transparent viewing region that enables testing of cell adhesion and imaging of cell morphology immediately before flow (Fig. 11A, B), at various time points during flow (Fig. 11C), and after flow (Fig. 11D). These experiments are illustrated with human umbilical cord blood-derived endothelial progenitor cells (EPCs) and porcine EPCs. This method is also applicable to other adherent cell types, e.g. smooth muscle cells (SMCs) or fibroblasts. The chamber and all parts of the circuit are easily sterilized with steam autoclaving. In contrast to other chambers, e.g. microfluidic chambers, large numbers of cells (> 1 million depending on cell size) can be recovered after the flow experiment under sterile conditions for cell culture or other experiments, e.g. DNA or RNA extraction, or immunohistochemistry (Fig. 11E), or scanning electron microscopy. The shear stress can be adjusted by varying the flow rate of the perfusate, the fluid viscosity, or the channel height and width. The latter can reduce fluid volume or cell needs while ensuring that one-dimensional flow is maintained. It is not necessary to measure chamber height between experiments, since the chamber height does not depend on the use of gaskets, which greatly increases the ease of multiple experiments. Furthermore, the circuit design easily enables the collection of perfusate samples for analysis and/or quantification of metabolites secreted by cells under fluid shear stress exposure, e.g. nitric oxide (Fig. 12).

  12. Alternative flow cytometry strategies to analyze stem cells and cell death in planarians.

    PubMed

    Peiris, Tanuja Harshani; García-Ojeda, Marcos E; Oviedo, Néstor J

    2016-04-01

    Planarians possess remarkable stem cell populations that continuously support cellular turnover and are instrumental in the regeneration of tissues upon injury. Cellular turnover and tissue regeneration in planarians rely on the proper integration of local and systemic signals that regulate cell proliferation and cell death. Thus, understanding the signals controlling cellular proliferation and cell death in planarians could provide valuable insights for maintenance of adult body homeostasis and the biology of regeneration. Flow cytometry techniques have been utilized widely to identify, isolate, and characterize planarian stem cell populations. We developed alternative flow cytometry strategies that reduce the number of reagents and the time of sample preparation to analyze stem cells and cell death in planarians. The sensitivity of these methods is validated with functional studies using RNA interference and treatment with  γ irradiation or stressful conditions that are known to trigger cell death. Altogether, we provide a community resource intended to minimize adverse effects during ex vivo studies of stem cells and cell death in planarians.

  13. Alternative flow cytometry strategies to analyze stem cells and cell death in planarians

    PubMed Central

    Peiris, Tanuja Harshani; García‐Ojeda, Marcos E.

    2016-01-01

    Abstract Planarians possess remarkable stem cell populations that continuously support cellular turnover and are instrumental in the regeneration of tissues upon injury. Cellular turnover and tissue regeneration in planarians rely on the proper integration of local and systemic signals that regulate cell proliferation and cell death. Thus, understanding the signals controlling cellular proliferation and cell death in planarians could provide valuable insights for maintenance of adult body homeostasis and the biology of regeneration. Flow cytometry techniques have been utilized widely to identify, isolate, and characterize planarian stem cell populations. We developed alternative flow cytometry strategies that reduce the number of reagents and the time of sample preparation to analyze stem cells and cell death in planarians. The sensitivity of these methods is validated with functional studies using RNA interference and treatment with  γ irradiation or stressful conditions that are known to trigger cell death. Altogether, we provide a community resource intended to minimize adverse effects during ex vivo studies of stem cells and cell death in planarians. PMID:27307993

  14. The Analysis of Cell Cycle, Proliferation, and Asymmetric Cell Division by Imaging Flow Cytometry.

    PubMed

    Filby, Andrew; Day, William; Purewal, Sukhveer; Martinez-Martin, Nuria

    2016-01-01

    Measuring cellular DNA content by conventional flow cytometry (CFC) and fluorescent DNA-binding dyes is a highly robust method for analysing cell cycle distributions within heterogeneous populations. However, any conclusions drawn from single-parameter DNA analysis alone can often be confounded by the asynchronous nature of cell proliferation. We have shown that by combining fluorescent DNA stains with proliferation tracking dyes and antigenic staining for mitotic cells one can elucidate the division history and cell cycle position of any cell within an asynchronously dividing population. Furthermore if one applies this panel to an imaging flow cytometry (IFC) system then the spatial information allows resolution of the four main mitotic phases and the ability to study molecular distributions within these populations. We have employed such an approach to study the prevalence of asymmetric cell division (ACD) within activated immune cells by measuring the distribution of key fate determining molecules across the plane of cytokinesis in a high-throughput, objective, and internally controlled manner. Moreover the ability to perform high-resolution, temporal dissection of the cell division process lends itself perfectly to investigating the influence chemotherapeutic agents exert on the proliferative capacity of transformed cell lines. Here we describe the method in detail and its application to both ACD and general cell cycle analysis. PMID:27460238

  15. Synergistic effect of high-affinity binding and flow preconditioning on endothelial cell adhesion.

    PubMed

    Mathur, Anshu B; Truskey, George A; Reichert, William M

    2003-01-01

    The current study examined whether the combined introduction of high-affinity avidin-biotin bonds and fibronectin-integrin bonds (i.e., dual ligand treatment) would further augment the adhesion of flow-preconditioned endothelial cells to model substrates via contributions to the actin cytoskeleton and the formation of focal contacts. Human umbilical vein endothelial cells (HUVEC) were grown under static conditions or exposed to a flow-preconditioning regimen for 24 h. Cell retention was determined by exposure to 75 dynes/cm(2). The combination of flow preconditioning and the dual ligand treatment yielded higher cell retention under flow compared to the cells adherent via fibronectin-integrin bonds only. This increase in adhesion strength correlated with a greater focal contact area. Elongation of the HUVEC occurred after exposure to flow preconditioning; however, orientation of dual ligand adherent cells was restricted due to the presence of the high-affinity ligand. Flow-preconditioned cells showed increased stress fiber formation compared to nonconditioned cells although the stress fibers per cell for flow-preconditioned cells were the same on both the ligand systems employed. The results indicate that enhanced adhesion strength is due to a combination of increased focal contact area, stress fiber formation, and cell alignment. PMID:12483708

  16. Experimental and theoretical study of dispersion in potential flows

    NASA Astrophysics Data System (ADS)

    Eames, Ian; Woods, Andy

    1998-11-01

    We examine tracer dispersion in a potential flow, where the velocity field may be writted in terms of the gradient of a scalar function. A range of relevant environmental flows, such as the flow through porous media or thin cracks, may be described to leading order as potential flows. In such flows, tracer dispersion is significantly affected by diverging and converging streamlines which may occur due to variations in permeability, crack thickness or the thickness of a porous layer. We build upon previous studies of dispersion in potential flows in order to examine the effect of geometry of porous inclusions or the spatial variation of crack thickness on plume dispersion. Complementary experimental work is also present using a Hele-Shaw cell, which consists of a uniform viscid flow between two rigid plates. By introducing various shaped obstacles of prescribed thickness between the plates, we are able to study the effect of geometry and permeability on plume dispersion. Here the streamlines correspond to the flow past porous inclusions (and the magnetic field lines past paramagnetic materials), however the velocity field is not similar. These aspects of the flow are included in our analysis.

  17. Determinants of resting cerebral blood flow in sickle cell disease.

    PubMed

    Bush, Adam M; Borzage, Matthew T; Choi, Soyoung; Václavů, Lena; Tamrazi, Benita; Nederveen, Aart J; Coates, Thomas D; Wood, John C

    2016-09-01

    Stroke is common in children with sickle cell disease and results from an imbalance in oxygen supply and demand. Cerebral blood flow (CBF) is increased in patients with sickle cell disease to compensate for their anemia, but adequacy of their oxygen delivery has not been systematically demonstrated. This study examined the physiological determinants of CBF in 37 patients with sickle cell disease, 38 ethnicity matched control subjects and 16 patients with anemia of non-sickle origin. Cerebral blood flow was measured using phase contrast MRI of the carotid and vertebral arteries. CBF increased inversely to oxygen content (r(2)  = 0.69, P < 0.0001). Brain oxygen delivery, the product of CBF and oxygen content, was normal in all groups. Brain composition, specifically the relative amounts of grey and white matter, was the next strongest CBF predictor, presumably by influencing cerebral metabolic rate. Grey matter/white matter ratio and CBF declined monotonically until the age of 25 in all subjects, consistent with known maturational changes in brain composition. Further CBF reductions were observed with age in subjects older than 35 years of age, likely reflecting microvascular aging. On multivariate regression, CBF was independent of disease state, hemoglobin S, hemoglobin F, reticulocyte count and cell free hemoglobin, suggesting that it is regulated similarly in patients and control subjects. In conclusion, sickle cell disease patients had sufficient oxygen delivery at rest, but accomplish this only by marked increases in their resting CBF, potentially limiting their ability to further augment flow in response to stress. Am. J. Hematol. 91:912-917, 2016. © 2016 Wiley Periodicals, Inc. PMID:27263497

  18. Two-Phase Flow within Geological Flow Analogies--A Computational Study

    SciTech Connect

    Crandall, D.M.; Ahmadi, G.; Smith, D.H.; Ferer, M.V.; Richards, M.; Bromhal, G.S.

    2006-10-01

    Displacement of a viscous fluid in heterogeneous geological media by a less viscous one does not evacuate 100% of the defending fluid due to capillary and viscous fingering. This is of importance in geological flows that are encountered in secondary oil recovery and carbon dioxide sequestration in saturated brine fields. Hele-Shaw and pore/throat cells are commonly used to study this in the labratory. Numerical simulations of this flow phenomenon with pore-throat models have been prevalent for over two decades. This current work solves the full Navier-Stokes equations of conservation within random pore-throat geometries with varying properties to study the resulting flow properties. Verification of the solution method is performed by comparison of the model predictions with the available experimental data in the literature. Experimental flows in a pore-throat cell with a known geometrical structure are shown to be in good agreement with the model. Dynamic comparisons to a computational pore-throat model have been shown to be in good agreement as well. There are also additional two-phase immiscible flow patterns that can be identified from the current solutions for which the corresponding laboratory counter part or the pore-throat model predictions are not available. The identification of these flow patterns may allow more accurate modeling of fluid displacement on the reservoir scale.

  19. Activation of β–catenin Signaling in MLO-Y4 Osteocytic Cells versus 2T3 Osteoblastic Cells by Fluid Flow Shear Stress and PGE2: Implications for the Study of Mechanosensation in Bone

    PubMed Central

    Kamel, Mohamed A.; Picconi, Jason L.; Lara-Castillo, Nuria; Johnson, Mark L.

    2010-01-01

    The osteocyte is hypothesized to be the mechanosensory cell in bone. However, osteoblastic cell models have been most commonly used to investigate mechanisms of mechanosensation in bone. Therefore, we sought to determine if differences might exist between osteocytic and osteoblastic cell models relative to the activation of β-catenin signaling in MLO-Y4 osteocytic, 2T3 osteoblastic and primary neonatal calvarial cells (NCCs) in response to pulsatile fluid flow shear stress (PFFSS). β–catenin nuclear translocation was observed in MLO-Y4 cells at 2 and 16 dynes/cm2 PFFSS, but only at 16 dynes/cm2 in the 2T3 or NCC cultures. MLO-Y4 cells released high amounts of PGE2 into the media at all levels of PFFSS (2–24 dynes/cm2) and we observed a biphasic pattern of relative to the level of PFFSS. In contrast PGE2 release by 2T3 cells was only detected during 16 and 24 dynes/cm2 PFFSS starting at >1 hour and never reached the levels produced by MLO-Y4 cells. Exogenously added PGE2 was able to induce β–catenin nuclear translocation in all cells suggesting that the differences between the cell lines observed for β–catenin nuclear translocation was associated with the differences in PGE2 production. To investigate a possible mechanism for the differences in PGE2 release by MLO-Y4 and 2T3 cells we examined the regulation of Ptgs2 (Cox-2) gene expression by PFFSS. 2T3 cell Ptgs2 mRNA levels at both 0 and 24 hours after 2 hours of PFFSS showed biphasic increases with peaks at 4 and 24 dynes/cm2 and 24 hour levels were higher than 0 hour levels. MLO-Y4 cell Ptgs2 expression was similarly biphasic; however at 24 hours post flow Ptgs2 mRNA levels were lower. Our data suggest significant differences in the sensitivity and kinetics of the response mechanisms of 2T3 and neonatal calvarial osteoblastic versus MLO-Y4 osteocytic cells to PFFSS. Furthermore our data support a role for PGE2 in mediating the activation of β–catenin signaling in response to fluid flow shear

  20. Research development of designing flow cells for optical absorption detectors.

    PubMed

    Yang, Sandong; Tang, Tao; Li, Tong; Wang, Fengyun; Hao, Qingli

    2016-02-01

    The optical absorption detector is one of the most commonly used detectors for high performance liquid chromatography (HPLC). As a core part of this kind of detector, the designs of flow cells, where light passes through samples for acquiring samples information, will affect the performance of a detector. In order to enhance the signal to noise ratio of detectors and reduce the bands broadening that come from flow cells, it is necessary to design a flow cell with a longer optical path length and a less cell volume while maintaining the luminous flux. However the limitations of the machining capacity make it difficult to increase the optical path length, reduce the cell volume and keep or increase the luminous flux simultaneously. It is a challenge to optimize the designing and machining of flow cells so as to improve the performance of detectors. This review discusses the development of designing flow cells based on the detection principle in some aspects of increasing the optical path length, reducing the cell volume, taking the advantages of total reflection and so on. At the same time, some of the designs are illustrated in detail. These various ideas and structures are significant references for designing flow cells and developing optical absorption detectors. PMID:27382716

  1. Actin polymerization and intracellular solvent flow in cell surface blebbing

    PubMed Central

    1995-01-01

    The cortical actin gel of eukaryotic cells is postulated to control cell surface activity. One type of protrusion that may offer clues to this regulation are the spherical aneurysms of the surface membrane known as blebs. Blebs occur normally in cells during spreading and alternate with other protrusions, such as ruffles, suggesting similar protrusive machinery is involved. We recently reported that human melanoma cell lines deficient in the actin filament cross-linking protein, ABP-280, show prolonged blebbing, thus allowing close study of blebs and their dynamics. Blebs expand at different rates of volume increase that directly predict the final size achieved by each bleb. These rates decrease as the F-actin concentration of the cells increase over time after plating on a surface, but do so at lower concentrations in ABP-280 expressing cells. Fluorescently labeled actin and phalloidin injections of blebbing cells indicate that a polymerized actin structure is not present initially, but appears later and is responsible for stopping further bleb expansion. Therefore, it is postulated that blebs occur when the fluid-driven expansion of the cell membrane is sufficiently rapid to initially outpace the local rate of actin polymerization. In this model, the rate of intracellular solvent flow driving this expansion decreases as cortical gelation is achieved, whether by factors such as ABP-280, or by concentrated actin polymers alone, thereby leading to decreased size and occurrence of blebs. Since the forces driving bleb extension would always be present in a cell, this process may influence other cell protrusions as well. PMID:7790356

  2. Refinement Of Hexahedral Cells In Euler Flow Computations

    NASA Technical Reports Server (NTRS)

    Melton, John E.; Cappuccio, Gelsomina; Thomas, Scott D.

    1996-01-01

    Topologically Independent Grid, Euler Refinement (TIGER) computer program solves Euler equations of three-dimensional, unsteady flow of inviscid, compressible fluid by numerical integration on unstructured hexahedral coordinate grid refined where necessary to resolve shocks and other details. Hexahedral cells subdivided, each into eight smaller cells, as needed to refine computational grid in regions of high flow gradients. Grid Interactive Refinement and Flow-Field Examination (GIRAFFE) computer program written in conjunction with TIGER program to display computed flow-field data and to assist researcher in verifying specified boundary conditions and refining grid.

  3. Single chamber fuel cells: Flow geometry, rate and composition considerations

    SciTech Connect

    Stefan, Ionel C.; Jacobson, Craig P.; Visco, Steven J.; De Jonghe, Lutgard C.

    2003-11-17

    Four different single chamber fuel cell designs were compared using propane-air gas mixtures. Gas flow around the electrodes has a significant influence on the open circuit voltage and the power density of the cell. The strong influence of flow geometry is likely due to its effect on gas composition, particularly on the oxygen chemical potential at the two electrodes as a result of gas mixing. The chamber design which exposes the cathode first to the inlet gas was found to yield the best performance at lower flow rates, while the open tube design with the electrodes equally exposed to the inlet gas worked best at higher flow rates.

  4. Numerical Simulation of Sickle Cell Blood Flow in the Microcirculation

    NASA Astrophysics Data System (ADS)

    Berger, Stanley A.; Carlson, Brian E.

    2001-11-01

    A numerical simulation of normal and sickle cell blood flow through the transverse arteriole-capillary microcirculation is carried out to model the dominant mechanisms involved in the onset of vascular stasis in sickle cell disease. The transverse arteriole-capillary network is described by Strahler's network branching method, and the oxygen and blood transport in the capillaries is modeled by a Krogh cylinder analysis utilizing Lighthill's lubrication theory, as developed by Berger and King. Poiseuille's law is used to represent blood flow in the arterioles. Applying this flow and transport model and utilizing volumetric flow continuity at each network bifurcation, a nonlinear system of equations is obtained, which is solved iteratively using a steepest descent algorithm coupled with a Newton solver. Ten different networks are generated and flow results are calculated for normal blood and sickle cell blood without and with precapillary oxygen loss. We find that total volumetric blood flow through the network is greater in the two sickle cell blood simulations than for normal blood owing to the anemia associated with sickle cell disease. The percentage of capillary blockage in the network increases dramatically with decreasing pressure drop across the network in the sickle cell cases while there is no blockage when normal blood flows through simulated networks. It is concluded that, in sickle cell disease, without any vasomotor dilation response to decreasing oxygen concentrations in the blood, capillary blockage will occur in the microvasculature even at average pressure drops across the transverse arteriole-capillary networks.

  5. Tension of red blood cell membrane in simple shear flow

    NASA Astrophysics Data System (ADS)

    Omori, T.; Ishikawa, T.; Barthès-Biesel, D.; Salsac, A.-V.; Imai, Y.; Yamaguchi, T.

    2012-11-01

    When a red blood cell (RBC) is subjected to an external flow, it is deformed by the hydrodynamic forces acting on its membrane. The resulting elastic tensions in the membrane play a key role in mechanotransduction and govern its rupture in the case of hemolysis. In this study, we analyze the motion and deformation of an RBC in a simple shear flow and the resulting elastic tensions on the membrane. The large deformation of the red blood cell is modelled by coupling a finite element method to solve the membrane mechanics and a boundary element method to solve the flows of the internal and external liquids. Depending on the capillary number Ca, ratio of the viscous to elastic forces, we observe three kinds of RBC motion: tumbling at low Ca, swinging at larger Ca, and breathing at the transitions. In the swinging regime, the region of the high principal tensions periodically oscillates, whereas that of the high isotropic tensions is almost unchanged. Due to the strain-hardening property of the membrane, the deformation is limited but the membrane tension increases monotonically with the capillary number. We have quantitatively compared our numerical results with former experimental results. It indicates that a membrane isotropic tension O(10-6 N/m) is high enough for molecular release from RBCs and that the typical maximum membrane principal tension for haemolysis would be O(10-4 N/m). These findings are useful to clarify not only the membrane rupture but also the mechanotransduction of RBCs.

  6. Lymphatic vessel development: fluid flow and valve-forming cells.

    PubMed

    Kume, Tsutomu

    2015-08-01

    Hemodynamic forces regulate many aspects of blood vessel disease and development, including susceptibility to atherosclerosis and remodeling of primary blood vessels into a mature vascular network. Vessels of the lymphatic circulatory system are also subjected to fluid flow-associated forces, but the molecular and cellular mechanisms by which these forces regulate the formation and maintenance of lymphatic vessels remain largely uncharacterized. This issue of the JCI includes two articles that begin to address how fluid flow influences lymphatic vessel development and function. Sweet et al. demonstrate that lymph flow is essential for the remodeling of primary lymphatic vessels, for ensuring the proper distribution of smooth muscle cells (SMCs), and for the development and maturation of lymphatic valves. Kazenwadel et al. show that flow-induced lymphatic valve development is initiated by the upregulation of GATA2, which has been linked to lymphedema in patients with Emberger syndrome. Together, these observations and future studies inspired by these results have potential to lead to the development of strategies for the treatment of lymphatic disorders.

  7. Human red blood cells deformed under thermal fluid flow.

    PubMed

    Foo, Ji-Jinn; Chan, Vincent; Feng, Zhi-Qin; Liu, Kuo-Kang

    2006-03-01

    The flow-induced mechanical deformation of a human red blood cell (RBC) during thermal transition between room temperature and 42.0 degrees C is interrogated by laser tweezer experiments. Based on the experimental geometry of the deformed RBC, the surface stresses are determined with the aid of computational fluid dynamics simulation. It is found that the RBC is more deformable while heating through 37.0 degrees C to 42.0 degrees C, especially at a higher flow velocity due to a thermal-fluid effect. More importantly, the degree of RBC deformation is irreversible and becomes softer, and finally reaches a plateau (at a uniform flow velocity U > 60 microm s(-1)) after the heat treatment, which is similar to a strain-hardening dominated process. In addition, computational simulated stress is found to be dependent on the progression of thermotropic phase transition. Overall, the current study provides new insights into the highly coupled temperature and hydrodynamic effects on the biomechanical properties of human erythrocyte in a model hydrodynamic flow system.

  8. Surface deformation and shear flow in ligand mediated cell adhesion.

    PubMed

    Sircar, Sarthok; Roberts, Anthony J

    2016-10-01

    We present a unified, multiscale model to study the attachment/detachment dynamics of two deforming, charged, near spherical cells, coated with binding ligands and subject to a slow, homogeneous shear flow in a viscous, ionic fluid medium. The binding ligands on the surface of the cells experience both attractive and repulsive forces in an ionic medium and exhibit finite resistance to rotation via bond tilting. The microscale drag forces and couples describing the fluid flow inside the small separation gap between the cells, are calculated using a combination of methods in lubrication theory and previously published numerical results. For a selected range of material and fluid parameters, a hysteretic transition of the sticking probability curves (i.e., the function [Formula: see text]) between the adhesion phase (when [Formula: see text]) and the fragmentation phase (when [Formula: see text]) is attributed to a nonlinear relation between the total nanoscale binding forces and the separation gap between the cells. We show that adhesion is favoured in highly ionic fluids, increased deformability of the cells, elastic binders and a higher fluid shear rate (until a critical threshold value of shear rate is reached). Within a selected range of critical shear rates, the continuation of the limit points (i.e., the turning points where the slope of [Formula: see text] changes sign) predict a bistable region, indicating an abrupt switching between the adhesion and the fragmentation regimes. Although, bistability in the adhesion-fragmentation phase diagram of two deformable, charged cells immersed in an ionic aqueous environment has been identified by some in vitro experiments, but until now, has not been quantified theoretically.

  9. Detection of circulating immune complexes by Raji cell assay: comparison of flow cytometric and radiometric methods

    SciTech Connect

    Kingsmore, S.F.; Crockard, A.D.; Fay, A.C.; McNeill, T.A.; Roberts, S.D.; Thompson, J.M.

    1988-01-01

    Several flow cytometric methods for the measurement of circulating immune complexes (CIC) have recently become available. We report a Raji cell flow cytometric assay (FCMA) that uses aggregated human globulin (AHG) as primary calibrator. Technical advantages of the Raji cell flow cytometric assay are discussed, and its clinical usefulness is evaluated in a method comparison study with the widely used Raji cell immunoradiometric assay. FCMA is more precise and has greater analytic sensitivity for AHG. Diagnostic sensitivity by the flow cytometric method is superior in systemic lupus erythematosus (SLE), rheumatoid arthritis, and vasculitis patients: however, diagnostic specificity is similar for both assays, but the reference interval of FCMA is narrower. Significant correlations were found between CIC levels obtained with both methods in SLE, rheumatoid arthritis, and vasculitis patients and in longitudinal studies of two patients with cerebral SLE. The Raji cell FCMA is recommended for measurement of CIC levels to clinical laboratories with access to a flow cytometer.

  10. Laser rastering flow cytometry: fast cell counting and identification

    NASA Astrophysics Data System (ADS)

    Vacca, G.; Junnarkar, M. R.; Goldblatt, N. R.; Yee, M. W.; Van Slyke, B. M.; Briese, T. C.

    2009-02-01

    We describe the concept of laser rastering flow cytometry, where a rapidly scanning laser beam allows counting and classification of cells at much higher rates than currently possible. Modifications to existing flow cytometers to implement the concept include an acousto-optic deflector, fast analog-to-digital conversion, and a two-step digital-signal-processing scheme that handles the high data rates and provides key assay information. Results are shown that prove the concept, demonstrating the ability to resolve closely spaced cells and to measure cells at rates more than an order of magnitude faster than on conventional flow-cytometer-based hematology analyzers.

  11. Fundamentals of free flux flow: proposed studies

    NASA Astrophysics Data System (ADS)

    Alexander, J. A.; Gafarov, O.; Gapud, A. A.; Wu, J. Z.

    2013-03-01

    Although much is known about free flux flow (FFF) in superconductors - in which pinning is insignificant compared to interactions between quantized vortices - there still remain questions concerning fundamental dynamics. Building on our previous work in correlating FFF with vortex core size (PRB 80, 134524), we propose three new studies examining more deeply the normal state in the vortex core and interactions between vortices. A correlation between scattering inside cores and the viscosity of FFF has not been explicitly determined; this may be investigated by probing the effect of scattering centers created by proton irradiation. Using results of previous irradiation work, one could control the extent of normal state scattering while monitoring effects on FFF. Questions also exist concerning vortex motion in channels with widths approaching that of individual vortices - as determined solely by inter-vortex interactions. Studies have suggested that flux flow through constrictions could imitate ``jamming'' in the collective motion of grains: Under certain conditions, it is possible for grains to form a barrier, blocking flow. More than just qualitatively comparing flux flow and granular flow to find evidence of jamming, we propose a new experiment for quantitatively modeling flux jamming by realizing the flux flow equivalent of granular jamming in a ``hopper''. In the same way, we also propose a FFF equivalent of another granular-flow phenomenon, ``non-Newtonian'' fluids, where rapid shear causes jamming. Funded by NSF-RUI grant, DMR-0907038.

  12. Cerebral blood flow in sickle cell cerebrovascular disease

    SciTech Connect

    Huttenlocher, P.R.; Moohr, J.W.; Johns, L.; Brown, F.D.

    1984-05-01

    Cerebral blood flow (CBF) has been studied by the xenon-133 (/sup 133/Xe) inhalation method in 16 children with suspected sickle cell cerebrovascular disease. Abnormalities consisting of decreases in total, hemispheral, or regional CBF were found in 17 of 26 studies. Eleven studies performed immediately after stroke, transient ischemic attack, or depression of state of alertness showed abnormalities. In addition to confirming regional cerebrovascular insufficiency in children with stroke due to major cerebral artery occlusion, the method detected diffuse decrease in CBF in children with stupor, coma, and seizures who had normal angiographic findings. In contrast, six of seven studies obtained after exchange transfusion or during maintenance on hypertransfusion therapy showed normal findings. The difference between results in patients with acute neurologic disturbances and those receiving transfusion therapy was statistically significant (P less than .005). The data indicate that the /sup 133/Xe method reliably demonstrates cerebrovascular impairment in sickle cell disease. They also suggest that CBF changes in patients with sickle cell disease can be reversed by exchange transfusion and by hypertransfusion therapy. The /sup 133/Xe CBF method may be useful for following up children with sickle cell disease who are at high risk for recurrent stroke.

  13. Protein kinase G inhibits flow-induced Ca2+ entry into collecting duct cells.

    PubMed

    Du, Juan; Wong, Wei-Yan; Sun, Lei; Huang, Yu; Yao, Xiaoqiang

    2012-07-01

    The renal cortical collecting duct (CCD) contributes to the maintenance of K(+) homeostasis by modulating renal K(+) secretion. Cytosolic Ca(2+) ([Ca(2+)](i)) mediates flow-induced K(+) secretion in the CCD, but the mechanisms regulating flow-induced Ca(2+) entry into renal epithelial cells are not well understood. Here, we found that atrial natriuretic peptide, nitric oxide, and cyclic guanosine monophosphate (cGMP) act through protein kinase G (PKG) to inhibit flow-induced increases in [Ca(2+)](i) in M1-CCD cells. Coimmunoprecipitation, double immunostaining, and functional studies identified heteromeric TRPV4-P2 channels as the mediators of flow-induced Ca(2+) entry into M1-CCD cells and HEK293 cells that were coexpressed with both TRPV4 and TRPP2. In these HEK293 cells, introducing point mutations at two putative PKG phosphorylation sites on TRPP2 abolished the ability of cGMP to inhibit flow-induced Ca(2+) entry. In addition, treating M1-CCD cells with fusion peptides that compete with the endogenous PKG phosphorylation sites on TRPP2 also abolished the cGMP-mediated inhibition of the flow-induced Ca(2+) entry. Taken together, these data suggest that heteromeric TRPV4-P2 channels mediate the flow-induced entry of Ca(2+) into collecting duct cells. Furthermore, substances such as atrial natriuretic peptide and nitric oxide, which increase cGMP, abrogate flow-induced Ca(2+) entry through PKG-mediated inhibition of these channels. PMID:22518003

  14. Rapid Flow Analysis Studies with Spectroscopic Detectors.

    NASA Astrophysics Data System (ADS)

    Thalib, Amlius

    A rapid flow analysis study based on segmented flow and flow injection principles is described in this thesis. The main objective of this study was to establish the response characteristics in continuous flow analysis systems in order to improve sampling rates with several types of spectroscopic detectors. It was found from flame photometric studies that non-segmented flowing streams are applicable to rapid flow analysis with automatic sample aspiration. Calcium was used as a typical example and determined at sampling rates up to 360 h('-1) with a detection limit of 0.05 mg L(' -1). A rapid flow system is reported using direct aspiration for AAS analysis with both manual injection and automatic aspiration techniques, and found to give sampling rates of up to 600-720 samples h('-1). Speed of analysis was reduced by about 50% when using an external peristaltic pump in the flow system design, due to increased sample dispersion. A novel aspect of a rapid flow injection approach reported with ICPAES detection includes the method of injecting samples via a peristaltic pump with simultaneous computer data processing. Determination of serum cations (Na, K, Ca, Mg and Fe) was demonstrated as an example of an application of the technique at sampling rates of 240 h('-1). Precision and detection limits for 13 elements in a single standard solution are reported. The use of automated aspiration sampling is also reported in this method for comparison. Further studies on flow characteristics were carried out by a combination of the rapid flow system with very short sampling times as low as 2 seconds using UV-visible spectrophotometric detection. Analysis of human blood serum samples was used as an example where total protein and inorganic phosphate were determined at sampling rates of 240 h('-1) and 360 h('-1) respectively. The novel aspects of the results from these studies include the very rapid sample throughput developed with simple and inexpensive experimental approaches in

  15. Flow over periodic hills: an experimental study

    NASA Astrophysics Data System (ADS)

    Rapp, Ch.; Manhart, M.

    2011-07-01

    Two-dimensional flow over periodically arranged hills was investigated experimentally in a water channel. Two-dimensional particle image velocimetry (PIV) and one-dimensional laser Doppler anemometry (LDA) measurements were undertaken at four Reynolds numbers ({5,600} le Re le {37,000}). Two-dimensional PIV field measurements were thoroughly validated by means of point-by-point 1D LDA measurements at certain positions of the flow. A detailed study of the periodicity and the homogeneity was undertaken, which demonstrates that the flow can be regarded as two-dimensional and periodic for Re ge {10,000}. We found a decreasing reattachment length with increasing Reynolds number. This is connected to a higher momentum in the near-wall zone close to flow separation which comes from the velocity speed up above the obstacle. This leads to a velocity overshoot directly above the hill crest which increases with Reynolds number as the inner layer depth decreases. The flow speed up above that layer is independent of the Reynolds number which supports the assumption of inviscid flow disturbance in the outer layer usually made in asymptotic theory for flow over small hills.

  16. Microfluidic-based single cell trapping using a combination of stagnation point flow and physical barrier

    NASA Astrophysics Data System (ADS)

    Yu, Miao; Chen, Zongzheng; Xiang, Cheng; Liu, Bo; Xie, Handi; Qin, Kairong

    2016-06-01

    Single cell trapping in vitro by microfluidic device is an emerging approach for the study of the relationship between single cells and their dynamic biochemical microenvironments. In this paper, a hydrodynamic-based microfluidic device for single cell trapping is designed using a combination of stagnation point flow and physical barrier. The microfluidic device overcomes the weakness of the traditional ones, which have been only based upon either stagnation point flows or physical barriers, and can conveniently load dynamic biochemical signals to the trapped cell. In addition, it can connect with a programmable syringe pump and a microscope to constitute an integrated experimental system. It is experimentally verified that the microfluidic system can trap single cells in vitro even under flow disturbance and conveniently load biochemical signals to the trapped cell. The designed micro-device would provide a simple yet effective experimental platform for further study of the interactions between single cells and their microenvironments.

  17. Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses.

    PubMed Central

    Davey, H M; Kell, D B

    1996-01-01

    The most fundamental questions such as whether a cell is alive, in the sense of being able to divide or to form a colony, may sometimes be very hard to answer, since even axenic microbial cultures are extremely heterogeneous. Analyses that seek to correlate such things as viability, which is a property of an individual cell, with macroscopic measurements of culture variables such as ATP content, respiratory activity, and so on, must inevitably fail. It is therefore necessary to make physiological measurements on individual cells. Flow cytometry is such a technique, which allows one to analyze cells rapidly and individually and permits the quantitative analysis of microbial heterogeneity. It therefore offers many advantages over conventional measurements for both routine and more exploratory analyses of microbial properties. While the technique has been widely applied to the study of mammalian cells, is use in microbiology has until recently been much more limited, largely because of the smaller size of microbes and the consequently smaller optical signals obtainable from them. Since these technical barriers no longer hold, flow cytometry with appropriate stains has been used for the rapid discrimination and identification of microbial cells, for the rapid assessment of viability and of the heterogeneous distributions of a wealth of other more detailed physiological properties, for the analysis of antimicrobial drug-cell interactions, and for the isolation of high-yielding strains of biotechnological interest. Flow cytometric analyses provide an abundance of multivariate data, and special methods have been devised to exploit these. Ongoing advances mean that modern flow cytometers may now be used by nonspecialists to effect a renaissance in our understanding of microbial heterogeneity. PMID:8987359

  18. A cell counting/sorting system incorporated with a microfabricated flow cytometer chip

    NASA Astrophysics Data System (ADS)

    Yang, Sung-Yi; Hsiung, Suz-Kai; Hung, Yung-Ching; Chang, Chen-Min; Liao, Teh-Lu; Lee, Gwo-Bin

    2006-07-01

    Flow cytometry is a popular technique for counting and sorting individual cells. This study presents and demonstrates a new cell counting/sorting system integrated with several essential components including a micromachined flow cytometer chip device, an optical detection system and a data analysis and control system to achieve the functions of cell sample injection, optical signal detection and cell collection. By using MEMS technology, we have integrated several microfluidic components such as micro pneumatic pumps/valves onto a polymer-based chip device. Three pneumatic micropumps are used to provide the hydrodynamic driving force for both sample and sheath flows such that hydrodynamic flow focusing can be achieved, and a micro flow switch device comprising three pneumatic microvalves located downstream of the micro sample flow channel is used for cell collection. Cell samples of human lung cancer cells labelled with commercially available fluorescent dyes have been detected and collected successfully utilizing the developed device. The real-time image of dye-labelled cell samples being excited and detected can be monitored and observed through the LCD panel by a custom designed CCD/APD holder and moving stage. Finally, micro flow switch devices were used to successfully sort the cells into the desired outlet channel, and the counting results of the specific cell samples were monitored through the counting panel. The current study focuses on the setup of the overall system. The proposed flow cytometer system has several advantages such as portability, low cost and easy operation process. The size of the system is 37 cm × 16 cm × 18 cm and the weight is 3.5 kg. The error rate of counting and sorting was 1.5% and 2%, respectively. The sorting frequency of the microvalve device is calculated to be 120 cells min-1. The developed microfluidic chip device could be a promising tool for cell-based application fields such as profiling, counting and sorting.

  19. Electrochemical cell for rebalancing REDOX flow system

    NASA Technical Reports Server (NTRS)

    Thaller, L. H. (Inventor)

    1979-01-01

    An electrically rechargeable REDOX cell or battery system including one of more rebalancing cells is described. Each rebalancing cell is divided into two chambers by an ion permeable membrane. The first chamber is fed with gaseous hydrogen and a cathode fluid which is circulated through the cathode chamber of the REDOX cell is also passed through the second chamber of the rebalancing cell. Electrochemical reactions take place on the surface of insert electrodes in the first and second chambers to rebalance the electrochemical capacity of the anode and cathode fluids of the REDOX system.

  20. Methods for improved resolution of flow electrophoresis cells

    NASA Technical Reports Server (NTRS)

    Mccreight, L. R.; Fogal, G. L.

    1974-01-01

    First method involves remote adjusting of zeta potential. Second approach sandwiches two conducting metal plates between opposite cell walls and thin insulating layer. Third method forces buffer to flow in direction opposite particle streams.

  1. High-throughput magnetic flow sorting of human cells selected on the basis of magnetophoretic mobility

    NASA Astrophysics Data System (ADS)

    Reece, Lisa M.; Sanders, Lehanna; Kennedy, David; Guernsey, Byron; Todd, Paul; Leary, James F.

    2010-02-01

    We have shown the potential of a new method for optimizing the separation of human stem cell subsets from peripheral blood based on a novel cell labeling technique that leverages the capabilities of a new commercially available high speed magnetic cell sorting system (IKOTECH LLC, New Albany, IN). This new system sorts cells in a continuously flowing manner using a Quadrupole Magnetic cell Sorter (QMS). The sorting mechanism is based upon the magnetophoretic mobility of the cells, a property related to the relative binding distributions of magnetic particles per cell, as determined by the utilization of a Magnetic Cell Tracking Velocimeter (MCTV). KG-1 cells were competitively labeled with anti-CD34 magnetic beads and anti-CD34 FITC to obtain an optimal level of magnetophoretic mobility as visualized by the MCTV for high throughput sort recovery in the QMS. In QMS sorting, the concept of split-flow thin channel (SPLITT) separation technology is applied by having a sample stream enter a vertical annular flow channel near the channel's interior wall followed by another sheath flow entering near the exterior wall. The two flows are initially separated by a flow splitter. They pass through the bore of a Halbach permanent quadrupole magnet assembly, which draws magnetized cells outward and deflects them into a positive outflow, while negative cells continue straight out via the inner flow lamina. QMS sorts cells based upon their magnetophoretic mobility, or the velocity of a cell per unit ponderomotive force, the counterpart of fluorescence intensity in flow cytometry. The magnetophoretic mobility distribution of a cell population, measured by automated MCTV, is used as input data for the algorithmic control of sample, sheath, and outlet flow velocities of the QMS. In this study, the relative binding distributions of magnetic particles per cell were determined by MCTV using novel sorting and sizing algorithms. The resulting mobility histograms were used to set the QMS

  2. Low cost, radial flow, solid oxide fuel cell

    NASA Astrophysics Data System (ADS)

    Petrik, M. A.

    The Interscience Radial Flow (IRF) SOFC is designed to minimize problems in high-temperature operation and for low-cost fabrication. The cell has planar, non-sintered construction, uses particulate materials to form porous electrodes, and has internal radial flow. The object of this phase was to demonstrate feasibility of multi-cell stack operation. Performance milestone was 15% DC HHV efficiency with hydrogen at greater than 50 mW/sq cm over 100 h.

  3. A study of thin liquid sheet flows

    NASA Technical Reports Server (NTRS)

    Chubb, Donald L.; Calfo, Frederick D.; Mcconley, Marc W.; Mcmaster, Matthew S.; Afjeh, Abdollah A.

    1993-01-01

    This study was a theoretical and experimental investigation of thin liquid sheet flows in vacuum. A sheet flow created by a narrow slit of width, W, coalesces to a point at a distance, L, as a result of surface tension forces acting at the sheet edges. As the flow coalesces, the fluid accumulates in the sheet edges. The observed triangular shape of the sheet agrees with the calculated triangular result. Experimental results for L/W as a function of Weber number, We, agree with the calculated result, L/W = the sq. root of 8We. The edge cross sectional shape is found to oscillate from elliptic to 'cigar' like to 'peanut' like and then back to elliptic in the flow direction. A theoretical one-dimensional model was developed that yielded only elliptic solutions for the edge cross section. At the points where the elliptic shapes occur, there is agreement between theory and experiment.

  4. Comparative Study of Airfoil Flow Separation Criteria

    NASA Astrophysics Data System (ADS)

    Laws, Nick; Kahouli, Waad; Epps, Brenden

    2015-11-01

    Airfoil flow separation impacts a multitude of applications including turbomachinery, wind turbines, and bio-inspired micro-aerial vehicles. In order to achieve maximum performance, some devices operate near the edge of flow separation, and others use dynamic flow separation advantageously. Numerous criteria exist for predicting the onset of airfoil flow separation. This talk presents a comparative study of a number of such criteria, with emphasis paid to speed and accuracy of the calculations. We evaluate the criteria using a two-dimensional unsteady vortex lattice method, which allows for rapid analysis (on the order of seconds instead of days for a full Navier-Stokes solution) and design of optimal airfoil geometry and kinematics. Furthermore, dynamic analyses permit evaluation of dynamic stall conditions for enhanced lift via leading edge vortex shedding, commonly present in small flapping-wing flyers such as the bumblebee and hummingbird.

  5. Trace metal leaching behavior studied through the use of parametric modeling of water borne soil particles fractionated with a split-flow thin cell.

    PubMed

    Magnuson, Matthew L; Kelty, Keith C; Kelty, Catherine A

    2002-10-15

    Leaching of particle-bound metals affects the ability of settling ponds and other engineered structures to remove metallic pollutants, and leaching behavior is related to particle size. In this investigation, water borne soil particles were leached and fractionated with a split-flow thin cell, and the metal loadings were quantified as a function of particle size. For comparison of the metal-loading curves, different empirical modeling procedures were investigated to convert the data to a precise functional form suitable for quantitative comparison of changes in differential loading as a function of particle size. Results of this investigation are presented for a soil sample before and after leaching caused by simulated acid rain conditions. Following simulated acid rain leaching, the shape of the differential distribution curves change, and these changes reflect the particle size mediated leaching behavior. For the soil used in this demonstration, simulated acid rain leaching shifted the differential loading toward larger particle sizes, and the magnitude of the shift varied significantly among the metals. Because settling rate decreases as the square of particle size, this could potentially affect management decisions for settling ponds receiving these particles. The high precision afforded by the analysis allows the development of insight into the leaching mechanisms through comparing "partial" acid rain leaching with "total recoverable" leaching by EPA Method 3050.

  6. Improving Viability of Stem Cells During Syringe Needle Flow Through the Design of Hydrogel Cell Carriers

    PubMed Central

    Aguado, Brian A.; Mulyasasmita, Widya; Su, James; Lampe, Kyle J.

    2012-01-01

    Cell transplantation is a promising therapy for a myriad of debilitating diseases; however, current delivery protocols using direct injection result in poor cell viability. We demonstrate that during the actual cell injection process, mechanical membrane disruption results in significant acute loss of viability at clinically relevant injection rates. As a strategy to protect cells from these damaging forces, we hypothesize that cell encapsulation within hydrogels of specific mechanical properties will significantly improve viability. We use a controlled in vitro model of cell injection to demonstrate success of this acute protection strategy for a wide range of cell types including human umbilical vein endothelial cells (HUVEC), human adipose stem cells, rat mesenchymal stem cells, and mouse neural progenitor cells. Specifically, alginate hydrogels with plateau storage moduli (G′) ranging from 0.33 to 58.1 Pa were studied. A compliant crosslinked alginate hydrogel (G′=29.6 Pa) yielded the highest HUVEC viability, 88.9%±5.0%, while Newtonian solutions (i.e., buffer only) resulted in 58.7%±8.1% viability. Either increasing or decreasing the hydrogel storage modulus reduced this protective effect. Further, cells within noncrosslinked alginate solutions had viabilities lower than media alone, demonstrating that the protective effects are specifically a result of mechanical gelation and not the biochemistry of alginate. Experimental and theoretical data suggest that extensional flow at the entrance of the syringe needle is the main cause of acute cell death. These results provide mechanistic insight into the role of mechanical forces during cell delivery and support the use of protective hydrogels in future clinical stem cell injection studies. PMID:22011213

  7. Flow field measurements in the cell culture unit

    NASA Technical Reports Server (NTRS)

    Walker, Stephen; Wilder, Mike; Dimanlig, Arsenio; Jagger, Justin; Searby, Nancy

    2002-01-01

    The cell culture unit (CCU) is being designed to support cell growth for long-duration life science experiments on the International Space Station (ISS). The CCU is a perfused loop system that provides a fluid environment for controlled cell growth experiments within cell specimen chambers (CSCs), and is intended to accommodate diverse cell specimen types. Many of the functional requirements depend on the fluid flow field within the CSC (e.g., feeding and gas management). A design goal of the CCU is to match, within experimental limits, all environmental conditions, other than the effects of gravity on the cells, whether the hardware is in microgravity ( micro g), normal Earth gravity, or up to 2g on the ISS centrifuge. In order to achieve this goal, two steps are being taken. The first step is to characterize the environmental conditions of current 1g cell biology experiments being performed in laboratories using ground-based hardware. The second step is to ensure that the design of the CCU allows the fluid flow conditions found in 1g to be replicated from microgravity up to 2g. The techniques that are being used to take these steps include flow visualization, particle image velocimetry (PIV), and computational fluid dynamics (CFD). Flow visualization using the injection of dye has been used to gain a global perspective of the characteristics of the CSC flow field. To characterize laboratory cell culture conditions, PIV is being used to determine the flow field parameters of cell suspension cultures grown in Erlenmeyer flasks on orbital shakers. These measured parameters will be compared to PIV measurements in the CSCs to ensure that the flow field that cells encounter in CSCs is within the bounds determined for typical laboratory experiments. Using CFD, a detailed simulation is being developed to predict the flow field within the CSC for a wide variety of flow conditions, including microgravity environments. Results from all these measurements and analyses of the

  8. Fibrosarcoma versus fibromatoses and cellular nodular fasciitis. A comparative study of their proliferative activity using proliferating cell nuclear antigen, DNA flow cytometry, and p53.

    PubMed

    Oshiro, Y; Fukuda, T; Tsuneyoshi, M

    1994-07-01

    We analyzed the proliferative activities, immunoreactivity of the p53 protein, and aneuploidy in patients with benign and malignant fibrous lesions, including 19 with nodular fasciitis (cellular type) (6-88 years old, mean 42.9), 11 with abdominal fibromatoses (22-74 years old, mean 37.9), 13 with extraabdominal fibromatoses (2-38 years old, mean 19.5), and 23 with fibrosarcomas (adult type: 16-71 years old, mean 47.3; infantile type: 3 months to 9 years, mean 2.9) using immunohistochemistry to determine proliferating cell nuclear antigen (PC10) and p53 protein (CM1) as well as performing DNA flow cytometry. The proliferating cell nuclear antigen (PCNA) score was measured as the ratio of PCNA-positive nuclear size/total nuclear size determined by an image analysis computer system. The distribution pattern of the PCNA-positive cells was uneven in each instance of nodular fasciitis, in contrast to the distribution in abdominal fibromatosis, extraabdominal fibromatosis, and fibrosarcoma. Both fibrosarcoma (28.4 +/- 20.0) and nodular fasciitis (33.6 +/- 20.9) exhibited a larger value and a greater variation in the PCNA score than did either abdominal (13.5 +/- 14.5) or extraabdominal fibromatosis (19.9 +/- 21.5). Abdominal fibromatosis exhibited a smaller value and less variation in the score. In short, the PCNA score did not correlate with the malignant potential. The proliferative index (S + G2 + M fraction) in fibrosarcoma was significantly higher than in either nodular fasciitis or abdominal fibromatosis. Aneuploidy was detected in five cases (26%) of fibrosarcoma, while six (26%) fibrosarcomas showed p53 positivity. Furthermore, p53-positive patients had a worse survival (0.01 < p < 0.05), and p53 positivity correlated with the proliferative index (p < 0.01). In conclusion, the PCNA score simply indicates the proliferative activity independent of malignant potential. On the other hand, p53 positivity, proliferative index, and aneuploidy are all indicators of

  9. Computational cell analysis for label-free detection of cell properties in a microfluidic laminar flow.

    PubMed

    Zhang, Alex Ce; Gu, Yi; Han, Yuanyuan; Mei, Zhe; Chiu, Yu-Jui; Geng, Lina; Cho, Sung Hwan; Lo, Yu-Hwa

    2016-06-20

    Although a flow cytometer, being one of the most popular research and clinical tools for biomedicine, can analyze cells based on the cell size, internal structures such as granularity, and molecular markers, it provides little information about the physical properties of cells such as cell stiffness and physical interactions between the cell membrane and fluid. In this paper, we propose a computational cell analysis technique using cells' different equilibrium positions in a laminar flow. This method utilizes a spatial coding technique to acquire the spatial position of the cell in a microfluidic channel and then uses mathematical algorithms to calculate the ratio of cell mixtures. Most uniquely, the invented computational cell analysis technique can unequivocally detect the subpopulation of each cell type without labeling even when the cell type shows a substantial overlap in the distribution plot with other cell types, a scenario limiting the use of conventional flow cytometers and machine learning techniques. To prove this concept, we have applied the computation method to distinguish live and fixed cancer cells without labeling, count neutrophils from human blood, and distinguish drug treated cells from untreated cells. Our work paves the way for using computation algorithms and fluidic dynamic properties for cell classification, a label-free method that can potentially classify over 200 types of human cells. Being a highly cost-effective cell analysis method complementary to flow cytometers, our method can offer orthogonal tests in companion with flow cytometers to provide crucial information for biomedical samples. PMID:27163941

  10. Upward swimming of a sperm cell in shear flow

    NASA Astrophysics Data System (ADS)

    Omori, Toshihiro; Ishikawa, Takuji

    2016-03-01

    Mammalian sperm cells are required to swim over long distances, typically around 1000-fold their own length. They must orient themselves and maintain a swimming motion to reach the ovum, or egg cell. Although the mechanism of long-distance navigation is still unclear, one possible mechanism, rheotaxis, was reported recently. This work investigates the mechanism of the rheotaxis in detail by simulating the motions of a sperm cell in shear flow adjacent to a flat surface. A phase diagram was developed to show the sperm's swimming motion under different shear rates, and for varying flagellum waveform conditions. The results showed that, under shear flow, the sperm is able to hydrodynamically change its swimming direction, allowing it to swim upwards against the flow, which suggests that the upward swimming of sperm cells can be explained using fluid mechanics, and this can then be used to further understand physiology of sperm cell navigation.

  11. Upward swimming of a sperm cell in shear flow.

    PubMed

    Omori, Toshihiro; Ishikawa, Takuji

    2016-03-01

    Mammalian sperm cells are required to swim over long distances, typically around 1000-fold their own length. They must orient themselves and maintain a swimming motion to reach the ovum, or egg cell. Although the mechanism of long-distance navigation is still unclear, one possible mechanism, rheotaxis, was reported recently. This work investigates the mechanism of the rheotaxis in detail by simulating the motions of a sperm cell in shear flow adjacent to a flat surface. A phase diagram was developed to show the sperm's swimming motion under different shear rates, and for varying flagellum waveform conditions. The results showed that, under shear flow, the sperm is able to hydrodynamically change its swimming direction, allowing it to swim upwards against the flow, which suggests that the upward swimming of sperm cells can be explained using fluid mechanics, and this can then be used to further understand physiology of sperm cell navigation. PMID:27078385

  12. Upward swimming of a sperm cell in shear flow.

    PubMed

    Omori, Toshihiro; Ishikawa, Takuji

    2016-03-01

    Mammalian sperm cells are required to swim over long distances, typically around 1000-fold their own length. They must orient themselves and maintain a swimming motion to reach the ovum, or egg cell. Although the mechanism of long-distance navigation is still unclear, one possible mechanism, rheotaxis, was reported recently. This work investigates the mechanism of the rheotaxis in detail by simulating the motions of a sperm cell in shear flow adjacent to a flat surface. A phase diagram was developed to show the sperm's swimming motion under different shear rates, and for varying flagellum waveform conditions. The results showed that, under shear flow, the sperm is able to hydrodynamically change its swimming direction, allowing it to swim upwards against the flow, which suggests that the upward swimming of sperm cells can be explained using fluid mechanics, and this can then be used to further understand physiology of sperm cell navigation.

  13. Adhesion behavior of endothelial progenitor cells to endothelial cells in simple shear flow

    NASA Astrophysics Data System (ADS)

    Gong, Xiao-Bo; Li, Yu-Qing; Gao, Quan-Chao; Cheng, Bin-Bin; Shen, Bao-Rong; Yan, Zhi-Qiang; Jiang, Zong-Lai

    2011-12-01

    The adhesion of endothelial progenitor cells (EPCs) on endothelial cells (ECs) is one of the critical physiological processes for the regenesis of vascular vessels and the prevention of serious cardiovascular diseases. Here, the rolling and adhesion behavior of EPCs on ECs was studied numerically. A two-dimensional numerical model was developed based on the immersed boundary method for simulating the rolling and adhesion of cells in a channel flow. The binding force arising from the catch bond of a receptor and ligand pair was modeled with stochastic Monte Carlo method and Hookean spring model. The effect of tumor necrosis factor alpha (TNF- α) on the expression of the number of adhesion molecules in ECs was analyzed experimentally. A flow chamber system with CCD camera was set up to observe the top view of the rolling of EPCs on the substrate cultivated with ECs. Numerical results prove that the adhesion of EPC on ECs is closely related to membrane stiffness of the cell and shear rate of the flow. It also suggests that the adhesion force between EPC and EC by P-selectin glycoprotein ligand-1 only is not strong enough to bond the cell onto vessel walls unless contributions of other catch bond are considered. Experimental results demonstrate that TNF- α enhanced the expressions of VCAM, ICAM, P-selectin and E-selectin in ECs, which supports the numerical results that the rolling velocity of EPC on TNF- α treated EC substrate decreases obviously compared with its velocity on the untreated one. It is found that because the adhesion is affected by both the rolling velocity and the deformability of the cell, an optimal stiffness of EPC may exist at a given shear rate of flow for achieving maximum adhesion rates.

  14. Dynamic modes of red blood cells in oscillatory shear flow

    NASA Astrophysics Data System (ADS)

    Noguchi, Hiroshi

    2010-06-01

    The dynamics of red blood cells (RBCs) in oscillatory shear flow was studied using differential equations of three variables: a shape parameter, the inclination angle θ , and phase angle ϕ of the membrane rotation. In steady shear flow, three types of dynamics occur depending on the shear rate and viscosity ratio. (i) tank-treading (TT): ϕ rotates while the shape and θ oscillate. (ii) tumbling (TB): θ rotates while the shape and ϕ oscillate. (iii) intermediate motion: both ϕ and θ rotate synchronously or intermittently. In oscillatory shear flow, RBCs show various dynamics based on these three motions. For a low shear frequency with zero mean shear rate, a limit-cycle oscillation occurs, based on the TT or TB rotation at a high or low shear amplitude, respectively. This TT-based oscillation well explains recent experiments. In the middle shear amplitude, RBCs show an intermittent or synchronized oscillation. As shear frequency increases, the vesicle oscillation becomes delayed with respect to the shear oscillation. At a high frequency, multiple limit-cycle oscillations coexist. The thermal fluctuations can induce transitions between two orbits at very low shear amplitudes. For a high mean shear rate with small shear oscillation, the shape and θ oscillate in the TT motion but only one attractor exists even at high shear frequencies. The measurement of these oscillatory modes is a promising tool for quantifying the viscoelasticity of RBCs, synthetic capsules, and lipid vesicles.

  15. Rapid Cell Population Identification in Flow Cytometry Data*

    PubMed Central

    Aghaeepour, Nima; Nikolic, Radina; Hoos, Holger H.; Brinkman, Ryan R.

    2011-01-01

    We have developed flowMeans, a time-efficient and accurate method for automated identification of cell populations in flow cytometry (FCM) data based on K-means clustering. Unlike traditional K-means, flowMeans can identify concave cell populations by modelling a single population with multiple clusters. flowMeans uses a change point detection algorithm to determine the number of sub-populations, enabling the method to be used in high throughput FCM data analysis pipelines. Our approach compares favourably to manual analysis by human experts and current state-of-the-art automated gating algorithms. flowMeans is freely available as an open source R package through Bioconductor. PMID:21182178

  16. Fluid flow plate for decreased density of fuel cell assembly

    DOEpatents

    Vitale, Nicholas G.

    1999-01-01

    A fluid flow plate includes first and second outward faces. Each of the outward faces has a flow channel thereon for carrying respective fluid. At least one of the fluids serves as reactant fluid for a fuel cell of a fuel cell assembly. One or more pockets are formed between the first and second outward faces for decreasing density of the fluid flow plate. A given flow channel can include one or more end sections and an intermediate section. An interposed member can be positioned between the outward faces at an interface between an intermediate section, of one of the outward faces, and an end section, of that outward face. The interposed member can serve to isolate the reactant fluid from the opposing outward face. The intermediate section(s) of flow channel(s) on an outward face are preferably formed as a folded expanse.

  17. Combustor air flow control method for fuel cell apparatus

    DOEpatents

    Clingerman, Bruce J.; Mowery, Kenneth D.; Ripley, Eugene V.

    2001-01-01

    A method for controlling the heat output of a combustor in a fuel cell apparatus to a fuel processor where the combustor has dual air inlet streams including atmospheric air and fuel cell cathode effluent containing oxygen depleted air. In all operating modes, an enthalpy balance is provided by regulating the quantity of the air flow stream to the combustor to support fuel cell processor heat requirements. A control provides a quick fast forward change in an air valve orifice cross section in response to a calculated predetermined air flow, the molar constituents of the air stream to the combustor, the pressure drop across the air valve, and a look up table of the orifice cross sectional area and valve steps. A feedback loop fine tunes any error between the measured air flow to the combustor and the predetermined air flow.

  18. Fuel cell with metal screen flow-field

    DOEpatents

    Wilson, M.S.; Zawodzinski, C.

    1998-08-25

    A polymer electrolyte membrane (PEM) fuel cell is provided with electrodes supplied with a reactant on each side of a catalyzed membrane assembly (CMA). The fuel cell includes a metal mesh defining a rectangular flow-field pattern having an inlet at a first corner and an outlet at a second corner located on a diagonal from the first corner, wherein all flow paths from the inlet to the outlet through the square flow field pattern are equivalent to uniformly distribute the reactant over the CMA. In a preferred form of metal mesh, a square weave screen forms the flow-field pattern. In a particular characterization of the present invention, a bipolar plate electrically connects adjacent fuel cells, where the bipolar plate includes a thin metal foil having an anode side and a cathode side; a first metal mesh on the anode side of the thin metal foil; and a second metal mesh on the cathode side of the thin metal foil. In another characterization of the present invention, a cooling plate assembly cools adjacent fuel cells, where the cooling plate assembly includes an anode electrode and a cathode electrode formed of thin conducting foils; and a metal mesh flow field there between for distributing cooling water flow over the electrodes to remove heat generated by the fuel cells. 11 figs.

  19. Fuel cell with metal screen flow-field

    DOEpatents

    Wilson, Mahlon S.; Zawodzinski, Christine

    2001-01-01

    A polymer electrolyte membrane (PEM) fuel cell is provided with electrodes supplied with a reactant on each side of a catalyzed membrane assembly (CMA). The fuel cell includes a metal mesh defining a rectangular flow-field pattern having an inlet at a first corner and an outlet at a second corner located on a diagonal from the first corner, wherein all flow paths from the inlet to the outlet through the square flow field pattern are equivalent to uniformly distribute the reactant over the CMA. In a preferred form of metal mesh, a square weave screen forms the flow-field pattern. In a particular characterization of the present invention, a bipolar plate electrically connects adjacent fuel cells, where the bipolar plate includes a thin metal foil having an anode side and a cathode side; a first metal mesh on the anode side of the thin metal foil; and a second metal mesh on the cathode side of the thin metal foil. In another characterization of the present invention, a cooling plate assembly cools adjacent fuel cells, where the cooling plate assembly includes an anode electrode and a cathode electrode formed of thin conducting foils; and a metal mesh flow field therebetween for distributing cooling water flow over the electrodes to remove heat generated by the fuel cells.

  20. Fuel cell with metal screen flow-field

    DOEpatents

    Wilson, Mahlon S.; Zawodzinski, Christine

    1998-01-01

    A polymer electrolyte membrane (PEM) fuel cell is provided with electrodes supplied with a reactant on each side of a catalyzed membrane assembly (CMA). The fuel cell includes a metal mesh defining a rectangular flow-field pattern having an inlet at a first corner and an outlet at a second corner located on a diagonal from the first corner, wherein all flow paths from the inlet to the outlet through the square flow field pattern are equivalent to uniformly distribute the reactant over the CMA. In a preferred form of metal mesh, a square weave screen forms the flow-field pattern. In a particular characterization of the present invention, a bipolar plate electrically connects adjacent fuel cells, where the bipolar plate includes a thin metal foil having an anode side and a cathode side; a first metal mesh on the anode side of the thin metal foil; and a second metal mesh on the cathode side of the thin metal foil. In another characterization of the present invention, a cooling plate assembly cools adjacent fuel cells, where the cooling plate assembly includes an anode electrode and a cathode electrode formed of thin conducting foils; and a metal mesh flow field therebetween for distributing cooling water flow over the electrodes to remove heat generated by the fuel cells.

  1. New all-vanadium redox flow cell

    NASA Astrophysics Data System (ADS)

    Skyllas-Kazacos, M.; Rychcik, M.; Robins, R. G.; Fane, A. G.; Green, M. A.

    1986-05-01

    A laboratory-scale cell was constructed to test the performance of V(II)/V(III) and V(IV)/V(V) half-cells in an all-vanadium redox battery. Graphite plates were used as electrodes, and the membrane was manufactured from a sulfonated polyehylene anion-selective material. The average charging efficiency of the cell was over 90 percent. Stability tests on the reduced and oxidized electrolytes, measured over the temperature range of -5 C to 60 C, showed no accelerated decomposition at high temperatures and no crystallization at the lower temperatures. After prolonged usage, however, a slow deterioration of the positive electrode and the membrane was observed.

  2. Full dynamics of a red blood cell in shear flow.

    PubMed

    Dupire, Jules; Socol, Marius; Viallat, Annie

    2012-12-18

    At the cellular scale, blood fluidity and mass transport depend on the dynamics of red blood cells in blood flow, specifically on their deformation and orientation. These dynamics are governed by cellular rheological properties, such as internal viscosity and cytoskeleton elasticity. In diseases in which cell rheology is altered genetically or by parasitic invasion or by changes in the microenvironment, blood flow may be severely impaired. The nonlinear interplay between cell rheology and flow may generate complex dynamics, which remain largely unexplored experimentally. Under simple shear flow, only two motions, "tumbling" and "tank-treading," have been described experimentally and relate to cell mechanics. Here, we elucidate the full dynamics of red blood cells in shear flow by coupling two videomicroscopy approaches providing multidirectional pictures of cells, and we analyze the mechanical origin of the observed dynamics. We show that contrary to common belief, when red blood cells flip into the flow, their orientation is determined by the shear rate. We discuss the "rolling" motion, similar to a rolling wheel. This motion, which permits the cells to avoid energetically costly deformations, is a true signature of the cytoskeleton elasticity. We highlight a hysteresis cycle and two transient dynamics driven by the shear rate: an intermittent regime during the "tank-treading-to-flipping" transition and a Frisbee-like "spinning" regime during the "rolling-to-tank-treading" transition. Finally, we reveal that the biconcave red cell shape is highly stable under moderate shear stresses, and we interpret this result in terms of stress-free shape and elastic buckling. PMID:23213229

  3. Sonochemical destruction of chloroform by using low frequency ultrasound in batch and flow cell.

    PubMed

    Thangavadivel, Kandasamy; Megharaj, Mallavarapu; Smart, Roger St C; Lesniewski, Peter J; Naidu, Ravi

    2010-01-01

    Ultrasound assisted environmental remediation is emerging as a viable technology to remove organic pollutants. In this study, the potential of low frequency ultrasound (20 kHz) to remediate chloroform contaminated waters (demineralised water and groundwater) in batch and flow cell treatment was evaluated. The results show that approx. 8 mg/L of chloroform was completely mineralized within 60 min in batch as well as flow cell treatments in both waters. However, flow cell treatment was superior to the batch mode for demineralised water in contrast to the groundwater for which there was no appreciable difference between batch and flow cell modes during initial 30 min. The presence of dissolved organic carbon, higher total dissolved solid content and any other co-contaminants might have contributed to the slower rate of chloroform destruction in the groundwater compared to the demineralised water. This study demonstrates the potential of low frequency ultrasound for remediation of chloroform contaminated waters.

  4. Pockels-effect cell for gas-flow simulation

    NASA Technical Reports Server (NTRS)

    Weimer, D.

    1982-01-01

    A Pockels effect cell using a 75 cu cm DK*P crystal was developed and used as a gas flow simulator. Index of refraction gradients were produced in the cell by the fringing fields of parallel plate electrodes. Calibration curves for the device were obtained for index of refraction gradients in excess of .00025 m.

  5. Design of a miniature flow cell for in situ x-ray imaging of redox flow batteries

    NASA Astrophysics Data System (ADS)

    Jervis, Rhodri; Brown, Leon D.; Neville, Tobias P.; Millichamp, Jason; Finegan, Donal P.; Heenan, Thomas M. M.; Brett, Dan J. L.; Shearing, Paul R.

    2016-11-01

    Flow batteries represent a possible grid-scale energy storage solution, having many advantages such as scalability, separation of power and energy capabilities, and simple operation. However, they can suffer from degradation during operation and the characteristics of the felt electrodes are little understood in terms of wetting, compression and pressure drops. Presented here is the design of a miniature flow cell that allows the use of x-ray computed tomography (CT) to study carbon felt materials in situ and operando, in both lab-based and synchrotron CT. Through application of the bespoke cell it is possible to observe felt fibres, electrolyte and pore phases and therefore enables non-destructive characterisation of an array of microstructural parameters during the operation of flow batteries. Furthermore, we expect this design can be readily adapted to the study of other electrochemical systems.

  6. Margination of White Blood Cells in Microcapillary Flow

    NASA Astrophysics Data System (ADS)

    Fedosov, Dmitry A.; Fornleitner, Julia; Gompper, Gerhard

    2012-01-01

    Margination of white blood cells (WBCs) towards vessel walls is an essential precondition for their efficient adhesion to the vascular endothelium. We perform numerical simulations with a two-dimensional blood flow model to investigate the dependence of WBC margination on hydrodynamic interactions of blood cells with the vessel walls, as well as on their collective behavior and deformability. We find WBC margination to be optimal in intermediate ranges of red blood cell (RBC) volume fractions and flow rates, while, beyond these ranges, it is substantially attenuated. RBC aggregation enhances WBC margination, while WBC deformability reduces it. These results are combined in state diagrams, which identify WBC margination for a wide range of flow and cell suspension conditions.

  7. Development of bipolar plates with different flow channel configurations for fuel cells

    NASA Astrophysics Data System (ADS)

    Boddu, Rajesh; Marupakula, Uday Kumar; Summers, Benjamin; Majumdar, Pradip

    Bipolar plates include separate gas flow channels for anode and cathode electrodes of a fuel cell. These gases flow channels supply reactant gasses as well as remove products from the cathode side of the fuel cell. Fluid flow, heat and mass transport processes in these channels have significant effect on fuel cell performance, particularly to the mass transport losses. The design of the bipolar plates should minimize plate thickness for low volume and mass. Additionally, contact faces should provide a high degree of surface uniformity for low thermal and electrical contact resistances. Finally, the flow fields should provide for efficient heat and mass transport processes with reduced pressure drops. In this study, bipolar plates with different serpentine flow channel configurations are analyzed using computational fluid dynamics modeling. Flow characteristics including variation of pressure in the flow channel across the bipolar plate are presented. Pressure drop characteristics for different flow channel designs are compared. Results show that with increased number of parallel channels and smaller sizes, a more effective contact surface area along with decreased pressured drop can be achieved. Correlations of such entrance region coefficients will be useful for the PEM fuel cell simulation model to evaluate the affects of the bipolar plate design on mass transfer loss and hence on the total current and power density of the fuel cell.

  8. Basic flow structure in saccular aneurysms: a flow visualization study.

    PubMed

    Steiger, H J; Poll, A; Liepsch, D; Reulen, H J

    1987-01-01

    Basic flow patterns were investigated in a set of glass aneurysm models by means of flow visualization methods. Dye injection and streaming double refraction were used to visualize flow. The circulation inside lateral aneurysms arising at a 90 degree angle from a straight parent conduit could not be visualized by the dye-injection technique but could be demonstrated by streaming double refraction. The inflow was seen to arise from the downstream lip of the orifice and to project to the dome of the aneurysm. Backflow to the parent conduit took place along the walls of the aneurysm. In aneurysms located at bifurcations, flow characteristics depended on the geometry of the bifurcation and the flow ratio between the branches. Relatively little intra-aneurysmal flow was demonstrated in side branch-related aneurysms arising distal to an asymmetric 90 degrees bifurcation of the type encountered at the junction of the internal carotid and posterior communicating arteries. Stagnation of flow at the neck and little intra-aneurysmal circulation were found with terminal aneurysms of the basilar bifurcation type if the outflow through the branches was symmetric. With asymmetric outflow, however, or if the axis of the aneurysm did not coincide with that of the afferent vessel, an active rotation developed in these aneurysms. The size of the aneurysm had no influence on the basic pattern of intra-aneurysmal circulation. The use of pulsatile perfusion did not significantly alter the basic flow patterns observed with steady flow. Locally disturbed laminar flow was observed in certain models at physiological Reynold's numbers, but there were no signs of fully developed turbulence.

  9. Experimental study of turbulent axisymmetric cavity flow

    NASA Astrophysics Data System (ADS)

    Lee, D. H.; Sung, H. J.

    1994-08-01

    An experimental study is made of turbulent axisymmetric cavity flow. The flow configuration consists of a sudden expansion and contraction pipe joint. In using the LDV system, in an effort to minimize refraction of laser beams at the curved interface, a refraction correction formula for the Reynolds shear stress is devised. Three values of the cavity length ( L = 300, 600 and 900 mm) are chosen, and the cavity height ( H) is fixed at 55 mm. Both open and closed cavities are considered. Special attention is given to the critical case L = 600 mm, where the cavity length L is nearly equal to the reattachment length of the flow. The Reynolds number, based on the inlet diameter ( D = 110 mm) is 73,000. Measurement data are presented for the static wall pressure, mean velocity profiles, vorticity thickness distributions, and turbulence quantities.

  10. Detection of circulating breast cancer cells using photoacoustic flow cytometry

    NASA Astrophysics Data System (ADS)

    Bhattacharyya, Kiran

    According to the American Cancer Society, more than 200,000 new cases of breast cancer are expected to be diagnosed this year. Moreover, about 40,000 women died from breast cancer last year alone. As breast cancer progresses in an individual, it can transform from a localized state to a metastatic one with multiple tumors distributed through the body, not necessarily contained within the breast. Metastasis is the spread of cancer through the body by circulating tumor cells (CTCs) which can be found in the blood and lymph of the diagnosed patient. Diagnosis of a metastatic state by the discovery of a secondary tumor can often come too late and hence, significantly reduce the patient's chance of survival. There is a current need for a CTC detection method which would diagnose metastasis before the secondary tumor occurs or reaches a size resolvable by current imaging systems. Since earlier detection would improve prognosis, this study proposes a method of labeling of breast cancer cells for detection with a photoacoustic flow cytometry system as a model for CTC detection in human blood. Gold nanoparticles and fluorescent polystyrene nanoparticles are proposed as contrast agents for T47D, the breast cancer cell line of choice. The labeling, photoacoustic detection limit, and sensitivity are first characterized and then applied to a study to show detection from human blood.

  11. Liquid chromatography/Fourier transform IR spectrometry interface flow cell

    DOEpatents

    Johnson, Charles C.; Taylor, Larry T.

    1986-01-01

    A zero dead volume (ZDV) microbore high performance liquid chromatography (.mu.HPLC)/Fourier transform infrared (FTIR) interface flow cell includes an IR transparent crystal having a small diameter bore therein through which a sample liquid is passed. The interface flow cell further includes a metal holder in combination with a pair of inner, compressible seals for directly coupling the thus configured spectrometric flow cell to the outlet of a .mu.HPLC column end fitting to minimize the transfer volume of the effluents exiting the .mu.HPLC column which exhibit excellent flow characteristics due to the essentially unencumbered, open-flow design. The IR beam passes transverse to the sample flow through the circular bore within the IR transparent crystal, which is preferably comprised of potassium bromide (KBr) or calcium fluoride (CaF.sub.2), so as to minimize interference patterns and vignetting encountered in conventional parallel-plate IR cells. The long IR beam pathlength and lensing effect of the circular cross-section of the sample volume in combination with the refractive index differences between the solvent and the transparent crystal serve to focus the IR beam in enhancing sample detection sensitivity by an order of magnitude.

  12. Liquid chromatography/Fourier transform IR spectrometry interface flow cell

    DOEpatents

    Johnson, C.C.; Taylor, L.T.

    1985-01-04

    A zero dead volume (ZDV) microbore high performance liquid chromatography (..mu.. HPLC)/Fourier transform infrared (FTIR) interface flow cell includes an IR transparent crystal having a small diameter bore therein through which a sample liquid is passed. The interface flow cell further includes a metal holder in combination with a pair of inner, compressible seals for directly coupling the thus configured spectrometric flow cell to the outlet of a ..mu.. HPLC column end fitting to minimize the transfer volume of the effluents exiting the ..mu.. HPLC column which exhibit excellent flow characteristics due to the essentially unencumbered, open-flow design. The IR beam passes transverse to the sample flow through the circular bore within the IR transparent crystal, which is preferably comprised of potassium bromide (KBr) or calcium fluoride (CaF/sub 2/), so as to minimize interference patterns and vignetting encountered in conventional parallel-plate IR cells. The long IR beam pathlength and lensing effect of the circular cross-section of the sample volume in combination with the refractive index differences between the solvent and the transparent crystal serve to focus the IR beam in enhancing sample detection sensitivity by an order of magnitude.

  13. Numerical study of secondary flows in curved ducts

    SciTech Connect

    Hur, N.

    1988-01-01

    The secondary flow of an incompressible viscous fluid in a curved duct with and without sidewall heating is studied by using a finite-volume method. It is known that for low Dean numbers, the secondary flow is characterized by a pair of counter-rotating vortices. This study shows that, as the Dean number is increased, the secondary-flow structure evolves into a double vortex pair for low-aspect-ratio ducts and roll cells for ducts of high aspect ratio. A stability diagram is obtained in the domain of curvature ratio and Reynolds number. It is found that, for ducts of high curvature, the onset of instability depends on the Dean number and the curvature ratio while, for ducts of small curvature, the onset can be characterized by the Dean number alone. A comparison with the available theoretical and experimental results indicates good agreement. When there is sidewall heating, the interaction between the centrifugal and the buoyancy forces characterizes the secondary flow structure. As the Grashof number is increased, the friction factor can decrease due to the transition from centrifugally dominant flow to buoyancy-dominated flow. Also, for curved ducts the inertial effect dominates and the heat transfer may be enhanced at lower Grashof numbers.

  14. How cells flow in the spreading of cellular aggregates

    PubMed Central

    Beaune, Grégory; Stirbat, Tomita Vasilica; Khalifat, Nada; Cochet-Escartin, Olivier; Garcia, Simon; Gurchenkov, Vasily Valérïévitch; Murrell, Michael P.; Dufour, Sylvie; Cuvelier, Damien; Brochard-Wyart, Françoise

    2014-01-01

    Like liquid droplets, cellular aggregates, also called “living droplets,” spread onto adhesive surfaces. When deposited onto fibronectin-coated glass or polyacrylamide gels, they adhere and spread by protruding a cellular monolayer (precursor film) that expands around the droplet. The dynamics of spreading results from a balance between the pulling forces exerted by the highly motile cells at the periphery of the film, and friction forces associated with two types of cellular flows: (i) permeation, corresponding to the entry of the cells from the aggregates into the film; and (ii) slippage as the film expands. We characterize these flow fields within a spreading aggregate by using fluorescent tracking of individual cells and particle imaging velocimetry of cell populations. We find that permeation is limited to a narrow ring of width ξ (approximately a few cells) at the edge of the aggregate and regulates the dynamics of spreading. Furthermore, we find that the subsequent spreading of the monolayer depends heavily on the substrate rigidity. On rigid substrates, the migration of the cells in the monolayer is similar to the flow of a viscous liquid. By contrast, as the substrate gets softer, the film under tension becomes unstable with nucleation and growth of holes, flows are irregular, and cohesion decreases. Our results demonstrate that the mechanical properties of the environment influence the balance of forces that modulate collective cell migration, and therefore have important implications for the spreading behavior of tissues in both early development and cancer. PMID:24835175

  15. Principles of bone marrow processing and progenitor cell/mononuclear cell concentrate collection in a continuous flow blood cell separation system.

    PubMed

    Hester, J P; Rondón, G; Huh, Y O; Lauppe, M J; Champlin, R E; Deisseroth, A B

    1995-08-01

    The application of continuous flow apheresis technology to processing bone marrow for collection of the mononuclear progenitor cell population appears to follow the same principles as collection of mononuclear cells from peripheral blood. Unlike peripheral blood, however, where mobilization of cells from extravascular sites during the procedures contributes significantly to the final cell yield, the entire quantity of progenitor cells available for recovery from marrow is present in the original marrow when it is pooled. The process then becomes one of attempting optimal recovery of the cells of interest while excluding contaminating erythrocytes and cells of the myeloid series. This study reports the development of a protocol for recovery of MNC, CD33+, CD34+, and CD34+/DR- cells from harvested marrow for autologous and allogeneic transplants using a continuous flow blood cell separator, the variables influencing the recovery of the cells of interest and the clinical response to infusion of the processed cells.

  16. Flow rate and humidification effects on a PEM fuel cell performance and operation

    NASA Astrophysics Data System (ADS)

    Guvelioglu, Galip H.; Stenger, Harvey G.

    A new algorithm is presented to integrate component balances along polymer electrolyte membrane fuel cell (PEMFC) channels to obtain three-dimensional results from a detailed two-dimensional finite element model. The analysis studies the cell performance at various hydrogen flow rates, air flow rates and humidification levels. This analysis shows that hydrogen and air flow rates and their relative humidity are critical to current density, membrane dry-out, and electrode flooding. Uniform current densities along the channels are known to be critical for thermal management and fuel cell life. This approach, of integrating a detailed two-dimensional across-the-channel model, is a promising method for fuel cell design due to its low computational cost compared to three-dimensional computational fluid dynamics models, its applicability to a wide range of fuel cell designs, and its ease of extending to fuel cell stack models.

  17. Three-dimensional cell culture model for measuring the effects of interstitial fluid flow on tumor cell invasion.

    PubMed

    Tchafa, Alimatou M; Shah, Arpit D; Wang, Shafei; Duong, Melissa T; Shieh, Adrian C

    2012-07-25

    fluid flow on cells in vitro and quantifies its effects on invasion (Figure 1). This method has been published in multiple studies to measure the effects of fluid flow on stromal and cancer cell invasion (13-15, 17). By changing the matrix composition, cell type, and cell concentration, this method can be applied to other diseases and physiological systems to study the effects of interstitial flow on cellular processes such as invasion, differentiation, proliferation, and gene expression.

  18. Numerical Study of Tip Vortex Flows

    NASA Technical Reports Server (NTRS)

    Dacles-Mariani, Jennifer; Hafez, Mohamed

    1998-01-01

    This paper presents an overview and summary of the many different research work related to tip vortex flows and wake/trailing vortices as applied to practical engineering problems. As a literature survey paper, it outlines relevant analytical, theoretical, experimental and computational study found in literature. It also discusses in brief some of the fundamental aspects of the physics and its complexities. An appendix is also included. The topics included in this paper are: 1) Analytical Vortices; 2) Experimental Studies; 3) Computational Studies; 4) Wake Vortex Control and Management; 5) Wake Modeling; 6) High-Lift Systems; 7) Issues in Numerical Studies; 8) Instabilities; 9) Related Topics; 10) Visualization Tools for Vertical Flows; 11) Further Work Needed; 12) Acknowledgements; 13) References; and 14) Appendix.

  19. Simultaneous measurement of flow over and transmigration through a cultured endothelial cell layer

    NASA Astrophysics Data System (ADS)

    Lambert, Lori; Pipinos, Iraklis; Baxter, Timothy; MacTaggart, Jason; Karniadakis, George; Moormeier, Derek; Bayles, Kenneth; Wei, Timothy

    2013-11-01

    This talk focuses on the methodologies associated with the integration of temporally and spatially resolved μPIV measurements of flow over live endothelial cells with measurements of chemical transport through and across the cells. The ultimate goal of the study is to examine and model the transport and transmigration of key agents responsible for the formation of atherosclerotic plaques. Flow over endothelial cells cultured in a microchannel was measured using μPIV. By making measurements in a number of planes parallel to the wall, key dynamic quantities such as shear and pressure distributions, along with surface topography could be computed from the flow measurements. Experiments were conducted in a 65 μm × 65 μm cross section microchannel at shear rates up to 20 dynes/cm2. Changes in cell conformation as a function of time after flow was started were examined. The deposition and transmigration of LDL was also examined using fluorescent-tagged LDL molecules.

  20. New optical configuration for flow cytometric sorting of aspherical cells

    NASA Astrophysics Data System (ADS)

    Sharpe, John C.; Schaare, Peter N.; Kuennemeyer, Rainer

    1997-05-01

    The orthogonal axes of illumination, flow, and detection in conventional sorting flow cytometers can limit accuracy or throughput when making fluorescence measurements on a spherical cells. A new radially symmetric optical configuration has been designed to overcome these problems. Both illumination and fluorescence collection are performed by a single optical element which encircles the sample stream flow axis. Unlike existing epi-illumination flow cytometer designs, these optics are compatible with electrostatic sorting. The resolution of this system is currently being evaluated for DNA chromosome content measurement with an ultimate goal of separation of X- and Y- chromosome-bearing mammalian spermatozoa. We describe the new optical configuration and present preliminary results of instrument performance. Comparison with a conventional orthogonal optical geometry is made using fluorescent microspheres, chicken red blood cells and chinchilla sperm.

  1. Good Cell, Bad Cell: Flow Cytometry Reveals T-cell Subsets Important in HIV Disease

    PubMed Central

    Chattopadhyay, Pratip K.; Roederer, Mario

    2010-01-01

    Flow cytometry is a key technology in the study of HIV disease. In this article, we review various cellular markers that can be measured in the setting of pathogenesis or vaccination studies, including markers of activation, differentiation, senescence, immune suppression, and function. In addition, we discuss important considerations for making these measurements. Finally, we examine how flow cytometry studies have taught researchers about the disease process, and the potential for flow cytometry technology to guide treatment decisions and evaluate vaccine candidates in the future. PMID:20583275

  2. Comparative study of pressure-flow parameters.

    PubMed

    Eri, Lars M; Wessel, Nicolai; Tysland, Ole; Berge, Viktor

    2002-01-01

    Methods for quantification of bladder outlet obstruction (BOO) are still controversial. Parameters such as detrusor opening pressure (p(det.open)), maximum detrusor pressure (p(det.max)), minimum voiding pressure (p(det.min.void)), and detrusor pressure at maximum flow rate (P(det.Qmax)) separate obstructed from nonobstructed patients to some extent, but two nomograms, the Abrams-Griffiths nomogram and the linearized passive urethral resistance relation (LinPURR), are more accepted for this purpose, along with the urethral resistance algorithm. In this retrospective, methodologic study, we evaluated the properties of these parameters with regard to test-retest reproducibility and ability to detect a moderate (pharmacologic) and a pronounced (surgical) relief of bladder outlet obstruction. We studied the pressure-flow charts of 42 patients who underwent 24 weeks of androgen suppressive therapy, 42 corresponding patients who received placebo, and 30 patients who had prostate surgery. The patients performed repeat void pressure-flow examinations before and after treatment or placebo. The various parameters were compared. Among the bladder pressure parameters, P(det.Qmax) seemed to have some advantages, supporting the belief that it is the most relevant detrusor pressure parameter to include in nomograms to quantify BOO. In assessment of a large decrease in urethral resistance, such as after TURp, resistance parameters that are based on maximum flow rate as well as detrusor pressure are preferable. PMID:11948710

  3. B-1 AFT Nacelle Flow Visualization Study

    NASA Technical Reports Server (NTRS)

    Celniker, Robert

    1975-01-01

    A 2-month program was conducted to perform engineering evaluation and design tasks to prepare for visualization and photography of the airflow along the aft portion of the B-1 nacelles and nozzles during flight test. Several methods of visualizing the flow were investigated and compared with respect to cost, impact of the device on the flow patterns, suitability for use in the flight environment, and operability throughout the flight. Data were based on a literature search and discussions with the test personnel. Tufts were selected as the flow visualization device in preference to several other devices studied. A tuft installation pattern has been prepared for the right-hand aft nacelle area of B-1 air vehicle No.2. Flight research programs to develop flow visualization devices other than tufts for use in future testing are recommended. A design study was conducted to select a suitable motion picture camera, to select the camera location, and to prepare engineering drawings sufficient to permit installation of the camera. Ten locations on the air vehicle were evaluated before the selection of the location in the horizontal stabilizer actuator fairing. The considerations included cost, camera angle, available volume, environmental control, flutter impact, and interference with antennas or other instrumentation.

  4. Diffusional solute flux during osmotic water flow across the human red cell membrane.

    PubMed

    Brahm, J; Galey, W R

    1987-05-01

    The effect of solvent drag on the unidirectional efflux of labeled water, urea, and chloride from human red cells was studied by means of the continuous flow tube method under conditions of osmotic equilibrium and net volume flow. Solvent (water) flow out of cells was created by mixing cells equilibrated in 100 mM salt solution with a 200-mM or 250-mM salt solution, while flow of water into cells was obtained by equilibrating the cells in the higher concentration and mixing them with the 100-mM solution. Control experiments constitute measurements of efflux of [14C]ethanol in normal cells and 3H2O in cells treated with p-chloromercuribenzosulfonate under the conditions described above. In both instances, the solute is known to penetrate the membrane through nonporous pathways. As anticipated, the tracer flux of neither urea nor chloride showed any dependence on net solvent flow, regardless of the direction. If one assumes the recently reported reflection coefficient for urea of 0.7, the urea tracer flux should change by at least 24% under volume flow conditions. Since such changes would be easily detected with our method, we conclude that the pathways for water, for urea, and for chloride are functionally separated. PMID:3037007

  5. Endothelial Cell Layer Subjected to Impinging Flow Mimicking the Apex of an Arterial Bifurcation

    PubMed Central

    Szymanski, Michael P.; Metaxa, Eleni; Meng, Hui; Kolega, John

    2008-01-01

    Little is known about endothelial responses to the impinging flow hemodynamics that occur at arterial bifurcation apices, where intracranial aneurysms usually form. Such hemodynamic environments are characterized by high wall shear stress (WSS >40 dynes/cm2) and high wall shear stress gradients (WSSG >300 dynes/cm3). In this study, confluent bovine aortic endothelial cells were exposed to impinging flow in a T-shaped chamber designed to mimic a bifurcation. After 24−72 h under flow, cells around the stagnation point maintained polygonal shapes but cell density was reduced, whereas cells in adjacent downstream regions exposed to very high WSS and WSSG were elongated, aligned parallel to flow, and at higher density. Such behavior was not blocked by inhibiting proliferation, indicating that cells migrated downstream from the stagnation point in response to impinging flow. Furthermore, although the area of highest cell density moved downstream and away from the impingement point over time, it never moved beyond the WSS maximum. The accumulation of cells upstream of maximal WSS and downstream of maximal WSSG suggests that positive WSSG is responsible for the observed migration. These results demonstrate a unique endothelial response to aneurysm-promoting flow environments at bifurcation apices. PMID:18654851

  6. Flow Cytometric Analysis of T, B, and NK Cells Antigens in Patients with Mycosis Fungoides.

    PubMed

    Yazıcı, Serkan; Bülbül Başkan, Emel; Budak, Ferah; Oral, Barbaros; Adim, Şaduman Balaban; Ceylan Kalin, Zübeyde; Özkaya, Güven; Aydoğan, Kenan; Saricaoğlu, Hayriye; Tunali, Şükran

    2015-01-01

    We retrospectively analyzed the clinicopathological correlation and prognostic value of cell surface antigens expressed by peripheral blood mononuclear cells in patients with mycosis fungoides (MF). 121 consecutive MF patients were included in this study. All patients had peripheral blood flow cytometry as part of their first visit. TNMB and histopathological staging of the cases were retrospectively performed in accordance with International Society for Cutaneous Lymphomas/European Organization of Research and Treatment of Cancer (ISCL/EORTC) criteria at the time of flow cytometry sampling. To determine prognostic value of cell surface antigens, cases were divided into two groups as stable and progressive disease. 17 flow cytometric analyses of 17 parapsoriasis (PP) and 11 analyses of 11 benign erythrodermic patients were included as control groups. Fluorescent labeled monoclonal antibodies were used to detect cell surface antigens: T cells (CD3(+), CD4(+), CD8(+), TCRαβ(+), TCRγδ(+), CD7(+), CD4(+)CD7(+), CD4(+)CD7(-), and CD71(+)), B cells (HLA-DR(+), CD19(+), and HLA-DR(+)CD19(+)), NKT cells (CD3(+)CD16(+)CD56(+)), and NK cells (CD3(-)CD16(+)CD56(+)). The mean value of all cell surface antigens was not statistically significant between parapsoriasis and MF groups. Along with an increase in cases of MF stage statistically significant difference was found between the mean values of cell surface antigens. Flow cytometric analysis of peripheral blood cell surface antigens in patients with mycosis fungoides may contribute to predicting disease stage and progression. PMID:26788525

  7. Red blood cell clustering in Poiseuille microcapillary flow

    NASA Astrophysics Data System (ADS)

    Tomaiuolo, Giovanna; Lanotte, Luca; Ghigliotti, Giovanni; Misbah, Chaouqi; Guido, Stefano

    2012-05-01

    Red blood cells (RBC) flowing in microcapillaries tend to associate into clusters, i.e., small trains of cells separated from each other by a distance comparable to cell size. This process is usually attributed to slower RBCs acting to create a sequence of trailing cells. Here, based on the first systematic investigation of collective RBC flow behavior in microcapillaries in vitro by high-speed video microscopy and numerical simulations, we show that RBC size polydispersity within the physiological range does not affect cluster stability. Lower applied pressure drops and longer residence times favor larger RBC clusters. A limiting cluster length, depending on the number of cells in a cluster, is found by increasing the applied pressure drop. The insight on the mechanism of RBC clustering provided by this work can be applied to further our understanding of RBC aggregability, which is a key parameter implicated in clotting and thrombus formation.

  8. MEMS-based flow cytometry: microfluidics-based cell identification system by fluorescent imaging.

    PubMed

    Wu, W K; Liang, C K; Huang, J Z

    2004-01-01

    This study utilizes MEMS technology to realize a novel low-cost microfluidics-based biochip system for flow-type cell handling. Powered by vacuum pump, the microfluidic driving system enables cells to move in order one by one in the biochip by an effect of sheath flow prefocus. Then, cells are guided to a fluorescent inspection region where two detection tasks such as cell image identification and cell counting are conducted. Currently, the glass-based biochip has been manufactured and all the related devices have been well set up in our laboratory. With this proposed prototype system, typical results about cell separation of yeast cell and PC-3 cell are available and their separated images are also presented, respectively. PMID:17270801

  9. Cross-flow electrochemical reactor cells, cross-flow reactors, and use of cross-flow reactors for oxidation reactions

    DOEpatents

    Balachandran, Uthamalingam; Poeppel, Roger B.; Kleefisch, Mark S.; Kobylinski, Thaddeus P.; Udovich, Carl A.

    1994-01-01

    This invention discloses cross-flow electrochemical reactor cells containing oxygen permeable materials which have both electron conductivity and oxygen ion conductivity, cross-flow reactors, and electrochemical processes using cross-flow reactor cells having oxygen permeable monolithic cores to control and facilitate transport of oxygen from an oxygen-containing gas stream to oxidation reactions of organic compounds in another gas stream. These cross-flow electrochemical reactors comprise a hollow ceramic blade positioned across a gas stream flow or a stack of crossed hollow ceramic blades containing a channel or channels for flow of gas streams. Each channel has at least one channel wall disposed between a channel and a portion of an outer surface of the ceramic blade, or a common wall with adjacent blades in a stack comprising a gas-impervious mixed metal oxide material of a perovskite structure having electron conductivity and oxygen ion conductivity. The invention includes reactors comprising first and second zones seprated by gas-impervious mixed metal oxide material material having electron conductivity and oxygen ion conductivity. Prefered gas-impervious materials comprise at least one mixed metal oxide having a perovskite structure or perovskite-like structure. The invention includes, also, oxidation processes controlled by using these electrochemical reactors, and these reactions do not require an external source of electrical potential or any external electric circuit for oxidation to proceed.

  10. Circulation times of cancer cells by in vivo flow cytometry

    NASA Astrophysics Data System (ADS)

    Zhang, Li; Li, Yan; Gu, Zhengqin; Chen, Tong; Wang, Cheng; Wei, Xunbin

    2012-03-01

    Liver cancer is one of the most common malignancies in the world, with approximately 1,000,000 cases reported every year. Hepatocellular carcinoma may metastasize to lung, bones, kidney, and many other organs. Surgical resection, liver transplantation, chemotherapy and radiation therapy are the foundation of current HCC therapies. However the outcomes are poor: the survival rate is almost zero for metastatic HCC patients. Molecular mechanisms of HCC metastasis need to be understood better and new therapies must be developed. A recently developed "in vivo flow cytometer" combined with real-time confocal fluorescence imaging are used to assess spreading and the circulation kinetics of liver tumor cells. The in vivo flow cytometer has the capability to detect and quantify continuously the number and flow characteristics of fluorescently labeled cells in vivo in real time without extracting blood sample. We have measured the depletion kinetics of two related human HCC cell lines, high-metastatic HCCLM3 cells and low-metastatic HepG2 cells, which were from the same origin and obtained by repetitive screenings in mice. >60% HCCLM3 cells are depleted within the first hour. Interestingly, the low-metastatic HepG2 cells possess noticeably slower depletion kinetics. In comparison, <40% HepG2 cells are depleted within the first hour. The differences in depletion kinetics might provide insights into early metastasis processes.

  11. Performance study of a data flow architecture

    NASA Technical Reports Server (NTRS)

    Adams, George

    1985-01-01

    Teams of scientists studied data flow concepts, static data flow machine architecture, and the VAL language. Each team mapped its application onto the machine and coded it in VAL. The principal findings of the study were: (1) Five of the seven applications used the full power of the target machine. The galactic simulation and multigrid fluid flow teams found that a significantly smaller version of the machine (16 processing elements) would suffice. (2) A number of machine design parameters including processing element (PE) function unit numbers, array memory size and bandwidth, and routing network capability were found to be crucial for optimal machine performance. (3) The study participants readily acquired VAL programming skills. (4) Participants learned that application-based performance evaluation is a sound method of evaluating new computer architectures, even those that are not fully specified. During the course of the study, participants developed models for using computers to solve numerical problems and for evaluating new architectures. These models form the bases for future evaluation studies.

  12. Cortical Flow-Driven Shapes of Nonadherent Cells

    NASA Astrophysics Data System (ADS)

    Callan-Jones, A. C.; Ruprecht, V.; Wieser, S.; Heisenberg, C. P.; Voituriez, R.

    2016-01-01

    Nonadherent polarized cells have been observed to have a pearlike, elongated shape. Using a minimal model that describes the cell cortex as a thin layer of contractile active gel, we show that the anisotropy of active stresses, controlled by cortical viscosity and filament ordering, can account for this morphology. The predicted shapes can be determined from the flow pattern only; they prove to be independent of the mechanism at the origin of the cortical flow, and are only weakly sensitive to the cytoplasmic rheology. In the case of actin flows resulting from a contractile instability, we propose a phase diagram of three-dimensional cell shapes that encompasses nonpolarized spherical, elongated, as well as oblate shapes, all of which have been observed in experiment.

  13. Gradient isolator for flow field of fuel cell assembly

    DOEpatents

    Ernst, William D.

    1999-01-01

    Isolator(s) include isolating material and optionally gasketing material strategically positioned within a fuel cell assembly. The isolating material is disposed between a solid electrolyte and a metal flow field plate. Reactant fluid carried by flow field plate channel(s) forms a generally transverse electrochemical gradient. The isolator(s) serve to isolate electrochemically a portion of the flow field plate, for example, transversely outward from the channel(s), from the electrochemical gradient. Further, the isolator(s) serve to protect a portion of the solid electrolyte from metallic ions.

  14. Gradient isolator for flow field of fuel cell assembly

    DOEpatents

    Ernst, W.D.

    1999-06-15

    Isolator(s) include isolating material and optionally gasketing material strategically positioned within a fuel cell assembly. The isolating material is disposed between a solid electrolyte and a metal flow field plate. Reactant fluid carried by flow field plate channel(s) forms a generally transverse electrochemical gradient. The isolator(s) serve to isolate electrochemically a portion of the flow field plate, for example, transversely outward from the channel(s), from the electrochemical gradient. Further, the isolator(s) serve to protect a portion of the solid electrolyte from metallic ions. 4 figs.

  15. Single-cell trapping and selective treatment via co-flow within a microfluidic platform.

    PubMed

    Benavente-Babace, A; Gallego-Pérez, D; Hansford, D J; Arana, S; Pérez-Lorenzo, E; Mujika, M

    2014-11-15

    Lab on a chip (LOC) systems provide interesting and low-cost solutions for key studies and applications in the biomedical field. Along with microfluidics, these microdevices make single-cell manipulation possible with high spatial and temporal resolution. In this work we have designed, fabricated and characterized a versatile and inexpensive microfluidic platform for on-chip selective single-cell trapping and treatment using laminar co-flow. The combination of co-existing laminar flow manipulation and hydrodynamic single-cell trapping for selective treatment offers a cost-effective solution for studying the effect of novel drugs on single-cells. The operation of the whole system is experimentally simple, highly adaptable and requires no specific equipment. As a proof of concept, a cytotoxicity study of ethanol in isolated hepatocytes is presented. The developed microfluidic platform controlled by means of co-flow is an attractive and multipurpose solution for the study of new substances of high interest in cell biology research. In addition, this platform will pave the way for the study of cell behavior under dynamic and controllable fluidic conditions providing information at the individual cell level. Thus, this analysis device could also hold a great potential to easily use the trapped cells as sensing elements expanding its functionalities as a cell-based biosensor with single-cell resolution. PMID:24907537

  16. Axial compressor middle stage secondary flow study

    NASA Technical Reports Server (NTRS)

    Wagner, J. H.; Dring, R. P.; Joslyn, H. D.

    1983-01-01

    This report describes an experimental investigation of the secondary flow within and aft of an axial compressor model with thick endwall boundary layers. The objective of the study was to obtain detailed aerodynamic and trace gas concentration traverse data aft of a well documented isolated rotor for the ultimate purpose of improving the design phases of compressor development based on an improved physical understanding of secondary flow. It was determined from the flow visualization, aerodynamic, and trace gas concentration results that the relative unloading of the midspan region of the airfoil inhibitied a fullspan separation at high loading preventing the massive radial displacement of the hub corner stall to the tip. Radial distribution of high and low total pressure fluid influenced the magnitude of the spanwise distribution of loss, such that, there was a general decreases in loss near the hub to the extent that for the least loaded case a negative loss (increase in total pressure) was observed. The ability to determine the spanwise distribution of blockage was demonstrated. Large blockage was present in the endwall regions due to the corner stall and tip leakage with little blockage in the core flow region. Hub blockage was found to increase rapidly with loading.

  17. Experimental Study of Flow in a Bifurcation

    NASA Astrophysics Data System (ADS)

    Fresconi, Frank; Prasad, Ajay

    2003-11-01

    An instability known as the Dean vortex occurs in curved pipes with a longitudinal pressure gradient. A similar effect is manifest in the flow in a converging or diverging bifurcation, such as those found in the human respiratory airways. The goal of this study is to characterize secondary flows in a bifurcation. Particle image velocimetry (PIV) and laser-induced fluorescence (LIF) experiments were performed in a clear, plastic model. Results show the strength and migration of secondary vortices. Primary velocity features are also presented along with dispersion patterns from dye visualization. Unsteadiness, associated with a hairpin vortex, was also found at higher Re. This work can be used to assess the dispersion of particles in the lung. Medical delivery systems and pollution effect studies would profit from such an understanding.

  18. Cytosolic Calcium Measurements in Renal Epithelial Cells by Flow Cytometry

    PubMed Central

    Lee, Wing-Kee; Dittmar, Thomas

    2014-01-01

    A variety of cellular processes, both physiological and pathophysiological, require or are governed by calcium, including exocytosis, mitochondrial function, cell death, cell metabolism and cell migration to name but a few. Cytosolic calcium is normally maintained at low nanomolar concentrations; rather it is found in high micromolar to millimolar concentrations in the endoplasmic reticulum, mitochondrial matrix and the extracellular compartment. Upon stimulation, a transient increase in cytosolic calcium serves to signal downstream events. Detecting changes in cytosolic calcium is normally performed using a live cell imaging set up with calcium binding dyes that exhibit either an increase in fluorescence intensity or a shift in the emission wavelength upon calcium binding. However, a live cell imaging set up is not freely accessible to all researchers. Alternative detection methods have been optimized for immunological cells with flow cytometry and for non-immunological adherent cells with a fluorescence microplate reader. Here, we describe an optimized, simple method for detecting changes in epithelial cells with flow cytometry using a single wavelength calcium binding dye. Adherent renal proximal tubule epithelial cells, which are normally difficult to load with dyes, were loaded with a fluorescent cell permeable calcium binding dye in the presence of probenecid, brought into suspension and calcium signals were monitored before and after addition of thapsigargin, tunicamycin and ionomycin. PMID:25407650

  19. Measuring Cell Death by Propidium Iodide Uptake and Flow Cytometry.

    PubMed

    Crowley, Lisa C; Scott, Adrian P; Marfell, Brooke J; Boughaba, Jeanne A; Chojnowski, Grace; Waterhouse, Nigel J

    2016-01-01

    Propidium iodide (PI) is a small fluorescent molecule that binds to DNA but cannot passively traverse into cells that possess an intact plasma membrane. PI uptake versus exclusion can be used to discriminate dead cells, in which plasma membranes become permeable regardless of the mechanism of death, from live cells with intact membranes. PI is excited by wavelengths between 400 and 600 nm and emits light between 600 and 700 nm, and is therefore compatible with lasers and photodetectors commonly available in flow cytometers. This protocol for PI staining can be used to quantitate cell death in most modern research facilities and universities. PMID:27371595

  20. Reduction of Europium in a Redox Flow Cell

    NASA Astrophysics Data System (ADS)

    Lu, Daluh; Horng, Jiin-Shiung; Tung, Chia-Pao

    1988-05-01

    An electrolytic cell similar to the iron I chromium redox flow cell was used to investigate the reduction of europium. The cell contains two compartments partitioned by an anion exchange membrane, which is permeable to chloride ions. The anolyte is ferrous chloride which is oxidized to ferric form at the anode. Rare-earth chloride prepared from Taiwan black monazite is fed as the catholyte. The reduction of europium was tested in two connected cells at 20 and 45°C. All of Eu3+ can be reduced at 45°C, and 72% of the europium can be recovered in sulfate form. In oxide form, purity is about 84%.

  1. Cell-based flow cytometry assay to measure cytotoxic activity.

    PubMed

    Noto, Alessandra; Ngauv, Pearline; Trautmann, Lydie

    2013-12-17

    Cytolytic activity of CD8+ T cells is rarely evaluated. We describe here a new cell-based assay to measure the capacity of antigen-specific CD8+ T cells to kill CD4+ T cells loaded with their cognate peptide. Target CD4+ T cells are divided into two populations, labeled with two different concentrations of CFSE. One population is pulsed with the peptide of interest (CFSE-low) while the other remains un-pulsed (CFSE-high). Pulsed and un-pulsed CD4+ T cells are mixed at an equal ratio and incubated with an increasing number of purified CD8+ T cells. The specific killing of autologous target CD4+ T cells is analyzed by flow cytometry after coculture with CD8+ T cells containing the antigen-specific effector CD8+ T cells detected by peptide/MHCI tetramer staining. The specific lysis of target CD4+ T cells measured at different effector versus target ratios, allows for the calculation of lytic units, LU₃₀/10(6) cells. This simple and straightforward assay allows for the accurate measurement of the intrinsic capacity of CD8+ T cells to kill target CD4+ T cells.

  2. Study of argon-oxygen flowing afterglow

    NASA Astrophysics Data System (ADS)

    Mazánková, V.; Trunec, D.; Navrátil, Z.; Raud, J.; Krčma, F.

    2016-06-01

    The reaction kinetics in argon-oxygen flowing afterglow (post-discharge) was studied using NO titration and optical emission spectroscopy. The flowing DC post-discharge in argon-oxygen mixture was created in a quartz tube at the total gas pressure of 1000 Pa and discharge power of 90 W. The O(3P) atom concentration was determined by NO titration at different places along the flow tube. The optical emission spectra were also measured along the flow tube. Argon spectral lines, oxygen lines at 777 nm and 844.6 nm and atmospheric A-band of {{\\text{O}}2} were identified in the spectra. Rotational temperature of {{\\text{O}}2} was determined from the oxygen atmospheric A-band and also the outer wall temperature of the flow tube was measured by a thermocouple and by an IR thermometer. A zero-dimensional kinetic model for the reactions in the afterglow was developed. This model allows the time dependencies of particle concentrations and of gas temperature to be calculated. The wall recombination probability for O(3P) atoms {γ\\text{O≤ft(\\text{P}\\right)}}=≤ft(1.63+/- 0.06\\right)× {{10}-3} and wall deactivation probability for {{\\text{O}}2} (b {{}1}Σ\\text{g}+ ) molecules {γ{{\\text{O}2}≤ft(\\text{b}\\right)}}=≤ft(1.7+/- 0.1\\right)× {{10}-3} were determined from the fit of model results to experimental data. Sensitivity analysis was applied for the analysis of kinetic model in order to reveal the most important reactions in the model. The calculated gas temperature increases in the afterglow and then decreases at later afterglow times after reaching the maximum. This behavior is in good agreement with the spatial rotational temperature dependence. A similar trend was also observed at outer wall temperature measurement.

  3. Study of argon–oxygen flowing afterglow

    NASA Astrophysics Data System (ADS)

    Mazánková, V.; Trunec, D.; Navrátil, Z.; Raud, J.; Krčma, F.

    2016-06-01

    The reaction kinetics in argon–oxygen flowing afterglow (post-discharge) was studied using NO titration and optical emission spectroscopy. The flowing DC post-discharge in argon–oxygen mixture was created in a quartz tube at the total gas pressure of 1000 Pa and discharge power of 90 W. The O(3P) atom concentration was determined by NO titration at different places along the flow tube. The optical emission spectra were also measured along the flow tube. Argon spectral lines, oxygen lines at 777 nm and 844.6 nm and atmospheric A-band of {{\\text{O}}2} were identified in the spectra. Rotational temperature of {{\\text{O}}2} was determined from the oxygen atmospheric A-band and also the outer wall temperature of the flow tube was measured by a thermocouple and by an IR thermometer. A zero-dimensional kinetic model for the reactions in the afterglow was developed. This model allows the time dependencies of particle concentrations and of gas temperature to be calculated. The wall recombination probability for O(3P) atoms {γ\\text{O≤ft(\\text{P}\\right)}}=≤ft(1.63+/- 0.06\\right)× {{10}-3} and wall deactivation probability for {{\\text{O}}2} (b {{}1}Σ\\text{g}+ ) molecules {γ{{\\text{O}2}≤ft(\\text{b}\\right)}}=≤ft(1.7+/- 0.1\\right)× {{10}-3} were determined from the fit of model results to experimental data. Sensitivity analysis was applied for the analysis of kinetic model in order to reveal the most important reactions in the model. The calculated gas temperature increases in the afterglow and then decreases at later afterglow times after reaching the maximum. This behavior is in good agreement with the spatial rotational temperature dependence. A similar trend was also observed at outer wall temperature measurement.

  4. Transient studies of capillary-induced flow

    NASA Technical Reports Server (NTRS)

    Reagan, M. K.; Bowman, W. J.

    1993-01-01

    This paper presents the numerical and experimental results of a study performed on the transient rise of fluid in a capillary tube. The capillary tube problem provides an excellent mechanism from which to launch an investigation into the transient flow of a fluid in a porous wick structure where capillary forces must balance both adverse gravitational effects and frictional losses. For the study, a capillary tube, initially charged with a small volume of water, was lowered into a pool of water. The behavior of the column of fluid during the transient that followed as more water entered the tube from the pool was both numerically and experimentally studied.

  5. A study of intermittent flow in downward inclined pipes

    SciTech Connect

    Yang, J.; Sarica, C.; Chen, X.; Brill, J.P.

    1996-12-31

    The downward simultaneous flow of gas and liquid is often encountered in hilly terrain pipelines and steam injection wells. Most of the available methods for predicting the behavior of gas-liquid flow in pipes have been developed for horizontal and upward inclined pipes. In this study, co-current steady state slug flow in downward inclined pipes is investigated, experimentally and theoretically. A series of slug flow experiments are conducted with an air-kerosene system in a 2-in. diameter, 75-ft long pipe installed on an inclinable structure. Liquid holdup and pressure drop measurements are obtained for downward inclination angles from 0{degree} to {minus}90{degree} at different flow conditions. Correlations for slug flow characteristics are obtained based on the experimental data. A mechanistic model based on a unit cell approach has been proposed for the prediction of the detailed slug structure, and subsequently the pressure gradient. Fully developed slug flow could not be observed from {minus}50{degree} to {minus}90{degree}. A correlation was obtained for slug liquid holdup, and an analytical model and a correlation were developed for slug translational velocity. The lognormal distribution was found to best fit all the experimental slug length data. Equations for mean and design slug length were derived from the lognormal distribution function for inclination angles ranging from 0{degree} to {minus}30{degree}. A slug frequency correlation was also developed. The model can be used to predict intermittent flow behavior in downward inclined pipes. The correlations for slug liquid holdup, slug translational velocity, and slug length and frequency are closure relationships applicable to any model. Slug frequency information is also imperative for erosion and corrosion rate predictions.

  6. Performance on ETL 1 kW redox flow cell

    NASA Astrophysics Data System (ADS)

    Nozaki, K.; Kaneko, H.; Negishi, A.; Ozawa, T.

    A 1 kW - 3 kWh redox flow cell, in which 96 bipolar cells with the apparent electrode area of 432 sq cm are involved, has been developed and tested in Electrotechnical Laboratory. The rated output current and voltage are 26 A and 43 V. To elucidate fundamental aspects of the anolyte, polarography and spectroscopy were applied as well as observations with a miniaturized redox flow cell, and influence of the complex species in the anolyte on the cell performance has been recognized. During the charge and discharge cycles the anolyte was continuously monitored by the spectroscopy or controlled potential coulometry, while a voltammetric detector was applied for monitoring the catholyte. Further screening of carbon fiber electrode materials has been continued after the previous presentation, and among more than 70 varieties were found a few kinds of carbon fiber, with which the target performance can be achieved.

  7. Integration of flow studies for robust selection of mechanoresponsive genes.

    PubMed

    Maimari, Nataly; Pedrigi, Ryan M; Russo, Alessandra; Broda, Krysia; Krams, Rob

    2016-03-01

    Blood flow is an essential contributor to plaque growth, composition and initiation. It is sensed by endothelial cells, which react to blood flow by expressing > 1000 genes. The sheer number of genes implies that one needs genomic techniques to unravel their response in disease. Individual genomic studies have been performed but lack sufficient power to identify subtle changes in gene expression. In this study, we investigated whether a systematic meta-analysis of available microarray studies can improve their consistency. We identified 17 studies using microarrays, of which six were performed in vivo and 11 in vitro. The in vivo studies were disregarded due to the lack of the shear profile. Of the in vitro studies, a cross-platform integration of human studies (HUVECs in flow cells) showed high concordance (> 90 %). The human data set identified > 1600 genes to be shear responsive, more than any other study and in this gene set all known mechanosensitive genes and pathways were present. A detailed network analysis indicated a power distribution (e. g. the presence of hubs), without a hierarchical organisation. The average cluster coefficient was high and further analysis indicated an aggregation of 3 and 4 element motifs, indicating a high prevalence of feedback and feed forward loops, similar to prokaryotic cells. In conclusion, this initial study presented a novel method to integrate human-based mechanosensitive studies to increase its power. The robust network was large, contained all known mechanosensitive pathways and its structure revealed hubs, and a large aggregate of feedback and feed forward loops. PMID:26842798

  8. Interstitial flows promote an amoeboid cell phenotype and motility of breast cancer cells

    NASA Astrophysics Data System (ADS)

    Tung, Chih-Kuan; Huang, Yu Ling; Zheng, Angela; Wu, Mingming

    2015-03-01

    Lymph nodes, the drainage systems for interstitial flows, are clinically known to be the first metastatic sites of many cancer types including breast and prostate cancers. Here, we demonstrate that breast cancer cell morphology and motility is modulated by interstitial flows in a cell-ECM adhesion dependent manner. The average aspect ratios of the cells are significantly lower (or are more amoeboid like) in the presence of the flow in comparison to the case when the flow is absent. The addition of exogenous adhesion molecules within the extracellular matrix (type I collagen) enhances the overall aspect ratio (or are more mesenchymal like) of the cell population. Using measured cell trajectories, we find that the persistence of the amoeboid cells (aspect ratio less than 2.0) is shorter than that of mesenchymal cells. However, the maximum speed of the amoeboid cells is larger than that of mesenchymal cells. Together these findings provide the novel insight that interstitial flows promote amoeboid cell morphology and motility and highlight the plasticity of tumor cell motility in response to its biophysical environment. Supported by NIH Grant R21CA138366.

  9. An analytical study of reduced-gravity flow dynamics

    NASA Technical Reports Server (NTRS)

    Bradshaw, R. D.; Kramer, J. L.; Zich, J. L.

    1976-01-01

    Addition of surface tension forces to a marker-and-cell code and the performance of four incompressible fluid simulations in reduced gravity, were studied. This marker-and-cell code has a variable grid capability with arbitrary curved boundaries and time dependent acceleration fields. The surface tension logic includes a spline fit of surface marker particles as well as contact angle logic for straight and curved wall boundaries. Three types of flow motion were simulated with the improved code: impulsive settling in a model Centaur LH2 tank, continuous settling in a model and full scale Centaur LO2 tank and mixing in a Centaur LH2 tank. The impulsive settling case confirmed a drop tower analysis which indicated more orderly fluid collection flow patterns with this method providing a potential savings in settling propellants. In the LO2 tank, fluid collection and flow simulation into the thrust barrel were achieved. The mixing simulation produced good results indicating both the development of the flow field and fluid interface behavior.

  10. Investigations on proton exchange membrane fuel cells with different configurations and flow fields

    NASA Astrophysics Data System (ADS)

    Kazim, Ayoub Mohamed

    In this study, two mathematical models are developed. The first one is a simple mathematical approach that computes all transport and electrochemical parameters inside the different layers of a fuel cell regardless of its configuration. Through heat and mass transfer analogy, convective mass transfer coefficients at different Reynolds number are determined for both concentric cylindrical and conventional proton exchange membrane (PEM) fuel cells. Concentrations of oxygen and hydrogen are then determined at each layer of the fuel cell using steady-state diffusion analysis. The concentration equations are solved together with the electrochemical equations inside the fuel cell, to obtain the fuel cell voltage and power density. The results from this simple approach compared well with the existing numerical and experimental results. The second mathematical model is to study PEM fuel cell with conventional and non-conventional namely interdigitated flow fields. Through proper handling of the boundary conditions at the gas diffusion/catalyst layer interface, the numerical solution of the model resulted in the profiles of transport and electrochemical parameters in the cathode. Parameters such as pressure distribution, velocity profile, oxygen concentration, molar flux, current density, polarization and overall power density at different cell over-potentials in both flow fields were determined. The results demonstrates the superiority of interdigitated flow field over the conventional type in terms of overall performance and illustrated the importance of the convective term of the species equation in enhancing the reaction rates, leading to a significant improvement in the fuel cell performance. The effects of different parameters, such as cathode porosity, inlet oxygen mole fraction, and operating pressure on fuel cell performance have been studied using this 2-D mathematical model. Finally, a simple efficiency and economical analysis was formulated and implemented on

  11. An optical absorption cell with vapor cross flow.

    NASA Technical Reports Server (NTRS)

    Hendrickson, P. E.; Walls, W. L.; Broersma, S.

    1973-01-01

    Description of a water vapor cross flow system that simulates meteorological conditions and effectively curbs any disturbing effects of walls and vacuum connections in an optical absorption cell. Vapor equilibrium is established within 30 min. A 6.3 micron infrared beam traverses the pressure, temperature, and humidity controlled vapor column. The effect of these thermodynamic parameters can be examined.

  12. Dynamic deformability of sickle red blood cells in microphysiological flow

    PubMed Central

    Alapan, Y.; Matsuyama, Y.; Little, J. A.; Gurkan, U. A.

    2016-01-01

    In sickle cell disease (SCD), hemoglobin molecules polymerize intracellularly and lead to a cascade of events resulting in decreased deformability and increased adhesion of red blood cells (RBCs). Decreased deformability and increased adhesion of sickle RBCs lead to blood vessel occlusion (vaso-occlusion) in SCD patients. Here, we present a microfluidic approach integrated with a cell dimensioning algorithm to analyze dynamic deformability of adhered RBC at the single-cell level in controlled microphysiological flow. We measured and compared dynamic deformability and adhesion of healthy hemoglobin A (HbA) and homozygous sickle hemoglobin (HbS) containing RBCs in blood samples obtained from 24 subjects. We introduce a new parameter to assess deformability of RBCs: the dynamic deformability index (DDI), which is defined as the time-dependent change of the cell’s aspect ratio in response to fluid flow shear stress. Our results show that DDI of HbS-containing RBCs were significantly lower compared to that of HbA-containing RBCs. Moreover, we observed subpopulations of HbS containing RBCs in terms of their dynamic deformability characteristics: deformable and non-deformable RBCs. Then, we tested blood samples from SCD patients and analyzed RBC adhesion and deformability at physiological and above physiological flow shear stresses. We observed significantly greater number of adhered non-deformable sickle RBCs than deformable sickle RBCs at flow shear stresses well above the physiological range, suggesting an interplay between dynamic deformability and increased adhesion of RBCs in vaso-occlusive events. PMID:27437432

  13. Fluid flow releases fibroblast growth factor-2 from human aortic smooth muscle cells

    NASA Technical Reports Server (NTRS)

    Rhoads, D. N.; Eskin, S. G.; McIntire, L. V.

    2000-01-01

    This study tested the hypothesis that fluid shear stress regulates the release of fibroblast growth factor (FGF)-2 from human aortic smooth muscle cells. FGF-2 is a potent mitogen that is involved in the response to vascular injury and is expressed in a wide variety of cell types. FGF-2 is found in the cytoplasm of cells and outside cells, where it associates with extracellular proteoglycans. To test the hypothesis that shear stress regulates FGF-2 release, cells were exposed to flow, and FGF-2 amounts were measured from the conditioned medium, pericellular fraction (extracted by heparin treatment), and cell lysate. Results from the present study show that after 15 minutes of shear stress at 25 dyne/cm(2) in a parallel-plate flow system, a small but significant fraction (17%) of the total FGF-2 was released from human aortic smooth muscle cells. FGF-2 levels in the circulating medium increased 10-fold over medium from static controls (P<0.01). A 50% increase in FGF-2 content versus control (P<0.01) was found in the pericellular fraction (extracted by heparin treatment). Furthermore, a significant decrease in FGF-2 was detected in the cell lysate, indicating that FGF-2 was released from inside the cell. Cell permeability studies with fluorescent dextran were performed to examine whether transient membrane disruption caused FGF-2 release. Flow cytometry detected a 50% increase in mean fluorescence of cells exposed to 25 dyne/cm(2) versus control cells. This indicates that the observed FGF-2 release from human aortic smooth muscle cells is likely due to transient membrane disruption on initiation of flow.

  14. Design, evaluation, and application of continuous-flow cells for organic electrochemical synthesis. Final report

    SciTech Connect

    Nobe, K.

    1982-09-30

    Two examples of the oxidation and reduction of aldehydes as a paired synthesis have been studied. These are model systems for potential energy savings in organic electrochemical synthesis. Both are indirect processes; the reduction via alkali metal amalgam and the oxidation via hypobromite. One, using furfural as the substrate, has proved unsuccessful due to the oxidation of its reduction products with the electrogenerated oxidant, bromine. The other paired synthesis, using glucose, has been operated successfully in two types of parallel plate flow cells and two types of porous, packed bed flow cells. To date, the optimum electrode materials and operating conditions for the glucose paired reaction, as determined by product yields and current efficiencies, are an amalgamated zinc cathode, a graphite anode, an initial glucose concentration of 0.8M, a 0.8M NaBr supporting electrolyte and an electrolyte flow rate of 0.8 1/min. Both constant current (10 mA/cm/sup 2/) and constant cathode potential (-2.10V vs SCE) electrolyses were performed under the above conditions. Electrolyses carried out in the parallel plate flow cell and the packed bed flow cell in which the current and electrolyte flow are parallel to one another gave comparable results. Lower current efficiencies were obtained in the porous, packed bed flow cell with perpendicular current and flow. The reasons for the poorer results in the perpendicular configuration cell are not known at this time and further experimentation is required with this system. The current efficiencies and yields of both the oxidation reaction (the production of gluconic acid) and the reduction reaction (the formation of sorbitol) in the packed bed cells were found to be dependent on solution pH.

  15. A Study of Blood Flow and of Aggregation of Blood Cells Under Conditions of Zero Gravity: Its Relevance to the Occlusive Diseases and Cancer

    NASA Technical Reports Server (NTRS)

    Dintenfass, L.

    1985-01-01

    The objectives of this program are: (1) to determine whether the size of red cell aggregates, kinetics and morphology of these aggregates are influenced by near-zero gravity; (2) whether viscosity, especially at low shear rate, is afflicted by near-zero gravity (the latter preventing sedimentation of red cells); (3) whether the actual shape of red cells changes; and (4) whether blood samples obtained from different donors (normal and patients suffering from different disorders) react in the same manner to near-zero gravity.

  16. Effect of the glycocalyx layer on transmission of interstitial flow shear stress to embedded cells.

    PubMed

    Tarbell, John M; Shi, Zhong-Dong

    2013-01-01

    In this paper, a simple theoretical model is developed to describe the transmission of force from interstitial fluid flow to the surface of a cell covered by a proteoglycan / glycoprotein layer (glycocalyx) and embedded in an extracellular matrix. Brinkman equations are used to describe flow through the extracellular matrix and glycocalyx layers and the solid mechanical stress developed in the glycocalyx by the fluid flow loading is determined. Using reasonable values for the Darcy permeability of extracellular matrix and glycocalyx layers and interstitial flow velocity, we are able to estimate the fluid and solid shear stresses imposed on the surface of embedded vascular, cartilage and tumor cells in vivo and in vitro. The principal finding is that the surface solid stress is typically one to two orders of magnitude larger than the surface fluid stress. This indicates that interstitial flow shear stress can be sensed by the cell surface glycocalyx, supporting numerous recent observations that interstitial flow can induce mechanotransduction in embedded cells. This study may contribute to understanding of interstitial flow-related mechanobiology in embryogenesis, tumorigenesis, tissue physiology and diseases and has implications in tissue engineering.

  17. Red blood cell aggregation and dissociation in shear flows simulated by lattice Boltzmann method.

    PubMed

    Zhang, Junfeng; Johnson, Paul C; Popel, Aleksander S

    2008-01-01

    In this paper we develop a lattice Boltzmann algorithm to simulate red blood cell (RBC) behavior in shear flows. The immersed boundary method is employed to incorporate the fluid-membrane interaction between the flow field and deformable cells. The cell membrane is treated as a neo-Hookean viscoelastic material and a Morse potential is adopted to model the intercellular interaction. Utilizing the available mechanical properties of RBCs, multiple cells have been studied in shear flows using a two-dimensional approximation. These cells aggregate and form a rouleau under the action of intercellular interaction. The equilibrium configuration is related to the interaction strength. The end cells exhibit concave shapes under weak interaction and convex shapes under strong interaction. In shear flows, such a rouleau-like aggregate will rotate or be separated, depending on the relative strengths of the intercellular interaction and hydrodynamic viscous forces. These behaviors are qualitatively similar to experimental observations and show the potential of this numerical scheme for future studies of blood flow in microvessels. PMID:17888442

  18. Malignant human cell transformation of Marcellus Shale gas drilling flow back water.

    PubMed

    Yao, Yixin; Chen, Tingting; Shen, Steven S; Niu, Yingmei; DesMarais, Thomas L; Linn, Reka; Saunders, Eric; Fan, Zhihua; Lioy, Paul; Kluz, Thomas; Chen, Lung-Chi; Wu, Zhuangchun; Costa, Max; Zelikoff, Judith

    2015-10-01

    The rapid development of high-volume horizontal hydraulic fracturing for mining natural gas from shale has posed potential impacts on human health and biodiversity. The produced flow back waters after hydraulic stimulation are known to carry high levels of saline and total dissolved solids. To understand the toxicity and potential carcinogenic effects of these wastewaters, flow back waters from five Marcellus hydraulic fracturing oil and gas wells were analyzed. The physicochemical nature of these samples was analyzed by inductively coupled plasma mass spectrometry and scanning electron microscopy/energy dispersive X-ray spectroscopy. A cytotoxicity study using colony formation as the endpoint was carried out to define the LC50 values of test samples using human bronchial epithelial cells (BEAS-2B). The BEAS-2B cell transformation assay was employed to assess the carcinogenic potential of the samples. Barium and strontium were among the most abundant metals in these samples and the same metals were found to be elevated in BEAS-2B cells after long-term treatment. BEAS-2B cells treated for 6weeks with flow back waters produced colony formation in soft agar that was concentration dependent. In addition, flow back water-transformed BEAS-2B cells show better migration capability when compared to control cells. This study provides information needed to assess the potential health impact of post-hydraulic fracturing flow back waters from Marcellus Shale natural gas mining.

  19. Red Blood Cell Aggregation and Dissociation in Shear Flows Simulated by Lattice Boltzmann Method

    PubMed Central

    Zhang, Junfeng; Johnson, Paul C.; Popel, Aleksander S.

    2008-01-01

    In this paper we develop a lattice Boltzmann algorithm to simulate red blood cell (RBC) behavior in shear flows. The immersed boundary method is employed to incorporate the fluid-membrane interaction between the flow field and deformable cells. The cell membrane is treated as a neo-Hookean viscoelastic material and a Morse potential is adopted to model the intercellular interaction. Utilizing the available mechanical properties of RBCs, multiple cells have been studied in shear flows using a two-dimensional approximation. These cells aggregate and form a rouleau under the action of intercellular interaction. The equilibrium configuration is related to the interaction strength. The end cells exhibit concave shapes under weak interaction and convex shapes under strong interaction. In shear flows, such a rouleau-like aggregate will rotate or be separated, depending on the relative strengths of the intercellular interaction and hydrodynamic viscous forces. These behaviors are qualitatively similar to experimental observations and show the potential of this numerical scheme for future studies of blood flow in microvessels. PMID:17888442

  20. Malignant human cell transformation of Marcellus Shale gas drilling flow back water.

    PubMed

    Yao, Yixin; Chen, Tingting; Shen, Steven S; Niu, Yingmei; DesMarais, Thomas L; Linn, Reka; Saunders, Eric; Fan, Zhihua; Lioy, Paul; Kluz, Thomas; Chen, Lung-Chi; Wu, Zhuangchun; Costa, Max; Zelikoff, Judith

    2015-10-01

    The rapid development of high-volume horizontal hydraulic fracturing for mining natural gas from shale has posed potential impacts on human health and biodiversity. The produced flow back waters after hydraulic stimulation are known to carry high levels of saline and total dissolved solids. To understand the toxicity and potential carcinogenic effects of these wastewaters, flow back waters from five Marcellus hydraulic fracturing oil and gas wells were analyzed. The physicochemical nature of these samples was analyzed by inductively coupled plasma mass spectrometry and scanning electron microscopy/energy dispersive X-ray spectroscopy. A cytotoxicity study using colony formation as the endpoint was carried out to define the LC50 values of test samples using human bronchial epithelial cells (BEAS-2B). The BEAS-2B cell transformation assay was employed to assess the carcinogenic potential of the samples. Barium and strontium were among the most abundant metals in these samples and the same metals were found to be elevated in BEAS-2B cells after long-term treatment. BEAS-2B cells treated for 6weeks with flow back waters produced colony formation in soft agar that was concentration dependent. In addition, flow back water-transformed BEAS-2B cells show better migration capability when compared to control cells. This study provides information needed to assess the potential health impact of post-hydraulic fracturing flow back waters from Marcellus Shale natural gas mining. PMID:26210350

  1. Malignant human cell transformation of Marcellus shale gas drilling flow back water

    PubMed Central

    Yao, Yixin; Chen, Tingting; Shen, Steven S.; Niu, Yingmei; DesMarais, Thomas L; Linn, Reka; Saunders, Eric; Fan, Zhihua; Lioy, Paul; Kluz, Thomas; Chen, Lung-Chi; Wu, Zhuangchun; Costa, Max

    2015-01-01

    The rapid development of high-volume horizontal hydraulic fracturing for mining natural gas from shale has posed potential impacts on human health and biodiversity. The produced flow back waters after hydraulic stimulation is known to carry high levels of saline and total dissolved solids. To understand the toxicity and potential carcinogenic effects of these waste waters, flow back water from five Marcellus hydraulic fracturing oil and gas wells were analyzed. The physicochemical nature of these samples was analyzed by inductively coupled plasma mass spectrometry and scanning electron microscopy / energy dispersive X-ray spectroscopy. A cytotoxicity study using colony formation as the endpoint was carried out to define the LC50 values of test samples using human bronchial epithelial cells (BEAS-2B). The BEAS-2B cell transformation assay was employed to assess the carcinogenic potential of the samples. Barium and strontium were among the most abundant metals in these samples and the same metals were found elevated in BEAS-2B cells after long-term treatment. BEAS-2B cells treated for 6 weeks with flow back waters produced colony formation in soft agar that was concentration dependant. In addition, flow back water-transformed BEAS-2B cells show a better migration capability when compared to control cells. This study provides information needed to assess the potential health impact of post-hydraulic fracturing flow back waters from Marcellus Shale natural gas mining. PMID:26210350

  2. Flow bioreactor design for quantitative measurements over endothelial cells using micro-particle image velocimetry

    NASA Astrophysics Data System (ADS)

    Leong, Chia Min; Voorhees, Abram; Nackman, Gary B.; Wei, Timothy

    2013-04-01

    Mechanotransduction in endothelial cells (ECs) is a highly complex process through which cells respond to changes in hemodynamic loading by generating biochemical signals involving gene and protein expression. To study the effects of mechanical loading on ECs in a controlled fashion, different in vitro devices have been designed to simulate or replicate various aspects of these physiological phenomena. This paper describes the design, use, and validation of a flow chamber which allows for spatially and temporally resolved micro-particle image velocimetry measurements of endothelial surface topography and stresses over living ECs immersed in pulsatile flow. This flow chamber also allows the study of co-cultures (i.e., ECs and smooth muscle cells) and the effect of different substrates (i.e., coverslip and/or polyethylene terepthalate (PET) membrane) on cellular response. In this report, the results of steady and pulsatile flow on fixed endothelial cells seeded on PET membrane and coverslip, respectively, are presented. Surface topography of ECs is computed from multiple two-dimensional flow measurements. The distributions of shear stress and wall pressure on each individual cell are also determined and the importance of both types of stress in cell remodeling is highlighted.

  3. Flow bioreactor design for quantitative measurements over endothelial cells using micro-particle image velocimetry.

    PubMed

    Leong, Chia Min; Voorhees, Abram; Nackman, Gary B; Wei, Timothy

    2013-04-01

    Mechanotransduction in endothelial cells (ECs) is a highly complex process through which cells respond to changes in hemodynamic loading by generating biochemical signals involving gene and protein expression. To study the effects of mechanical loading on ECs in a controlled fashion, different in vitro devices have been designed to simulate or replicate various aspects of these physiological phenomena. This paper describes the design, use, and validation of a flow chamber which allows for spatially and temporally resolved micro-particle image velocimetry measurements of endothelial surface topography and stresses over living ECs immersed in pulsatile flow. This flow chamber also allows the study of co-cultures (i.e., ECs and smooth muscle cells) and the effect of different substrates (i.e., coverslip and∕or polyethylene terepthalate (PET) membrane) on cellular response. In this report, the results of steady and pulsatile flow on fixed endothelial cells seeded on PET membrane and coverslip, respectively, are presented. Surface topography of ECs is computed from multiple two-dimensional flow measurements. The distributions of shear stress and wall pressure on each individual cell are also determined and the importance of both types of stress in cell remodeling is highlighted.

  4. Method for Studying Microbial Biofilms in Flowing-Water Systems

    PubMed Central

    Pedersen, Karsten

    1982-01-01

    A method for the study of microbial biofilms in flowing-water systems was developed with special reference to the flow conditions in electrochemical concentration cells. Seawater was circulated in a semiclosed flow system through biofilm reactors (3 cm s−1) with microscope cover slips arranged in lamellar piles parallel with the flow. At fixed time intervals cover slips with their biofilm were removed from the pile, stained with crystal violet, and mounted on microscope slides. The absorbances of the slides were measured at 590 nm and plotted against time to give microbial biofilm development. From calibration experiments a staining time of 1 min and a rinse time of 10 min in a tap water flow (3 cm s−1) were considered sufficient. When an analysis of variance was performed on biofilm development data, 78% of the total variance was found to be due to random natural effects; the rest could be explained by experimental effects. The absorbance values correlated well with protein N, dry weight, and organic weight in two biofilm experiments, one with a biofilm with a high (75%) and one with a low (∼25%, normal) inorganic content. Comparisons of regression lines revealed that the absorbance of the stained biofilms was an estimate closely related to biofilm dry weight. PMID:16345929

  5. A critical review of two-phase flow in gas flow channels of proton exchange membrane fuel cells

    NASA Astrophysics Data System (ADS)

    Anderson, Ryan; Zhang, Lifeng; Ding, Yulong; Blanco, Mauricio; Bi, Xiaotao; Wilkinson, David P.

    Water management in PEM fuel cells has received extensive attention due to its key role in fuel cell performance. The unavoidable water, from humidified gas streams and electrochemical reaction, leads to gas-liquid two-phase flow in the flow channels of fuel cells. The presence of two-phase flow increases the complexity in water management in PEM fuel cells, which remains a challenging hurdle in the commercialization of this technology. Unique water emergence from the gas diffusion layer, which is different from conventional gas-liquid two-phase flow where water is introduced from the inlet together with the gas, leads to different gas-liquid flow behaviors, including pressure drop, flow pattern, and liquid holdup along flow field channels. These parameters are critical in flow field design and fuel cell operation and therefore two-phase flow has received increasing attention in recent years. This review emphasizes gas-liquid two-phase flow in minichannels or microchannels related to PEM fuel cell applications. In situ and ex situ experimental setups have been utilized to visualize and quantify two-phase flow phenomena in terms of flow regime maps, flow maldistribution, and pressure drop measurements. Work should continue to make the results more relevant for operating PEM fuel cells. Numerical simulations have progressed greatly, but conditions relevant to the length scales and time scales experienced by an operating fuel cell have not been realized. Several mitigation strategies exist to deal with two-phase flow, but often at the expense of overall cell performance due to parasitic power losses. Thus, experimentation and simulation must continue to progress in order to develop a full understanding of two-phase flow phenomena so that meaningful mitigation strategies can be implemented.

  6. Electrotaxis of oral squamous cell carcinoma cells in a multiple-electric-field chip with uniform flow field

    PubMed Central

    Tsai, Hsieh-Fu; Peng, Shih-Wei; Wu, Chun-Ying; Chang, Hui-Fang; Cheng, Ji-Yen

    2012-01-01

    We report a new design of microfluidic chip (Multiple electric Field with Uniform Flow chip, MFUF chip) to create multiple electric field strengths (EFSs) while providing a uniform flow field simultaneously. MFUF chip was fabricated from poly-methyl methacrylates (PMMA) substrates by using CO2 laser micromachining. A microfluidic network with interconnecting segments was utilized to de-couple the flow field and the electric field (EF). Using our special design, different EFSs were obtained in channel segments that had an identical cross-section and therefore a uniform flow field. Four electric fields with EFS ratio of 7.9:2.8:1:0 were obtained with flow velocity variation of only 7.8% CV (coefficient of variation). Possible biological effect of shear force can therefore be avoided. Cell behavior under three EFSs and the control condition, where there is no EF, was observed in a single experiment. We validated MFUF chip performance using lung adenocarcinoma cell lines and then used the chip to study the electrotaxis of HSC-3, an oral squamous cell carcinoma cell line. The MFUF chip has high throughput capability for studying the EF-induced cell behavior under various EFSs, including the control condition (EFS = 0). PMID:24009650

  7. Detection, isolation, and capture of circulating breast cancer cells with photoacoustic flow cytometry

    NASA Astrophysics Data System (ADS)

    Bhattacharyya, Kiran; Njoroge, Martin; Goldschmidt, Benjamin S.; Gaffigan, Brian; Rood, Kyle; Viator, John A.

    2013-03-01

    According to the CDC, breast cancer is the most common cancer and the second leading cause of cancer related deaths among women. Metastasis, or the presence of secondary tumors caused by the spread of cancer cells via the circulatory or lymphatic systems, significantly worsens the prognosis of any breast cancer patient. In this study, a technique is developed to detect circulating breast cancer cells in human blood using a photoacoustic flow cytometry method. A Q-switched laser with a 5 ns pulse at 532 nm is used to interrogate thousands of cells with one pulse as they flow through the beam path. Cells which are pigmented, either naturally or artificially, emit an ultrasound wave as a result of the photoacoustic (PA) effect. Breast cancer cells are targeted with chromophores through immunochemistry in order to provide pigment. After which, the device is calibrated to demonstrate a single-cell detection limit. Cultured breast cancer cells are added to whole blood to reach a biologically relevant concentration of about 25-45 breast cancer cells per 1 mL of blood. An in vitro photoacoustic flow cytometer is used to detect and isolate these cells followed by capture with the use of a micromanipulator. This method can not only be used to determine the disease state of the patient and the response to therapy, it can also be used for genetic testing and in vitro drug trials since the circulating cell can be captured and studied.

  8. Space shuttle orbiter flow visualization study. [water tunnel study of vortex flow during atmospheric entry

    NASA Technical Reports Server (NTRS)

    Lorincz, D. J.

    1980-01-01

    The vortex flows generated at subsonic speed during the final portion of atmospheric reentry were defined using a 0.01 scale model of the orbiter in a diagnostic water tunnel. Flow visualization photographs were obtained over an angle-of-attack range to 40 deg and sideslip angles up to 10 deg. The vortex flow field development, vortex path, and vortex breakdown characteristics were determined as a function of angle-of-attack at zero sideslip. Vortex flows were found to develop on the highly swept glove, on the wing, and on the upper surface of the fuselage. No significant asymmetries were observed at zero sideslip in the water tunnel tests. The sensitivity of the upper surface vortex flow fields to variations in sideslip angle was also studied. The vortex formed on the glove remained very stable in position above the wing up through the 10 deg of sideslip tested. There was a change in the vortex lifts under sideslip due to effective change in leading-edge sweep angles. Asymmetric flow separation occurred on the upper surface of the fuselage at small sideslip angles. The influence of vortex flow fields in sideslip on the lateral/ directional characteristics of the orbiter is discussed.

  9. Mathematical analysis of mis-estimation of cell subsets in flow cytometry: viability staining revisited.

    PubMed

    Petrunkina, A M; Harrison, R A P

    2011-05-31

    Many research projects in cell biology now use flow cytometry for analysis or for isolation of specific cell types. In such studies, cell viability is obviously a crucial issue. However, many studies appear to rely upon light-scattering characteristics to identify and gate out non-viable cells, despite the fact that reliable identification of such cells can only be achieved through staining with impermeable fluorescent nuclear dyes such as propidium iodide or 7-amino actinomycin. In this paper we apply mathematical analysis to the theoretical problem of quantifying cell sub-populations labeled with two or more fluorescent markers, comparing situations in which dead cells have been identified with those in which cell viability has not been assessed. We demonstrate that in all cases in which dead cells are present within the population, percentages of live sub-populations in different subsets are mis-estimated. In cases where the pattern of marker expression differs greatly between live and dead cells, or where the proportion of dead cells is high, this mis-estimation will be aggravated; the subsets pattern will therefore be biased in a population selected only on the basis of light-scatter behavior. The importance of accurately detecting and gating out dead cells is illustrated by an experimental example accompanying the mathematical analysis. To conclude, identification of dead cells by means of viability stains should be an absolute routine in practical flow cytometry, so as to avoid mis-estimation in sorting or analysis.

  10. Fuel cell with interdigitated porous flow-field

    DOEpatents

    Wilson, Mahlon S.

    1997-01-01

    A polymer electrolyte membrane (PEM) fuel cell is formed with an improved system for distributing gaseous reactants to the membrane surface. A PEM fuel cell has an ionic transport membrane with opposed catalytic surfaces formed thereon and separates gaseous reactants that undergo reactions at the catalytic surfaces of the membrane. The fuel cell may also include a thin gas diffusion layer having first and second sides with a first side contacting at least one of the catalytic surfaces. A macroporous flow-field with interdigitated inlet and outlet reactant channels contacts the second side of the thin gas diffusion layer for distributing one of the gaseous reactants over the thin gas diffusion layer for transport to an adjacent one of the catalytic surfaces of the membrane. The porous flow field may be formed from a hydrophilic material and provides uniform support across the backside of the electrode assembly to facilitate the use of thin backing layers.

  11. Fuel cell with interdigitated porous flow-field

    DOEpatents

    Wilson, M.S.

    1997-06-24

    A polymer electrolyte membrane (PEM) fuel cell is formed with an improved system for distributing gaseous reactants to the membrane surface. A PEM fuel cell has an ionic transport membrane with opposed catalytic surfaces formed thereon and separates gaseous reactants that undergo reactions at the catalytic surfaces of the membrane. The fuel cell may also include a thin gas diffusion layer having first and second sides with a first side contacting at least one of the catalytic surfaces. A macroporous flow-field with interdigitated inlet and outlet reactant channels contacts the second side of the thin gas diffusion layer for distributing one of the gaseous reactants over the thin gas diffusion layer for transport to an adjacent one of the catalytic surfaces of the membrane. The porous flow field may be formed from a hydrophilic material and provides uniform support across the backside of the electrode assembly to facilitate the use of thin backing layers. 9 figs.

  12. Gold recycling; a materials flow study

    USGS Publications Warehouse

    Amey, Earle B.

    2000-01-01

    This materials flow study includes a description of trends in consumption, loss, and recycling of gold-containing materials in the United States in 1998 in order to illustrate the extent to which gold is presently being recycled and to identify recycling trends. The quantity of gold recycled, as a percent of the apparent supply of gold, was estimated to be about 30 percent. Of the approximately 446 metric tons of gold refined in the United States in 1998, the fabricating and industrial use losses were 3 percent.

  13. Flow field studies on yawed, stranded cables

    NASA Astrophysics Data System (ADS)

    Batill, S. M.; Nelson, R. C.; Nebres, J. V.

    A study of the flowfield near yawed, stranded cables was conducted in order to investigate the mechanisms associated with the generation of both steady and unsteady fluid forces on the cables. Rigid cable models and a circular cylinder were tested in a wind tunnel at four different cable angles over a Reynolds number range from 6000 to 14,600 based on the nominal cable diameter. The smoke-wire and the kerosene smoke flow visualization techniques were used to qualitatively evaluate the flowfields associated with each cable geometry.

  14. Flow cytometry analysis of cell cycle and specific cell synchronization with butyrate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Synchronized cells have been invaluable in many kinds of cell cycle and cell proliferation studies. Butyrate induces cell cycle arrest and apoptosis in MDBK cells. The possibility of using butyrate-blocked cells to obtain synchronized cells was explored and the properties of butyrate-induced cell ...

  15. In Vivo Monitoring of Multiple Circulating Cell Populations Using Two-photon Flow Cytometry

    PubMed Central

    Tkaczyk, Eric R.; Zhong, Cheng Frank; Ye, Jing Yong; Myc, Andrzej; Thomas, Thommey; Cao, Zhengyi; Duran-Struuck, Raimon; Luker, Kathryn E.; Luker, Gary D.; Norris, Theodore B.; Baker, James R.

    2008-01-01

    To detect and quantify multiple distinct populations of cells circulating simultaneously in the blood of living animals, we developed a novel optical system for two-channel, two-photon flow cytometry in vivo. We used this system to investigate the circulation dynamics in live animals of breast cancer cells with low (MCF-7) and high (MDA-MB-435) metastatic potential, showing for the first time that two different populations of circulating cells can be quantified simultaneously in the vasculature of a single live mouse. We also non-invasively monitored a population of labeled, circulating red blood cells for more than two weeks, demonstrating that this technique can also quantify the dynamics of abundant cells in the vascular system for prolonged periods of time. These data are the first in vivo application of multichannel flow cytometry utilizing two-photon excitation, which will greatly enhance our capability to study circulating cells in cancer and other disease processes. PMID:19221581

  16. In Vivo Monitoring of Multiple Circulating Cell Populations Using Two-photon Flow Cytometry.

    PubMed

    Tkaczyk, Eric R; Zhong, Cheng Frank; Ye, Jing Yong; Myc, Andrzej; Thomas, Thommey; Cao, Zhengyi; Duran-Struuck, Raimon; Luker, Kathryn E; Luker, Gary D; Norris, Theodore B; Baker, James R

    2008-02-15

    To detect and quantify multiple distinct populations of cells circulating simultaneously in the blood of living animals, we developed a novel optical system for two-channel, two-photon flow cytometry in vivo. We used this system to investigate the circulation dynamics in live animals of breast cancer cells with low (MCF-7) and high (MDA-MB-435) metastatic potential, showing for the first time that two different populations of circulating cells can be quantified simultaneously in the vasculature of a single live mouse. We also non-invasively monitored a population of labeled, circulating red blood cells for more than two weeks, demonstrating that this technique can also quantify the dynamics of abundant cells in the vascular system for prolonged periods of time. These data are the first in vivo application of multichannel flow cytometry utilizing two-photon excitation, which will greatly enhance our capability to study circulating cells in cancer and other disease processes.

  17. Granular mass flows and Coulomb's friction in shear cell experiments: Implications for geophysical flows

    NASA Astrophysics Data System (ADS)

    Cagnoli, B.; Manga, M.

    2004-12-01

    Granular mass flows of rock fragments are studied in the lab by means of a high-speed video camera at 2000 frames per second. These granular flows are generated using beds of pumice fragments positioned on a rough rotating disk, whose angular velocity is controlled by a motor. The experimental apparatus allows an understanding of the arrangement of the particles in granular mass flows with relatively small and relatively large values of the Savage number (the Savage number represents the ratio between grain collision stresses and gravitational grain contact stresses). In particular, these flows develop a basal layer of agitated and colliding particles underneath a relatively rigid upper layer. Our experimental results suggest the validity, on average, of the Coulomb's relationship between shear and normal forces at the base of granular mass flows irrespective of their Savage number value. In Coulomb's equation the shear stresses do not depend on the shear rate. We expect the Coulomb friction law to be valid also in moving pyroclastic flows. Our experiments suggest that the collisions and subsequent comminution of pumice fragments in moving pyroclastic flows could provide ash for the overriding ash clouds. In our experiments the amount of ash generated by particle-particle and particle-boundary interactions increases as the value of the Savage number increases. In nature, part of this ash may also simply move toward the base of the flows because of kinetic sieving.

  18. Progress Towards a Cartesian Cut-Cell Method for Viscous Compressible Flow

    NASA Technical Reports Server (NTRS)

    Berger, Marsha; Aftosmis, Michael J.

    2012-01-01

    We present preliminary development of an approach for simulating high Reynolds number steady compressible flow in two space dimensions using a Cartesian cut-cell finite volume method. We consider both laminar and turbulent flow with both low and high cell Reynolds numbers near the wall. The approach solves the full Navier-Stokes equations in all cells, and uses a wall model to address the resolution requirements near boundaries and to mitigate mesh irregularities in cut cells. We present a quadratic wall model for low cell Reynolds numbers. At high cell Reynolds numbers, the quadratic is replaced with a newly developed analytic wall model stemming from solution of a limiting form of the Spalart-Allmaras turbulence model which features a forward evaluation for flow velocity and exactly matches characteristics of the SA turbulence model in the field. We develop multigrid operators which attain convergence rates similar to inviscid multigrid. Investigations focus on preliminary verification and validation of the method. Flows over flat plates and compressible airfoils show good agreement with both theoretical results and experimental data. Mesh convergence studies on sub- and transonic airfoil flows show convergence of surface pressures with wall spacings as large as approx.0.1% chord. With the current analytic wall model, one or two additional refinements near the wall are required to obtain mesh converged values of skin friction.

  19. Flow cells as quasi-ideal systems for biofouling simulation of industrial piping systems.

    PubMed

    Teodósio, Joana S; Silva, Filipe C; Moreira, Joana M R; Simões, Manuel; Melo, Luís F; Alves, Manuel A; Mergulhão, Filipe J

    2013-09-01

    Semi-circular flow cells are often used to simulate the formation of biofilms in industrial pipes with circular section because their planar surface allows easy sampling using coupons. Computational fluid dynamics was used to assess whether the flow in pipe systems can be emulated by the semi-circular flow cells that are used to study biofilm formation. The results show that this is the case for Reynolds numbers (Re) ranging from 10 to 1000 and 3500 to 10,000. A correspondence involving the friction factor was obtained in order to correlate any semi-circular flow cell to any circular pipe for Re between 10 and 100,000. The semi-circular flow cell was then used to assess experimentally the effect of Reynolds number (Re = 4350 and 6720) on planktonic cell concentration and biofilm formation using Escherichia coli JM109 (DE3). Lower planktonic cell concentrations and thicker biofilms (>1.2 mm) were obtained with the lower Re.

  20. A study of grout flow pattern analysis

    SciTech Connect

    Lee, S. Y.; Hyun, S.

    2013-01-10

    A new disposal unit, designated as Salt Disposal Unit no. 6 (SDU6), is being designed for support of site accelerated closure goals and salt nuclear waste projections identified in the new Liquid Waste System plan. The unit is cylindrical disposal vault of 380 ft diameter and 43 ft in height, and it has about 30 million gallons of capacity. Primary objective was to develop the computational model and to perform the evaluations for the flow patterns of grout material in SDU6 as function of elevation of grout discharge port, and slurry rheology. A Bingham plastic model was basically used to represent the grout flow behavior. A two-phase modeling approach was taken to achieve the objective. This approach assumes that the air-grout interface determines the shape of the accumulation mound. The results of this study were used to develop the design guidelines for the discharge ports of the Saltstone feed materials in the SDU6 facility. The focusing areas of the modeling study are to estimate the domain size of the grout materials radially spread on the facility floor under the baseline modeling conditions, to perform the sensitivity analysis with respect to the baseline design and operating conditions such as elevation of discharge port, discharge pipe diameter, and grout properties, and to determine the changes in grout density as it is related to grout drop height. An axi-symmetric two-phase modeling method was used for computational efficiency. Based on the nominal design and operating conditions, a transient computational approach was taken to compute flow fields mainly driven by pumping inertia and natural gravity. Detailed solution methodology and analysis results are discussed here.

  1. Bodies in flowing plasmas - Laboratory studies

    NASA Technical Reports Server (NTRS)

    Stone, N. H.; Samir, U.

    1981-01-01

    A brief review of early rudimentary laboratory studies of bodies in flowing, rarefied plasmas is presented (e.g., Birkeland, 1908), along with a discussion of more recent parametric studies conducted in steady plasma wind tunnels, which includes the study by Hall et al. (1964), in which a strong ion density enhancement in the center of the ion void created downstream from the body was observed. Good agreement was found between the experimental results and theoretical calculations which omit ion thermal motion. Examples in which in situ data on the interaction between satellites and the ionospheric plasma have been elucidated by the laboratory results are presented, and include evidence for a midwake axial ion peak, and ion current density in the near-wake region. The application of the ionospheric laboratory to basic space plasma physics is discussed, and its application to some types of solar system plasma phenomena is illustrated.

  2. Measuring NAPL-Water Interfacial Areas to Evaluate the Effectiveness of In-Situ Chemical Oxidation for DNAPL-Contaminated Source Zones: A Two-Dimensional Flow Cell Study

    NASA Astrophysics Data System (ADS)

    Li, M.; Brusseau, M. L. L.; Yan, N.; Wan, L.

    2015-12-01

    In-situ chemical oxidation (ISCO) using persulfate was employed to remediate a flow cell contaminated with a model dense nonaqueous-phase liquid (DNAPL), trichloroethene (TCE). The flow cell was packed homogeneously with 359 μm diameter natural sand. Dyed TCE DNAPL was naturally distributed in the flow cell. Fe2+-activated persulfate (5 mM) was used for ISCO. Interfacial partitioning tracer tests (IPTT) were conducted before and after ISCO to measure NAPL-water interfacial area, using sodium dodecyl benzenesulfonate (SDBS, 35mg/L) as the tracer. The change in interfacial area was examined as influenced by ISCO remediation. The interfacial areas measured for this two-dimensional system are compared to previously reported values obtained from one-dimensional column experiments.

  3. Couette flows of a granular monolayer: An experimental study

    SciTech Connect

    Elliott, K.E.; Ahmadi, G.; Kvasnak, W.

    1995-03-01

    An experimental study concerning rapid flows of granular materials in a two dimensional planar granular Couette flow apparatus is performed. The device is capable of generating particulate flows in grain-inertia regime at different shearing rates and solid volume fractions. Multi-color spherical glass particles are sheared across an annular test-section for several wall angular velocities. A video recorder is used to record the motion of particles, and consecutive images are stored and analyzed by an image processing technique for evaluating individual grain velocities. Experimental data for the mean velocity, the root mean-square fluctuation velocity components and the solid volume fraction profile are obtained. The resulting mean velocity profiles have a roughly linear variation for the range of solid volume fractions and shear rates studied. The solid volume fraction profiles exhibit nonuniform variations with the highest concentration occuring near the center of the shearing cell. The RMS-fluctuation velocities are roughly constant, with the streamwise fluctuation being somewhat larger than the cross-stream direction. The experimentally measured flow properties are in reasonable agreement with the earlier theoretical and simulation results.

  4. Analysis on the design and property of flow field plates of innovative direct methanol fuel cell.

    PubMed

    Chang, Ho; Kao, Mu-Jung; Chen, Chih-Hao; Kuo, Chin-Guo; Lee, Kuang-Ying

    2014-10-01

    The paper uses technology of lithography process to etch flow fields on single side of a printed circuit board (PCB), and combines flow field plate with collector plate to make innovative anode flow field plates and cathode flow field plates required in direct methanol fuel cell (DMFC), and meanwhile makes membrane electrode assembly (MEA) and methanol fuel plate. The flow field plates are designed to be in the form of serpentine flow field. The paper measured the assembled DMFC to achieve the overall efficiency of DMFC under the conditions of different screw torques and different concentration, flow rate and temperature of methanol. Experimental results show that when the flow field width of flow field plate is 1 mm, the screw torque is 16 kgf/cm, and the concentration, flow rate and temperature of methanol-water are 1 M, 180 ml/h and 50 degrees C respectively, the prepared DMFC can have better power density of 5.5 mW/cm2, 5.4 mW/cm2, 11.2 mW/cm2 and 11.8 mW/cm2. Besides, the volume of the DMFC designed and assembled by the study is smaller than the generally existing DMFC by 40%. PMID:25942924

  5. Simultaneous measurement of human hematopoietic stem and progenitor cells in blood using multicolor flow cytometry.

    PubMed

    Cimato, Thomas R; Furlage, Rosemary L; Conway, Alexis; Wallace, Paul K

    2016-09-01

    Hematopoietic stem cells are the source of all inflammatory cell types. Discovery of specific cell surface markers unique to human hematopoietic stem (HSC) and progenitor (HSPC) cell populations has facilitated studies of their development from stem cells to mature cells. The specific marker profiles of HSCs and HSPCs can be used to understand their role in human inflammatory diseases. The goal of this study is to simultaneously measure HSCs and HSPCs in normal human venous blood using multicolor flow cytometry. Our secondary aim is to determine how G-CSF mobilization alters the quantity of each HSC and HSPC population. Here we show that cells within the CD34+ fraction of human venous blood contains cells with the same cell surface markers found in human bone marrow samples. Mobilization with G-CSF significantly increases the quantity of total CD34+ cells, blood borne HSCs, multipotent progenitors, common myeloid progenitors, and megakaryocyte erythroid progenitors as a percentage of total MNCs analyzed. The increase in blood borne common lymphoid and granulocyte macrophage progenitors with G-CSF treatment did not reach significance. G-CSF treatment predominantly increased the numbers of HSCs and multipotent progenitors in the total CD34+ cell population; common myeloid progenitors and megakaryocyte erythroid progenitors were enriched relative to total MNCs analyzed, but not relative to total CD34+ cells. Our findings illustrate the utility of multicolor flow cytometry to quantify circulating HSCs and HSPCs in venous blood samples from human subjects. © 2016 International Clinical Cytometry Society. PMID:26663713

  6. Flow structures and red blood cell dynamics in arteriole of dilated or constricted cross section.

    PubMed

    Gambaruto, Alberto M

    2016-07-26

    Vessel with 'circular' or 'star-shaped' cross sections are studied, representing respectively dilated or constricted cases where endothelial cells smoothly line or bulge into the lumen. Computational haemodynamics simulations are carried out on idealised periodic arteriole-sized vessels, with red blood cell 'tube' hematocrit value=24%. A further simulation of a single red blood cell serves for comparison purposes. The bulk motion of the red blood cells reproduces well-known effects, including the presence of a cell-free layer and the apparent shear-thinning non-Newtonian rheology. The velocity flow field is analysed in a Lagrangian reference frame, relative to any given red blood cell, hence removing the bulk coaxial motion and highlighting instead the complex secondary flow patterns. An aggregate formation becomes apparent, continuously rearranging and dynamic, brought about by the inter-cellular fluid mechanics interactions and the deformability properties of the cells. The secondary flow field induces a vacillating radial migration of the red blood cells. At different radial locations, the red blood cells express different residence times, orientation and shape. The shear stresses exerted by the flow on the vessel wall are influenced by the motion of red blood cells, despite the presence of the cell-free layer. Spatial (and temporal) variations of wall shear stress patters are observed, especially for the 'circular' vessel. The 'star-shaped' vessel bears considerable stress at the protruding endothelial cell crests, where the stress vectors are coaxially aligned. The bulging endothelial cells hence regularise the transmission of stresses on the vessel wall. PMID:26822224

  7. Flow structures and red blood cell dynamics in arteriole of dilated or constricted cross section.

    PubMed

    Gambaruto, Alberto M

    2016-07-26

    Vessel with 'circular' or 'star-shaped' cross sections are studied, representing respectively dilated or constricted cases where endothelial cells smoothly line or bulge into the lumen. Computational haemodynamics simulations are carried out on idealised periodic arteriole-sized vessels, with red blood cell 'tube' hematocrit value=24%. A further simulation of a single red blood cell serves for comparison purposes. The bulk motion of the red blood cells reproduces well-known effects, including the presence of a cell-free layer and the apparent shear-thinning non-Newtonian rheology. The velocity flow field is analysed in a Lagrangian reference frame, relative to any given red blood cell, hence removing the bulk coaxial motion and highlighting instead the complex secondary flow patterns. An aggregate formation becomes apparent, continuously rearranging and dynamic, brought about by the inter-cellular fluid mechanics interactions and the deformability properties of the cells. The secondary flow field induces a vacillating radial migration of the red blood cells. At different radial locations, the red blood cells express different residence times, orientation and shape. The shear stresses exerted by the flow on the vessel wall are influenced by the motion of red blood cells, despite the presence of the cell-free layer. Spatial (and temporal) variations of wall shear stress patters are observed, especially for the 'circular' vessel. The 'star-shaped' vessel bears considerable stress at the protruding endothelial cell crests, where the stress vectors are coaxially aligned. The bulging endothelial cells hence regularise the transmission of stresses on the vessel wall.

  8. A study of temporal estaurine flow dynamics

    NASA Technical Reports Server (NTRS)

    Mairs, R. L.; Clark, D. K.

    1972-01-01

    Multispectral photography,infrared imagery, image enhancement, and oceanographic, radiometric, and meteorological data were used in the study of temporal estuarine flow dynamics, nearshore circulation, and the resulting dispersal of suspended and dissolved substances introduced from the continent. Repetitive multispectral photography, IR imagery, total radiance and irradiance, water surface temperatures, salinity, total suspended solids, visibility, current velocity, winds, dye implants, and high contrast image enhancement were used to observe and describe water mass boundaries in the nearshore zone and to attempt to establish on what repetitive scale these coastal features should be observed to better understand their behavior. Water mass variability patterns, seen naturally and with the use of dyes, along the North Carolina coast and in the Chesapeake Bay are being studied as synoptic data on the basic dynamics of circulation, flushing, and mixing in coastal waters.

  9. Parallel flow in hele-shaw cells with ferrofluids

    PubMed

    Miranda; Widom

    2000-02-01

    Parallel flow in a Hele-Shaw cell occurs when two immiscible liquids flow with relative velocity parallel to the interface between them. The interface is unstable due to a Kelvin-Helmholtz type of instability in which fluid flow couples with inertial effects to cause an initial small perturbation to grow. Large amplitude disturbances form stable solitons. We consider the effects of applied magnetic fields when one of the two fluids is a ferrofluid. The dispersion relation governing mode growth is modified so that the magnetic field can destabilize the interface even in the absence of inertial effects. However, the magnetic field does not affect the speed of wave propogation for a given wave number. We note that the magnetic field creates an effective interaction between the solitons. PMID:11046508

  10. Effect of blood flow parameters on flow patterns at arterial bifurcations--studies in models.

    PubMed

    Liepsch, D W

    1990-01-01

    Atherosclerotic lesions are found primarily at arterial bends and bifurcations. Flow disturbances at these anatomic sites play a major role in atherogenesis. How hemodynamic factors such as vessel geometry, the pulsatile nature of blood flow, vessel wall elasticity and the non-Newtonian flow behavior of blood influence the flow field at these sites must be clarified. We have performed fundamental studies using a birefringent solution in a simplified rigid 90 degree T-bifurcation and pulsatile flow. The velocity distribution was measured with a laser Doppler anemometer. Flow in an elastic abdominal aorta model has been visualized using magnetic resonance imaging. In both flow studies, zones with negative velocity were found. These model measurements demonstrate that no flow parameter can be neglected. Further detailed studies are necessary to examine the interaction between fluid dynamic and cellular surface properties. PMID:2404201

  11. Recent Advances in Hot Flow Anomaly Studies

    NASA Astrophysics Data System (ADS)

    Zhang, H.

    2015-12-01

    Hot flow anomalies (HFAs) are events observed near planetary bow shocks that are characterized by greatly heated solar wind plasmas and substantial flow deflection. HFAs are universal phenomena that have been observed near the bow shock of Earth, Venus, Mars, and Saturn. HFAs are not stable structures and they evolve with time. Statistical study shows that both ion and electron spectra can be used to classify young and mature HFAs. HFAs were also classified into four categories ("-+", "+-", "M", and "W") according their dynamic pressure profile. Most "W" type HFAs are mature HFAs (according to ion spectra) and most "-+" and "+-" type HFAs are young HFAs. Half of the "M" type HFAs are mature HFAs. Superposed epoch analysis result shows that variations of plasma parameters and magnetic field of mature HFAs are more dramatic than those of young HFAs, except for temperature. "M" and "W" type HFAs may be the later evolution stages of "-+" and "+-" type HFAs; on the other hand, four categories of HFAs may be due to the fact that the spacecraft crossed an HFA structure along different paths.

  12. A simple microfluidic device for the deformability assessment of blood cells in a continuous flow.

    PubMed

    Rodrigues, Raquel O; Pinho, Diana; Faustino, Vera; Lima, Rui

    2015-12-01

    Blood flow presents several interesting phenomena in microcirculation that can be used to develop microfluidic devices capable to promote blood cells separation and analysis in continuous flow. In the last decade there have been numerous microfluidic studies focused on the deformation of red blood cells (RBCs) flowing through geometries mimicking microvessels. In contrast, studies focusing on the deformation of white blood cells (WBCs) are scarce despite this phenomenon often happens in the microcirculation. In this work, we present a novel integrative microfluidic device able to perform continuous separation of a desired amount of blood cells, without clogging or jamming, and at the same time, capable to assess the deformation index (DI) of both WBCs and RBCs. To determine the DI of both WBCs and RBCs, a hyperbolic converging microchannel was used, as well as a suitable image analysis technique to measure the DIs of these blood cells along the regions of interest. The results show that the WBCs have a much lower deformability than RBCs when subjected to the same in vitro flow conditions, which is directly related to their cytoskeleton and nucleus contents. The proposed strategy can be easily transformed into a simple and inexpensive diagnostic microfluidic system to simultaneously separate and assess blood cells deformability. PMID:26482154

  13. High Throughput Label Free Measurement of Cancer Cell Adhesion Kinetics Under Hemodynamic Flow

    PubMed Central

    Spencer, Adrianne; Baker, Aaron B.

    2016-01-01

    The kinetics of receptor-mediated cell adhesion to extracellular matrix and adherent cell monolayers plays a key role in many physiological and pathological processes including cancer metastasis. Within this process the presence of fluidic shear forces is a key regulator of binding equilibrium and kinetics of cell adhesion. Current techniques to examine the kinetics of cell adhesion are either performed in the absence of flow or are low throughput, limiting their application to pharmacological compound screening or the high throughput investigation of biological mechanisms. We developed a high throughput flow device that applies flow in a multi-well format and interfaced this system with electric cell-substrate impedance sensing (ECIS) system to allow label free detection of cell adhesion. We demonstrate that this combined system is capable of making real time measurements of cancer cell adhesion to extracellular matrix and immobilized platelets. In addition, we examined the dependence of the kinetics of binding of cancer cells on the level of shear stress and in the presence of small molecule inhibitors to adhesion-related pathways. This versatile system is broadly adaptable to the high throughput study of cell adhesion kinetics for many applications including drug screening and the investigation of the mechanisms of cancer metastasis. PMID:26816215

  14. Coded illumination for motion-blur free imaging of cells on cell-phone based imaging flow cytometer

    NASA Astrophysics Data System (ADS)

    Saxena, Manish; Gorthi, Sai Siva

    2014-10-01

    Cell-phone based imaging flow cytometry can be realized by flowing cells through the microfluidic devices, and capturing their images with an optically enhanced camera of the cell-phone. Throughput in flow cytometers is usually enhanced by increasing the flow rate of cells. However, maximum frame rate of camera system limits the achievable flow rate. Beyond this, the images become highly blurred due to motion-smear. We propose to address this issue with coded illumination, which enables recovery of high-fidelity images of cells far beyond their motion-blur limit. This paper presents simulation results of deblurring the synthetically generated cell/bead images under such coded illumination.

  15. A dual fluorescence flow cytometric analysis of bacterial adherence to mammalian host cells

    PubMed Central

    Hara-Kaonga, Bochiwe; Pistole, Thomas G.

    2009-01-01

    Flow cytometry has provided a powerful tool for analyzing bacteria-host cell associations. Established approaches have used bacteria, labeled either directly with fluorochromes or indirectly with fluorescently conjugated antibodies, to detect these associations. Although useful, these techniques are unable consistently to include all host cells in the analysis while excluding free, aggregated bacteria. This study describes a new flow cytometry method of assessing bacterial adherence to host cells based on direct fluorescent labeling of both bacteria and host cells. Eukaryotic host cells were labeled with PKH-26, a red fluorescent dye, and bacteria were labeled with fluorescein isothiocyanate, a green fluorescent dye. The red host cells were gated and the mean green fluorescence intensity (MFI) of these red cells was determined. We used MFI values obtained from control samples (unlabeled and labeled host cells with unlabeled bacteria) to eliminate contributions due to autofluorescence. The final MFI values represent fluorescence of host cells resulting from the adherent bacteria. Because all red fluorescent cells are analyzed, this method includes all the eukaryotic cells for analysis but excludes all free or aggregated bacteria that are not bound to target cells. PMID:17222473

  16. A dual fluorescence flow cytometric analysis of bacterial adherence to mammalian host cells.

    PubMed

    Hara-Kaonga, Bochiwe; Pistole, Thomas G

    2007-04-01

    Flow cytometry has provided a powerful tool for analyzing bacteria-host cell associations. Established approaches have used bacteria, labeled either directly with fluorochromes or indirectly with fluorescently conjugated antibodies, to detect these associations. Although useful, these techniques are consistently unable to include all host cells in the analysis while excluding free, aggregated bacteria. This study describes a new flow cytometry method of assessing bacterial adherence to host cells based on direct fluorescent labeling of both bacteria and host cells. Eukaryotic host cells were labeled with PKH-26, a red fluorescent dye, and bacteria were labeled with fluorescein isothiocyanate, a green fluorescent dye. The red host cells were gated and the mean green fluorescence intensity (MFI) of these red cells was determined. We used MFI values obtained from control samples (unlabeled and labeled host cells with unlabeled bacteria) to eliminate contributions due to autofluorescence. The final MFI values represent fluorescence of host cells resulting from the adherent bacteria. Because all red fluorescent cells are analyzed, this method includes all the eukaryotic cells for analysis but excludes all free or aggregated bacteria that are not bound to target cells.

  17. Detection of Apoptotic Versus Autophagic Cell Death by Flow Cytometry.

    PubMed

    Sica, Valentina; Maiuri, M Chiara; Kroemer, Guido; Galluzzi, Lorenzo

    2016-01-01

    Different modes of regulated cell death (RCD) can be initiated by distinct molecular machineries and their morphological manifestations can be difficult to discriminate. Moreover, cells responding to stress often activate an adaptive response centered around autophagy, and whether such a response is cytoprotective or cytotoxic cannot be predicted based on morphological parameters only. Molecular definitions are therefore important to understand various RCD subroutines from a mechanistic perspective. In vitro, various forms of RCD including apoptosis and autophagic cell death can be easily discriminated from each other with assays that involve chemical or pharmacological interventions targeting key components of either pathway. Here, we detail a straightforward method to discriminate apoptosis from autophagic cell death by flow cytometry, based on the broad-spectrum caspase inhibitor Z-VAD-fmk and the genetic inhibition of ATG5.

  18. Endothelial cell alignment as a result of anisotropic strain and flow induced shear stress combinations

    PubMed Central

    Sinha, Ravi; Le Gac, Séverine; Verdonschot, Nico; van den Berg, Albert; Koopman, Bart; Rouwkema, Jeroen

    2016-01-01

    Endothelial cells (ECs) are continuously exposed in vivo to cyclic strain and shear stress from pulsatile blood flow. When these stimuli are applied in vitro, ECs adopt an appearance resembling their in vivo state, most apparent in their alignment (perpendicular to uniaxial strain and along the flow). Uniaxial strain and flow perpendicular to the strain, used in most in vitro studies, only represent the in vivo conditions in straight parts of vessels. The conditions present over large fractions of the vasculature can be better represented by anisotropic biaxial strains at various orientations to flow. To emulate these biological complexities in vitro, we have developed a medium-throughput device to screen for the effects on cells of variously oriented anisotropic biaxial strains and flow combinations. Upon the application of only strains for 24 h, ECs (HUVECs) aligned perpendicular to the maximum principal strain and the alignment was stronger for a higher maximum:minimum principal strain ratio. A 0.55 Pa shear stress, when applied alone or with strain for 24 h, caused cells to align along the flow. Studying EC response to such combined physiological mechanical stimuli was not possible with existing platforms and to our best knowledge, has not been reported before. PMID:27404382

  19. Applications of fixed-time flow cytometry in cell biology

    NASA Astrophysics Data System (ADS)

    Tarnok, Attila

    1998-04-01

    Flow cytometric (FCM) measurement of intracellular free calcium [Ca2+]i, transients is usually done by two methods: (a) after a short prerun period to assess the baseline the measurement is stopped, stimulus is added and the measurement continued or (b) stimulus is injected during measurement and the sample pressure briefly increased to deliver cells rapidly to the detection point. In (a) measurement of very short transients [Ca2+]i is impeded by the lag time between stimulus addition and restart of acquisition. In (b) response of pressure sensitive cells is hard to analyze. Furthermore, (a) and (b) do not allow to quantify and sort rare responders. A simple Fixed- Time device has been developed. Ca2+ sensitive fluorescent dye labeled cells and a stimulus are placed in different vials. Both fluids are forced by the same pressure through tubing that merges into a T-junction where they mix and are delivered through a connecting tube to the FCM: [Ca2+]i is measured at a certain time after stimulation that is adjusted by sample flow rate and length of the connecting tube. With Fixed-Time, the pressure sensitive neuronal NH15-CA2 cell was analyzed. Furthermore, rare neurotransmitter responsive fibroblast from normal and transfected cultures were sorted and cloned and their dose response characterized. The results demonstrate that fixed- time FCM is an important tool for the analysis of the cells physiology and the preparation of responders.

  20. An experimental study of the elastic theory for granular flows

    NASA Astrophysics Data System (ADS)

    Guo, Tongtong; Campbell, Charles S.

    2016-08-01

    This paper reports annular shear cell measurements granular flows with an eye towards experimentally confirming the flow regimes laid out in the elastic theory of granular flow. Tests were carried out on four different kinds of plastic spherical particles under both constant volume flows and constant applied stress flows. In particular, observations were made of the new regime in that model, the elastic-inertial regime, and the predicted transitions between the elastic-inertial and both the elastic-quasistatic and pure inertial regimes.

  1. An experimental study of separated flow on a finite wing

    NASA Technical Reports Server (NTRS)

    Winkelmann, A. E.

    1981-01-01

    The flow field associated with the formation of a mushroom shaped trailing edge stall cell on a low-aspect-ratio (AR = 4.0) wing was investigated in a series of low speed wind tunnel tests (Reynolds number based on 15.2 cm chord = 480,000). Flow field surveys of the separation bubble and wake of a partially stalled and fully stalled wing were completed using a hot-wire probe, a split-film probe, and a directional sensitive pressure probe. A new color video display technique was developed to display the flow field survey data. Photographs were obtained of surface oil flow patterns and smoke flow visualization

  2. Monitoring circulating apoptotic cells by in-vivo flow cytometry

    NASA Astrophysics Data System (ADS)

    Wei, Xunbin; Tan, Yuan; Chen, Yun; Zhang, Li; Li, Yan; Liu, Guangda; Wu, Bin; Wang, Chen

    2008-02-01

    Chemotherapies currently constitute one main venue of cancer treatment. For a large number of adult and elderly patients, however, treatment options are poor. These patients may suffer from disease that is resistant to conventional chemotherapy or may not be candidates for curative therapies because of advanced age or poor medical conditions. To control disease in these patients, new therapies must be developed that are selectively targeted to unique characteristics of tumor cell growth and metastasis. A reliable early evaluation and prediction of response to the chemotherapy is critical to its success. Chemotherapies induce apoptosis in tumor cells and a portion of such apoptotic cancer cells may be present in the circulation. However, the fate of circulating tumor cells is difficult to assess with conventional methods that require blood sampling. We report the in situ measurement of circulating apoptotic cells in live animals using in vivo flow cytometry, a novel method that enables real-time detection and quantification of circulating cells without blood extraction. Apoptotic cells are rapidly cleared from the circulation with a half-life of ~10 minutes. Real-time monitoring of circulating apoptotic cells can be useful for detecting early changes in disease processes, as well as for monitoring response to therapeutic intervention.

  3. Tracking Immune Cell Proliferation and Cytotoxic Potential Using Flow Cytometry

    PubMed Central

    Tario, Joseph D.; Muirhead, Katharine A.; Pan, Dalin; Munson, Mark E.; Wallace, Paul K.

    2015-01-01

    In the second edition of this series, we described the use of cell tracking dyes in combination with tetramer reagents and traditional phenotyping protocols to monitor levels of proliferation and cytokine production in antigen-specific CD8+ T cells. In particular, we illustrated how tracking dye fluorescence profiles could be used to ascertain the precursor frequencies of different subsets in the T-cell pool that are able to bind tetramer, synthesize cytokines, undergo antigen-driven proliferation, and/or carry out various combinations of these functional responses. Analysis of antigen-specific proliferative responses represents just one of many functions that can be monitored using cell tracking dyes and flow cytometry. In this third edition, we address issues to be considered when combining two different tracking dyes with other phenotypic and viability probes for the assessment of cytotoxic effector activity and regulatory T-cell functions. We summarize key characteristics of and differences between general protein- and membrane-labeling dyes, discuss determination of optimal staining concentrations, and provide detailed labeling protocols for both dye types. Examples of the advantages of two-color cell tracking are provided in the form of protocols for (a) independent enumeration of viable effector and target cells in a direct cytotoxicity assay and (b) simultaneous monitoring of proliferative responses in effector and regulatory T cells. PMID:21116982

  4. Mechanical response of tumor cells flowing through a microfluidic capillary

    NASA Astrophysics Data System (ADS)

    Khan, Zeina S.; Kamyabi, Nabiollah; Hussain, Fazle; Vanapalli, Siva A.

    2014-03-01

    Circulating tumor cells, the primary cause of cancer metastasis, are transported throughout the body to distant organs by blood flow. Despite the importance of cell transport and deformability in the vasculature for cancer metastasis, quantitative understanding of the hydrodynamic interactions between the cells and the blood vessel walls is lacking. Using a model microfluidic capillary of rectangular cross-section with an on-chip manometer coupled with high speed video imaging, we quantitatively investigate the hydrodynamic behavior via the cell excess pressure drop. By characterizing our device with simple model systems including viscous drops and soft elastic particles, we find that the excess pressure drop shows no apparent dependence on elastic modulus or interfacial tension, but depends significantly on internal viscosity for moderate confinements and shear stresses within the physiological range of 1-10 Pa. This suggests that the metastatic potential of circulating cells can be characterized by the effective viscosity. We test this hypothesis with several tumor cell lines and find that the effective cell viscosity determined from excess pressure drop measurements can be used to differentiate highly from lowly invasive cells.

  5. Flow distribution and maximum current density studies in redox flow batteries with a single passage of the serpentine flow channel

    NASA Astrophysics Data System (ADS)

    Ke, Xinyou; Alexander, J. Iwan D.; Prahl, Joseph M.; Savinell, Robert F.

    2014-12-01

    Flow batteries show promise for very large-scale stationary energy storage such as needed for the grid and renewable energy implementation. In recent years, researchers and developers of redox flow batteries (RFBs) have found that electrode and flow field designs of PEM fuel cell (PEMFC) technology can increase the power density and consequently push down the cost of flow battery stacks. In this paper we present a macroscopic model of a typical PEMFC-like RFB electrode-flow field design. The model is a layered system comprised of a single passage of a serpentine flow channel and a parallel underlying porous electrode (or porous layer). The effects of the inlet volumetric flow rate, permeability of the porous layer, thickness of the porous layer and thickness of the flow channel on the flow penetration into the porous layer are investigated. The maximum current density corresponding to stoichiometry is estimated to be 377 mA cm-2 and 724 mA cm-2, which compares favorably with experiments of ∼400 mA cm-2 and ∼750 mA cm-2, for a single layer and three layers of the carbon fiber paper, respectively.

  6. RhizoFlowCell system reveals early effects of micropollutants on aquatic plant rhizosphere.

    PubMed

    Mynampati, Kalyan Chakravarthy; Lee, Yong Jian; Wijdeveld, Arjan; Reuben, Sheela; Samavedham, Lakshminarayanan; Kjelleberg, Staffan; Swarup, Sanjay

    2015-12-01

    In aquatic systems, one of the non-destructive ways to quantify toxicity of contaminants to plants is to monitor changes in root exudation patterns. In aquatic conditions, monitoring and quantifying such changes are currently challenging because of dilution of root exudates in water phase and lack of suitable instrumentation to measure them. Exposure to pollutants would not only change the plant exudation, but also affect the microbial communities that surround the root zone, thereby changing the metabolic profiles of the rhizosphere. This study aims at developing a device, the RhizoFlowCell, which can quantify metabolic response of plants, as well as changes in the microbial communities, to give an estimate of the stress to which the rhizosphere is exposed. The usefulness of RhizoFlowCell is demonstrated using naphthalene as a test pollutant. Results show that RhizoFlowCell system is useful in quantifying the dynamic metabolic response of aquatic rhizosphere to determine ecosystem health.

  7. RhizoFlowCell system reveals early effects of micropollutants on aquatic plant rhizosphere.

    PubMed

    Mynampati, Kalyan Chakravarthy; Lee, Yong Jian; Wijdeveld, Arjan; Reuben, Sheela; Samavedham, Lakshminarayanan; Kjelleberg, Staffan; Swarup, Sanjay

    2015-12-01

    In aquatic systems, one of the non-destructive ways to quantify toxicity of contaminants to plants is to monitor changes in root exudation patterns. In aquatic conditions, monitoring and quantifying such changes are currently challenging because of dilution of root exudates in water phase and lack of suitable instrumentation to measure them. Exposure to pollutants would not only change the plant exudation, but also affect the microbial communities that surround the root zone, thereby changing the metabolic profiles of the rhizosphere. This study aims at developing a device, the RhizoFlowCell, which can quantify metabolic response of plants, as well as changes in the microbial communities, to give an estimate of the stress to which the rhizosphere is exposed. The usefulness of RhizoFlowCell is demonstrated using naphthalene as a test pollutant. Results show that RhizoFlowCell system is useful in quantifying the dynamic metabolic response of aquatic rhizosphere to determine ecosystem health. PMID:26386206

  8. Visualization studies of turbulent transition flows in a porous medium

    NASA Technical Reports Server (NTRS)

    Bilardo, V. J.

    1983-01-01

    Results are reported for flow-visualization studies of the flow regimes of water passing through a porous medium consisting of cylindrical glass and plexiglas rods arranged in a complex and fixed three-dimensional geometry. The Reynolds number (Re) varied from 50 to 700; the flow was visualized by injecting a 5% potassium permanganate dye solution into the pores and photographing the resulting dye streaklines with both a still camera and a movie camera. The results indicate that four distinct flow regimes exist in the porous medium: (1) Darcy or creeping flow up to Re = 3; (2) steady inertia-dominated laminar flow for Re = 3-150; (3) unsteady transitional laminar flow for Re = 150-250; and (4) fully turbulent flow for Re greater than 250. It is concluded that a laminar wake instability mechanism typical of the external flow about bluff bodies may be responsible for the overall transition from laminar to turbulent flow in porous media.

  9. Flow Cytometric Analysis of T, B, and NK Cells Antigens in Patients with Mycosis Fungoides

    PubMed Central

    Yazıcı, Serkan; Bülbül Başkan, Emel; Budak, Ferah; Oral, Barbaros; Adim, Şaduman Balaban; Ceylan Kalin, Zübeyde; Özkaya, Güven; Aydoğan, Kenan; Saricaoğlu, Hayriye; Tunali, Şükran

    2015-01-01

    We retrospectively analyzed the clinicopathological correlation and prognostic value of cell surface antigens expressed by peripheral blood mononuclear cells in patients with mycosis fungoides (MF). 121 consecutive MF patients were included in this study. All patients had peripheral blood flow cytometry as part of their first visit. TNMB and histopathological staging of the cases were retrospectively performed in accordance with International Society for Cutaneous Lymphomas/European Organization of Research and Treatment of Cancer (ISCL/EORTC) criteria at the time of flow cytometry sampling. To determine prognostic value of cell surface antigens, cases were divided into two groups as stable and progressive disease. 17 flow cytometric analyses of 17 parapsoriasis (PP) and 11 analyses of 11 benign erythrodermic patients were included as control groups. Fluorescent labeled monoclonal antibodies were used to detect cell surface antigens: T cells (CD3+, CD4+, CD8+, TCRαβ+, TCRγδ+, CD7+, CD4+CD7+, CD4+CD7−, and CD71+), B cells (HLA-DR+, CD19+, and HLA-DR+CD19+), NKT cells (CD3+CD16+CD56+), and NK cells (CD3−CD16+CD56+). The mean value of all cell surface antigens was not statistically significant between parapsoriasis and MF groups. Along with an increase in cases of MF stage statistically significant difference was found between the mean values of cell surface antigens. Flow cytometric analysis of peripheral blood cell surface antigens in patients with mycosis fungoides may contribute to predicting disease stage and progression. PMID:26788525

  10. Flow microfluorometric analysis of phagocyte degranulation in bacteria-infected whole human blood cell cultures

    NASA Astrophysics Data System (ADS)

    Kravtsov, Alexander L.; Bobyleva, Elena V.; Grebenyukova, Tatyana P.; Kuznetsov, Oleg S.; Kulyash, Youri V.

    2002-07-01

    A quantitative flow microfluorometric method was used to study the intensity of human blood phagocyte degranulation in response to viable staphylococcus aureus or Yersinia pestis cells. Microorganisms were added directly to defibrinated whole blood. Uninfected and infected blood samples were incubated at 37 degrees C to 8 h. The results were recorded in dynamics after the staining of whole blood with acridine orange solution. Lymphocytes with a low azurophilic granule per cell content were discriminated from phagocytes by the measurement of single cell red cytoplasmic granule fluorescence. 30,000 cells in each sample were examined. S. aureus cells caused a dose-dependent decrease in the number of phagocytes having a high red cytoplasmic fluorescence intensity and a corresponding increase in the weakly fluorescence cell population. In the presence of an initial S. aureus-to-phagocyte ratio more than 1:1, degranulation was measured after 3 h of incubation and to 8 h the percentage of degranulated phagocytes was at least 100 percent Y. pestis cells grown for 48 h at 28 degrees C caused at same condition as the degranulation only about 50 percent of cells. Y.pestis EV cells preincubated in broth for 12 h at 37 degrees C did no stimulate the phahocyte degranulation. The results of these studies suggest that analysis of cell populations via flow microfluorimeter technology may be a powerful tool in analysis bacterial infection.

  11. Improved Method for Bacterial Cell Capture after Flow Cytometry Cell Sorting ▿

    PubMed Central

    Guillebault, D.; Laghdass, M.; Catala, P.; Obernosterer, I.; Lebaron, P.

    2010-01-01

    Fixed cells with different nucleic acid contents and scatter properties (low nucleic acid [LNA], high nucleic acid 1 [HNA1], and HNA2) were sorted by flow cytometry (FCM). For each sort, 10,000 cells were efficiently captured on poly-l-lysine-coated microplates, resulting in efficient and reproducible PCR amplification. PMID:20817799

  12. Studies of fluid instabilities in flows of lava and debris

    NASA Technical Reports Server (NTRS)

    Fink, Jonathan H.

    1987-01-01

    At least two instabilities have been identified and utilized in lava flow studies: surface folding and gravity instability. Both lead to the development of regularly spaced structures on the surfaces of lava flows. The geometry of surface folds have been used to estimate the rheology of lava flows on other planets. One investigation's analysis assumed that lava flows have a temperature-dependent Newtonian rheology, and that the lava's viscosity decreased exponentially inward from the upper surface. The author reviews studies by other investigators on the analysis of surface folding, the analysis of Taylor instability in lava flows, and the effect of surface folding on debris flows.

  13. Commodity Flow Study - Clark County, Nevada, USA

    SciTech Connect

    Conway, S.Ph.D.; Navis, I.

    2008-07-01

    The United States Department of Energy has designated Clark County, Nevada as an 'Affected Unit of Local Government' due to the potential for impacts by activities associated with the Yucca Mountain High Level Nuclear Waste Repository project. Urban Transit, LLC has led a project team of transportation including experts from the University of Nevada Las Vegas Transportation Research Center to conduct a hazardous materials community flow study along Clark County's rail and truck corridors. In addition, a critical infrastructure analysis has also been carried out in order to assess the potential impacts of transportation within Clark County of high level nuclear waste and spent nuclear fuel to a proposed repository 90 miles away in an adjacent county on the critical infrastructure in Clark County. These studies were designed to obtain information relating to the transportation, identification and routing of hazardous materials through Clark County. Coordinating with the United States Department of Energy, the U.S. Department of Agriculture, the U. S. Federal Highway Administration, the Nevada Department of Transportation, and various other stakeholders, these studies and future research will examine the risk factors along the entire transportation corridor within Clark County and provide a context for understanding the additional vulnerability associated with shipping spent fuel through Clark County. (authors)

  14. A high-order adaptive Cartesian cut-cell method for simulation of compressible viscous flow over immersed bodies

    NASA Astrophysics Data System (ADS)

    Muralidharan, Balaji; Menon, Suresh

    2016-09-01

    A new adaptive finite volume conservative cut-cell method that is third-order accurate for simulation of compressible viscous flows is presented. A high-order reconstruction approach using cell centered piecewise polynomial approximation of flow quantities, developed in the past for body-fitted grids, is now extended to the Cartesian based cut-cell method. It is shown that the presence of cut-cells of very low volume results in numerical oscillations in the flow solution near the embedded boundaries when standard small cell treatment techniques are employed. A novel cell clustering approach for polynomial reconstruction in the vicinity of the small cells is proposed and is shown to achieve smooth representation of flow field quantities and their derivatives on immersed interfaces. It is further shown through numerical examples that the proposed clustering method achieves the design order of accuracy and is fairly insensitive to the cluster size. Results are presented for canonical flow past a single cylinder and a sphere at different flow Reynolds numbers to verify the accuracy of the scheme. Investigations are then performed for flow over two staggered cylinders and the results are compared with prior data for the same configuration. All the simulations are carried out with both quadratic and cubic reconstruction, and the results indicate a clear improvement with the cubic reconstruction. The new cut-cell approach with cell clustering is able to predict accurate results even at relatively low resolutions. The ability of the high-order cut-cell method in handling sharp geometrical corners and narrow gaps is also demonstrated using various examples. Finally, three-dimensional flow interactions between a pair of spheres in cross flow is investigated using the proposed cut-cell scheme. The results are shown to be in excellent agreement with past studies, which employed body-fitted grids for studying this complex case.

  15. Optical analysis of nanomaterial-cell interactions: flow cytometry and digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Mues, Sarah; Antunovic, Jan; Ossig, Rainer; Kemper, Björn; Schnekenburger, Jürgen

    2015-05-01

    The in vitro cytotoxicity assessment of engineered nanoparticles commonly involves the measurement of different endpoints like the formation of reactive oxygen species, cell viability or cell death. Usually these parameters are determined by optical readouts of enzymatically converted substrates that often interfere with the tested nanomaterials. Using cell viability (WST-8) and cell death (LDH) as parameter we have initially investigated the toxic effects of spherical (NM 300) and rod shaped (NM 302) silver nanomaterials with a matrix of four cell lines representing different functions: lung and kidney epithelial cells, macrophages and fibroblasts. In addition, we have used a label-free flow cytometer configuration to investigate interactions of particles and macrophages by side scatter signal analysis. Finally, we explored digital holographic microscopy (DHM) for multimodal label-free analysis of nanomaterial toxicity. Quantitative DHM phase images were analyzed for cell thickness, volume, density, dry mass and refractive index. We could demonstrate that silver spheres lead to more cytotoxic effects than rods in all four examined cell lines and both assay. Exemplarily a dose dependent interaction increase of cells with NM 300 and NM 302 analyzed by flow cytometry is shown. Furthermore, we found that the refractive index of cells is influenced by incubation with NM 300 in a decreasing manner. A 24 hours time-lapse measurement revealed a dose dependent decrease of dry mass and surface area development indicating reduced cell viability and cell death. Our results demonstrate digital holographic microscopy and flow cytometry as valuable label-free tools for nanomaterial toxicity and cell interaction studies.

  16. Studies on pressure losses and flow rate optimization in vanadium redox flow battery

    NASA Astrophysics Data System (ADS)

    Tang, Ao; Bao, Jie; Skyllas-Kazacos, Maria

    2014-02-01

    Premature voltage cut-off in the operation of the vanadium redox flow battery is largely associated with the rise in concentration overpotential at high state-of-charge (SOC) or state-of-discharge (SOD). The use of high constant volumetric flow rate will reduce concentration overpotential, although potentially at the cost of consuming excessive pumping energy which in turn lowers system efficiency. On the other hand, any improper reduction in flow rate will also limit the operating SOC and lead to deterioration in battery efficiency. Pressure drop losses are further exacerbated by the need to reduce shunt currents in flow battery stacks that requires the use of long, narrow channels and manifolds. In this paper, the concentration overpotential is modelled as a function of flow rate in an effort to determine an appropriate variable flow rate that can yield high system efficiency, along with the analysis of pressure losses and total pumping energy. Simulation results for a 40-cell stack under pre-set voltage cut-off limits have shown that variable flow rates are superior to constant flow rates for the given system design and the use of a flow factor of 7.5 with respect to the theoretical flow rate can reach overall high system efficiencies for different charge-discharge operations.

  17. Studying tumor metastasis by in vivo imaging and flow cytometer

    NASA Astrophysics Data System (ADS)

    Wei, Xunbin; Guo, Jin; Liu, Guangda; Li, Yan; Chen, Yun; Zhang, Li; Tan, Yuan; Chen, Tong; Gu, Zhenqin; Wang, Chen

    2009-02-01

    Liver cancer is one of the most common malignancies in the world, with approximately 1,000,000 cases reported every year. This ranges from 15,000 cases in the United States to more than a 250,000 in China. About 80% of people with primary liver cancer are male. Although two-thirds of people have advanced liver disease when they seek medical help, one third of the patients have cancer that has not progressed beyond the liver. Primary liver cancer (hepatocellular carcinoma, or HCC) is associated with liver cirrhosis 60-80% of the time. HCC may metastasize to the lung, bones, kidney, and many other organs. Surgical resection, liver transplantation, chemotherapy and radiation therapy are the foundation of current HCC therapies. However the outcomes are poor-the survival rate is almost zero for metastatic HCC patients. Molecular mechanisms of HCC metastasis need to be understood better and new therapies must be developed to selectively target to unique characteristics of HCC cell growth and metastasis. We have developed the "in vivo microscopy" to study the mechanisms that govern liver tumor cell spread through the microenvironment in vivo in real-time confocal near-infrared fluorescence imaging. A recently developed "in vivo flow cytometer" and optical imaging are used to assess liver tumor cell spreading and the circulation kinetics of liver tumor cells. A real-time quantitative monitoring of circulating liver tumor cells by the in vivo flow cytometer will be useful to assess the effectiveness of the potential therapeutic interventions.

  18. Mimicking the Interfacial Dynamics of Flowing White Blood Cells

    NASA Astrophysics Data System (ADS)

    Santore, Maria

    2015-03-01

    The rolling of particles on surfaces, facilitated by hydrodynamic forces combined with localized surface interactions of the appropriate strengths, spatial arrangements, and ranges, is a technologically useful means of transporting and manipulating particles. One's intuition for the rolling of a marble or a car tire cannot be extrapolated down to microparticle length scales because the microparticle interactions are dominated by electrostatic, van der Waals, and hydrogen bonding interactions rather than a friction that depends on an imposed normal force. Indeed, our microparticle rolling systems are inspired by the rolling of white blood cells on the inner walls of venules as part of the innate immune response: Selectin molecules engage with their counterparts on the opposing surfaces to slow cell motion relative to that for freely flowing cells. In the resulting rolling signature, ligand-receptor binding and crack closing on the front of the cell are balanced with molecular dis-bonding and crack opening at the rear. The contact region is relatively static, allowing other interactions (for instance signaling) to occur for a finite duration. Thus, achieving particle rolling in synthetic systems is important because it facilitates particle-surface interactions in a continuous nonfouling fashion where the contact surface is continually renewed. In developing a synthetic model for this system, we employ polymers to modify flowing particles and /or planar collectors, producing heterogeneous interfaces which can support rolling or produce other motion signatures such as skipping, arrest, or free flow. We identify, in the synthetic system, combinations of variables that produce rolling and demonstrate how the distinction between rolling and arrest is not a simple matter of the adhesion strength between the particles and the collector. Rolling is a cooperative process and the coordination of binding in one location with dis-bonding in another requires appropriate length

  19. Modeling of gaseous flows within proton exchange membrane fuel cells

    SciTech Connect

    Weisbrod, K.R.; Vanderborgh, N.E.; Grot, S.A.

    1996-12-31

    Development of a comprehensive mechanistic model has been helpful to understand PEM fuel cell performance. Both through-the-electrode and down-the-channel models have been developed to support our experimental effort to enhance fuel cell design and operation. The through-the-electrode model was described previously. This code describes the known transport properties and dynamic processes that occur within a membrane and electrode assembly. Key parameters include transport through the backing layers, water diffusion and electroosmotic transport in the membrane, and reaction electrochemical kinetics within the cathode catalyst layer. In addition, two geometric regions within the cathode layer are represented, the first region below saturation and second with liquid water present. Although processes at high gas stoichiometry are well represented by more simple codes, moderate stoichiometry processes require a two dimensional representation that include the gaseous composition and temperature along flow channel. Although usually PEM hardware utilizes serpentine flow channels, this code does not include such geometric features and thus the flow can be visualized along a single channel.

  20. Improved Flow-Field Structures for Direct Methanol Fuel Cells

    SciTech Connect

    Gurau, Bogdan

    2013-05-31

    The direct methanol fuel cell (DMFC) is ideal if high energy-density liquid fuels are required. Liquid fuels have advantages over compressed hydrogen including higher energy density and ease of handling. Although state-of-the-art DMFCs exhibit manageable degradation rates, excessive fuel crossover diminishes system energy and power density. Although use of dilute methanol mitigates crossover, the concomitant lowering of the gross fuel energy density (GFED) demands a complex balance-of-plant (BOP) that includes higher flow rates, external exhaust recirculation, etc. An alternative approach is redesign of the fuel delivery system to accommodate concentrated methanol. NuVant Systems Inc. (NuVant) will maximize the GFED by design and assembly of a DMFC that uses near neat methanol. The approach is to tune the diffusion of highly concentrated methanol (to the anode catalytic layer) to the back-diffusion of water formed at the cathode (i.e. in situ generation of dilute methanol at the anode layer). Crossover will be minimized without compromising the GFED by innovative integration of the anode flow-field and the diffusion layer. The integrated flow-field-diffusion-layers (IFDLs) will widen the current and potential DMFC operating ranges and enable the use of cathodes optimized for hydrogen-air fuel cells.

  1. A Multichannel Dampened Flow System for Studies on Shear Stress-Mediated Mechanotransduction

    PubMed Central

    Voyvodic, Peter L.; Min, Daniel; Baker, Aaron B.

    2012-01-01

    Shear stresses are powerful regulators of cellular function and potent mediators of the development of vascular disease. We have designed and optimized a system allowing the application of flow to cultured cells in a multichannel format. By using a multichannel peristaltic pump, flow can be driven continuously in the system for long-term studies in multiple isolated flow loops. A key component of the system is a dual-chamber pulse dampener that removes the pulsatility of the flow without the need for having an open system or elevated reservoir. We optimized the design parameters of the pulse dampening chambers for the maximum reduction in flow pulsation while minimizing the fluid needed for each isolated flow channel. Human umbilical vein endothelial cells (HUVECs) were exposed to steady and pulsatile shear stress using the system. We found that cells under steady flow had a marked increased production of eNOS and formation of actin stress fibers in comparison to those under pulsatile flow conditions. Overall, the results confirm the utility of the device as a practical means to apply shear stress to cultured cells in the multichannel format and provide steady, long term flow to microfluidic devices. PMID:22836694

  2. A multichannel dampened flow system for studies on shear stress-mediated mechanotransduction.

    PubMed

    Voyvodic, Peter L; Min, Daniel; Baker, Aaron B

    2012-09-21

    Shear stresses are powerful regulators of cellular function and potent mediators of the development of vascular disease. We have designed and optimized a system allowing the application of flow to cultured cells in a multichannel format. By using a multichannel peristaltic pump, flow can be driven continuously in the system for long-term studies in multiple isolated flow loops. A key component of the system is a dual-chamber pulse dampener that removes the pulsatility of the flow without the need for having an open system or elevated reservoir. We optimized the design parameters of the pulse dampening chambers for the maximum reduction in flow pulsation while minimizing the fluid needed for each isolated flow channel. Human umbilical vein endothelial cells (HUVECs) were exposed to steady and pulsatile shear stress using the system. We found that cells under steady flow had a marked increased production of eNOS and formation of actin stress fibers in comparison to those under pulsatile flow conditions. Overall, the results confirm the utility of the device as a practical means to apply shear stress to cultured cells in the multichannel format and provide steady, long term flow to microfluidic devices.

  3. Microfluidic Impedance Flow Cytometry Enabling High-Throughput Single-Cell Electrical Property Characterization

    PubMed Central

    Chen, Jian; Xue, Chengcheng; Zhao, Yang; Chen, Deyong; Wu, Min-Hsien; Wang, Junbo

    2015-01-01

    This article reviews recent developments in microfluidic impedance flow cytometry for high-throughput electrical property characterization of single cells. Four major perspectives of microfluidic impedance flow cytometry for single-cell characterization are included in this review: (1) early developments of microfluidic impedance flow cytometry for single-cell electrical property characterization; (2) microfluidic impedance flow cytometry with enhanced sensitivity; (3) microfluidic impedance and optical flow cytometry for single-cell analysis and (4) integrated point of care system based on microfluidic impedance flow cytometry. We examine the advantages and limitations of each technique and discuss future research opportunities from the perspectives of both technical innovation and clinical applications. PMID:25938973

  4. DNA cell-cycle analysis of cervical cancer by flow cytometry using simultaneous cytokeratin labelling for identification of tumour cells.

    PubMed

    Kimmig, R; Kapsner, T; Spelsberg, H; Untch, M; Hepp, H

    1995-01-01

    DNA ploidy and cell-cycle distribution were determined by flow cytometry in fresh tumour tissue of 53 cervical carcinomas. Epithelial cells were labelled by a fluorescein-isothiocyanate-conjugated cytokeratin antibody (CK6, CK18) to study the influence of contaminating stromal and inflammatory cells on results of cell-cycle analysis of tumour cells. Without identification of cytokeratin-positive cells 30/53 (57%) tumours were found to be DNA-aneuploid compared to 43/53 (81%) after gating for cytokeratin. Only 7 of 15 DNA-multiploid tumours could be detected without cytokeratin staining. In addition, cytokeratin-negative cells, which are found in all tumours, can be used as an internal standard for the calculation of ploidy and for quality control (coefficient of variation, linearity) of each individual sample. Cell-cycle analysis revealed significantly higher S-phase and G2M-phase fractions in cytokeratin-gated compared to ungated samples (13.1% versus 10.0% and 8.0% versus 5.4%; P < 0.001). This difference was more pronounced in DNA-diploid than DNA-aneuploid tumours. In conclusion, about 30% of DNA-aneuploid tumours could only be detected after cytokeratin labelling of epithelial cells. Owing to the identification of cytokeratin-positive cells the influence of non-tumoural cell elements on cell-cycle analysis was reduced markedly. Therefore, in cervical cancer, cytokeratin labelling can optimize both the determination of DNA ploidy and cell-cycle analysis.

  5. In vivo photoacoustic flow cytometry for monitoring of circulating single cancer cells and contrast agents

    NASA Astrophysics Data System (ADS)

    Zharov, Vladimir P.; Galanzha, Ekaterina I.; Shashkov, Evgeny V.; Khlebtsov, Nicolai G.; Tuchin, Valery V.

    2006-12-01

    A new photoacoustic flow cytometry was developed for real-time detection of circulating cells, nanoparticles, and contrast agents in vivo. Its capability, integrated with photothermal and optical clearing methods, was demonstrated using a near-infrared tunable laser to characterize the in vivo kinetics of Indocyanine Green alone and single cancer cells labeled with gold nanorods and Indocyanine Green in the vasculature of the mouse ear. In vivo applications are discussed, including selective nanophotothermolysis of metastatic squamous cells, label-free detection of melanoma cells, study of pharmokinetics, and immune response to apoptotic and necrotic cells, with potential translation to humans. The threshold sensitivity is estimated as one cancer cell in the background of 107 normal blood cells.

  6. Computational modeling of air-breathing microfluidic fuel cells with flow-over and flow-through anodes

    NASA Astrophysics Data System (ADS)

    Zhang, Biao; Ye, Ding-ding; Sui, Pang-Chieh; Djilali, Ned; Zhu, Xun

    2014-08-01

    A three-dimensional computational model for air-breathing microfluidic fuel cells (AMFCs) with flow-over and flow-through anodes is developed. The coupled multiphysics phenomena of fluid flow, species transport and electrochemical reactions are resolved numerically. The model has been validated against experimental data using an in-house AMFC prototype with a flow-through anode. Characteristics of fuel transfer and fuel crossover for both types of anodes are investigated. The model results reveal that the fuel transport to the flow-over anode is intrinsically limited by the fuel concentration boundary layer. Conversely, fuel transport for the flow-through anode is convectively enhanced by the permeate flow, and no concentration boundary layer is observed. An unexpected additional advantage of the flow-through anode configuration is lower parasitic (crossover) current density than the flow-over case at practical low flow rates. Cell performance of the flow-through case is found to be limited by reaction kinetics. The present model provides insights into the fuel transport and fuel crossover in air-breathing microfluidic fuel cells and provides guidance for further design and operation optimization.

  7. Basic studies of microstructure of combusting turbulent flows

    NASA Astrophysics Data System (ADS)

    Hussain, Fazle

    1991-03-01

    The goal is to develop a state-of-the-art measurement technique, Holographic Particle Displacement Velocimetry (HPV), which can provide instantaneous velocities everywhere in the flow field simultaneously. Another goal is to use the power of supercomputers to simulate 3D flows with heat release to study the physics of combusting turbulent flows. Computations suffer from limited flow times and Reynolds number but can provide flow properties in more detail than possible by any existing experimental techniques. Moreover, numerical simulations can provide quantities almost impossible to measure experimentally. This article discusses efforts to develop the holographic particle displacement velocimetry system and results of direct numerical numerical simulations of combusting flows.

  8. Extension of dynamics of granular flow methodology to cell biology

    NASA Astrophysics Data System (ADS)

    Kummer, A.; Ocone, R.

    2003-04-01

    In a previous paper (J. Non-Newtonian Fluid Mech. 76 (1998) 5), the analogy between the methodology typical of the dynamics of polymeric liquids and those used in granular flow theory was investigated. It was shown that such a methodology could be successfully extended to granular flow, and then it was speculated on the possibility of extending it to diverse areas. In this paper two important conclusions are reached. Firstly we show that the methodology behind the statistical theories (which starting from the microstructural element eventually leads to the formulation of constitutive equations (AICHE Symposium Series, Vol. 93, 1997, p. 103)) can be extended to an apparently completely different field, namely cell biology. We then show that classical thermodynamics, as applied to epigenetic systems, presents limitations which can be overcome following an axiomatic thermodynamic route (J. Rheol. 37 (1993) 727).

  9. Ultrasonic flow imaging system: A feasibility study

    SciTech Connect

    Sheen, S.H.; Lawrence, W.P.; Chien, H.T.; Raptis, A.C.

    1991-09-01

    This report examines the feasibility and potential problems in developing a real-time ultrasonic flow imaging instrument for on-line monitoring of mixed-phased flows such as coal slurries. State-of-the-art ultrasonic imaging techniques are assessed for this application. Reflection and diffraction tomographies are proposed for further development, including image-reconstruction algorithms and parallel processing systems. A conventional ultrasonic C-scan technique is used to demonstrate the feasibility of imaging the particle motion in a solid/water flow. 13 refs., 11 figs.

  10. Correlation between DNAPL distribution area and dissolved concentration in surfactant enhanced aquifer remediation effluent: A two-dimensional flow cell study.

    PubMed

    Wu, Bin; Li, Huiying; Du, Xiaoming; Zhong, Lirong; Yang, Bin; Du, Ping; Gu, Qingbao; Li, Fasheng

    2016-02-01

    During the process of surfactant enhanced aquifer remediation (SEAR), free phase dense non-aqueous phase liquid (DNAPL) may be mobilized and spread. The understanding of the impact of DNAPL spreading on the SEAR remediation is not sufficient with its positive effect infrequently mentioned. To evaluate the correlation between DNAPL spreading and remediation efficiency, a two-dimensional sandbox apparatus was used to simulate the migration and dissolution process of 1,2-DCA (1,2-dichloroethane) DNAPL in SEAR. Distribution area of DNAPL in the sandbox was determined by digital image analysis and correlated with effluent DNAPL concentration. The results showed that the effluent DNAPL concentration has significant positive linear correlation with the DNAPL distribution area, indicating the mobilization of DNAPL could improve remediation efficiency by enlarging total NAPL-water interfacial area for mass transfer. Meanwhile, the vertical migration of 1,2-DCA was limited within the boundary of aquifer in all experiments, implying that by manipulating injection parameters in SEAR, optimal remediation efficiency can be reached while the risk of DNAPL vertical migration is minimized. This study provides a convenient visible and quantitative method for the optimization of parameters for SEAR project, and an approach of rapid predicting the extent of DNAPL contaminant distribution based on the dissolved DNAPL concentration in the extraction well.

  11. A modular cell culture device for generating arrays of gradients using stacked microfluidic flows

    PubMed Central

    Sip, Christopher G.; Bhattacharjee, Nirveek; Folch, Albert

    2011-01-01

    Microfluidics has become increasingly important for the study of biochemical cues because it enables exquisite spatiotemporal control of the microenvironment. Well-characterized, stable, and reproducible generation of biochemical gradients is critical for understanding the complex behaviors involved in many biological phenomena. Although many microfluidic devices have been developed which achieve these criteria, the ongoing challenge for these platforms is to provide a suitably benign and physiologically relevant environment for cell culture in a user-friendly format. To achieve this paradigm, microfluidic designs must consider the full scope of cell culture from substrate preparation, cell seeding, and long-term maintenance to properly observe gradient sensing behavior. In addition, designs must address the challenges associated with altered culture conditions and shear forces in flow-based devices. With this consideration, we have designed and characterized a microfluidic device based on the principle of stacked flows to achieve highly stable gradients of diffusible molecules over large areas with extremely low shear forces. The device utilizes a benign vacuum sealing strategy for reversible application to pre-established cell cultures. We apply this device to an existing culture of breast cancer cells to demonstrate the negligible effect of its shear flow on migratory behavior. Lastly, we extend the stacked-flow design to demonstrate its scalable architecture with a prototype device for generating an array of combinatorial gradients. PMID:21799716

  12. Modeling and Simulation of Mucus Flow in Human Bronchial Epithelial Cell Cultures - Part I: Idealized Axisymmetric Swirling Flow.

    PubMed

    Vasquez, Paula A; Jin, Yuan; Palmer, Erik; Hill, David; Forest, M Gregory

    2016-08-01

    A multi-mode nonlinear constitutive model for mucus is constructed directly from micro- and macro-rheology experimental data on cell culture mucus, and a numerical algorithm is developed for the culture geometry and idealized cilia driving conditions. This study investigates the roles that mucus rheology, wall effects, and HBE culture geometry play in the development of flow profiles and the shape of the air-mucus interface. Simulations show that viscoelasticity captures normal stress generation in shear leading to a peak in the air-mucus interface at the middle of the culture and a depression at the walls. Linear and nonlinear viscoelastic regimes can be observed in cultures by varying the hurricane radius and mean rotational velocity. The advection-diffusion of a drug concentration dropped at the surface of the mucus flow is simulated as a function of Peclet number. PMID:27494700

  13. Modeling and Simulation of Mucus Flow in Human Bronchial Epithelial Cell Cultures – Part I: Idealized Axisymmetric Swirling Flow

    PubMed Central

    Vasquez, Paula A.; Jin, Yuan; Palmer, Erik; Hill, David; Forest, M. Gregory

    2016-01-01

    A multi-mode nonlinear constitutive model for mucus is constructed directly from micro- and macro-rheology experimental data on cell culture mucus, and a numerical algorithm is developed for the culture geometry and idealized cilia driving conditions. This study investigates the roles that mucus rheology, wall effects, and HBE culture geometry play in the development of flow profiles and the shape of the air-mucus interface. Simulations show that viscoelasticity captures normal stress generation in shear leading to a peak in the air-mucus interface at the middle of the culture and a depression at the walls. Linear and nonlinear viscoelastic regimes can be observed in cultures by varying the hurricane radius and mean rotational velocity. The advection-diffusion of a drug concentration dropped at the surface of the mucus flow is simulated as a function of Peclet number. PMID:27494700

  14. Experimental study of streaming flows associated with ultrasonic levitators

    NASA Astrophysics Data System (ADS)

    Trinh, E. H.; Robey, J. L.

    1994-11-01

    Steady-state acoustic streaming flow patterns have been observed during the operation of a variety of resonant single-axis ultrasonic levitators in a gaseous environment and in the 20-37 kHz frequency range. Light sheet illumination and scattering from smoke particles have revealed primary streaming flows which display different characteristics at low and high sound pressure levels. Secondary macroscopic streaming cells around levitated samples are superimposed on the primary streaming flow pattern generated by the standing wave. These recorded flows are quite reproducible, and are qualitatively the same for a variety of levitator physical geometries. An onset of flow instability can also be recorded in nonisothermal systems, such as levitated spot-heated samples when the resonance conditions are not exactly satisfied. A preliminary qualitative interpretation of these experimental results is presented in terms of the superposition of three discrete sets of circulation cells operating on different spatial scales. These relevant length scales are the acoustic wavelength, the levitated sample size, and finally the acoustic boundary layer thickness. This approach fails, however, to explain the streaming flow-field morphology around liquid drops levitated on Earth. Observation of the interaction between the flows cells and the levitated samples also suggests the existence of a steady-state torque induced by the streaming flows.

  15. Effect of Flow on Gene Regulation in Smooth Muscle Cells and Macromolecular Transport Across Endothelial Cell Monolayers

    NASA Technical Reports Server (NTRS)

    McIntire, Larry V.; Wagner, John E.; Papadaki, Maria; Whitson, Peggy A.; Eskin, Suzanne G.

    1996-01-01

    Endothelial cells line all of the vessels of the circulatory system, providing a non-thrombogenic conduit for blood flow; they regulate many complex functions in the vasculature, such as coagulation, fibrinolysis, platelet aggregation, vessel tone and growth, and leukocyte traffic; and they form the principal barrier to transport of substances between the blood and the surrounding tissue space. The permeability of endothelial cell changes with environmental stimuli; shear stress, in particular, applied either in vivo, or in vitro, induces changes in protein expression and secretion of vasoactive factors by endothelial cells. The ability to study the effects of shear on the macromolecular permeability of the cerebral vasculature is particularly important, since in no other place is the barrier function of the endothelium more important than in the brain. The endothelial cells of this organ have developed special barrier properties that keep the cerebral system from experiencing any drastic change in composition; together with glial cells, they form the blood brain barrier (BBB). We have studied the effect of flow on bovine BBB using flow chambers and tissue culture systems.

  16. Proliferation and differentiation of oligodendrocyte progenitor cells induced from rat embryonic neural precursor cells followed by flow cytometry.

    PubMed

    Lü, He-Zuo; Wang, Yan-Xia; Li, Ying; Fu, Sai-Li; Hang, Qin; Lu, Pei-Hua

    2008-08-01

    Previous studies have shown that a cell-intrinsic timer might determine when oligodendrocyte progenitor cells (OPCs) isolated from the central nervous system (CNS) stop dividing and initiate differentiation in a defined environment. In this report, the proliferation and differentiation of OPCs induced from neural precursor cells (NPCs) were analyzed by flow cytometry combined with carboxyfluorescein diacetate succinimidyl ester labeling and propidium iodide staining, respectively. When OPCs were cultured in OPC-medium, more than 30% of cells were in S- and G2/M-phases, and continuously self-renewed without differentiation. After exposure to thyroid hormone, there was an obvious decrease in the fraction of cells in both S- and G2/M-phases (<10%). Furthermore, the OPCs no longer proliferated, but differentiated into oligodendrocytes. The dynamic proliferation and differentiation characteristics of OPCs induced from NPCs and analyzed by flow cytometry were similar to those of OPCs isolated from the CNS and analyzed by other methods. These studies indicated that the proliferation and differentiation of OPCs can be followed simply and rapidly by flow cytometry. PMID:18473382

  17. Photothermal imaging of moving cells in lymph and blood flow in vivo

    NASA Astrophysics Data System (ADS)

    Zharov, Vladimir P.; Galanzha, Ekaterina I.; Tuchin, Valery V.

    2004-07-01

    The in vivo capabilities of a new, integrated optical system for studying lymph and blood flow were explored, including imaging of moving red and white blood cells. This system combined transmission microscopy with different dual-beam photothermal (PT) techniques, such as PT imaging, PT thermolens method, and PT deflection velocimetry. All of these PT techniques are based on irradiation of rat mesenteric microvessels with a short laser pulse and on detection of temperature-dependent variations of the refractive index with a second, probe laser beam. In general, the concept of in vivo PT flow cytometry was developed, with a focus on real-time monitoring of moving blood cells in their natural states without labeling (e.g., fluorescent), including obtaining PT images of the cells and determining their flow velocity and response to different interventions. Preliminary experiments revealed many potential applications of this integrated system: (1) quantitation of lymph and blood flow without probes; (2) imaging of moving red and white blood cells; (3) visualization and tracking of PT nanoprobes and sensitizers; (4) comparison of laser-tissue interactions in vivo and in vitro, especially optimization of laser treatment of vascular lesions (port-wine stains, lymphatic malformations, etc.); and (5) determination of the link between in vitro and in vivo cytotoxicity studies.

  18. The art and science of flow control - case studies using flow visualization methods

    NASA Astrophysics Data System (ADS)

    Alvi, F. S.; Cattafesta, L. N., III

    2010-04-01

    Active flow control (AFC) has been the focus of significant research in the last decade. This is mainly due to the potentially substantial benefits it affords. AFC applications range from the subsonic to the supersonic (and beyond) regime for both internal and external flows. These applications are wide and varied, such as controlling flow transition and separation over various external components of the aircraft to active management of separation and flow distortion in engine components and over turbine and compressor blades. High-speed AFC applications include control of flow oscillations in cavity flows, supersonic jet screech, impinging jets, and jet-noise control. In this paper we review some of our recent applications of AFC through a number of case studies that illustrate the typical benefits as well as limitations of present AFC methods. The case studies include subsonic and supersonic canonical flowfields such as separation control over airfoils, control of supersonic cavity flows and impinging jets. In addition, properties of zero-net mass-flux (ZNMF) actuators are also discussed as they represent one of the most widely studied actuators used for AFC. In keeping with the theme of this special issue, the flowfield properties and their response to actuation are examined through the use of various qualitative and quantitative flow visualization methods, such as smoke, shadowgraph, schlieren, planar-laser scattering, and Particle image velocimetry (PIV). The results presented here clearly illustrate the merits of using flow visualization to gain significant insight into the flow and its response to AFC.

  19. Cytoplasm dynamics and cell motion: two-phase flow models.

    PubMed

    Alt, W; Dembo, M

    1999-03-01

    The motion of amoeboid cells is characterized by cytoplasmic streaming and by membrane protrusions and retractions which occur even in the absence of interactions with a substratum. Cell translocation requires, in addition, a transmission mechanism wherein the power produced by the cytoplasmic engine is applied to the substratum in a highly controlled fashion through specific adhesion proteins. Here we present a simple mechano-chemical model that tries to capture the physical essence of these complex biomolecular processes. Our model is based on the continuum equations for a viscous and reactive two-phase fluid model with moving boundaries, and on force balance equations that average the stochastic interactions between actin polymers and membrane proteins. In this paper we present a new derivation and analysis of these equations based on minimization of a power functional. This derivation also leads to a clear formulation and classification of the kinds of boundary conditions that should be specified at free surfaces and at the sites of interaction of the cell and the substratum. Numerical simulations of a one-dimensional lamella reveal that even this extremely simplified model is capable of producing several typical features of cell motility. These include periodic 'ruffle' formation, protrusion-retraction cycles, centripetal flow and cell-substratum traction forces. PMID:10204394

  20. CFD study on electrolyte distribution in redox flow batteries

    NASA Astrophysics Data System (ADS)

    Bortolin, S.; Toninelli, P.; Maggiolo, D.; Guarnieri, M.; Del, D., Col

    2015-11-01

    The most important component in a redox flow battery (RFB) cell is the MEA (membrane electrode assembly), a sandwich consisting of two catalyzed electrodes with an interposed polymeric membrane. In order to allow electrolyte flow toward the electroactive sites, the electrodes have a porous structure that can be obtained with carbon base materials such as carbon felts. The RFB cell is closed by two plates containing the distribution flow channels. Considering that a uniform electrolyte distribution in the reaction region is a prerequisite for high-efficiency operation, the flow pattern is an important parameter to be investigated for the optimization of the cell. In the present work, the effect of different channels patterns on the electrolyte distribution and on the pressure drop is numerically investigated. Three-dimensional simulations have been carried out with ANSYS Fluent code and four different layouts have been considered. Calculations have been performed both in the distribution channels and in the felt porous region.

  1. Minimizing ultraviolet noise due to mis-matches between detector flow cell and post column mobile phase temperatures in supercritical fluid chromatography: effect of flow cell design.

    PubMed

    Berger, Terry A

    2014-10-17

    A mis-match between the post-column mobile phase temperature and the UV detector flow cell temperature can cause significant UV noise in supercritical fluid chromatography (SFC). Deviations as little as 5 °C can increase noise as much as 5 times, making the detector unsuited for trace analysis. Two approaches were used to minimize this noise. When a flow cell was in direct thermal contact (metal on metal) with the detector optical bench, the mobile phase temperature was actively controlled to the measured flow cell temperature, by using one of the heat exchangers (HX) in the column compartment. However, with some older, but still widely used flow cell designs, this required repeated, hourly monitoring of the flow cell temperature and repeated manual adjustment of the heat exchanger temperature, due to thermal drift. Flow cell design had a strong influence on susceptibility to this thermally induced noise. Thermally insulating the flow cell from the optical bench made some cells much less susceptible to such thermally induced noise. Five different flow cells, some insulated, some un-insulated, were evaluated. Most had a truncated conical flow path, but one had a cylindrical flow path. Using either approach, the ASTM noise, with a 10mm, 13 μL conical flow cell, could be optimized to ≈0.007 mAU at 2.5 Hz, in SFC, which is very near the 0.006 mAU manufacturer's specification for HPLC. The insulated version of this flow cell required far less optimization, compared to the un-insulated version. At 150 bar, an experimental 3mm, 2 μL flow cell, with only one side insulated, yielded noise slightly too high (≈0.16-0.18 mAU) for trace analysis, at 80 Hz. However, at 200 bar, noise at 80 Hz was <0.06 mAU, which should allow quantification of a 1 mAU tall trace component with a signal to noise ratio (S/N) >10. Even partially un-insulated, this flow cell design was much less susceptible to thermally induced noise. Further insulating this flow cell design failed to improve

  2. Minimizing ultraviolet noise due to mis-matches between detector flow cell and post column mobile phase temperatures in supercritical fluid chromatography: effect of flow cell design.

    PubMed

    Berger, Terry A

    2014-10-17

    A mis-match between the post-column mobile phase temperature and the UV detector flow cell temperature can cause significant UV noise in supercritical fluid chromatography (SFC). Deviations as little as 5 °C can increase noise as much as 5 times, making the detector unsuited for trace analysis. Two approaches were used to minimize this noise. When a flow cell was in direct thermal contact (metal on metal) with the detector optical bench, the mobile phase temperature was actively controlled to the measured flow cell temperature, by using one of the heat exchangers (HX) in the column compartment. However, with some older, but still widely used flow cell designs, this required repeated, hourly monitoring of the flow cell temperature and repeated manual adjustment of the heat exchanger temperature, due to thermal drift. Flow cell design had a strong influence on susceptibility to this thermally induced noise. Thermally insulating the flow cell from the optical bench made some cells much less susceptible to such thermally induced noise. Five different flow cells, some insulated, some un-insulated, were evaluated. Most had a truncated conical flow path, but one had a cylindrical flow path. Using either approach, the ASTM noise, with a 10mm, 13 μL conical flow cell, could be optimized to ≈0.007 mAU at 2.5 Hz, in SFC, which is very near the 0.006 mAU manufacturer's specification for HPLC. The insulated version of this flow cell required far less optimization, compared to the un-insulated version. At 150 bar, an experimental 3mm, 2 μL flow cell, with only one side insulated, yielded noise slightly too high (≈0.16-0.18 mAU) for trace analysis, at 80 Hz. However, at 200 bar, noise at 80 Hz was <0.06 mAU, which should allow quantification of a 1 mAU tall trace component with a signal to noise ratio (S/N) >10. Even partially un-insulated, this flow cell design was much less susceptible to thermally induced noise. Further insulating this flow cell design failed to improve

  3. Localization and relative quantification of carbon nanotubes in cells with multispectral imaging flow cytometry.

    PubMed

    Marangon, Iris; Boggetto, Nicole; Ménard-Moyon, Cécilia; Luciani, Nathalie; Wilhelm, Claire; Bianco, Alberto; Gazeau, Florence

    2013-01-01

    Carbon-based nanomaterials, like carbon nanotubes (CNTs), belong to this type of nanoparticles which are very difficult to discriminate from carbon-rich cell structures and de facto there is still no quantitative method to assess their distribution at cell and tissue levels. What we propose here is an innovative method allowing the detection and quantification of CNTs in cells using a multispectral imaging flow cytometer (ImageStream, Amnis). This newly developed device integrates both a high-throughput of cells and high resolution imaging, providing thus images for each cell directly in flow and therefore statistically relevant image analysis. Each cell image is acquired on bright-field (BF), dark-field (DF), and fluorescent channels, giving access respectively to the level and the distribution of light absorption, light scattered and fluorescence for each cell. The analysis consists then in a pixel-by-pixel comparison of each image, of the 7,000-10,000 cells acquired for each condition of the experiment. Localization and quantification of CNTs is made possible thanks to some particular intrinsic properties of CNTs: strong light absorbance and scattering; indeed CNTs appear as strongly absorbed dark spots on BF and bright spots on DF with a precise colocalization. This methodology could have a considerable impact on studies about interactions between nanomaterials and cells given that this protocol is applicable for a large range of nanomaterials, insofar as they are capable of absorbing (and/or scattering) strongly enough the light. PMID:24378540

  4. Flow Cytometric Method for the Detection of Flavonoids in Cell Lines.

    PubMed

    Grootaert, Charlotte; Gonzales, Gerard Bryan; Vissenaekens, Hanne; Van de Wiele, Tom; Raes, Katleen; Smagghe, Guy; Van Camp, John

    2016-09-01

    Here, we describe an easy-to-use flow cytometric method using diphenylboric acid 2-amino ethyl ester (DPBA) stain for the detection of flavonoids in cells from human/animal origin. Flavonoid bioavailability and bioactivity depend on structure, conjugation and the cell type to which they are presented. We have studied cellular uptake of five flavonoids with different structures and conjugation forms. First, parameters including fixation method, technical and batch variability, and concentration were optimized. Second, uptake of two aglycones-quercetin and hesperetin-and their corresponding glycosides-rutin and hesperidin-in Caco-2 cells was compared. Third, the aglycone quercetin, glycoside rutin, and glucuronide baicalin were added to the Caco-2, HepG2, and CHO-K1 cell lines at 1, 10, and 20 µM concentrations and cellular uptake was measured after 1, 4, and 7 h. We conclude that quercetin was taken up by cells in a dose-dependent way, and that HepG2 cells had the highest uptake factors, followed by CHO-K1 and Caco-2 cells. Confocal microscopy showed cell type-dependent localization of quercetin in the cell membrane and cytoplasm. No uptake of flavonoid glycosides was detected. This flow cytometric method can be used for future research unravelling mechanisms behind flavonoid bioactivity in health and disease at the cellular level. PMID:27280551

  5. Numerical study of transient flow phenomena in shock tunnels

    NASA Technical Reports Server (NTRS)

    Tokarcik-Polsky, Susan; Cambier, Jean-Luc

    1994-01-01

    Computational fluid dynamics (CFD) was used to study some transient flow features that can occur during the startup process of a shoch tunnel. The investigation concentrated on two areas: (1) the flow near the endwall of the driven tube during shock reflection and (2) the transient flow in the nozzle. The driven tube calculations were inviscid and focused on the study of a vortex system that was seen to form at the driven tube's axis of symmetry. The nozzle flow calculations examined viscous and inviscid effects during nozzle startup. The CFD solutions of the nozzle flows were compared with experimental data to demonstrate the effectiveness of the numerical analysis.

  6. Colloidal Suspensions in Shear Flow : a Real Space Study

    NASA Astrophysics Data System (ADS)

    Derks, D.

    2006-09-01

    We investigate the effect of shear flow on the microstructure of colloidal suspensions by means of microscopy. Systems of nearly equally sized particles are used, whose interactions and phase behavior are predominantly determined by their size and shape, and can further be tuned by the addition of polymers. Recently, a new type of shear cell was developed to study flowing suspensions in real space. The key property of this setup is the counter-rotating principle of the cone and plate, opening up the possibility to create a stationary layer in the bulk of the cell. In Chapter 2, we elaborate on the details of this setup and its performance. Fluorescence confocal microscopy is used to visualize the sheared suspension, and allows imaging of individual particles in the bulk in the stationary plane for a prolonged time. This way, the particle positions in a layer of, for example, a sheared colloidal crystal can be tracked. The particle dynamics in colloidal crystals in shear flow are the subject of Chapter 3. Here, the particles interact through a (nearly) hard sphere potential. Apart from the alignment of the crystal in the shear field and the collective zigzag motion, which had also been deduced from early scattering experiments, we find that random particle displacements increase with shear rate. Those increased fluctuations result in shear induced melting when their mean square displacement has reached about 13 % of the particle separation. Apart from hard spheres, we investigate mixtures of colloids and polymers in shear flow. The polymers cause an effective attraction between the spheres, leading to phase separation into a colloid rich (polymer poor) and a colloid poor (polymer rich) phase at sufficiently high colloid and polymer concentration. In Chapter 4, we study the demixing process in the (spinodal) two-phase region of the phase diagram. The system is quenched from an initially almost homogeneous state at very high shear rate to a low shear rate. A spinodal

  7. Single cell kinase signaling assay using pinched flow coupled droplet microfluidics.

    PubMed

    Ramji, Ramesh; Wang, Ming; Bhagat, Ali Asgar S; Tan Shao Weng, Daniel; Thakor, Nitish V; Teck Lim, Chwee; Chen, Chia-Hung

    2014-05-01

    Droplet-based microfluidics has shown potential in high throughput single cell assays by encapsulating individual cells in water-in-oil emulsions. Ordering cells in a micro-channel is necessary to encapsulate individual cells into droplets further enhancing the assay efficiency. This is typically limited due to the difficulty of preparing high-density cell solutions and maintaining them without cell aggregation in long channels (>5 cm). In this study, we developed a short pinched flow channel (5 mm) to separate cell aggregates and to form a uniform cell distribution in a droplet-generating platform that encapsulated single cells with >55% encapsulation efficiency beating Poisson encapsulation statistics. Using this platform and commercially available Sox substrates (8-hydroxy-5-(N,N-dimethylsulfonamido)-2-methylquinoline), we have demonstrated a high throughput dynamic single cell signaling assay to measure the activity of receptor tyrosine kinases (RTKs) in lung cancer cells triggered by cell surface ligand binding. The phosphorylation of the substrates resulted in fluorescent emission, showing a sigmoidal increase over a 12 h period. The result exhibited a heterogeneous signaling rate in individual cells and showed various levels of drug resistance when treated with the tyrosine kinase inhibitor, gefitinib. PMID:24926389

  8. Single cell kinase signaling assay using pinched flow coupled droplet microfluidics

    PubMed Central

    Ramji, Ramesh; Wang, Ming; Bhagat, Ali Asgar S.; Tan Shao Weng, Daniel; Thakor, Nitish V.; Teck Lim, Chwee; Chen, Chia-Hung

    2014-01-01

    Droplet-based microfluidics has shown potential in high throughput single cell assays by encapsulating individual cells in water-in-oil emulsions. Ordering cells in a micro-channel is necessary to encapsulate individual cells into droplets further enhancing the assay efficiency. This is typically limited due to the difficulty of preparing high-density cell solutions and maintaining them without cell aggregation in long channels (>5 cm). In this study, we developed a short pinched flow channel (5 mm) to separate cell aggregates and to form a uniform cell distribution in a droplet-generating platform that encapsulated single cells with >55% encapsulation efficiency beating Poisson encapsulation statistics. Using this platform and commercially available Sox substrates (8-hydroxy-5-(N,N-dimethylsulfonamido)-2-methylquinoline), we have demonstrated a high throughput dynamic single cell signaling assay to measure the activity of receptor tyrosine kinases (RTKs) in lung cancer cells triggered by cell surface ligand binding. The phosphorylation of the substrates resulted in fluorescent emission, showing a sigmoidal increase over a 12 h period. The result exhibited a heterogeneous signaling rate in individual cells and showed various levels of drug resistance when treated with the tyrosine kinase inhibitor, gefitinib. PMID:24926389

  9. A modular segmented-flow platform for 3D cell cultivation.

    PubMed

    Lemke, Karen; Förster, Tobias; Römer, Robert; Quade, Mandy; Wiedemeier, Stefan; Grodrian, Andreas; Gastrock, Gunter

    2015-07-10

    In vitro 3D cell cultivation is promised to equate tissue in vivo more realistically than 2D cell cultivation corresponding to cell-cell and cell-matrix interactions. Therefore, a scalable 3D cultivation platform was developed. This platform, called pipe-based bioreactors (pbb), is based on the segmented-flow technology: aqueous droplets are embedded in a water-immiscible carrier fluid. The droplet volumes range from 60 nL to 20 μL and are used as bioreactors lined up in a tubing like pearls on a string. The modular automated platform basically consists of several modules like a fluid management for a high throughput droplet generation for self-assembly or scaffold-based 3D cell cultivation, a storage module for incubation and storage, and an analysis module for monitoring cell aggregation and proliferation basing on microscopy or photometry. In this report, the self-assembly of murine embryonic stem cells (mESCs) to uniformly sized embryoid bodies (EBs), the cell proliferation, the cell viability as well as the influence on the cell differentiation to cardiomyocytes are described. The integration of a dosage module for medium exchange or agent addition will enable pbb as long-term 3D cell cultivation system for studying stem cell differentiation, e.g. cardiac myogenesis or for diagnostic and therapeutic testing in personalized medicine.

  10. Shape transitions of fluid vesicles and red blood cells in capillary flows

    PubMed Central

    Noguchi, Hiroshi; Gompper, Gerhard

    2005-01-01

    The dynamics of fluid vesicles and red blood cells (RBCs) in cylindrical capillary flow is studied by using a three-dimensional mesoscopic simulation approach. As flow velocity increases, a model RBC is found to transit from a nonaxisymmetric discocyteto an axisymmetric parachute shape (coaxial with the flow axis), while a fluid vesicle is found to transit from a discocyte to a prolate ellipsoid. Both shape transitions reduce the flow resistance. The critical velocities of the shape transitions are linearly dependent on the bending rigidity and on the shear modulus of the membrane. Slipper-like shapes of the RBC model are observed around the transition velocities. Our results are in good agreement with experiments on RBCs. PMID:16186506

  11. Shape transitions of fluid vesicles and red blood cells in capillary flows

    NASA Astrophysics Data System (ADS)

    Noguchi, Hiroshi; Gompper, Gerhard

    2005-10-01

    The dynamics of fluid vesicles and red blood cells (RBCs) in cylindrical capillary flow is studied by using a three-dimensional mesoscopic simulation approach. As flow velocity increases, a model RBC is found to transit from a nonaxisymmetric discocyteto an axisymmetric parachute shape (coaxial with the flow axis), while a fluid vesicle is found to transit from a discocyte to a prolate ellipsoid. Both shape transitions reduce the flow resistance. The critical velocities of the shape transitions are linearly dependent on the bending rigidity and on the shear modulus of the membrane. Slipper-like shapes of the RBC model are observed around the transition velocities. Our results are in good agreement with experiments on RBCs. mesoscale hydrodynamics simulation | microfluidics | shape transformations

  12. Measurement of Flow Pattern Within a Rotating Stall Cell in an Axial Compressor

    NASA Technical Reports Server (NTRS)

    Lepicovsky, Jan; Braunscheidel, Edward P.

    2006-01-01

    Effective active control of rotating stall in axial compressors requires detailed understanding of flow instabilities associated with this compressor regime. Newly designed miniature high frequency response total and static pressure probes as well as commercial thermoanemometric probes are suitable tools for this task. However, during the rotating stall cycle the probes are subjected to flow direction changes that are far larger than the range of probe incidence acceptance, and therefore probe data without a proper correction would misrepresent unsteady variations of flow parameters. A methodology, based on ensemble averaging, is proposed to circumvent this problem. In this approach the ensemble averaged signals acquired for various probe setting angles are segmented, and only the sections for probe setting angles close to the actual flow angle are used for signal recombination. The methodology was verified by excellent agreement between velocity distributions obtained from pressure probe data, and data measured with thermoanemometric probes. Vector plots of unsteady flow behavior during the rotating stall regime indicate reversed flow within the rotating stall cell that spreads over to adjacent rotor blade channels. Results of this study confirmed that the NASA Low Speed Axial Compressor (LSAC) while in a rotating stall regime at rotor design speed exhibits one stall cell that rotates at a speed equal to 50.6 percent of the rotor shaft speed.

  13. Apheresis experience with a continuous flow cell separator.

    PubMed

    Norol, F; Aubert, C; Scotto, F; Duedari, N

    1988-01-01

    The Dideco Vivacell separator is a continuous-flow centrifugation system that has only recently been used for cytapheresis. The authors' experience with this separator in 451 plateletpheresis and 164 leukapheresis procedures is presented. Platelet collection provided high platelet yields (9.53 +/- 2.85 X 10(11) with a collection efficiency of 74 +/- 14 percent for about 6 liters of total blood processed. Functional integrity was confirmed by normal in vitro tests (aggregation and response to hypotonic stress) and good in vivo recovery (55%). In leukapheresis, white cell yields were high (3.42 +/- 1.2 X 10(10) with 85 percent polymorphonuclear neutrophil cells. Their oxidative metabolism functions (generation of free oxygen radicals), investigated by chemiluminescence, were increased over donor values. Donor reactions, all of the mild citrate type, were rare.

  14. Flow cytometric measurement of pollutant stresses on algal cells

    SciTech Connect

    Berglund, D.L.; Eversman, S.

    1988-03-01

    The lichen Usnea fulvoreagens (Raes). Raes. was treated with four pH levels (5.5, 4.5, 3.5, and 2.5) of simulated acid rain (sulfuric acid, nitric acid, and a 1:1 combination of both) and automobile exhaust. The samples were dissociated and analyzed by a Becton-Dickinson FACS 440 flow cytometer. Analyses included measurement of chlorophyll autofluorescence and fluorescence due to uptake of fluorescein diacetate (FDA) and calcofluor white M2R (CFW). Cell parameters measured were esterase activity (FDA), membrane permeability (FDA, CFW), and intracellular pH (FDA). Mean fluorescence intensity from FDA staining and numbers of events were incorporated with autofluorescence information to produce a stress index of relative cell stress. Results indicated that highly stressed samples (lower pH treatments and greater exposure to exhaust) exhibited a low stress index of FDA fluorescence.Au

  15. Flow cytometric measurement of pollutant stresses on algal cells.

    PubMed

    Berglund, D L; Eversman, S

    1988-03-01

    The lichen Usnea fulvoreagens (Räs). Räs. was treated with four pH levels (5.5, 4.5, 3.5, and 2.5) of simulated acid rain (sulfuric acid, nitric acid, and a 1:1 combination of both) and automobile exhaust. The samples were dissociated and analyzed by a Becton-Dickinson FACS 440 flow cytometer. Analyses included measurement of chlorophyll autofluorescence and fluorescence due to uptake of fluorescein diacetate (FDA) and calcofluor white M2R (CFW). Cell parameters measured were esterase activity (FDA), membrane permeability (FDA, CFW), and intracellular pH (FDA). Mean fluorescence intensity from FDA staining and numbers of events were incorporated with autofluorescence information to produce a "stress index" of relative cell stress. Results indicated that highly stressed samples (lower pH treatments and greater exposure to exhaust) exhibited a low "stress index" of FDA fluorescence.

  16. Flow-induced clustering and alignment of vesicles and red blood cells in microcapillaries

    PubMed Central

    McWhirter, J. Liam; Noguchi, Hiroshi; Gompper, Gerhard

    2009-01-01

    The recent development of microfluidic devices allows the investigation and manipulation of individual liquid microdroplets, capsules, and cells. The collective behavior of several red blood cells (RBCs) or microcapsules in narrow capillaries determines their flow-induced morphology, arrangement, and effective viscosity. Of fundamental interest here is the relation between the flow behavior and the elasticity and deformability of these objects, their long-range hydrodynamic interactions in microchannels, and thermal membrane undulations. We study these mechanisms in an in silico model, which combines a particle-based mesoscale simulation technique for the fluid hydrodynamics with a triangulated-membrane model. The 2 essential control parameters are the volume fraction of RBCs (the tube hematocrit, HT), and the flow velocity. Our simulations show that already at very low HT, the deformability of RBCs implies a flow-induced cluster formation above a threshold flow velocity. At higher HT values, we predict 3 distinct phases: one consisting of disordered biconcave-disk-shaped RBCs, another with parachute-shaped RBCs aligned in a single file, and a third with slipper-shaped RBCs arranged as 2 parallel interdigitated rows. The deformation-mediated clustering and the arrangements of RBCs and microcapsules are relevant for many potential applications in physics, biology, and medicine, such as blood diagnosis and cell sorting in microfluidic devices. PMID:19369212

  17. An Improved Flow Cytometry Method For Precise Quantitation Of Natural-Killer Cell Activity

    NASA Technical Reports Server (NTRS)

    Crucian, Brian; Nehlsen-Cannarella, Sandra; Sams, Clarence

    2006-01-01

    The ability to assess NK cell cytotoxicity using flow cytometry has been previously described and can serve as a powerful tool to evaluate effector immune function in the clinical setting. Previous methods used membrane permeable dyes to identify target cells. The use of these dyes requires great care to achieve optimal staining and results in a broad spectral emission that can make multicolor cytometry difficult. Previous methods have also used negative staining (the elimination of target cells) to identify effector cells. This makes a precise quantitation of effector NK cells impossible due to the interfering presence of T and B lymphocytes, and the data highly subjective to the variable levels of NK cells normally found in human peripheral blood. In this study an improved version of the standard flow cytometry assay for NK activity is described that has several advantages of previous methods. Fluorescent antibody staining (CD45FITC) is used to positively identify target cells in place of membranepermeable dyes. Fluorescent antibody staining of target cells is less labor intensive and more easily reproducible than membrane dyes. NK cells (true effector lymphocytes) are also positively identified by fluorescent antibody staining (CD56PE) allowing a simultaneous absolute count assessment of both NK cells and target cells. Dead cells are identified by membrane disruption using the DNA intercalating dye PI. Using this method, an exact NK:target ratio may be determined for each assessment, including quantitation of NK target complexes. Backimmunoscatter gating may be used to track live vs. dead Target cells via scatter properties. If desired, NK activity may then be normalized to standardized ratios for clinical comparisons between patients, making the determination of PBMC counts or NK cell percentages prior to testing unnecessary. This method provides an exact cytometric determination of NK activity that highly reproducible and may be suitable for routine use in the

  18. Large eddy simulation study of spanwise spacing effects on secondary flows in turbulent channel flow

    NASA Astrophysics Data System (ADS)

    Aliakbarimiyanmahaleh, Mohammad; Anderson, William

    2015-11-01

    The structure of turbulent flow over a complex topography composed of streamwise-aligned rows of cones with varying spanwise spacing, s is studied with large-eddy simulation (LES). Similar to the experimental study of Vanderwel and Ganapathisubramani, 2015: J. Fluid Mech., we investigate the relationship between secondary flow and s, for 0 . 25 <= s / δ <= 5 . For cases with s / δ > 2 , domain-scale rollers freely exist. These had previously been called ``turbulent secondary flows'' (Willingham et al., 2014: Phys. Fluids; Barros and Christensen, 2014: J. Fluid Mech.; Anderson et al., 2015: J. Fluid Mech.), but closer inspection of the statistics indicates these are a turbulent tertiary flow: they only remain ``anchored'' to the conical roughness elements for s / δ > 2 . For s / δ < 2 , turbulent tertiary flows are prevented from occupying the domain by virtue of proximity to adjacent, counter-rotating tertiary flows. Turbulent secondary flows are associated with the conical roughness elements. These turbulent secondary flows emanate from individual conical topographic elements and set the roughness sublayer depth. The turbulent secondary flows remain intact for large and small spacing. For s / δ < 1 , a mean tertiary flow is not present. This work was supported by the Air Force Office of Sci. Research, Young Inv. Program (PM: Dr. R. Ponnoppan and Ms. E. Montomery) under Grant # FA9550-14-1-0394. Computational resources were provided by the Texas Adv. Comp. Center at the Univ. of Texas.

  19. CFD Validation Studies for Hypersonic Flow Prediction

    NASA Technical Reports Server (NTRS)

    Gnoffo, Peter A.

    2001-01-01

    A series of experiments to measure pressure and heating for code validation involving hypersonic, laminar, separated flows was conducted at the Calspan-University at Buffalo Research Center (CUBRC) in the Large Energy National Shock (LENS) tunnel. The experimental data serves as a focus for a code validation session but are not available to the authors until the conclusion of this session. The first set of experiments considered here involve Mach 9.5 and Mach 11.3 N2 flow over a hollow cylinder-flare with 30 degree flare angle at several Reynolds numbers sustaining laminar, separated flow. Truncated and extended flare configurations are considered. The second set of experiments, at similar conditions, involves flow over a sharp, double cone with fore-cone angle of 25 degrees and aft-cone angle of 55 degrees. Both sets of experiments involve 30 degree compressions. Location of the separation point in the numerical simulation is extremely sensitive to the level of grid refinement in the numerical predictions. The numerical simulations also show a significant influence of Reynolds number on extent of separation. Flow unsteadiness was easily introduced into the double cone simulations using aggressive relaxation parameters that normally promote convergence.

  20. CFD Validation Studies for Hypersonic Flow Prediction

    NASA Technical Reports Server (NTRS)

    Gnoffo, Peter A.

    2001-01-01

    A series of experiments to measure pressure and heating for code validation involving hypersonic, laminar, separated flows was conducted at the Calspan-University at Buffalo Research Center (CUBRC) in the Large Energy National Shock (LENS) tunnel. The experimental data serves as a focus for a code validation session but are not available to the authors until the conclusion of this session. The first set of experiments considered here involve Mach 9.5 and Mach 11.3 N, flow over a hollow cylinder-flare with 30 deg flare angle at several Reynolds numbers sustaining laminar, separated flow. Truncated and extended flare configurations are considered. The second set of experiments, at similar conditions, involves flow over a sharp, double cone with fore-cone angle of 25 deg and aft-cone angle of 55 deg. Both sets of experiments involve 30 deg compressions. Location of the separation point in the numerical simulation is extremely sensitive to the level of grid refinement in the numerical predictions. The numerical simulations also show a significant influence of Reynolds number on extent of separation. Flow unsteadiness was easily introduced into the double cone simulations using aggressive relaxation parameters that normally promote convergence.

  1. Numerical study of eccentric Couette Taylor flows and effect of eccentricity on flow patterns

    NASA Astrophysics Data System (ADS)

    Shu, C.; Wang, L.; Chew, Y. T.; Zhao, N.

    2004-10-01

    In this study, the differential quadrature (DQ) method was used to simulate the eccentric Couette Taylor vortex flow in an annulus between two eccentric cylinders with rotating inner cylinder and stationary outer cylinder. An approach combining the SIMPLE (semi-implicit method for pressure-linked equations) and DQ discretization on a non-staggered mesh was proposed to solve the time-dependent, three-dimensional incompressible Navier Stokes equations in the primitive variable form. The eccentric steady Couette Taylor flow patterns were obtained from the solution of three-dimensional Navier Stokes equations. The reported numerical results for steady Couette flow were compared with those from Chou [1], and San and Szeri [2]. Very good agreement was achieved. For steady eccentric Taylor vortex flow, detailed flow patterns were obtained and analyzed. The effect of eccentricity on the eccentric Taylor vortex flow pattern was also studied.

  2. Flow cytometric quantification of tumour endothelial cells; an objective alternative for microvessel density assessment.

    PubMed

    Baeten, C I M; Wagstaff, J; Verhoeven, I C L; Hillen, H F P; Griffioen, A W

    2002-07-29

    Assessment of microvessel density by immunohistochemical staining is subject to a considerable inter-observer variation, and this has led to variability in correlation between microvessel density and clinical outcome in different studies. In order to improve the method of microvessel density measurement in tumour biopsies, we have developed a rapid, objective and quantitative method using flow cytometry on frozen tissues. Frozen tissue sections of archival tumour material were enzymatically digested. The single-cell suspension was stained for CD31 and CD34 for flow cytometry. The number of endothelial cells was quantified using light scatter- and fluorescence-characteristics. Tumour endothelial cells were detectable in a single cell suspension, and the percentage of endothelial cells detected in 32 colon carcinomas correlated highly (r=0.84, P<0.001) with the immunohistochemical assessment of microvessel density. Flow cytometric endothelial cells quantification was found to be more sensitive especially at lower levels of immunohistochemical microvessel density measurement. The current method was found to be applicable for various tumour types and has the major advantage that it provides a retrospective and quantitative approach to the angiogenic potential of tumours.

  3. Continuous-flow Ferrohydrodynamic Sorting of Particles and Cells in Microfluidic Devices

    PubMed Central

    Zhu, Taotao; Cheng, Rui; Lee, Sarah A.; Rajaraman, Eashwar; Eiteman, Mark A.; Querec, Troy D.; Unger, Elizabeth R.; Mao, Leidong

    2015-01-01

    A new sorting scheme based on ferrofluid hydrodynamics (ferrohydrodynamics) was used to separate mixtures of particles and live cells simultaneously. Two species of cells, including Escherichia coli and Saccharomyces cerevisiae, as well as fluorescent polystyrene microparticles were studied for their sorting throughput and efficiency. Ferrofluids are stable magnetic nanoparticles suspensions. Under external magnetic fields, magnetic buoyancy forces exerted on particles and cells lead to size-dependent deflections from their laminar flow paths and result in spatial separation. We report the design, modeling, fabrication and characterization of the sorting device. This scheme is simple, low-cost and label-free compared to other existing techniques. PMID:26430394

  4. Experimental Studies of the Interaction Between a Parallel Shear Flow and a Directionally-Solidifying Front

    NASA Technical Reports Server (NTRS)

    Zhang, Meng; Maxworthy, Tony

    1999-01-01

    It has long been recognized that flow in the melt can have a profound influence on the dynamics of a solidifying interface and hence the quality of the solid material. In particular, flow affects the heat and mass transfer, and causes spatial and temporal variations in the flow and melt composition. This results in a crystal with nonuniform physical properties. Flow can be generated by buoyancy, expansion or contraction upon phase change, and thermo-soluto capillary effects. In general, these flows can not be avoided and can have an adverse effect on the stability of the crystal structures. This motivates crystal growth experiments in a microgravity environment, where buoyancy-driven convection is significantly suppressed. However, transient accelerations (g-jitter) caused by the acceleration of the spacecraft can affect the melt, while convection generated from the effects other than buoyancy remain important. Rather than bemoan the presence of convection as a source of interfacial instability, Hurle in the 1960s suggested that flow in the melt, either forced or natural convection, might be used to stabilize the interface. Delves considered the imposition of both a parabolic velocity profile and a Blasius boundary layer flow over the interface. He concluded that fast stirring could stabilize the interface to perturbations whose wave vector is in the direction of the fluid velocity. Forth and Wheeler considered the effect of the asymptotic suction boundary layer profile. They showed that the effect of the shear flow was to generate travelling waves parallel to the flow with a speed proportional to the Reynolds number. There have been few quantitative, experimental works reporting on the coupling effect of fluid flow and morphological instabilities. Huang studied plane Couette flow over cells and dendrites. It was found that this flow could greatly enhance the planar stability and even induce the cell-planar transition. A rotating impeller was buried inside the

  5. TANK MIXING STUDY WITH FLOW RECIRCULATION

    SciTech Connect

    Lee, S.

    2014-06-25

    The primary objective of this work is to quantify the mixing time when two miscible fluids are mixed by one recirculation pump and to evaluate adequacy of 2.5 hours of pump recirculation to be considered well mixed in SRS tanks, JT-71/72. The work scope described here consists of two modeling analyses. They are the steady state flow pattern analysis during pump recirculation operation of the tank liquid and transient species transport calculations based on the initial steady state flow patterns. The modeling calculations for the mixing time are performed by using the 99% homogeneity criterion for the entire domain of the tank contents.

  6. Electrochemical Oscillations of Nickel Electrodissolution in an Epoxy-Based Microchip Flow Cell.

    PubMed

    Cioffi, Alexander G; Martin, R Scott; Kiss, István Z

    2011-08-01

    We investigate the nonlinear dynamics of transpassive electrodissolution of nickel in sulfuric acid in an epoxy-based microchip flow cell. We observed bistability, smooth, relaxation, and period-2 waveform current oscillations with external resistance attached to the electrode in the microfabricated electrochemical cell with 0.05 mm diameter Ni wire under potentiostatic control. Experiments with 1mm × 0.1 mm Ni electrode show spontaneous oscillations without attached external resistance; similar surface area electrode in macrocell does not exhibit spontaneous oscillations. Combined experimental and numerical studies show that spontaneous oscillation with the on-chip fabricated electrochemical cell occurs because of the unusually large ohmic potential drop due to the constrained current in the narrow flow channel. This large IR potential drop is expected to have an important role in destabilizing negative differential resistance electrochemical (e.g., metal dissolution and electrocatalytic) systems in on-chip integrated microfludic flow cells. The proposed experimental setup can be extendend to multi-electrode configurations; the epoxy-based substrate procedure thus holds promise in electroanalytical applications that require collector-generator multi-electrodes wires with various electrode sizes, compositions, and spacings as well as controlled flow conditions.

  7. Electrochemical Oscillations of Nickel Electrodissolution in an Epoxy-Based Microchip Flow Cell

    PubMed Central

    Cioffi, Alexander G.; Martin, R. Scott; Kiss, István Z.

    2011-01-01

    We investigate the nonlinear dynamics of transpassive electrodissolution of nickel in sulfuric acid in an epoxy-based microchip flow cell. We observed bistability, smooth, relaxation, and period-2 waveform current oscillations with external resistance attached to the electrode in the microfabricated electrochemical cell with 0.05 mm diameter Ni wire under potentiostatic control. Experiments with 1mm × 0.1 mm Ni electrode show spontaneous oscillations without attached external resistance; similar surface area electrode in macrocell does not exhibit spontaneous oscillations. Combined experimental and numerical studies show that spontaneous oscillation with the on-chip fabricated electrochemical cell occurs because of the unusually large ohmic potential drop due to the constrained current in the narrow flow channel. This large IR potential drop is expected to have an important role in destabilizing negative differential resistance electrochemical (e.g., metal dissolution and electrocatalytic) systems in on-chip integrated microfludic flow cells. The proposed experimental setup can be extendend to multi-electrode configurations; the epoxy-based substrate procedure thus holds promise in electroanalytical applications that require collector-generator multi-electrodes wires with various electrode sizes, compositions, and spacings as well as controlled flow conditions. PMID:21822407

  8. Noninvasive measurement of cerebrospinal fluid flow using an ultrasonic transit time flow sensor: a preliminary study.

    PubMed

    Pennell, Thomas; Yi, Juneyoung L; Kaufman, Bruce A; Krishnamurthy, Satish

    2016-03-01

    OBJECT Mechanical failure-which is the primary cause of CSF shunt malfunction-is not readily diagnosed, and the specific reasons for mechanical failure are not easily discerned. Prior attempts to measure CSF flow noninvasively have lacked the ability to either quantitatively or qualitatively obtain data. To address these needs, this preliminary study evaluates an ultrasonic transit time flow sensor in pediatric and adult patients with external ventricular drains (EVDs). One goal was to confirm the stated accuracy of the sensor in a clinical setting. A second goal was to observe the sensor's capability to record real-time continuous CSF flow. The final goal was to observe recordings during instances of flow blockage or lack of flow in order to determine the sensor's ability to identify these changes. METHODS A total of 5 pediatric and 11 adult patients who had received EVDs for the treatment of hydrocephalus were studied in a hospital setting. The primary EVD was connected to a secondary study EVD that contained a fluid-filled pressure transducer and an in-line transit time flow sensor. Comparisons were made between the weight of the drainage bag and the flow measured via the sensor in order to confirm its accuracy. Data from the pressure transducer and the flow sensor were recorded continuously at 100 Hz for a period of 24 hours by a data acquisition system, while the hourly CSF flow into the drip chamber was recorded manually. Changes in the patient's neurological status and their time points were noted. RESULTS The flow sensor demonstrated a proven accuracy of ± 15% or ± 2 ml/hr. The flow sensor allowed real-time continuous flow waveform data recordings. Dynamic analysis of CSF flow waveforms allowed the calculation of the pressure-volume index. Lastly, the sensor was able to diagnose a blocked catheter and distinguish between the blockage and lack of flow. CONCLUSIONS The Transonic flow sensor accurately measures CSF output within ± 15% or ± 2 ml

  9. Noninvasive measurement of cerebrospinal fluid flow using an ultrasonic transit time flow sensor: a preliminary study.

    PubMed

    Pennell, Thomas; Yi, Juneyoung L; Kaufman, Bruce A; Krishnamurthy, Satish

    2016-03-01

    OBJECT Mechanical failure-which is the primary cause of CSF shunt malfunction-is not readily diagnosed, and the specific reasons for mechanical failure are not easily discerned. Prior attempts to measure CSF flow noninvasively have lacked the ability to either quantitatively or qualitatively obtain data. To address these needs, this preliminary study evaluates an ultrasonic transit time flow sensor in pediatric and adult patients with external ventricular drains (EVDs). One goal was to confirm the stated accuracy of the sensor in a clinical setting. A second goal was to observe the sensor's capability to record real-time continuous CSF flow. The final goal was to observe recordings during instances of flow blockage or lack of flow in order to determine the sensor's ability to identify these changes. METHODS A total of 5 pediatric and 11 adult patients who had received EVDs for the treatment of hydrocephalus were studied in a hospital setting. The primary EVD was connected to a secondary study EVD that contained a fluid-filled pressure transducer and an in-line transit time flow sensor. Comparisons were made between the weight of the drainage bag and the flow measured via the sensor in order to confirm its accuracy. Data from the pressure transducer and the flow sensor were recorded continuously at 100 Hz for a period of 24 hours by a data acquisition system, while the hourly CSF flow into the drip chamber was recorded manually. Changes in the patient's neurological status and their time points were noted. RESULTS The flow sensor demonstrated a proven accuracy of ± 15% or ± 2 ml/hr. The flow sensor allowed real-time continuous flow waveform data recordings. Dynamic analysis of CSF flow waveforms allowed the calculation of the pressure-volume index. Lastly, the sensor was able to diagnose a blocked catheter and distinguish between the blockage and lack of flow. CONCLUSIONS The Transonic flow sensor accurately measures CSF output within ± 15% or ± 2 ml

  10. Measuring cell cycle progression kinetics with metabolic labeling and flow cytometry.

    PubMed

    Fleisig, Helen; Wong, Judy

    2012-01-01

    Precise control of the initiation and subsequent progression through the various phases of the cell cycle are of paramount importance in proliferating cells. Cell cycle division is an integral part of growth and reproduction and deregulation of key cell cycle components have been implicated in the precipitating events of carcinogenesis. Molecular agents in anti-cancer therapies frequently target biological pathways responsible for the regulation and coordination of cell cycle division. Although cell cycle kinetics tend to vary according to cell type, the distribution of cells amongst the four stages of the cell cycle is rather consistent within a particular cell line due to the consistent pattern of mitogen and growth factor expression. Genotoxic events and other cellular stressors can result in a temporary block of cell cycle progression, resulting in arrest or a temporary pause in a particular cell cycle phase to allow for instigation of the appropriate response mechanism. The ability to experimentally observe the behavior of a cell population with reference to their cell cycle progression stage is an important advance in cell biology. Common procedures such as mitotic shake off, differential centrifugation or flow cytometry-based sorting are used to isolate cells at specific stages of the cell cycle. These fractionated, cell cycle phase-enriched populations are then subjected to experimental treatments. Yield, purity and viability of the separated fractions can often be compromised using these physical separation methods. As well, the time lapse between separation of the cell populations and the start of experimental treatment, whereby the fractionated cells can progress from the selected cell cycle stage, can pose significant challenges in the successful implementation and interpretation of these experiments. Other approaches to study cell cycle stages include the use of chemicals to synchronize cells. Treatment of cells with chemical inhibitors of key

  11. 3D-printed and CNC milled flow-cells for chemiluminescence detection.

    PubMed

    Spilstead, Kara B; Learey, Jessica J; Doeven, Egan H; Barbante, Gregory J; Mohr, Stephan; Barnett, Neil W; Terry, Jessica M; Hall, Robynne M; Francis, Paul S

    2014-08-01

    Herein we explore modern fabrication techniques for the development of chemiluminescence detection flow-cells with features not attainable using the traditional coiled tubing approach. This includes the first 3D-printed chemiluminescence flow-cells, and a milled flow-cell designed to split the analyte stream into two separate detection zones within the same polymer chip. The flow-cells are compared to conventional detection systems using flow injection analysis (FIA) and high performance liquid chromatography (HPLC), with the fast chemiluminescence reactions of an acidic potassium permanganate reagent with morphine and a series of adrenergic phenolic amines. PMID:24881540

  12. Flow-through cell electroporation microchip integrating dielectrophoretic viable cell sorting.

    PubMed

    Wei, Zewen; Li, Xueming; Zhao, Deyao; Yan, Hao; Hu, Zhiyuan; Liang, Zicai; Li, Zhihong

    2014-10-21

    Microfluidics based continuous cell electroporation is an appealing approach for high-throughput cell transfection, but cell viability of existing methods is usually compromised by adverse electrical or hydrodynamic effects. Here we present the validation of a flow-through cell electroporation microchip, in which dielectrophoretic force was employed to sort viable cells. By integrating parallel electroporation electrodes and dielectrophoresis sorting electrodes together in a simple straight microfluidic channel, sufficient electrical pulses were applied for efficient electroporation, and a proper sinusoidal electrical field was subsequently utilized to exclude damaged cells by dielectrophoresis. Thus, the difficulties for seeking the fine balance between electrotransfection efficiency and cell viability were steered clear. After careful investigation and optimization of the DEP behaviors of electroporated cells, efficient electrotransfection of plasmid DNA was demonstrated in vulnerable neuron cells and several hard-to-transfect primary cell types with excellent cell viability. This microchip constitutes a novel way of continuous cell transfection to significantly improve the cell viability of existing methodologies.

  13. On conditional sampling for turbulent flow studies

    NASA Technical Reports Server (NTRS)

    Wang, F. C.

    1974-01-01

    The conditional sampling technique is analyzed as a weighted time average for turbulent flow. The various conditional averages are obtained by using different types of weighting functions. A second averaging relation is obtained between the conventional averages and the conditional averages. A few examples are given in which simplified expressions are used.

  14. Solution-Adaptive Cartesian Cell Approach for Viscous and Inviscid Flows

    NASA Technical Reports Server (NTRS)

    Coirier, William J.; Powell, Kenneth G.

    1996-01-01

    A Cartesian cell-based approach for adaptively refined solutions of the Euler and Navier-Stokes equations in two dimensions is presented. Grids about geometrically complicated bodies are generated automatically, by the recursive subdivision of a single Cartesian cell encompassing the entire flow domain. Where the resulting cells intersect bodies, polygonal cut cells are created using modified polygon-clipping algorithms. The grid is stored in a binary tree data structure that provides a natural means of obtaining cell-to-cell connectivity and of carrying out solution-adaptive mesh refinement. The Euler and Navier-Stokes equations are solved on the resulting grids using a finite volume formulation. The convective terms are upwinded: A linear reconstruction of the primitive variables is performed, providing input states to an approximate Riemann solver for computing the fluxes between neighboring cells. The results of a study comparing the accuracy and positivity of two classes of cell-centered, viscous gradient reconstruction procedures is briefly summarized. Adaptively refined solutions of the Navier-Stokes equations are shown using the more robust of these gradient reconstruction procedures, where the results computed by the Cartesian approach are compared to theory, experiment, and other accepted computational results for a series of low and moderate Reynolds number flows.

  15. An experimental study of flow around submerged grass vegetation

    NASA Astrophysics Data System (ADS)

    Lee, Julia; Mandre, Shreyas; Singh, Ravi

    2014-11-01

    Mixing of fluids through submerged vegetation caused by tidal currents facilitate various environmental and ecological transport processes. This fluid-vegetation interaction is believed to result from a Kelvin-Helmholtz instability from an inflection point in the flow profile. Recent studies suggest that flow in presence of grass can also become unstable due to shear instability of flow above the grass. We devise a two-dimensional lab scale analog of the fluid-vegetation interaction using ABS plastic filaments immersed in a soap film. We employ PIV of the surrounding flow to gain an understanding of the role of instabilities in the flow.

  16. Fabrication of versatile channel flow cells for quantitative electroanalysis using prototyping.

    PubMed

    Snowden, Michael E; King, Philip H; Covington, James A; Macpherson, Julie V; Unwin, Patrick R

    2010-04-15

    Here we demonstrate the use of microstereolithography (MSL), a 3D direct manufacturing technique, as a viable method to produce small-scale microfluidic components for electrochemical flow detection. The flow cell is assembled simply by resting the microfabricated component on the electrode of interest and securing with thread! This configuration allows the use of a wide range of electrode materials. Furthermore, our approach eliminates the need for additional sealing methods, such as adhesives, waxes, and screws, which have previously been deployed. In addition, it removes any issues associated with compression of the cell chamber. MSL allows a reduction of the dimensions of the channel geometry (and the resultant component) and, compared to most previously produced devices, it offers a high degree of flexibility in the design, reduced manufacture time, and high reliability. Importantly, the polymer utilized does not distort so that the cell maintains well-defined geometrical dimensions after assembly. For the studies herein the channel dimensions were 3 mm wide, 3.5 mm long, and 192 or 250 mum high. The channel flow cell dimensions were chosen to ensure that the substrate electrodes experienced laminar flow conditions, even with volume flow rates of up to 64 mL min(-1) (the limit of our pumping system). The steady-state transport-limited current response, for the oxidation of ferrocenylmethyl trimethylammonium hexaflorophosphate (FcTMA(+)), at gold and polycrystalline boron doped diamond (pBDD) band electrodes was in agreement with the Levich equation and/or finite element simulations of mass transport. We believe that this method of creating and using channel flow electrodes offers a wide range of new applications from electroanalysis to electrocatalysis.

  17. Primary cilia mechanics affects cell mechanosensation: A computational study.

    PubMed

    Khayyeri, Hanifeh; Barreto, Sara; Lacroix, Damien

    2015-08-21

    Primary cilia (PC) are mechanical cell structures linked to the cytoskeleton and are central to how cells sense biomechanical signals from their environment. However, it is unclear exactly how PC mechanics influences cell mechanosensation. In this study we investigate how the PC mechanical characteristics are involved in the mechanotransduction process whereby cilium deflection under fluid flow induces strains on the internal cell components that regulate the cell׳s mechanosensitive response. Our investigation employs a computational approach in which a finite element model of a cell consisting of a nucleus, cytoplasm, cortex, microtubules, actin bundles and a primary cilium was used together with a finite element representation of a flow chamber. Fluid-structure interaction analysis was performed by simulating perfusion flow of 1mm/s on the cell model. Simulations of cells with different PC mechanical characteristics, showed that the length and the stiffness of PC are responsible for the transmission of mechanical stimuli to the cytoskeleton. Fluid flow deflects the cilium, with the highest strains found at the base of the PC and in the cytoplasm. The PC deflection created further strains on the cell nucleus but did not influence microtubules and actin bundles significantly. Our results indicate that PC deflection under fluid flow stimulation transmits mechanical strain primarily to other essential organelles in the cytoplasm, such as the Golgi complex, that regulate cells' mechanoresponse. The simulations further suggest that cell mechanosensitivity can be altered by targeting PC length and rigidity.

  18. Using neural networks for high-speed blood cell classification in a holographic-microscopy flow-cytometry system

    NASA Astrophysics Data System (ADS)

    Schneider, B.; Vanmeerbeeck, G.; Stahl, R.; Lagae, L.; Bienstman, P.

    2015-03-01

    High-throughput cell sorting with flow cytometers is an important tool in modern clinical cell studies. Most cytometers use biomarkers that selectively bind to the cell, but induce significant changes in morphology and inner cell processes leading sometimes to its death. This makes label-based cell sorting schemes unsuitable for further investigation. We propose a label-free technique that uses a digital inline holographic microscopy for cell imaging and an integrated, optical neural network for high-speed classification. The perspective of dense integration makes it attractive to ultrafast, large-scale cell sorting. Network simulations for a ternary classification task (monocytes/granulocytes/lymphocytes) resulted in 89% accuracy.

  19. Motion of cells sedimenting on a solid surface in a laminar shear flow.

    PubMed Central

    Tissot, O; Pierres, A; Foa, C; Delaage, M; Bongrand, P

    1992-01-01

    Cell adhesion often occurs under dynamic conditions, as in flowing blood. A quantitative understanding of this process requires accurate knowledge of the topographical relationships between the cell membrane and potentially adhesive surfaces. This report describes an experimental study made on both the translational and rotational velocities of leukocytes sedimenting of a flat surface under laminar shear flow. The main conclusions are as follows: (a) Cells move close to the wall with constant velocity for several tens of seconds. (b) The numerical values of translational and rotational velocities are inconsistent with Goldman's model of a neutrally buoyant sphere in a laminar shear flow, unless a drag force corresponding to contact friction between cells and the chamber floor is added. The phenomenological friction coefficient was 7.4 millinewton.s/m. (c) Using a modified Goldman's theory, the width of the gap separating cells (6 microns radius) from the chamber floor was estimated at 1.4 micron. (d) It is shown that a high value of the cell-to-substrate gap may be accounted for by the presence of cell surface protrusions of a few micrometer length, in accordance with electron microscope observations performed on the same cell population. (e) In association with previously reported data (Tissot, O., C. Foa, C. Capo, H. Brailly, M. Delaage, and P. Bongrand. 1991. Biocolloids and Biosurfaces. In press), these results are consistent with the possibility that cell-substrate attachment be initiated by the formation of a single molecular bond, which might be considered as the rate limiting step. Images FIGURE 3 FIGURE 8 PMID:1540690

  20. Motion of cells sedimenting on a solid surface in a laminar shear flow.

    PubMed

    Tissot, O; Pierres, A; Foa, C; Delaage, M; Bongrand, P

    1992-01-01

    Cell adhesion often occurs under dynamic conditions, as in flowing blood. A quantitative understanding of this process requires accurate knowledge of the topographical relationships between the cell membrane and potentially adhesive surfaces. This report describes an experimental study made on both the translational and rotational velocities of leukocytes sedimenting of a flat surface under laminar shear flow. The main conclusions are as follows: (a) Cells move close to the wall with constant velocity for several tens of seconds. (b) The numerical values of translational and rotational velocities are inconsistent with Goldman's model of a neutrally buoyant sphere in a laminar shear flow, unless a drag force corresponding to contact friction between cells and the chamber floor is added. The phenomenological friction coefficient was 7.4 millinewton.s/m. (c) Using a modified Goldman's theory, the width of the gap separating cells (6 microns radius) from the chamber floor was estimated at 1.4 micron. (d) It is shown that a high value of the cell-to-substrate gap may be accounted for by the presence of cell surface protrusions of a few micrometer length, in accordance with electron microscope observations performed on the same cell population. (e) In association with previously reported data (Tissot, O., C. Foa, C. Capo, H. Brailly, M. Delaage, and P. Bongrand. 1991. Biocolloids and Biosurfaces. In press), these results are consistent with the possibility that cell-substrate attachment be initiated by the formation of a single molecular bond, which might be considered as the rate limiting step.

  1. New flow cytometric assays for monitoring cell-mediated cytotoxicity.

    PubMed

    Zaritskaya, Liubov; Shurin, Michael R; Sayers, Thomas J; Malyguine, Anatoli M

    2010-06-01

    The exact immunologic responses after vaccination that result in effective antitumor immunity have not yet been fully elucidated and the data from ex vivo T-cell assays have not yet defined adequate surrogate markers for clinical efficacy. A more detailed knowledge of the specific immune responses that correlate with positive clinical outcomes should help to develop better or novel strategies to effectively activate the immune system against tumors. Furthermore, clinically relevant material is often limited and, thus, precludes the ability to perform multiple assays. The two main assays currently used to monitor lymphocyte-mediated cytoxicity in cancer patients are the (51)Cr-release assay and IFN-gamma ELISpot assay. The former has a number of disadvantages, including low sensitivity, poor labeling and high spontaneous release of isotope from some tumor target cells. Additional problems with the (51)Cr-release assay include difficulty in obtaining autologous tumor targets, and biohazard and disposal problems for the isotope. The ELISpot assays do not directly measure cytotoxic activity and are, therefore, a surrogate marker of cyotoxic capacity of effector T cells. Furthermore, they do not assess cytotoxicity mediated by the production of the TNF family of death ligands by the cytotoxic cells. Therefore, assays that allow for the simultaneous measurement of several parameters may be more advantageous for clinical monitoring. In this respect, multifactor flow cytometry-based assays are a valid addition to the currently available immunologic monitoring assays. Use of these assays will enable detection and enumeration of tumor-specific cytotoxic T lymphocytes and their specific effector functions and any correlations with clinical responses. Comprehensive, multifactor analysis of effector cell responses after vaccination may help to detect factors that determine the success or failure of a vaccine and its immunological potency.

  2. NMR studies of multiphase flows II

    SciTech Connect

    Altobelli, S.A.; Caprihan, A.; Fukushima, E.

    1995-12-31

    NMR techniques for measurements of spatial distribution of material phase, velocity and velocity fluctuation are being developed and refined. Versions of these techniques which provide time average liquid fraction and fluid phase velocity have been applied to several concentrated suspension systems which will not be discussed extensively here. Technical developments required to further extend the use of NMR to the multi-phase flow arena and to provide measurements of previously unobtainable parameters are the focus of this report.

  3. Online recognition of the multiphase flow regime and study of slug flow in pipeline

    NASA Astrophysics Data System (ADS)

    Liejin, Guo; Bofeng, Bai; Liang, Zhao; Xin, Wang; Hanyang, Gu

    2009-02-01

    single sensor performance. Among various flow patterns of gas-liquid flow, slug flow occurs frequently in the petroleum, chemical, civil and nuclear industries. In the offshore oil and gas field, the maximum slug length and its statistical distribution are very important for the design of separator and downstream processing facility at steady state operations. However transient conditions may be encountered in the production, such as operational upsets, start-up, shut-down, pigging and blowdown, which are key operational and safety issues related to oil field development. So it is necessary to have an understanding the flow parameters under transient conditions. In this paper, the evolution of slug length along a horizontal pipe in gas-liquid flow is also studied in details and then an experimental study of flowrate transients in slug flow is provided. Also, the special gas-liquid flow phenomena easily encountered in the life span of offshore oil fields, called severe slugging, is studied experimentally and some results are presented.

  4. Blood flow in microvascular networks: A study in nonlinear biology

    PubMed Central

    Geddes, John B.; Carr, Russell T.; Wu, Fan; Lao, Yingyi; Maher, Meaghan

    2010-01-01

    Plasma skimming and the Fahraeus–Lindqvist effect are well-known phenomena in blood rheology. By combining these peculiarities of blood flow in the microcirculation with simple topological models of microvascular networks, we have uncovered interesting nonlinear behavior regarding blood flow in networks. Nonlinearity manifests itself in the existence of multiple steady states. This is due to the nonlinear dependence of viscosity on blood cell concentration. Nonlinearity also appears in the form of spontaneous oscillations in limit cycles. These limit cycles arise from the fact that the physics of blood flow can be modeled in terms of state dependent delay equations with multiple interacting delay times. In this paper we extend our previous work on blood flow in a simple two node network and begin to explore how topological complexity influences the dynamics of network blood flow. In addition we present initial evidence that the nonlinear phenomena predicted by our model are observed experimentally. PMID:21198135

  5. Hydraulic sand-model studies of miscible-fluid flow

    USGS Publications Warehouse

    Cahill, J.M.

    1973-01-01

    Hydraulic sand models are useful physical tools in the investigation of the transition zone that occurs between salt and fresh ground water in coastal aquifers. Such models are used to demonstrate the effects of transport mechanisms that influence the dynamic behavior and the shape of the transition zone. The techniques employed in obtaining in-place measurements of solute concentrations are generally the stumbling block in generating data for two-dimensional dispersion systems. Two in-place measurement techniques were used in the studies described: (1) conductivity probes when salt was used as a tracer; and (2) photoelectric cells when organic dye was used as a tracer. Results indicate that conductivity methods are more reliable; however, care must be exercised inasmuch as the probes tend to disturb the fluid flow.

  6. Dissipative particle dynamics simulations of deformation and aggregation of healthy and diseased red blood cells in a tube flow

    SciTech Connect

    Ye, Ting; Phan-Thien, Nhan Khoo, Boo Cheong; Lim, Chwee Teck

    2014-11-15

    In this paper, we report simulation results assessing the deformation and aggregation of mixed healthy and malaria-infected red blood cells (RBCs) in a tube flow. A three dimensional particle model based on Dissipative Particle Dynamics (DPD) is developed to predict the tube flow containing interacting cells. The cells are also modelled by DPD, with a Morse potential to characterize the cell-cell interaction. As validation tests, a single RBC in a tube flow and two RBCs in a static flow are simulated to examine the cell deformation and intercellular interaction, respectively. The study of two cells, one healthy and the other malaria-infected RBCs in a tube flow demonstrates that the malaria-infected RBC (in the leading position along flow direction) has different effects on the healthy RBC (in the trailing position) at the different stage of parasite development or at the different capillary number. With parasitic development, the malaria-infected RBC gradually loses its deformability, and in turn the corresponding trailing healthy RBC also deforms less due to the intercellular interaction. With increasing capillary number, both the healthy and malaria-infected RBCs are likely to undergo an axisymmetric motion. The minimum intercellular distance becomes small enough so that rouleaux is easily formed, i.e., the healthy and malaria-infected RBCs are difficultly disaggregated.

  7. The Fabrication of Flow Field Plates for Direct Methanol Fuel Cell Using Lithography and Radio Frequency Sputtering.

    PubMed

    Chang, Ho; Kao, Mu-Jung; Chen, Chih-Hao; Cho, Kun-Ching; Hsu, Chun-Yao; Chen, Zhi-Lun

    2015-08-01

    This study uses lithography to etch flow fields on a single side of a printed circuit board (PCB) and combines a flow field plate with a collector plate to make innovative anode flow field plates and cathode flow field plates for a direct methanol fuel cell (DMFC). TiO2 thin film is also sputtered on the anode flow field plate using radio frequency (RF) sputtering. The experimental results show that the prepared DMFC has a better maximum power density of 11.928 mW/cm2. Furthermore, when a TiO2 thin film is sputtered on the flow field plate of the assembled DMFC, the maximum power density is 14.426 mW/cm2, which is actually 21% more than that for a DMFC with no TiO2 thin film coated on the flow field plate.

  8. Arterial specification of endothelial cells derived from human induced pluripotent stem cells in a biomimetic flow bioreactor.

    PubMed

    Sivarapatna, Amogh; Ghaedi, Mahboobe; Le, Andrew V; Mendez, Julio J; Qyang, Yibing; Niklason, Laura E

    2015-01-01

    Endothelial cells (ECs) exist in different microenvironments in vivo, including under different levels of shear stress in arteries versus veins. Standard stem cell differentiation protocols to derive ECs and EC-subtypes from human induced pluripotent stem cells (hiPSCs) generally use growth factors or other soluble factors in an effort to specify cell fate. In this study, a biomimetic flow bioreactor was used to subject hiPSC-derived ECs (hiPSC-ECs) to shear stress to determine the impacts on phenotype and upregulation of markers associated with an anti-thrombotic, anti-inflammatory, arterial-like phenotype. The in vitro bioreactor system was able to efficiently mature hiPSC-ECs into arterial-like cells in 24 h, as demonstrated by qRT-PCR for arterial markers EphrinB2, CXCR4, Conexin40 and Notch1, as well protein-level expression of Notch1 intracellular domain (NICD). Furthermore, the exogenous addition of soluble factors was not able to fully recapitulate this phenotype that was imparted by shear stress exposure. The induction of these phenotypic changes was biomechanically mediated in the shear stress bioreactor. This biomimetic flow bioreactor is an effective means for the differentiation of hiPSC-ECs toward an arterial-like phenotype, and is amenable to scale-up for culturing large quantities of cells for tissue engineering applications.

  9. Arterial specification of endothelial cells derived from human induced pluripotent stem cells in a biomimetic flow bioreactor

    PubMed Central

    Sivarapatna, Amogh; Ghaedi, Mahboobe; Le, Andrew V.; Mendez, Julio J.; Qyang, Yibing; Niklason, Laura E.

    2015-01-01

    Endothelial cells (ECs) exist in different microenvironments in vivo, including under different levels of shear stress in arteries versus veins. Standard stem cell differentiation protocols to derive ECs and EC-subtypes from human induced pluripotent stem cells (hiPSCs) generally use growth factors or other soluble factors in an effort to specify cell fate. In this study, a biomimetic flow bioreactor was used to subject hiPSC-derived ECs (hiPSC-ECs) to shear stress to determine the impacts on phenotype and upregulation of markers associated with an anti-thrombotic, anti-inflammatory, arterial-like phenotype. The in vitro bioreactor system was able to efficiently mature hiPSC-ECs into arterial-like cells in 24 hours, as demonstrated by qRT-PCR for arterial markers EphrinB2, CXCR4, Conexin40 and Notch1, as well protein-level expression of Notch1 intracellular domain (NICD). Furthermore, the exogenous addition of soluble factors was not able to fully recapitulate this phenotype that was imparted by shear stress exposure. The induction of these phenotypic changes was biomechanically mediated in the shear stress bioreactor. This biomimetic flow bioreactor is an effective means for the differentiation of hiPSC-ECs toward an arterial-like phenotype, and is amenable to scale-up for culturing large quantities of cells for tissue engineering applications. PMID:25890758

  10. Liquid flow cells having graphene on nitride for microscopy

    DOEpatents

    Adiga, Vivekananda P.; Dunn, Gabriel; Zettl, Alexander K.; Alivisatos, A. Paul

    2016-09-20

    This disclosure provides systems, methods, and apparatus related to liquid flow cells for microscopy. In one aspect, a device includes a substrate having a first and a second oxide layer disposed on surfaces of the substrate. A first and a second nitride layer are disposed on the first and second oxide layers, respectively. A cavity is defined in the first oxide layer, the first nitride layer, and the substrate, with the cavity including a third nitride layer disposed on walls of the substrate and the second oxide layer that define the cavity. A channel is defined in the second oxide layer. An inlet port and an outlet port are defined in the second nitride layer and in fluid communication with the channel. A plurality of viewports is defined in the second nitride layer. A first graphene sheet is disposed on the second nitride layer covering the plurality of viewports.

  11. Solid oxide fuel cell having compound cross flow gas patterns

    DOEpatents

    Fraioli, A.V.

    1983-10-12

    A core construction for a fuel cell is disclosed having both parallel and cross flow passageways for the fuel and the oxidant gases. Each core passageway is defined by electrolyte and interconnect walls. Each electrolyte wall consists of cathode and anode materials sandwiching an electrolyte material. Each interconnect wall is formed as a sheet of inert support material having therein spaced small plugs of interconnect material, where cathode and anode materials are formed as layers on opposite sides of each sheet and are electrically connected together by the interconnect material plugs. Each interconnect wall in a wavy shape is connected along spaced generally parallel line-like contact areas between corresponding spaced pairs of generally parallel electrolyte walls, operable to define one tier of generally parallel flow passageways for the fuel and oxidant gases. Alternate tiers are arranged to have the passageways disposed normal to one another. Solid mechanical connection of the interconnect walls of adjacent tiers to the opposite sides of the common electrolyte wall therebetween is only at spaced point-like contact areas, 90 where the previously mentioned line-like contact areas cross one another.

  12. Solid oxide fuel cell having compound cross flow gas patterns

    DOEpatents

    Fraioli, Anthony V.

    1985-01-01

    A core construction for a fuel cell is disclosed having both parallel and cross flow passageways for the fuel and the oxidant gases. Each core passageway is defined by electrolyte and interconnect walls. Each electrolyte wall consists of cathode and anode materials sandwiching an electrolyte material. Each interconnect wall is formed as a sheet of inert support material having therein spaced small plugs of interconnect material, where cathode and anode materials are formed as layers on opposite sides of each sheet and are electrically connected together by the interconnect material plugs. Each interconnect wall in a wavy shape is connected along spaced generally parallel line-like contact areas between corresponding spaced pairs of generally parallel electrolyte walls, operable to define one tier of generally parallel flow passageways for the fuel and oxidant gases. Alternate tiers are arranged to have the passageways disposed normal to one another. Solid mechanical connection of the interconnect walls of adjacent tiers to the opposite sides of the common electrolyte wall therebetween is only at spaced point-like contact areas, 90 where the previously mentioned line-like contact areas cross one another.

  13. A flow boiling microchannel thermosyphon for fuel cell thermal management

    NASA Astrophysics Data System (ADS)

    Garrity, Patrick Thomas

    To provide a high power density thermal management system for proton exchange membrane (PEM) fuel cell applications, a passively driven thermal management system was assembled to operate in a closed loop two-phase thermosyphon. The system has two major components; a microchannel evaporator plate and a condenser. The microchannel evaporator plate was fabricated with 56 square channels that have a 1 mm x 1 mm cross section and are 115 mm long. Experiments were conducted with a liquid cooled condenser with heat flux as the control variable. Measurements of mass flow rate, temperature field, and pressure drop have been made for the thermosyphon loop. A model is developed to predict the system characteristics such as the temperature and pressure fields, flow rate, flow regime, heat transfer coefficient, and maximum heat flux. When the system is subjected to a heat load that exceeds the maximum heat flux, an unstable flow regime is observed that causes flow reversal and eventual dryout near the evaporator plate wall. This undesirable phenomenon is modeled based on a quasi-steady state assumption, and the model is capable of predicting the heat flux at the onset of instability for quasi-steady two-phase flow. Another focus of this work is the performance of the condenser portion of the loop, which will be air cooled in practice. The aim is to reduce air side thermal resistance and increase the condenser performance, which is accomplished with extended surfaces. A testing facility is assembled to observe the air side heat transfer performance of three aluminum foam samples and three modified carbon foam samples, used as extended surfaces. The aluminum foam samples have a bulk density of 216 kilograms per cubic meter with pore sizes of 0.5, 1, and 2 mm. The modified carbon foam samples have bulk densities of 284, 317, and 400 kilograms per cubic meter and machined flow passages of 3.2 mm. in diameter. Each sample is observed under forced convection with air velocity as the

  14. Innovative Flow Cytometry Allows Accurate Identification of Rare Circulating Cells Involved in Endothelial Dysfunction

    PubMed Central

    Boraldi, Federica; Bartolomeo, Angelica; De Biasi, Sara; Orlando, Stefania; Costa, Sonia; Cossarizza, Andrea; Quaglino, Daniela

    2016-01-01

    Introduction Although rare, circulating endothelial and progenitor cells could be considered as markers of endothelial damage and repair potential, possibly predicting the severity of cardiovascular manifestations. A number of studies highlighted the role of these cells in age-related diseases, including those characterized by ectopic calcification. Nevertheless, their use in clinical practice is still controversial, mainly due to difficulties in finding reproducible and accurate methods for their determination. Methods Circulating mature cells (CMC, CD45-, CD34+, CD133-) and circulating progenitor cells (CPC, CD45dim, CD34bright, CD133+) were investigated by polychromatic high-speed flow cytometry to detect the expression of endothelial (CD309+) or osteogenic (BAP+) differentiation markers in healthy subjects and in patients affected by peripheral vascular manifestations associated with ectopic calcification. Results This study shows that: 1) polychromatic flow cytometry represents a valuable tool to accurately identify rare cells; 2) the balance of CD309+ on CMC/CD309+ on CPC is altered in patients affected by peripheral vascular manifestations, suggesting the occurrence of vascular damage and low repair potential; 3) the increase of circulating cells exhibiting a shift towards an osteoblast-like phenotype (BAP+) is observed in the presence of ectopic calcification. Conclusion Differences between healthy subjects and patients with ectopic calcification indicate that this approach may be useful to better evaluate endothelial dysfunction in a clinical context. PMID:27560136

  15. Asymmetric energy flow in liquid alkylbenzenes: A computational study

    SciTech Connect

    Leitner, David M.; Pandey, Hari Datt

    2015-10-14

    Ultrafast IR-Raman experiments on substituted benzenes [B. C. Pein et al., J. Phys. Chem. B 117, 10898–10904 (2013)] reveal that energy can flow more efficiently in one direction along a molecule than in others. We carry out a computational study of energy flow in the three alkyl benzenes, toluene, isopropylbenzene, and t-butylbenzene, studied in these experiments, and find an asymmetry in the flow of vibrational energy between the two chemical groups of the molecule due to quantum mechanical vibrational relaxation bottlenecks, which give rise to a preferred direction of energy flow. We compare energy flow computed for all modes of the three alkylbenzenes over the relaxation time into the liquid with energy flow through the subset of modes monitored in the time-resolved Raman experiments and find qualitatively similar results when using the subset compared to all the modes.

  16. Improved Diffuse Fluorescence Flow Cytometer Prototype for High Sensitivity Detection of Rare Circulating Cells In Vivo

    NASA Astrophysics Data System (ADS)

    Pestana, Noah Benjamin

    Accurate quantification of circulating cell populations is important in many areas of pre-clinical and clinical biomedical research, for example, in the study of cancer metastasis or the immune response following tissue and organ transplants. Normally this is done "ex-vivo" by drawing and purifying a small volume of blood and then analyzing it with flow cytometry, hemocytometry or microfludic devices, but the sensitivity of these techniques are poor and the process of handling samples has been shown to affect cell viability and behavior. More recently "in vivo flow cytometry" (IVFC) techniques have been developed where fluorescently-labeled cells flowing in a small blood vessel in the ear or retina are analyzed, but the sensitivity is generally poor due to the small sampling volume. To address this, our group recently developed a method known as "Diffuse Fluorescence Flow Cytometry" (DFFC) that allows detection and counting of rare circulating cells with diffuse photons, offering extremely high single cell counting sensitivity. In this thesis, an improved DFFC prototype was designed and validated. The chief improvements were three-fold, i) improved optical collection efficiency, ii) improved detection electronics, and iii) development of a method to mitigate motion artifacts during in vivo measurements. In combination, these improvements yielded an overall instrument detection sensitivity better than 1 cell/mL in vivo, which is the most sensitive IVFC system reported to date. Second, development and validation of a low-cost microfluidic device reader for analysis of ocular fluids is described. We demonstrate that this device has equivalent or better sensitivity and accuracy compared a fluorescence microscope, but at an order-of-magnitude reduced cost with simplified operation. Future improvements to both instruments are also discussed.

  17. Fluid-flow-induced mesenchymal stem cell migration: role of focal adhesion kinase and RhoA kinase sensors.

    PubMed

    Riehl, Brandon D; Lee, Jeong Soon; Ha, Ligyeom; Lim, Jung Yul

    2015-03-01

    The study of mesenchymal stem cell (MSC) migration under flow conditions with investigation of the underlying molecular mechanism could lead to a better understanding and outcome in stem-cell-based cell therapy and regenerative medicine. We used peer-reviewed open source software to develop methods for efficiently and accurately tracking, measuring and processing cell migration as well as morphology. Using these tools, we investigated MSC migration under flow-induced shear and tested the molecular mechanism with stable knockdown of focal adhesion kinase (FAK) and RhoA kinase (ROCK). Under steady flow, MSCs migrated following the flow direction in a shear stress magnitude-dependent manner, as assessed by root mean square displacement and mean square displacement, motility coefficient and confinement ratio. Silencing FAK in MSCs suppressed morphology adaptation capability and reduced cellular motility for both static and flow conditions. Interestingly, ROCK silencing significantly increased migration tendency especially under flow. Blocking ROCK, which is known to reduce cytoskeletal tension, may lower the resistance to skeletal remodelling during the flow-induced migration. Our data thus propose a potentially differential role of focal adhesion and cytoskeletal tension signalling elements in MSC migration under flow shear.

  18. Calcium regulates the cell-to-cell water flow pathway in maize roots during variable water conditions.

    PubMed

    Wu, Yan; Liu, Xiaofang; Wang, Weifeng; Zhang, Suiqi; Xu, Bingcheng

    2012-09-01

    Soil water shortages can decrease root hydraulic conductivity and affect Ca uptake and movement through the plant. In this study, the effects of extra Ca(2+) applied in nutrient solution on the hydraulic properties of the whole roots (Lp(r)) and cortical cells (Lp(cell)) of maize (Zea mays L.) subjected to variable water conditions were investigated. Under well-watered conditions, extra Ca(2+) significantly increased the root Ca content, total root length, and lateral root number; however, it reduced the root cortical cell volume, Lp(r), and Lp(cell). Hg(2+) inhibition experiments suggested that extra Ca(2+) could reduce the contribution of the cell-to-cell water flow pathway. Osmotic stress (10% PEG6000) significantly decreased the cortical cell volume, Lp(r), and Lp(cell) in the control plants, but smaller decreases were observed in the extra Ca(2+) plants. The Hg(2+) treatment reduced the Lp(r) larger in the extra Ca(2+) plants (74.6%) than in the control plants (53.2%), suggesting a higher contribution of the cell-to-cell pathway. The larger Hg(2+) inhibition of the Lp(cell) in the extra Ca(2+) roots (67.2%) when compared to the controls (56.4%) indicated that extra Ca(2+) can mitigate the inhibition of aquaporin expression and/or activity levels via osmotic stress. After 2 d of rehydration, the extra Ca(2+) helped the Lp(r) and Lp(cell) to recover almost completely, but these properties only partially recovered in the control plants. In conclusion, extra Ca(2+) may adjust the contribution of cell-to-cell pathway by regulating the expression and/or activity levels of AQPs according to water availability; this regulation may weaken negative effects and optimize water use. PMID:22841977

  19. Modeling study of terminal transients of blood flow

    NASA Astrophysics Data System (ADS)

    Stiukhina, Elena S.; Postnov, Dmitry E.

    2016-04-01

    In spite of growing body of experimental and theoretical results on blood flow (BF) patterns under the continuously sustained circulation, much less is known about BF dynamics under the exceptional, but still important cases of venous or arterial occlusion used in medical probes. Since these conditions finally lead to complete or nearly complete stop of red blood cells (RBC) motion, we term it as TTBF, being the Terminal Transients of Blood Flow. An extreme case of such transients is the ultimate extinction of BF after the stopping of heart contractions, during which it is governed by gravitation, some vascular-originated propulsion mechanisms, and, possibly, by RBC aggregation. Quite little is known about this process, while reports the detectable post-mortal motion of mice RBC during at least 2 hours. In our work we present the modeling study of TTBF patterns due to gravitational forces. We present the minimalistic model configuration of vasculature in order to simulate what happens immediately after the pumping of blood has been stopped. Our main findings are concerned to reversal of arterial BF, as well as to duration and non-monotonicity of transients.

  20. High frequency of circulating HBcAg-specific CD8 T cells in hepatitis B infection: a flow cytometric analysis

    PubMed Central

    Matsumura, S; Yamamoto, K; Shimada, N; Okano, N; Okamoto, R; Suzuki, T; Hakoda, T; Mizuno, M; Higashi, T; Tsuji, T

    2001-01-01

    Viral antigen-specific T cells are important for virus elimination. We studied the hepatitis B virus (HBV)-specific T cell response using flow cytometry. Three phases of HBV infection were studied: Group A, HBeAg (+) chronic hepatitis; Group B, HBeAb (+) HBV carrier after seroconversion; and Group C, HBsAb (+) phase. Peripheral T cells were incubated with recombinant HB core antigen (HBcAg), and intracytoplasmic cytokines were analysed by flow cytometry. HBcAg-specific CD4 and CD8 T cells were identified in all three groups and the number of IFN-γpositive T cells was greater than TNF-α-positive T cells. The frequency of IFN-γ-positive CD4 and CD8 T cells was highest in Group C, compared with Groups A and B. No significant difference in the HBcAg-specific T cell response was observed between Group A and Group B. The HBcAg-specific CD8 T cell response was diminished by CD4 depletion, addition of antibody against human leucocyte antigen (HLA) class I, class II or CD40L. Cytokine-positive CD8 T cells without HBcAg stimulation were present at a high frequency (7 of 13 cases) in Group B, but were rare in other groups. HBcAg-specific T cells can be detected at high frequency by a sensitive flow cytometric analysis, and these cells are important for controlling HBV replication. PMID:11472405

  1. Study of unsteady flow conditions for slurry fuels

    SciTech Connect

    Ekmann, J.M.; Wildman, D.J.; Klinzing, G.E.

    1985-01-01

    During the past three years, transport characteristics of coal-water mixtures (CWMs) have been studied at the Pittsburgh Energy Technology Center. The effort has concentrated predominantly on studying flow conditions in straight horizontal and vertical sections, and to a lesser extent on studying the flow patterns around elbows of a one-inch-diameter loop and a two-inch-diameter loop. Steady-state flow was characterized for in-house prepared slurries and commercially prepared slurries. For lower concentrated slurries (55 wt % to 60 wt %) and coarse particle size distributions (50% finer than 75 microns), nonhomogeneous flow conditions were encountered across horizontal test sections. Since nonhomogeneous conditions existed in straight sections during steady-state flow, it was decided to further investigate flow conditions during changes in velocity (magnitude and direction). This paper concentrates on nonuniform flow conditions of two types. The first nonuniform flow condition arises from sudden increases in the magnitude of the flow velocity. Pressure measurements recorded at a fixed position in the vertical section of the two-inch-diameter loop during sudden changes in velocity can be analyzed via classic control theory to evaluate the dynamic properties of the CWM. The second nonuniform flow condition occurs as the CWM passes through a bend. Both long-radius bends and 90-degree elbows made of glass and steel have been studied. Pressure-loss data around the long-radius bends and elbows were analyzed with a modified version of the model developed by Ito for single-phase flow around bends. Flow patterns around glass bends and elbows were observed for slurries prepared of vinyl coating powder and water. They are described in an effort to increase understanding of the pressure-loss data. 8 refs., 11 figs., 1 tab.

  2. Studies in Transition and Time Varying Turbulent Flows

    NASA Technical Reports Server (NTRS)

    Grosch, Chester E.

    2004-01-01

    The research focused on two areas: (a) the dynamics of forced turbulent flows and (b) time filtered Large Eddy Simulations (TLES). The dynamics of turbulent flows arising from external forcing of the turbulence are poorly understood. In particular, here are many unanswered questions relating the basic dynamical balances and the existence or nonexistence of statistical equilibrium of forced turbulent flows. The research used direct numerical simulations to explore these questions. The properties of the temporally filtered Navier-Stokes equations were also studied.

  3. Circular Couette cell for two-dimensional turbulence experiments in sheared flow: initial results.

    NASA Astrophysics Data System (ADS)

    Ulmen, John V.; Fontana, Paul W.; Kearney-Fischer, Martin

    2006-03-01

    An experiment to study turbulence in quasi-two-dimensional flows with a controlled mean flow shear has been built. Experiments are underway to investigate the suppression of turbulent transport by sheared flow as seen in geostrophic flows and laboratory fusion plasmas. The apparatus, a circular Couette cell, uses a liquid film of dilute soap solution suspended freely in an annular channel with a rotating outer boundary. The channel is 7 cm wide with an average radius of 46.5 cm, and can be rotated at angular speeds exceeding 10 rad/s. Mean flow profiles will be presented showing the effect of air resistance on the flow; damping lengths on the order of 1 mm are observed. Turbulence is driven independently via electromagnetic forcing. The rate of turbulence injection can be varied continuously, and its spatial scale corresponds to the spatial frequency of an array of NdFeB magnets. Diagnostics include particle imaging velocimetry, two-point laser Doppler velocimetry, and thickness measurements via reflection interferometry. Initial results and plans for upcoming measurements will be presented.

  4. Cell-scale dynamic recycling and cortical flow of the actin–myosin cytoskeleton for rapid cell migration

    PubMed Central

    Yumura, Shigehiko; Itoh, Go; Kikuta, Yumi; Kikuchi, Takeomi; Kitanishi-Yumura, Toshiko; Tsujioka, Masatsune

    2013-01-01

    Summary Actin and myosin II play major roles in cell migration. Whereas pseudopod extension by actin polymerization has been intensively researched, less attention has been paid to how the rest of the actin cytoskeleton such as the actin cortex contributes to cell migration. In this study, cortical actin and myosin II filaments were simultaneously observed in migrating Dictyostelium cells under total internal reflection fluorescence microscopy. The cortical actin and myosin II filaments remained stationary with respect to the substratum as the cells advanced. However, fluorescence recovery after photobleaching experiments and direct observation of filaments showed that they rapidly turned over. When the cells were detached from the substratum, the actin and myosin filaments displayed a vigorous retrograde flow. Thus, when the cells migrate on the substratum, the cortical cytoskeleton firmly holds the substratum to generate the motive force instead. The present studies also demonstrate how myosin II localizes to the rear region of the migrating cells. The observed dynamic turnover of actin and myosin II filaments contributes to the recycling of their subunits across the whole cell and enables rapid reorganization of the cytoskeleton. PMID:23430058

  5. A fluid-structure interaction model to characterize bone cell stimulation in parallel-plate flow chamber systems.

    PubMed

    Vaughan, T J; Haugh, M G; McNamara, L M

    2013-04-01

    Bone continuously adapts its internal structure to accommodate the functional demands of its mechanical environment and strain-induced flow of interstitial fluid is believed to be the primary mediator of mechanical stimuli to bone cells in vivo. In vitro investigations have shown that bone cells produce important biochemical signals in response to fluid flow applied using parallel-plate flow chamber (PPFC) systems. However, the exact mechanical stimulus experienced by the cells within these systems remains unclear. To fully understand this behaviour represents a most challenging multi-physics problem involving the interaction between deformable cellular structures and adjacent fluid flows. In this study, we use a fluid-structure interaction computational approach to investigate the nature of the mechanical stimulus being applied to a single osteoblast cell under fluid flow within a PPFC system. The analysis decouples the contribution of pressure and shear stress on cellular deformation and for the first time highlights that cell strain under flow is dominated by the pressure in the PPFC system rather than the applied shear stress. Furthermore, it was found that strains imparted on the cell membrane were relatively low whereas significant strain amplification occurred at the cell-substrate interface. These results suggest that strain transfer through focal attachments at the base of the cell are the primary mediators of mechanical signals to the cell under flow in a PPFC system. Such information is vital in order to correctly interpret biological responses of bone cells under in vitro stimulation and elucidate the mechanisms associated with mechanotransduction in vivo.

  6. Optimum design of bipolar plates for separate air flow cooling system of PEM fuel cells stacks

    NASA Astrophysics Data System (ADS)

    Franco, Alessandro

    2015-12-01

    The paper discusses about thermal management of PEM fuel cells. The objective is to define criteria and guidelines for the design of the air flow cooling system of fuel cells stacks for different combination of power density, bipolar plates material, air flow rate, operating temperature It is shown that the optimization of the geometry of the channel permits interesting margins for maintaining the use of separate air flow cooling systems for high power density PEM fuel cells.

  7. Fluid Flow through a High Cell Density Fluidized-Bed during Centrifugal Bioreactor Culture

    PubMed Central

    Detzel, Christopher J.; Van Wie, Bernard J.; Ivory, Cornelius F.

    2010-01-01

    An increasing demand for products such as tissues, proteins, and antibodies from mammalian cell suspension cultures is driving interest in increasing production through high-cell density bioreactors. The centrifugal bioreactor (CCBR) retains cells by balancing settling forces with surface drag forces due to medium throughput and is capable of maintaining cell densities above 108 cells/mL. This article builds on a previous study where the fluid mechanics of an empty CCBR were investigated showing fluid flow is nonuniform and dominated by Coriolis forces, raising concerns about nutrient and cell distribution. In this article, we demonstrate that the previously reported Coriolis forces are still present in the CCBR, but masked by the presence of cells. Experimental dye injection observations during culture of 15 μm hybridoma cells show a continual uniform darkening of the cell bed, indicating the region of the reactor containing cells is well mixed. Simulation results also indicate the cell bed is well mixed during culture of mammalian cells ranging in size from 10 to 20 μm. However, simulations also allow for a slight concentration gradient to be identified and attributed to Coriolis forces. Experimental results show cell density increases from 0.16 to 0.26 when centrifugal force is doubled by increasing RPM from 650 to 920 at a constant inlet velocity of 6.5 cm/s; an effect also observed in the simulation. Results presented in this article indicate cells maintained in the CCBR behave as a high-density fluidized bed of cells providing a homogeneous environment to ensure optimal growth conditions. PMID:20205172

  8. A numerical analysis of forces exerted by laminar flow on spreading cells in a parallel plate flow chamber assay.

    PubMed

    Olivier, L A; Truskey, G A

    1993-10-01

    Exposure of spreading anchorage-dependent cells to laminar flow is a common technique to measure the strength of cell adhesion. Since cells protrude into the flow stream, the force exerted by the fluid on the cells is a function of cell shape. To assess the relationship between cell shape and the hydrodynamic force on adherent cells, we obtained numerical solutions of the velocity and stress fields around bovine aortic endothelial cells during various stages of spreading and calculated the force required to detach the cells. Morphometric parameters were obtained from light and scanning electron microscopy measurements. Cells were assumed to have a constant volume, but the surface area increased during spreading until the membrane was stretched taut. Two-dimensional models of steady flow were generated using the software packages ANSYS (mesh generation) and FIDAP (problem solution). The validity of the numerical results was tested by comparison with published results for a semicircle in contact with the surface. The drag force and torque were greatest for round cells making initial contact with the surface. During spreading, the drag force and torque declined by factors of 2 and 20, respectively. The calculated forces and moments were used in adhesion models to predict the wall shear stress at which the cells detached. Based upon published values for the bond force and receptor number, round cells should detach at shear stresses between 2.5 and 6 dyn/cm(2), whereas substantially higher stresses are needed to detach spreading and fully spread cells. Results from the simulations indicate that (1) the drag force varies little with cell shape whereas the torque is very sensitive to cell shape, and (2) the increase in the strength of adhesion during spreading is due to increased contact area and receptor densities within the contact area.

  9. Photoacoustic-fluorescence in vitro flow cytometry for quantification of absorption, scattering and fluorescence properties of the cells

    NASA Astrophysics Data System (ADS)

    Nedosekin, D. A.; Sarimollaoglu, M.; Foster, S.; Galanzha, E. I.; Zharov, V. P.

    2013-03-01

    Fluorescence flow cytometry is a well-established analytical tool that provides quantification of multiple biological parameters of cells at molecular levels, including their functional states, morphology, composition, proliferation, and protein expression. However, only the fluorescence and scattering parameters of the cells or labels are available for detection. Cell pigmentation, presence of non-fluorescent dyes or nanoparticles cannot be reliably quantified. Herewith, we present a novel photoacoustic (PA) flow cytometry design for simple integration of absorbance measurements into schematics of conventional in vitro flow cytometers. The integrated system allow simultaneous measurements of light absorbance, scattering and of multicolor fluorescence from single cells in the flow at rates up to 2 m/s. We compared various combinations of excitation laser sources for multicolor detection, including simultaneous excitation of PA and fluorescence using a single 500 kHz pulsed nanosecond laser. Multichannel detection scheme allows simultaneous detection of up to 8 labels, including 4 fluorescent tags and 4 PA colors. In vitro PA-fluorescence flow cytometer was used for studies of nanoparticles uptake and for the analysis of cell line pigmentation, including genetically encoded melanin expression in breast cancer cell line. We demonstrate that this system can be used for direct nanotoxicity studies with simultaneous quantification of nanoparticles content and assessment of cell viability using a conventional fluorescent apoptosis assays.

  10. Flow of a circulating tumor cell and red blood cells in microvessels

    NASA Astrophysics Data System (ADS)

    Takeishi, Naoki; Imai, Yohsuke; Yamaguchi, Takami; Ishikawa, Takuji

    2015-12-01

    Quantifying the behavior of circulating tumor cells (CTCs) in the blood stream is of fundamental importance for understanding metastasis. Here, we investigate the flow mode and velocity of CTCs interacting with red blood cells (RBCs) in various sized microvessels. The flow of leukocytes in microvessels has been described previously; a leukocyte forms a train with RBCs in small microvessels and exhibits margination in large microvessels. Important differences in the physical properties of leukocytes and CTCs result from size. The dimensions of leukocytes are similar to those of RBCs, but CTCs are significantly larger. We investigate numerically the size effects on the flow mode and the cell velocity, and we identify similarities and differences between leukocytes and CTCs. We find that a transition from train formation to margination occurs when (R -a ) /tR≈1 , where R is the vessel radius, a is the cell radius, and tR is the thickness of RBCs, but that the motion of RBCs differs from the case of leukocytes. Our results also show that the velocities of CTCs and leukocytes are larger than the average blood velocity, but only CTCs move faster than RBCs for microvessels of R /a ≈1.5 -2.0 . These findings are expected to be useful not only for understanding metastasis, but also for developing microfluidic devices.

  11. Hydrodynamic forces on a wall-bound leukocyte due to interactions with flowing red cells

    NASA Astrophysics Data System (ADS)

    Isfahani, Amir H. G.; Freund, Jonathan B.

    2011-11-01

    As part of both healthy and pathologically physiological mechanisms sphere-like white blood cells (leukocytes) adhere to the walls of small blood vessels. We use quantitative numerical simulations to compare the forces from flowing red blood cells on a wall-adhered leukocyte to a homogenized model of blood at the same flow conditions. We model the highly flexible red blood cells using a fast O (N log N) boundary integral formulation. These elastic membranes deform substantially but strongly resist surface dilatation. They enclose a higher than plasma viscosity hemoglobin solution. The no-slip condition is enforced on the stationary leukocyte as well as the vessel walls. Vessel diameters of 10 to 20 microns are studied. Different hematocrits, leukocyte shapes, and flow conditions are examined. In vessels comparable to the size of the cells, we show that the particulate character of blood significantly affects the magnitude of the forces that the leukocyte experiences, transiently increasing it well above the homogenized-blood prediction: for example, for a tube hematocrit of 25 % and a spherical protrusion with a diameter 0.75 that of the tube, the average forces are increased by about 40 % and the local forces by more than 100 % relative to those expected for a blood model homogenized by its effective viscosity.

  12. Pressure-driven occlusive flow of a confined red blood cell.

    PubMed

    Savin, Thierry; Bandi, M M; Mahadevan, L

    2016-01-14

    When red blood cells (RBCs) move through narrow capillaries in the microcirculation, they deform as they flow. In pathophysiological processes such as sickle cell disease and malaria, RBC motion and flow are severely restricted. To understand this threshold of occlusion, we use a combination of experiment and theory to study the motion of a single swollen RBC through a narrow glass capillary of varying inner diameter. By tracking the movement of the squeezed cell as it is driven by a controlled pressure drop, we measure the RBC velocity as a function of the pressure gradient as well as the local capillary diameter, and find that the effective blood viscosity in this regime increases with both decreasing RBC velocity and tube radius by following a power-law that depends upon the length of the confined cell. Our observations are consistent with a simple elasto-hydrodynamic model and highlight the role of lateral confinement in the occluded pressure-driven slow flow of soft confined objects. PMID:26497051

  13. Best practices for application of attachment cells to in vitro micronucleus assessment by flow cytometry.

    PubMed

    Bemis, Jeffrey C; Bryce, Steven M; Nern, Marlies; Raschke, Marian; Sutter, Andreas

    2016-01-01

    This work seeks to provide users with guidance on cell culture, treatment, processing and analytical conditions for achieving optimal performance of the in vitro micronucleus assay using the In Vitro MicroFlow(®) method. Experimental data are provided to support the advice described. The information provided covers specific topics or issues that are identified as critical to the methodology and thus is meant to work with instruction manuals, published papers and other references, and not as a replacement for these documents. The content is divided into several sections. Cell culture and treatment describes conditions for routine maintenance of cells as well as treatment with test articles. Preparation and processing of samples details steps found to be critical in execution of the procedure. Instrument parameters and analysis covers set-up of the flow cytometer and evaluation of the samples. General assay considerations and interpretation of results describes examination of data in terms of assay validity, viability and genotoxicity assessment. The goal is to educate users and enable them to design, conduct and interpret flow cytometric in vitro micronucleus (MN) studies. Readers should obtain an understanding of specific cell culture practices, options for assay formatting and execution and the information required to successfully integrate and validate the in vitro MN assay into their existing safety program. PMID:26774667

  14. Best practices for application of attachment cells to in vitro micronucleus assessment by flow cytometry.

    PubMed

    Bemis, Jeffrey C; Bryce, Steven M; Nern, Marlies; Raschke, Marian; Sutter, Andreas

    2016-01-01

    This work seeks to provide users with guidance on cell culture, treatment, processing and analytical conditions for achieving optimal performance of the in vitro micronucleus assay using the In Vitro MicroFlow(®) method. Experimental data are provided to support the advice described. The information provided covers specific topics or issues that are identified as critical to the methodology and thus is meant to work with instruction manuals, published papers and other references, and not as a replacement for these documents. The content is divided into several sections. Cell culture and treatment describes conditions for routine maintenance of cells as well as treatment with test articles. Preparation and processing of samples details steps found to be critical in execution of the procedure. Instrument parameters and analysis covers set-up of the flow cytometer and evaluation of the samples. General assay considerations and interpretation of results describes examination of data in terms of assay validity, viability and genotoxicity assessment. The goal is to educate users and enable them to design, conduct and interpret flow cytometric in vitro micronucleus (MN) studies. Readers should obtain an understanding of specific cell culture practices, options for assay formatting and execution and the information required to successfully integrate and validate the in vitro MN assay into their existing safety program.

  15. Stem cells and fluid flow drive cyst formation in an invertebrate excretory organ.

    PubMed

    Thi-Kim Vu, Hanh; Rink, Jochen C; McKinney, Sean A; McClain, Melainia; Lakshmanaperumal, Naharajan; Alexander, Richard; Sánchez Alvarado, Alejandro

    2015-06-09

    Cystic kidney diseases (CKDs) affect millions of people worldwide. The defining pathological features are fluid-filled cysts developing from nephric tubules due to defective flow sensing, cell proliferation and differentiation. The underlying molecular mechanisms, however, remain poorly understood, and the derived excretory systems of established invertebrate models (Caenorhabditis elegans and Drosophila melanogaster) are unsuitable to model CKDs. Systematic structure/function comparisons revealed that the combination of ultrafiltration and flow-associated filtrate modification that is central to CKD etiology is remarkably conserved between the planarian excretory system and the vertebrate nephron. Consistently, both RNA-mediated genetic interference (RNAi) of planarian orthologues of human CKD genes and inhibition of tubule flow led to tubular cystogenesis that share many features with vertebrate CKDs, suggesting deep mechanistic conservation. Our results demonstrate a common evolutionary origin of animal excretory systems and establish planarians as a novel and experimentally accessible invertebrate model for the study of human kidney pathologies.

  16. The intercell dynamics of T cells and dendritic cells in a lymph node-on-a-chip flow device.

    PubMed

    Moura Rosa, Patrícia; Gopalakrishnan, Nimi; Ibrahim, Hany; Haug, Markus; Halaas, Øyvind

    2016-10-01

    T cells play a central role in immunity towards cancer and infectious diseases. T cell responses are initiated in the T cell zone of the lymph node (LN), where resident antigen-bearing dendritic cells (DCs) prime and activate antigen-specific T cells passing by. In the present study, we investigated the T cell : DC interaction in a microfluidic device to understand the intercellular dynamics and physiological conditions in the LN. We show random migration of antigen-specific T cells onto the antigen-presenting DC monolayer independent of the flow direction with a mean T cell : DC dwell time of 12.8 min and a mean velocity of 6 μm min(-1). Furthermore, we investigated the antigen specific vs. unspecific attachment and detachment of CD8(+) and CD4(+) T cells to DCs under varying shear stress. In our system, CD4(+) T cells showed long stable contacts with APCs, whereas CD8(+) T cells presented transient interactions with DCs. By varying the shear stress from 0.01 to 100 Dyn cm(-2), it was also evident that there was a much stronger attachment of antigen-specific than unspecific T cells to stationary DCs up to 1-12 Dyn cm(-2). The mechanical force of the cell : cell interaction associated with the pMHC-TCR match under controlled tangential shear force was estimated to be in the range of 0.25-4.8 nN. Finally, upon performing attachment & detachment tests, there was a steady accumulation of antigen specific CD8(+) T cells and CD4(+) T cells on DCs at low shear stresses, which were released at a stress of 12 Dyn cm(-2). This microphysiological model provides new possibilities to recreate a controlled mechanical force threshold of pMHC-TCR binding, allowing the investigation of intercellular signalling of immune synapses and therapeutic targets for immunotherapy.

  17. Theory to Predict Shear Stress on Cells in Turbulent Blood Flow

    PubMed Central

    Morshed, Khandakar Niaz; Bark Jr., David; Forleo, Marcio; Dasi, Lakshmi Prasad

    2014-01-01

    Shear stress on blood cells and platelets transported in a turbulent flow dictates the fate and biological activity of these cells. We present a theoretical link between energy dissipation in turbulent flows to the shear stress that cells experience and show that for the case of physiological turbulent blood flow: (a) the Newtonian assumption is valid, (b) turbulent eddies are universal for the most complex of blood flow problems, and (c) shear stress distribution on turbulent blood flows is possibly universal. Further we resolve a long standing inconsistency in hemolysis between laminar and turbulent flow using the theoretical framework. This work demonstrates that energy dissipation as opposed to bulk shear stress in laminar or turbulent blood flow dictates local mechanical environment of blood cells and platelets universally. PMID:25171175

  18. Theory to predict shear stress on cells in turbulent blood flow.

    PubMed

    Morshed, Khandakar Niaz; Bark, David; Forleo, Marcio; Dasi, Lakshmi Prasad

    2014-01-01

    Shear stress on blood cells and platelets transported in a turbulent flow dictates the fate and biological activity of these cells. We present a theoretical link between energy dissipation in turbulent flows to the shear stress that cells experience and show that for the case of physiological turbulent blood flow: (a) the Newtonian assumption is valid, (b) turbulent eddies are universal for the most complex of blood flow problems, and (c) shear stress distribution on turbulent blood flows is possibly universal. Further we resolve a long standing inconsistency in hemolysis between laminar and turbulent flow using the theoretical framework. This work demonstrates that energy dissipation as opposed to bulk shear stress in laminar or turbulent blood flow dictates local mechanical environment of blood cells and platelets universally.

  19. Study flow diagrams in Cochrane systematic review updates: an adapted PRISMA flow diagram.

    PubMed

    Stovold, Elizabeth; Beecher, Deirdre; Foxlee, Ruth; Noel-Storr, Anna

    2014-05-29

    Cochrane systematic reviews are conducted and reported according to rigorous standards. A study flow diagram must be included in a new review, and there is clear guidance from the PRISMA statement on how to do this. However, for a review update, there is currently no guidance on how study flow diagrams should be presented. To address this, a working group was formed to find a solution and produce guidance on how to use these diagrams in review updates.A number of different options were devised for how these flow diagrams could be used in review updates, and also in cases where multiple searches for a review or review update have been conducted. These options were circulated to the Cochrane information specialist community for consultation and feedback. Following the consultation period, the working group refined the guidance and made the recommendation that for review updates an adapted PRISMA flow diagram should be used, which includes an additional box with the number of previously included studies feeding into the total. Where multiple searches have been conducted, the results should be added together and treated as one set of results.There is no existing guidance for using study flow diagrams in review updates. Our adapted diagram is a simple and pragmatic solution for showing the flow of studies in review updates.

  20. Localized Modeling of Biochemical and Flow Interactions during Cancer Cell Adhesion

    PubMed Central

    Behr, Julie; Gaskin, Byron; Fu, Changliang; Dong, Cheng; Kunz, Robert

    2015-01-01

    This work focuses on one component of a larger research effort to develop a simulation tool to model populations of flowing cells. Specifically, in this study a local model of the biochemical interactions between circulating melanoma tumor cells (TC) and substrate adherent polymorphonuclear neutrophils (PMN) is developed. This model provides realistic three-dimensional distributions of bond formation and attendant attraction and repulsion forces that are consistent with the time dependent Computational Fluid Dynamics (CFD) framework of the full system model which accounts local pressure, shear and repulsion forces. The resulting full dynamics model enables exploration of TC adhesion to adherent PMNs, which is a known participating mechanism in melanoma cell metastasis. The model defines the adhesion molecules present on the TC and PMN cell surfaces, and calculates their interactions as the melanoma cell flows past the PMN. Biochemical rates of reactions between individual molecules are determined based on their local properties. The melanoma cell in the model expresses ICAM-1 molecules on its surface, and the PMN expresses the β-2 integrins LFA-1 and Mac-1. In this work the PMN is fixed to the substrate and is assumed fully rigid and of a prescribed shear-rate dependent shape obtained from micro-PIV experiments. The melanoma cell is transported with full six-degrees-of-freedom dynamics. Adhesion models, which represent the ability of molecules to bond and adhere the cells to each other, and repulsion models, which represent the various physical mechanisms of cellular repulsion, are incorporated with the CFD solver. All models are general enough to allow for future extensions, including arbitrary adhesion molecule types, and the ability to redefine the values of parameters to represent various cell types. The model presented in this study will be part of a clinical tool for development of personalized medical treatment programs. PMID:26366568

  1. Study of low resistivity and high work function ITO films prepared by oxygen flow rates and N2O plasma treatment for amorphous/crystalline silicon heterojunction solar cells.

    PubMed

    Hussain, Shahzada Qamar; Oh, Woong-Kyo; Kim, Sunbo; Ahn, Shihyun; Le, Anh Huy Tuan; Park, Hyeongsik; Lee, Youngseok; Dao, Vinh Ai; Velumani, S; Yi, Junsin

    2014-12-01

    Pulsed DC magnetron sputtered indium tin oxide (ITO) films deposited on glass substrates with lowest resistivity of 2.62 x 10(-4) Ω x cm and high transmittance of about 89% in the visible wavelength region. We report the enhancement of ITO work function (Φ(ITO)) by the variation of oxygen (O2) flow rate and N2O surface plasma treatment. The Φ(ITO) increased from 4.43 to 4.56 eV with the increase in O2 flow rate from 0 to 4 sccm while surface treatment of N2O plasma further enhanced the ITO work function to 4.65 eV. The crystallinity of the ITO films improved with increasing O2 flow rate, as revealed by XRD analysis. The ITO work function was increased by the interfacial dipole resulting from the surface rich in O- ions and by the dipole moment formed at the ITO surface during N2O plasma treatment. The ITO films with high work functions can be used to modify the front barrier height in heterojunction with intrinsic thin layer (HIT) solar cells. PMID:25971043

  2. Cell-cycle distribution of urothelial tumour cells as measured by flow cytometry.

    PubMed Central

    Collste, L. G.; Darzynkiewicz, Z.; Traganos, F.; Sharpless, T. K.; Devonec, M.; Claps, M. L.; Whitmore, W. F.; Melamed, M. R.

    1979-01-01

    The fraction of cells in S + G2 + mitosis from 54 urothelial tumours was calculated by flow cytometry after acridine orange (AO) staining of cells obtained by bladder irrigation or biopsy. Fluorescence signals emitted by the AO-stained DNA and RNA of each cell were separated optically and measured for 5,000 cells per specimen. The patients were classified by the histology of their tumours and clinical data into 5 diagnostic categories: NED (no evidence of disease, but history of bladder tumour), 3; papilloma, 8; non-invasive papillary carcinoma, 8; carcinoma in situ, 17 and invasive carcinoma, 18. The fraction of cells with DNA values in S + G2 + M of the cell cycle varied between 7 and 57% of the total, with a wide range within each diagnostic category, but no statistically significant differences between the groups. The proportion of cells in S + G2 + M from an individual tumour was not correlated with histologic grade or clinical behaviour. The possibility that some tumour cells with DNA values above G1 level are quiescent cells arrested at S or G2 is discussed. PMID:526428

  3. Flow cytometric analysis of BDE 47 mediated injury to rainbow trout gill epithelial cells

    PubMed Central

    Shao, Jing; Dabrowski, Michael J.; White, Collin C.; Kavanagh, Terrance J.; Gallagher, Evan P.

    2012-01-01

    The polybrominated diphenyl ethers (PBDEs) are ubiquitous environmental contaminants whose residues are increasing in fish, wildlife and human tissues. However, relatively little is known regarding the mechanisms of cell injury caused by PBDE congeners in fish. In the present study, we employed flow cytometry-based analyses to understand the onset and mechanisms of cell injury in rainbow trout gill cells (RTgill-W1 cells) exposed to 2,2′,4,4′-tetrabromodiphenyl ether (BDE 47). Substantial optimization and validation for flow cytometry protocols were required during assay development for the trout gill cell line. Exposure to micromolar concentrations of BDE 47 elicited a significant loss in RTgill-W1 cell viability that was accompanied by a decrease in NAD(P)H autofluorescence, a marker associated with disruption of cellular redox status. This loss in NAD(P)H content was accompanied by a decrease in nonylacridine orange fluorescence, indicating mitochondrial membrane lipid peroxidation. Furthermore, low doses of BDE 47 altered cellular forward angle light scatter (FS, a measure of cell diameter or size) and side light scatter properties (SS, a measure of cellular internal complexity), consistent with the early stages of apoptosis. These changes were more pronounced at higher BDE 47 concentrations, which lead to an increase in the percentage of cells undergoing frank apoptosis as evidenced by sub-G1 DNA content. Apoptosis was also observed at a relatively low dose (3.2 μM) of BDE 47 if cells were exposed for an extended period of time (24 hr). Collectively, the results of these studies indicate that exposure of rainbow trout gill cells to BDE47 is associated with the induction of apoptosis likely originating from disruption of cellular redox status and mitochondrial oxidative injury. The current report extends observations in other species demonstrating that oxidative stress is an important mechanism of BDE 47 mediated cellular toxicity, and supports the use of

  4. Compound Lava Flow Fields on Planetary Surfaces: Hawaiian Analogue Studies

    NASA Astrophysics Data System (ADS)

    Crown, D. A.; Byrnes, J. M.; Ramsey, M. S.

    2002-12-01

    Quantitative, process-oriented analyses of planetary volcanism have primarily been based on analogue studies of single-lobed lava flows emplaced as discrete units. Comparative analyses of compound lava flow fields on the Earth and terrestrial planets are being conducted in order to include volcanic styles characterized by complex distributary systems, stratigraphic relationships, and emplacement histories. Field observations, differential Global Positioning Systems (dGPS) measurements, and visible, thermal, and radar remote sensing are being used to characterize Hawaiian lava flow fields and develop techniques for analyses of planetary flow fields using datasets with high spatial and/or spectral resolution, such as MOC and THEMIS. These terrestrial studies allow flow field surface morphology, topography, and lava textures as well as detailed maps of distributary networks to be used to examine flow field growth and development. Information on flow field evolution is provided by delineating relationships between remote sensing signatures, surface morphology, and lava transport processes and by identifying input parameters for flowfield emplacement models. Investigations of the Mauna Ulu (1969-1974) and Puu Oo (1983-present) flow fields (Kilauea Volcano, HI) have focused on understanding the nature of distributary networks at various scales in order to determine spatial and temporal variations in lava transport. Initial work at Mauna Ulu has included analyses of 1) the distribution, network morphometry, and volumetric significance of lava channels in the medial zone of the flow field, and 2) the distribution, lava texture, and volumetric significance of breakouts from surface conduits and subsurface storage. Analyses of the temporal evolution of individual conduit systems provide the basis for interpretation of complex patterns of overlapping surface units that characterize local flow stratigraphy. Reconstruction of lava transport networks and relationships to surface

  5. The Pilot Training Study: Personnel Flow and the PILOT Model.

    ERIC Educational Resources Information Center

    Mooz, W. E.

    The results of the Rand study of pilot flows and the computer-operated decision model, called the PILOT model, are described. The flows of pilots within the Air Force are caused by policies that require the career-development rotation of pilots from cockpit jobs to desk jobs, the maintenance of a supplement of pilots in excess of cockpit-related…

  6. New use for an old reagent: Cell cycle analysis of DNA content using flow cytometry in formamide treated cells.

    PubMed

    Carbonari, Maurizio

    2016-05-01

    Formamide has long been one of the most widely used reagents in the study of nucleic acids. However, the use of formamide for treating cells to be analyzed by flow cytometry is a recent development and is restricted to measuring telomere lengths by flow-FISH. In this field, we have published several papers in order to observe the effects of formamide treatment on cells at room temperature. We therefore discovered that, with suitable modifications, a short and simple incubation in this ionizing solvent facilitates cell cycle analysis by flow cytometry, equivalent or superior to that obtained with treatments in alcohol, acetone or detergent in hypotonic solution. Even using a bulky and problematic stain (low quantum efficiency and G-C base preference), such as 7-aminoactinomycin D (7-AAD) which, on the other hand, has the advantage of being excited at 488 nm and does not bind to the RNA, it is possible to obtain excellent coefficients of variation and (G2-M) mode/(G0-G1) mode ratios. These parameters, especially if stained cells are washed before acquisition, arrive at optimal values. It is noteworthy that the ability to wash the cells stained for DNA content analysis without affecting the stoichiometry of the staining has not been described elsewhere in the literature. With formamide treatment the doublets are practically absent, sample recovery is efficient, as well as the preservation of physical parameters, and the stained cells can be stored for at least 10 days at room temperature before acquisition. © 2016 International Society for Advancement of Cytometry. PMID:26866418

  7. Tracking and Measurement of the Motion of Blood Cells Using Optical Flow Methods

    PubMed Central

    Guo, Dongmin; Van de Ven, Anne L.; Zhou, Xiaobo

    2014-01-01

    The investigation of microcirculation is a critical task in biomedical and physiological research. In order to monitor human’s condition and develop effective therapies of some diseases, the microcirculation information, such as flow velocity and vessel density, must be evaluated in a noninvasive manner. As one of the tasks of microcirculation investigation, automatic blood cell tracking presents an effective approach to estimate blood flow velocity. Currently, the most common method for blood cell tracking is based on spatiotemporal image analysis, which has lots of limitations, such as the diameter of microvesssels cannot be too larger than blood cells or tracers, cells or tracers should have fixed velocity, and it requires the image with high qualification. In this paper, we propose an optical flow method for automatic cell tracking. The key algorithm of the method is to align an image to its neighbors in a large image collection consisting of a variety of scenes. Considering the method cannot solve the problems in all cases of cell movement, another optical flow method, SIFT (Scale Invariant Feature Transform) flow, is also presented. The experimental results show that both methods can track the cells accurately. Optical flow is specially robust to the case where the velocity of cell is unstable, while SIFT flow works well when there are large displacement of cell between two adjacent frames. Our proposed methods outperform other methods when doing in vivo cell tracking, which can be used to estimate the blood flow directly and help to evaluate other parameters in microcirculation. PMID:24058034

  8. Flow cytometric analysis of in vitro bluetongue virus infection of bovine blood mononuclear cells.

    PubMed

    Barratt-Boyes, S M; Rossitto, P V; Stott, J L; MacLachlan, N J

    1992-08-01

    Cultures of adherent and non-adherent bovine peripheral blood mononuclear (PBM) cells were inoculated with bluetongue virus (BTV) serotype 10. Some cultures of non-adherent cells were stimulated with interleukin 2 (IL-2) and concanavalin A for 24 h prior to virus inoculation. Cells were harvested at various intervals up to 72 h after inoculation. A panel of leukocyte differentiation antigen-specific monoclonal antibodies (MAbs), specific for bovine CD2, CD4 or CD8, monocytes and granulocytes, B cells, gamma delta T cells or the IL-2 receptor (IL-2r), was directly conjugated to fluorescein isothiocyanate, and a MAb specific for the BTV major core protein VP7 was directly conjugated to phycoerythrin. Cells were labelled with conjugated MAbs in single- and double-label immunofluorescence studies to identify specifically the BTV-infected cells in inoculated cultures. The viability of cells was determined by propidium iodide exclusion, and all analyses were done using flow cytometry. Productive infection of cultures of PBM cells was confirmed by virus titration. The data revealed a clear difference between subsets of bovine PBM cells in susceptibility to infection with BTV in vitro. Monocytes were readily infected with BTV, as were stimulated CD4+ cells, and infection was cytopathic to monocytes and stimulated lymphocytes. The proportion of infected cells decreased after 24 h and virus titres dropped markedly by 72 h in all cultures. CD4+ cells in cultures of unstimulated non-adherent cells inoculated with BTV showed increased expression of IL-2r. The possible relevance of these findings to the pathogenesis of BTV infection of cattle is discussed.

  9. Advancing the detection of maternal haematopoietic microchimeric cells in fetal immune organs in mice by flow cytometry

    PubMed Central

    Solano, Maria Emilia; Thiele, Kristin; Stelzer, Ina Annelies; Mittrücker, Hans-Willi; Arck, Petra Clara

    2014-01-01

    Maternal microchimerism, which occurs naturally during gestation in hemochorial placental mammals upon transplacental migration of maternal cells into the fetus, is suggested to significantly influence the fetal immune system. In our previous publication, we explored the sensitivity of quantitative polymerase chain reaction and flow cytometry to detect cellular microchimerism. With that purpose, we created mixed cells suspensions in vitro containing reciprocal frequencies of wild type cells and cells positive for enhanced green fluorescent protein or CD45.1+, respectively. Here, we now introduce the H-2 complex, which defines the major histocompatibility complex in mice and is homologous to HLA in human, as an additional target to detect maternal microchimerism among fetal haploidentical cells. We envision that this advanced approach to detect maternal microchimeric cells by flow cytometry facilitates the pursuit of phenotypic, gene expression and functional analysis of microchimeric cells in future studies. PMID:25483743

  10. A fluid–structure interaction model to characterize bone cell stimulation in parallel-plate flow chamber systems

    PubMed Central

    Vaughan, T. J.; Haugh, M. G.; McNamara, L. M.

    2013-01-01

    Bone continuously adapts its internal structure to accommodate the functional demands of its mechanical environment and strain-induced flow of interstitial fluid is believed to be the primary mediator of mechanical stimuli to bone cells in vivo. In vitro investigations have shown that bone cells produce important biochemical signals in response to fluid flow applied using parallel-plate flow chamber (PPFC) systems. However, the exact mechanical stimulus experienced by the cells within these systems remains unclear. To fully understand this behaviour represents a most challenging multi-physics problem involving the interaction between deformable cellular structures and adjacent fluid flows. In this study, we use a fluid–structure interaction computational approach to investigate the nature of the mechanical stimulus being applied to a single osteoblast cell under fluid flow within a PPFC system. The analysis decouples the contribution of pressure and shear stress on cellular deformation and for the first time highlights that cell strain under flow is dominated by the pressure in the PPFC system rather than the applied shear stress. Furthermore, it was found that strains imparted on the cell membrane were relatively low whereas significant strain amplification occurred at the cell–substrate interface. These results suggest that strain transfer through focal attachments at the base of the cell are the primary mediators of mechanical signals to the cell under flow in a PPFC system. Such information is vital in order to correctly interpret biological responses of bone cells under in vitro stimulation and elucidate the mechanisms associated with mechanotransduction in vivo. PMID:23365189

  11. Flow cytometric analysis of cell-surface and intracellular antigens in the diagnosis of acute leukemia.

    PubMed

    Paredes-Aguilera, R; Romero-Guzman, L; Lopez-Santiago, N; Burbano-Ceron, L; Camacho-Del Monte, O; Nieto-Martinez, S

    2001-10-01

    To evaluate the usefulness of flow cytometric detection of intracellular antigens (Ags) in establishing proper lineage affiliation and its contribution to the diagnosis of acute leukemia, we studied 100 consecutive patients in whom acute leukemia was diagnosed between January 1997 and July 1998. Immunological classification was assessed using a three-line panel of monoclonal antibodies for phenotypic characterization of leukemic blast cells as proposed at the First Latin American Consensus Conference for Flow Cytometric Immunophenotyping of Leukemia. We found 74 cases of B-cell lineage acute lymphoblastic leukemia (ALL), seven cases of T-cell ALL, and 19 cases of acute myeloid leukemia (AML). In this study cytoplasmic (cy) CD79a, cyCD22, cyCD3, and cyMPO were highly sensitive, specific B, T, and myeloid markers that were expressed in virtually all cases of B and T cell ALL and in all subtypes of AML. Applied in combination with immunophenotyping this knowledge led to improvement in diagnostic precision and refinement of immunological classification, ensuring the selection of the most appropriate therapy for the patients studied. In conclusion, intracellular Ags detection was of utmost importance in establishing correct lineage affiliation in cases lacking expression of B, T, or myeloid surface Ags or disclosing equivocal or ambiguous immunophenotypic features and in identifying biphenotypic acute leukemia. In combination with FAB morphology and immunophenotyping, we were able to reliably classify all patients with acute leukemia in this study.

  12. Characterizing dynamic hysteresis and fractal statistics of chaotic two-phase flow and application to fuel cells

    NASA Astrophysics Data System (ADS)

    Burkholder, Michael B.; Litster, Shawn

    2016-05-01

    In this study, we analyze the stability of two-phase flow regimes and their transitions using chaotic and fractal statistics, and we report new measurements of dynamic two-phase pressure drop hysteresis that is related to flow regime stability and channel water content. Two-phase flow dynamics are relevant to a variety of real-world systems, and quantifying transient two-phase flow phenomena is important for efficient design. We recorded two-phase (air and water) pressure drops and flow images in a microchannel under both steady and transient conditions. Using Lyapunov exponents and Hurst exponents to characterize the steady-state pressure fluctuations, we develop a new, measurable regime identification criteria based on the dynamic stability of the two-phase pressure signal. We also applied a new experimental technique by continuously cycling the air flow rate to study dynamic hysteresis in two-phase pressure drops, which is separate from steady-state hysteresis and can be used to understand two-phase flow development time scales. Using recorded images of the two-phase flow, we show that the capacitive dynamic hysteresis is related to channel water content and flow regime stability. The mixed-wettability microchannel and in-channel water introduction used in this study simulate a polymer electrolyte fuel cell cathode air flow channel.

  13. New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept

    PubMed Central

    Nava-Castro, Karen; Hernández-Bello, Romel; Muñiz-Hernández, Saé; Escobedo, Galileo; Morales-Montor, Jorge

    2011-01-01

    In parasitology, particularly in helminthes studies, several methods have been used to look for the expression of specific molecules, such as RT-PCR, western blot, 2D-electrophoresis, and microscopy, among others. However, these methods require homogenization of the whole helminth parasite, preventing evaluation of individual cells or specific cell types in a given parasite tissue or organ. Also, the extremely high interaction between helminthes and host cells (particularly immune cells) is an important point to be considered. It is really hard to obtain fresh parasites without host cell contamination. Then, it becomes crucial to determine that the analyzed proteins are exclusively from parasitic origin, and not a consequence of host cell contamination. Flow cytometry is a fluorescence-based technique used to evaluate the expression of extra-and intracellular proteins in different type cells, including protozoan parasites. It also allows the isolation and recovery of single-cell populations. Here, we describe a method to isolate and obtain purified helminthes cells. PMID:22187522

  14. NK cell isolation from liver biopsies: phenotypic and functional analysis of low cell numbers by flow cytometry.

    PubMed

    Li, Ning; Puga Yung, Gisella L; Pradier, Amandine; Toso, Christian; Giostra, Emiliano; Morard, Isabelle; Spahr, Laurent; Seebach, Jörg D

    2013-01-01

    Natural killer (NK) cells are considered to play a critical role in liver disease. However, the available numbers of intrahepatic lymphocytes (IHL) derived from liver biopsies (LB) for ex vivo analysis of intrahepatic NK cells is very limited; and the isolation method may hamper not only yields and viability, but also phenotype and function of IHL. The aim of the present study was therefore to (1) refine and evaluate the cell yields and viability of a modified isolation protocol from standard size needle LB; and (2) to test the effects of mechanical dissociation and enzymatic tissue digestion, as well as the analysis of very low cell numbers, on the phenotype and function of intrahepatic NK cells. Peripheral blood mononuclear cells (PBMC) and IHL, freshly isolated from the peripheral blood, LB (n = 11) or partial liver resections (n = 5), were used for phenotypic analysis by flow cytometry. NK cell function, i.e., degranulation and cytokine production, was determined by staining of CD107a and intracellular IFN-γ following in vitro stimulation. The mean weight of the LB specimens was 9.1 mg, and a mean number of 7,364 IHL/mg were obtained with a viability of >90%. Exposure of IHL and PBMC to 0.5 mg/ml collagenase IV and 0.02 mg/ml DNase I for 30 min did affect neither the viability, NK cell function, nor the percentages of CD56(+), NKp46(+), and CD16(+) NK cells, whereas the level of CD56 surface expression was reduced. The phenotype of LB-derived NK cells was reliably characterized by acquiring as few as 2,500 IHL per tube for flow cytometry. The functional assay of intrahepatic NK cells was miniaturized by culturing as few as 25,000 IHL in 25 μl (10(6)/ml) using 96-well V-bottom plates with IL-2 and IL-12 overnight, followed by a 4 h stimulation with K562 cells at a NK:K562 ratio of 1:1. In summary, we report reliable phenotypic and functional analyses of small numbers of intrahepatic NK cells isolated from LB specimens providing us with a

  15. A review of mathematical modeling of the zinc/bromine flow cell and battery

    NASA Astrophysics Data System (ADS)

    Evans, T. I.; White, R. E.

    1987-11-01

    Mathematical models which have been developed to study various aspects of the zinc/bromine cell and stack of cells are reviewed. Development of these macroscopic models begins with a material balance, a transport equation which includes a migration term for charged species in an electric field, and an electrode kinetic expression. Various types of models are discussed: partial differential equation models that can be used to predict current and potential distributions, an algebraic model that includes shunt currents and associated energy losses and can be used to determine the optimum resistivity of an electrolyte, and ordinary differential equation models that can be used to predict the energy efficiency of the cell as a function of the state of charge. These models have allowed researchers to better understand the physical phenomena occurring within parallel plate electrochemical flow reactors and have been instrumental in the improvement of the zinc/bromine cell design. Suggestions are made for future modeling work.

  16. Quantitative assessment of immune cells in the injured spinal cord tissue by flow cytometry: a novel use for a cell purification method.

    PubMed

    Nguyen, Hal X; Beck, Kevin D; Anderson, Aileen J

    2011-04-09

    Detection of immune cells in the injured central nervous system (CNS) using morphological or histological techniques has not always provided true quantitative analysis of cellular inflammation. Flow cytometry is a quick alternative method to quantify immune cells in the injured brain or spinal cord tissue. Historically, flow cytometry has been used to quantify immune cells collected from blood or dissociated spleen or thymus, and only a few studies have attempted to quantify immune cells in the injured spinal cord by flow cytometry using fresh dissociated cord tissue. However, the dissociated spinal cord tissue is concentrated with myelin debris that can be mistaken for cells and reduce cell count reliability obtained by the flow cytometer. We have advanced a cell preparation method using the OptiPrep gradient system to effectively separate lipid/myelin debris from cells, providing sensitive and reliable quantifications of cellular inflammation in the injured spinal cord by flow cytometry. As described in our recent study (Beck & Nguyen et al., Brain. 2010 Feb; 133 (Pt 2): 433-47), the OptiPrep cell preparation had increased sensitivity to detect cellular inflammation in the injured spinal cord, with counts of specific cell types correlating with injury severity. Critically, novel usage of this method provided the first characterization of acute and chronic cellular inflammation after SCI to include a complete time course for polymorphonuclear leukocytes (PMNs, neutrophils), macrophages/microglia, and T-cells over a period ranging from 2 hours to 180 days post-injury (dpi), identifying a surprising novel second phase of cellular inflammation. Thorough characterization of cellular inflammation using this method may provide a better understanding of neuroinflammation in the injured CNS, and reveal an important multiphasic component of neuroinflammation that may be critical for the design and implementation of rational therapeutic treatment strategies, including both

  17. Collection of peripheral blood stem cells using an automated discontinuous flow blood cell separator.

    PubMed

    Pierelli, L; Menichella, G; Paoloni, A; Teofili, L; Sica, S; Foddai, M L; Rossi, P L; Leone, G; Mango, G; Bizzi, B

    1990-01-01

    Twenty collections of peripheral blood stem cells were performed in 3 patients (2 NHL, 1 AML) using the Haemonetics V50S discontinuous flow blood cell separator. A modified lymphocyte collection protocol (Nebraska Surge) was used in all instances. Leukapheresis were performed after 1 or 2 courses of chemotherapy and started when peripheral blood leukocytes count reached 1 x 10e9/l and platelets count 80 x 10e9/l. A mean blood volume of 5.9 +/- 0.6 litres was processed per procedure and the mean yields for mononuclear cells, nucleated cells and CFU-GM were respectively 5.4 +/- 1.4 x 10e9, 4.9 +/- 1.6 x 10e9 and 128.5 +/- 182.3 x 10e4 per procedure. Haemonetics V50S had showed a mean collection efficiency for mononuclear cells of 67.5 +/- 5.0% per procedure. Results obtained are not significantly different from the ones obtained with an automated continuous flow separator even if extracorporeal circulation is consistently high in patients with a low hematocrit when the 250 ml Latham Bowl is used.

  18. Study of flow patterns in fume hood enclosures

    SciTech Connect

    Pathanjali, C.; Rahman, M.M.

    1996-12-31

    A three-dimensional model for flow inside a fume hood enclosure was developed and numerical computations were carried out to explore the flow pattern and possible path of contaminant transport under different operating conditions of the hood. Equations for the conservation of mass and momentum were solved for different flow rate and opening conditions in the hood. The face velocity was maintained constant at its rated value of 0.4 m/s. The flow was assumed to enter through the front window opening (positive x-direction) and leave the cupboard through an opening on the top of the hood (positive z-direction). The flow was assumed to be fully turbulent. The {kappa}-{var_epsilon} model was used for the prediction of turbulence. The flow pattern for different sash openings were studied. The flow patterns around an object located at the bottom of the hood was studied for different locations of the object. In addition, the effect of a person standing in front of the hood on the flow pattern was investigated. It was found that air entering the hood proceeds directly to the back wall, impinges it and turns upward toward the top wall and exits through the outlet. The flow finds its way around any object forming a recirculating region at its trailing surface. With an increase in the sash opening, the velocity becomes higher and the fluid traces the path to the outlet more quickly. The volume occupied by recirculating flow decreases with increase in sash opening. The computed flow patterns will be very useful to design experiments with optimum sash opening providing adequate disposal of contaminants with minimum use of conditioned air from inside the room.

  19. Experimental Studies of Low-Pressure Turbine Flows and Flow Control

    NASA Technical Reports Server (NTRS)

    Volino, Ralph J.

    2012-01-01

    This report summarizes research performed in support of the NASA Glenn Research Center (GRC) Low-Pressure Turbine (LPT) Flow Physics Program. The work was performed experimentally at the U.S. Naval Academy faculties. The geometry corresponded to "Pak B" LPT airfoil. The test section simulated LPT flow in a passage. Three experimental studies were performed: (a) Boundary layer measurements for ten baseline cases under high and low freestream turbulence conditions at five Reynolds numbers of 25,000, 50,000, 100,000, 200,000, and 300,000, based on passage exit velocity and suction surface wetted length; (b) Passive flow control studies with three thicknesses of two-dimensional bars, and two heights of three-dimensional circular cylinders with different spanwise separations, at same flow conditions as the 10 baseline cases; (c) Active flow control with oscillating synthetic (zero net mass flow) vortex generator jets, for one case with low freestream turbulence and a low Reynolds number of 25,000. The Passive flow control was successful at controlling the separation problem at low Reynolds numbers, with varying degrees of success from case to case and varying levels of impact at higher Reynolds numbers. The active flow control successfully eliminated the large separation problem for the low Reynolds number case. Very detailed data was acquired using hot-wire anemometry, including single and two velocity components, integral boundary layer quantities, turbulence statistics and spectra, turbulent shear stresses and their spectra, and intermittency, documenting transition, separation and reattachment. Models were constructed to correlate the results. The report includes a summary of the work performed and reprints of the publications describing the various studies.

  20. Flow distribution and pressure drop in parallel-channel configurations of planar fuel cells

    NASA Astrophysics Data System (ADS)

    Maharudrayya, S.; Jayanti, S.; Deshpande, A. P.

    Parallel-channel configurations for gas-distributor plates of planar fuel cells reduce the pressure drop, but give rise to the problem of severe flow maldistribution wherein some of the channels may be starved of the reactants. This study presents an analysis of the flow distribution through parallel-channel configurations. One-dimensional models based on mass and momentum balance equations in the inlet and exhaust gas headers are developed for Z- and U-type parallel-channel configurations. The resulting coupled ordinary differential equations are solved analytically to obtain closed-form solutions for the flow distribution in the individual channels and for the pressure drop over the entire distributor plate. The models have been validated by comparing the results with those obtained from three-dimensional computational fluid dynamics (CFD) simulations. Application of the models to typical fuel-cell distributor plates shows that severe maldistribution of flow may arise in certain cases and that this can be avoided by careful choice of the dimensions of the headers and the channels.

  1. On the oscillating motion of a red blood cell in bounded Poiseuille flows

    NASA Astrophysics Data System (ADS)

    Yu, Yao; Shi, Lingling; Glowinski, Roland; Pan, Tsorng-Whay

    2012-11-01

    Two motions of oscillation and vacillating breathing (swing) a RBC have been observed in bounded Poiseuille flows [Phys. Rev. E 85, 16307 (2012)]. To understand such motions, we have studied the motion of a neutrally buoyant rigid particle of the same shape in bounded Poiseuille flows and obtained that the equilibrium height of the mass center, the confined ratio of the long axis of the particle and the channel height, and the initial position of the particle are important factors for having such oscillating motion. But the crucial one is to have the particle interacting with Poiseuille flow with its mass center oscillating about the channel center. When the mass center is always away from the channel center, the particle just keep rotating. Since the mass center of the cell migrates to the channel center in bounded Poiseuille flow in the regime of low Reynolds number, the oscillating motion then is similar to the aforementioned motion as long as the cell keeps the shape of long body. This work is funded by the US NSF.

  2. NASA Non-Flow-Through PEM Fuel Cell System for Aerospace Applications

    NASA Technical Reports Server (NTRS)

    Araghi, Koorosh R.

    2011-01-01

    NASA is researching passive NFT Proton Exchange Membrane (PEM) fuel cell technologies for primary fuel cell power plants in air-independent applications. NFT fuel cell power systems have a higher power density than flow through systems due to both reduced parasitic loads and lower system mass and volume. Reactant storage still dominates system mass/volume considerations. NFT fuel cell stack testing has demonstrated equivalent short term performance to flow through stacks. More testing is required to evaluate long-term performance.

  3. Analysis of red blood cell deformation under fast shear flow for better estimation of hemolysis.

    PubMed

    Nakamura, Masanori; Bessho, Sadao; Wada, Shigeo

    2014-01-01

    We examined the deformation behavior of a red blood cell (RBC) in various flow fields to determine whether the extent of RBC deformation is correlated with the shear stress used as a hemolysis index. The RBC model was introduced to a simple shear flow (Couette flow) and to slightly complex flows (unsteady shear flows and stenosed flows). The RBC deformation was assessed by the maximum first principal strain over the RBC membrane and compared with the shear stress. Although the results were consistent under steady Couette flow, this was not the case under unsteady Couette flow or stenosed flow due to the viscoelastic nature of the RBC deformation caused by fluid forces. These results suggest that there is a limitation in accurately estimating the mechanical damage of RBCs solely from a macroscopic flow field, indicating the necessity of taking into account the dynamic deformation of RBCs to provide a better estimation of hemolysis.

  4. In-vitro study on haemodiluted blood flow in a sinusoidal microstenosis.

    PubMed

    Kang, M J; Ji, H-S; Lee, S J

    2010-01-01

    In-vitro experiments were carried out to investigate the haemodynamic and haemorheological behaviours of haemodiluted blood flow through a microstenosis using a micro-particle image velocimetry (PIV) technique. The micro-PIV system employed in this study consisted of a two-head neodymium:yttrium-aluminium-garnet (Nd:YAG) laser, a cooled charge-coupled device camera, and a delay generator. To simulate blood flow in a stenosed vascular vessel, a polydimethylsiloxane (PDMS) microchannel with a sinusoidal throat of 80 per cent severity was employed. The width and depth of the microchannel were 100 microm and 50 microm, respectively. To compare the flow characteristics in the microstenosis, the same experiments were repeated in a straight microchannel under the same flow conditions. Using a syringe pump, human blood with 5 per cent haematocrit was supplied into the microstenosis channel. The flow characteristics and transport of blood cells through the microstenosis were investigated with various flowrates. The mean velocity fields were nearly symmetric with respect to the channel centreline. In the contraction section, the oncoming blood flow was accelerated rapidly, and the maximum velocity at the throat was almost 4.99 times faster than that of the straight microchannel without stenosis. In the diffusion section, the blood cells show rolling, deformation, twisting, and tumbling motion due to the flow-choking characteristics at the stenotic region. The results from this study will provide useful basic data for comparison with those obtained by clinical researchers. PMID:20225454

  5. Study on flow instability and countermeasure in a draft tube with swirling flow

    NASA Astrophysics Data System (ADS)

    Nakashima, T.; Matsuzaka, R.; Miyagawa, K.; Yonezawa, K.; Tsujimoto, Y.

    2014-03-01

    The swirling flow in the draft tube of a Francis turbine can cause the flow instability and the cavitation surge and has a larger influence on hydraulic power operating system. In this paper, the cavitating flow with swirling flow in the diffuser was studied by the draft tube component experiment, the model Francis turbine experiment and the numerical simulation. In the component experiment, several types of fluctuations were observed, including the cavitation surge and the vortex rope behaviour by the swirling flow. While the cavitation surge and the vortex rope behaviour were suppressed by the aeration into the diffuser, the loss coefficient in the diffuser increased by the aeration. In the model turbine test the aeration decreased the efficiency of the model turbine by several percent. In the numerical simulation, the cavitating flow was studied using Scale-Adaptive Simulation (SAS) with particular emphasis on understanding the unsteady characteristics of the vortex rope structure. The generation and evolution of the vortex rope structures have been investigated throughout the diffuser using the iso-surface of vapor volume fraction. The pressure fluctuation in the diffuser by numerical simulation confirmed the cavitation surge observed in the experiment. Finally, this pressure fluctuation of the cavitation surge was examined and interpreted by CFD.

  6. Coupled thermal, electrical, and fluid flow analyses of AMTEC multitube cell with adiabatic side wall

    NASA Astrophysics Data System (ADS)

    Schock, A.; Or, C.; Noravian, H.

    1997-01-01

    The paper describes a novel OSC-generated methodology for analyzing the performance of multitube AMTEC (Alkali Metal Thermal-to-Electrical Conversion) cells, which are under development by AMPS (Advanced Modular Power Systems, Inc.) for the Air Force Phillips Laboratory (AFPL) and NASA's Jet Propulsion Laboratory (JPL), for possible application to the Pluto Express and other space missions. The OSC study was supported by the Department of Energy (DOE), and was strongly encouraged by JPL, AFPL, and AMPS. It resulted in an iterative procedure for the coupled solution of the interdependent thermal, electrical, and fluid flow differential and integral equations governing the performance of AMTEC cells and generators. The paper clarifies the OSC procedure by presenting detailed results of its application to an illustrative example of a converter cell with an adiabatic side wall, including the non-linear axial variation of temperature, pressure, open-circuit voltage, interelectrode voltage, current density, axial current, sodium mass flow, and power density. The next paper in these proceedings describes parametric results obtained by applying the same procedure to variations of the baseline adiabatic converter design, culminating in an OSC-recommended revised cell design. A subsequent paper in these proceedings extends the procedure to analyze a variety of OSC-designed radioisotope-heated generators employing non-adiabatic multitube AMTEC cells.

  7. Static magnetic fields affect capillary flow of red blood cells in striated skin muscle.

    PubMed

    Brix, Gunnar; Strieth, Sebastian; Strelczyk, Donata; Dellian, Marc; Griebel, Jürgen; Eichhorn, Martin E; Andrā, Wilfried; Bellemann, Matthias E

    2008-01-01

    Blood flowing in microvessels is one possible site of action of static magnetic fields (SMFs). We evaluated SMF effects on capillary flow of red blood cells (RBCs) in unanesthetized hamsters, using a skinfold chamber technique for intravital fluorescence microscopy. By this approach, capillary RBC velocities (v(RBC)), capillary diameters (D), arteriolar diameters (D(art)), and functional vessel densities (FVD) were measured in striated skin muscle at different magnetic flux densities. Exposure above a threshold level of about 500 mT resulted in a significant (P < 0.001) reduction of v(RBC) in capillaries as compared to the baseline value. At the maximum field strength of 587 mT, v(RBC) was reduced by more than 40%. Flow reduction was reversible when the field strength was decreased below the threshold level. In contrast, mean values determined at different exposure levels for the parameters D, D(art), and FVD did not vary by more than 5%. Blood flow through capillary networks is affected by strong SMFs directed perpendicular to the vessels. Since the influence of SMFs on blood flow in microvessels directed parallel to the field as well as on collateral blood supply could not be studied, our findings should be carefully interpreted with respect to the setting of safety guidelines.

  8. Some specific features of the NMR study of fluid flows

    NASA Astrophysics Data System (ADS)

    Davydov, V. V.

    2016-07-01

    Some specific features of studying fluid flows with a NMR spectrometer are considered. The consideration of these features in the NMR spectrometer design makes it possible to determine the relative concentrations of paramagnetic ions and measure the longitudinal and transverse relaxation times ( T 1 and T 2, respectively) in fluid flows with an error no larger than 0.5%. This approach allows one to completely avoid errors in determining the state of a fluid from measured relaxation constants T 1 and T 2, which is especially urgent when working with medical suspensions and biological solutions. The results of an experimental study of fluid flows are presented.

  9. Numerical studies of transverse curvature effects on transonic flow stability

    NASA Technical Reports Server (NTRS)

    Macaraeg, M. G.; Daudpota, Q. I.

    1992-01-01

    A numerical study of transverse curvature effects on compressible flow temporal stability for transonic to low supersonic Mach numbers is presented for axisymmetric modes. The mean flows studied include a similar boundary-layer profile and a nonsimilar axisymmetric boundary-layer solution. The effect of neglecting curvature in the mean flow produces only small quantitative changes in the disturbance growth rate. For transonic Mach numbers (1-1.4) and aerodynamically relevant Reynolds numbers (5000-10,000 based on displacement thickness), the maximum growth rate is found to increase with curvature - the maximum occurring at a nondimensional radius (based on displacement thickness) between 30 and 100.

  10. Chemostat flow cell system: an in vitro model for the evaluation of antiplaque agents.

    PubMed

    Herles, S; Olsen, S; Afflitto, J; Gaffar, A

    1994-11-01

    We developed an experimental in vitro model of dental plaque to assess the potential efficacy of antiplaque agents. The model used a chemostat, which provided a continuous source of 5 species of oral bacteria grown in an artificial "saliva-like" medium. This mixture was pumped through six flow cells, each containing two types of surfaces on which plaque formed and was subsequently measured. Formation of bacterial plaque on hydroxyapatite surfaces was assessed by measurement of the DNA and protein content of the plaque film. The amount of bacterial plaque formed on germanium surfaces was measured by attenuated total reflectance (ATR/FT-IR) spectroscopy. Plaque viability was also assessed by a fluorescent staining technique. The quantity of plaque formed on both types of surfaces gradually increased with the duration of flow (from 24 to 72 h) through the cells during a 72-hour experimental period. The flow cells were then pulsed with experimental treatment solutions for 30 s, twice daily. Parallel to results of human clinical studies, the model was capable of discriminating among water, a placebo mouthrinse, and an active antimicrobial mouthrinse formulation containing 0.03% triclosan. It therefore offers a valuable alternative to animal model testing and allows for more rapid evaluations under well-controlled experimental conditions. PMID:7983262

  11. Chemostat flow cell system: an in vitro model for the evaluation of antiplaque agents.

    PubMed

    Herles, S; Olsen, S; Afflitto, J; Gaffar, A

    1994-11-01

    We developed an experimental in vitro model of dental plaque to assess the potential efficacy of antiplaque agents. The model used a chemostat, which provided a continuous source of 5 species of oral bacteria grown in an artificial "saliva-like" medium. This mixture was pumped through six flow cells, each containing two types of surfaces on which plaque formed and was subsequently measured. Formation of bacterial plaque on hydroxyapatite surfaces was assessed by measurement of the DNA and protein content of the plaque film. The amount of bacterial plaque formed on germanium surfaces was measured by attenuated total reflectance (ATR/FT-IR) spectroscopy. Plaque viability was also assessed by a fluorescent staining technique. The quantity of plaque formed on both types of surfaces gradually increased with the duration of flow (from 24 to 72 h) through the cells during a 72-hour experimental period. The flow cells were then pulsed with experimental treatment solutions for 30 s, twice daily. Parallel to results of human clinical studies, the model was capable of discriminating among water, a placebo mouthrinse, and an active antimicrobial mouthrinse formulation containing 0.03% triclosan. It therefore offers a valuable alternative to animal model testing and allows for more rapid evaluations under well-controlled experimental conditions.

  12. Mechanotransduction in an extracted cell model: Fyn drives stretch- and flow-elicited PECAM-1 phosphorylation.

    PubMed

    Chiu, Yi-Jen; McBeath, Elena; Fujiwara, Keigi

    2008-08-25

    Mechanosensing followed by mechanoresponses by cells is well established, but the mechanisms by which mechanical force is converted into biochemical events are poorly understood. Vascular endothelial cells (ECs) exhibit flow- and stretch-dependent responses and are widely used as a model for studying mechanotransduction in mammalian cells. Platelet EC adhesion molecule 1 (PECAM-1) is tyrosine phosphorylated when ECs are exposed to flow or when PECAM-1 is directly pulled, suggesting that it is a mechanochemical converter. We show that PECAM-1 phosphorylation occurs when detergent-extracted EC monolayers are stretched, indicating that this phosphorylation is mechanically triggered and does not require the intact plasma membrane and soluble cytoplasmic components. Using kinase inhibitors and small interfering RNAs, we identify Fyn as the PECAM-1 kinase associated with the model. We further show that stretch- and flow-induced PECAM-1 phosphorylation in intact ECs is abolished when Fyn expression is down-regulated. We suggest that PECAM-1 and Fyn are essential components of a PECAM-1-based mechanosensory complex in ECs. PMID:18710921

  13. A numerical study of the Kernel-conformation transformation for transient viscoelastic fluid flows

    NASA Astrophysics Data System (ADS)

    Martins, F. P.; Oishi, C. M.; Afonso, A. M.; Alves, M. A.

    2015-12-01

    This work presents a numerical application of a generic conformation tensor transformation for simulating highly elastic flows of non-Newtonian fluids typically observed in computational rheology. In the Kernel-conformation framework [14], the conformation tensor constitutive law for a viscoelastic fluid is transformed introducing a generic tensor transformation function. The numerical stability of the application of the Kernel-conformation for highly elastic flows is ultimately related with the specific kernel function used in the matrix transformation, but also to the existence of singularities introduced either by flow geometry or by the characteristics of the constitutive equation. In this work, we implement this methodology in a free-surface Marker-And-Cell discretization methodology implemented in a finite differences method. The main contributions of this work are two fold: on one hand, we demonstrate the accuracy of this Kernel-conformation formulation using a finite differences method and free surfaces; on the other hand, we assess the numerical efficiency of specific kernel functions at high-Weissenberg number flows. The numerical study considers different viscoelastic fluid flow problems, including the Poiseuille flow in a channel, the lid-driven cavity flow and the die-swell free surface flow. The numerical results demonstrate the adequacy of this methodology for high Weissenberg number flows using the Oldroyd-B model.

  14. Analysis of cells in cerebrospinal fluid from patients with neurocysticercosis by means of flow cytometry.

    PubMed

    Bueno, E C; Vaz, A J; Oliveira, C A; Machado, L R; Livramento, J A; Mielli, S R; Ueda, M

    1999-06-15

    The events of the cellular immune response in neurocysticercosis (NC) are not fully understood. Studies of the CD3, CD3/CD4, CD3/CD8, CD45/CD19, and CD45/CD56 molecules and activation-related CD69 molecule in cells from the cerebrospinal fluid (CSF) and peripheral blood (PB) of patients with NC may provide a better elucidation of the inflammatory and immunological events occurring in this disease. Seven patients with NC and 3 individuals with other disorders were evaluated by a three-color flow cytometric method. CD69 was detected in a higher percentage of cells in all CSF samples from patients, but not in PB or CSF from the control group. The percentage of CD3+ cells did not differ significantly in CSF and PB cells from patients and controls. The predominance of CD3+CD8+ cells was observed in CSF from one patient and in PB from 2 patients, who were in stage III of the disease (inflammatory process). The percentage of CD45+CD19+ cells was higher in CSF than in PB from patients who presented anti-cysticercus antibodies in CSF. The percentage of CD45+CD56+ cells in CSF was higher than in PB, but this rate was similar to reference values reported by other authors. Our data suggest that the cytometric method applied to a larger number of CSF samples may provide a better understanding of the cell-mediated immune response involved in NC.

  15. Deformation of a single red blood cell in bounded Poiseuille flows

    NASA Astrophysics Data System (ADS)

    Shi, Lingling; Pan, Tsorng-Whay; Glowinski, Roland

    2012-01-01

    Deformation of a red blood cell (RBC) in bounded two-dimensional Poiseuille flows is studied by using an immersed boundary method (IBM). An elastic spring model is applied to simulate the skeleton structure of a RBC membrane. As a benchmarking test, the dynamical behavior of a single RBC under a simple shear flow has been validated. Then we focus on investigating the motion and the deformation of a single RBC in Poiseuille flows by varying the swelling ratio (s*), the initial angle of the long axis of the cell at the centerline (ϕ), the maximum velocity at the centerline of fluid flow (umax), the membrane bending stiffness of a RBC (kb), and the height of the microchannel (H). Two motions of oscillation and vacillating breathing (swing) of a RBC are observed in both narrow and wide channels. The strength of the vacillating-breathing motion depends on the degree of confinement and the value of umax. A RBC exhibits a strong vacillating-breathing motion as the degree of confinement is larger or the value of umax is higher. For the same degree of confinement, the vacillating-breathing motion appears to be relatively weaker but persists longer as the value of umax is lower. The continuation of shape change from the slippery to the parachute by varying the value of umax is obtained for the biconcave shape cell in a narrower channel. In particular, parachute shape and bulletlike shape, depending on the angle ϕ, coexist for the elliptic shape cell given initially with lower umax in a narrower channel.

  16. Cell contractility arising from topography and shear flow determines human mesenchymal stem cell fate

    PubMed Central

    Sonam, Surabhi; Sathe, Sharvari R.; Yim, Evelyn K.F.; Sheetz, Michael P.; Lim, Chwee Teck

    2016-01-01

    Extracellular matrix (ECM) of the human Mesenchymal Stem Cells (MSCs) influences intracellular tension and is known to regulate stem cell fate. However, little is known about the physiological conditions in the bone marrow, where external forces such as fluid shear stress, apart from the physical characteristics of the ECM, influence stem cell response. Here, we hypothesize that substrate topography and fluid shear stress alter the cellular contractile forces, influence the genetic expression of the stem cells and hence alter their lineage. When fluid shear stress was applied, human MSCs with higher contractility (seeded on 1 μm wells) underwent osteogenesis, whereas those with lower contractility (seeded on 2 μm gratings) remained multipotent. Compared to human MSCs seeded on gratings, those seeded on wells exhibited altered alignment and an increase in the area and number of focal adhesions. When actomyosin contractility was inhibited, human MSCs did not exhibit differentiation, regardless of the topographical feature they were being cultured on. We conclude that the stresses generated by the applied fluid flow impinge on cell contractility to drive the stem cell differentiation via the contractility of the stem cells. PMID:26879739

  17. Performance studies on an axial flow compressor stage

    NASA Astrophysics Data System (ADS)

    Sitaram, N.

    1986-12-01

    A low-speed, medium loaded axial flow compressor stage is studied experimentally and theoretically. The flow compressor facility, composed of an inlet guide vane row, a rotor blade row, and a stator blade row, and the principles of the streamline curvature method (SCM) and the Douglas-Neumann cascade program are described. The radial distribution of the flow properties, the rotor blade static pressure distribution, and the lift coefficient and relative flow angle derived experimentally and theoretically are compared. It is determined that there is good correlation between the experimental flow properties and the SCM data, the Douglas-Neumann cascade program and experimental rotor blade static pressure data, and the experimental and theoretical lift coefficients only in the midspan region. Modifications to the SCM and the Douglas-Neumann cascade program in order to improve their accuracy are discussed.

  18. An experimental study on downward slug flow in inclined pipes

    SciTech Connect

    Roumazeilles, P.M.; Yang, J.; Sarica, C.; Chen, X.T.; Wilson, J.F.; Brill, J.P.

    1996-08-01

    The downward simultaneous flow of gas and liquid is often encountered in hilly terrain pipelines and injection wells. Most of the methods for predicting pressure drop in gas/liquid, two-phase flow in pipes have been developed for either upward vertical or upward inclined pipes. This study experimentally investigated downward cocurrent slug flow in inclined pipes. A new test facility was designed and built to acquire data for the entire range of pipe inclination angles. A series of slug flow experiments was conducted in a 2-in. diameter, 65-ft long clear PVC pipe installed on an inclinable structure. Liquid holdup and pressure drop measurements were obtained for downward inclination angles from 0{degree} to {minus}30{degree} at different flow conditions. Translational velocity and liquid-slug holdup correlations were investigated based on the acquired data for different inclination angles.

  19. Nanowell-Trapped Charged Ligand-Bearing Nanoparticle Surfaces – A Novel Method of Enhancing Flow-Resistant Cell Adhesion

    PubMed Central

    Tran, Phat L.; Gamboa, Jessica R.; McCracken, Katherine E.; Riley, Mark R.

    2014-01-01

    Assuring cell adhesion to an underlying biomaterial surface is vital in implant device design and tissue engineering, particularly under circumstances where cells are subjected to potential detachment from overriding fluid flow. Cell-substrate adhesion is a highly regulated process involving the interplay of mechanical properties, surface topographic features, electrostatic charge, and biochemical mechanisms. At the nanoscale level the physical properties of the underlying substrate are of particular importance in cell adhesion. Conventionally, natural, pro-adhesive, and often thrombogenic, protein biomaterials are frequently utilized to facilitate adhesion. In the present study nanofabrication techniques are utilized to enhance the biological functionality of a synthetic polymer surface, polymethymethacrylate, with respect to cell adhesion. Specifically we examine the effect on cell adhesion of combining: 1. optimized surface texturing, 2. electrostatic charge and 3. cell adhesive ligands, uniquely assembled on the substrata surface, as an ensemble of nanoparticles trapped in nanowells. Our results reveal that the ensemble strategy leads to enhanced, more than simply additive, endothelial cell adhesion under both static and flow conditions. This strategy may be of particular utility for enhancing flow-resistant endothelialization of blood-contacting surfaces of cardiovascular devices subjected to flow-mediated shear. PMID:23225491

  20. Ground Based Studies of Thermocapillary Flows in Levitated Drops

    NASA Technical Reports Server (NTRS)

    Sadhal, Satwindar Singh; Trinh, Eugene H.

    1996-01-01

    Ground-based experiments together with analytical studies are presently being conducted for levitated drops. Both acoustic and electrostatic techniques are being employed to achieve levitation of drops in a gaseous environment. The scientific effort is principally on the thermal and the fluid phenomena associated with the local heating of levitated drops, both at 1-g and at low-g. In particular, the thermocapillary flow associated with local spot heating is being studied. Fairly stable acoustic levitation of drops has been achieved with some exceptions when random rotational motion of the drop persists. The flow visualization has been carried out by light scattering from smoke particles for the exterior flow and fluorescent tracer particles in the drop. The results indicate a lack of axial symmetry in the internal flow even though the apparatus and the heating are symmetric. The theoretical studies for the past year have included fundamental analyses of acoustically levitated spherical drops. The flow associated with a particle near the velocity antinode is being investigated by the singular perturbation technique. As a first step towards understanding the effect of the particle displacement from the antinode, the flow field about the node has been calculated for the first time. The effect of the acoustic field on the interior of a liquid drop has also been investigated. The results predict that the internal flow field is very weak.

  1. Analytical study of flow phenomena in SSME turnaround duct geometries

    NASA Astrophysics Data System (ADS)

    McLallin, K. L.

    1985-04-01

    The SSME fuel turbopump hot gas manifold was identified as a source of loss and flow distortion which significantly affects the performance and durability of both the drive turbine and the LOX injector area of the main combustion chamber. Two current SSME geometries were studied, the full power level (FPL) and the first manned orbital flight (FMOF) configuration. The effects of turnaround duct geometry on flow losses and distortions, by varying wall curvature and flow area variation in the 180 deg turnaround region were examined. The effects of the duct inlet flow phenomena such as the radial distortion of the inlet flow and inlet swirl level on turnaround duct performance were also investigated. It is shown that of the two current geometries, the FMOF configuration had lower pressure losses and generated less flow distortion, but had a small flow separation bubble at the 180 deg turnaround exit. It is found that by optimizing wall curvature and flow diffusion in the turnaround, improved duct performance can be achieved.

  2. Theoretical study on the constricted flow phenomena in arteries

    NASA Astrophysics Data System (ADS)

    Sen, S.; Chakravarty, S.

    2012-12-01

    The present study is dealt with the constricted flow characteristics of blood in arteries by making use of an appropriate mathematical model. The constricted artery experiences the generated wall shear stress due to flow disturbances in the presence of constriction. The disturbed flow in the stenosed arterial segment causes malfunction of the cardiovascular system leading to serious health problems in the form of heart attack and stroke. The flowing blood contained in the stenosed artery is considered to be non-Newtonian while the flow is treated to be two-dimensional. The present pursuit also accounts for the motion of the arterial wall and its effect on local fluid mechanics. The flow analysis applies the time-dependent, two-dimensional incompressible nonlinear Navier-Stokes equations for non-Newtonian fluid representing blood. An extensive quantitative analysis presented at the end of the paper based on large scale numerical computations of the quantities of major physiological significance enables one to estimate the constricted flow characteristics in the arterial system under consideration which deviates significantly from that of normal physiological flow conditions.

  3. Study on an undershot cross-flow water turbine

    NASA Astrophysics Data System (ADS)

    Nishi, Yasuyuki; Inagaki, Terumi; Li, Yanrong; Omiya, Ryota; Fukutomi, Junichiro

    2014-06-01

    This study aims to develop a water turbine suitable for ultra-low heads in open channels, with the end goal being the effective utilization of unutilized hydroelectric energy in agricultural water channels. We performed tests by applying a cross-flow runner to an open channel as an undershot water turbine while attempting to simplify the structure and eliminate the casing. We experimentally investigated the flow fields and performance of water turbines in states where the flow rate was constant for the undershot cross-flow water turbine mentioned above. In addition, we compared existing undershot water turbines with our undershot cross-flow water turbine after attaching a bottom plate to the runner. From the results, we were able to clarify the following. Although the effective head for cross-flow runners with no bottom plate was lower than those found in existing runners equipped with a bottom plate, the power output is greater in the high rotational speed range because of the high turbine efficiency. Also, the runner with no bottom plate differed from runners that had a bottom plate in that no water was being wound up by the blades or retained between the blades, and the former received twice the flow due to the flow-through effect. As a result, the turbine efficiency was greater for runners with no bottom plate in the full rotational speed range compared with that found in runners that had a bottom plate.

  4. Redox flow cell development and demonstration project, calendar year 1976

    NASA Technical Reports Server (NTRS)

    1977-01-01

    The major focus of the effort was the key technology issues that directly influence the fundamental feasibility of the overall redox concept. These issues were the development of a suitable semipermeable separator membrane for the system, the screening and study of candidate redox couples to achieve optimum cell performance, and the carrying out of systems analysis and modeling to develop system performance goals and cost estimates.

  5. Microscopic photometric quantification of stiffness and relaxation time of red blood cells in a flow chamber.

    PubMed

    Artmann, G M

    1995-01-01

    The Microscopic Photometric Monolayer Technique provides a tool to measure red blood cell (RBC) stiffness (resistance to elongation) and relaxation time. It combines many of the advantages of flow channel studies of point-attached RBCs with the simplicity, sensitivity and accuracy of photometric light transmission measurement. This technique allows the study of the effects of physicochemical factors on the elongation and relaxation time of the same cells within an average of four to five thousand cells adhered as a monolayer to glass. Further, the time course of physicochemical effects on cell membrane and wash-in/wash-out kinetics of interactions can be followed. An automated version of this technique was developed. A dense monolayer of point-attached RBCs was prepared at the bottom of a flow-chamber. A steady-state flow, with stepwise increases of flow rate, induced the RBC elongation. The light transmission perpendicular through the monolayer plane was measured photometrically. Photomicrographs compared with photometric results showed that the flow-induced bending and curvature change of RBC membrane was associated with the increase of light transmission. There was a linear correlation between the photometric index of elongation and the elongation taken from photomicrographs for shear stresses up to 0.75 Pa. A stiffness parameter, S (in Pa), was defined as the ratio of shear stress and elongation at a shear stress of 0.25 Pa. Following a sudden flow stoppage, the RBCs returned to their resting shape and the RBC relaxation time was measured. The stiffness-relaxation time product, V (in mPas), was calculated to provide an estimate of viscosity. Diamide treatment, known to stiffen RBCs, did result in dose-dependent decreases of elongation and relaxation time. With increasing temperature, the relaxation time decreased at a rate of -2.96 ms/K; the stiffness increased significantly at a rate of 0.0038 Pa/K, and the stiffness-relaxation time product decreased with -2

  6. Impacts of channel wettability and flow direction on liquid water transport in the serpentine flow field of a polymer electrolyte fuel cell

    NASA Astrophysics Data System (ADS)

    Nishida, Kosuke; Taniguchi, Ryo; Ishizaki, Yugo; Tsushima, Shohji; Hirai, Shuichiro

    2015-02-01

    Water plugging in cathode flow field of polymer electrolyte fuel cells (PEFCs) is one of the critical issues to be solved for high power density and high durability. To alleviate performance degradation due to water plugging, it is necessary to control liquid water transport inside gas channel and promote water removal. In this study, the in-situ visualization of liquid water behavior in the serpentine flow field of a specialized transparent PEFC was performed using a high-speed camera. Furthermore, the cell voltage and the differential pressure between the cathode inlet and outlet were simultaneously measured, and the effects of channel wettability and flow direction on the water transport and cell performance were investigated under operating conditions. It was found that the horizontal arrangement of the serpentine channel enhances liquid water removal from the cell without being disturbed by gravity. Especially, highly hydrophilic coating of the horizontally oriented serpentine channel effectively suppresses water plugging and reduces the differential pressure inside the cathode flow field.

  7. In vivo cell characteristic extraction and identification by photoacoustic flow cytography.

    PubMed

    He, Guo; Xu, Dong; Qin, Huan; Yang, Sihua; Xing, Da

    2015-10-01

    We present a photoacoustic flow cytography with fast cross-sectional (B-scan) imaging to precisely identify specific cells in vivo. The B-scan imaging speed of the system is up to 200 frame/s with a lateral resolution of 1.5 μm, which allows to dynamically image the flowing cells within the microvascular. The shape, size and photoacoustic intensity of the target cells are extracted from streaming images and integrated into a standard pattern to distinguish cell types. Circulating red blood cells and melanoma cells in blood vessels are simultaneously identified on melanoma-bearing mouse model. The results demonstrate that in vivo photoacoustic flow cytography can provide cells characteristics analysis and cell type's visual identification, which will be applied for noninvasively monitoring circulating tumor cells (CTCs) and analyzing hematologic diseases.

  8. Multiscale modeling of blood flow: from single cells to blood rheology.

    PubMed

    Fedosov, Dmitry A; Noguchi, Hiroshi; Gompper, Gerhard

    2014-04-01

    Mesoscale simulations of blood flow, where the red blood cells are described as deformable closed shells with a membrane characterized by bending rigidity and stretching elasticity, have made much progress in recent years to predict the flow behavior of blood cells and other components in various flows. To numerically investigate blood flow and blood-related processes in complex geometries, a highly efficient simulation technique for the plasma and solutes is essential. In this review, we focus on the behavior of single and several cells in shear and microcapillary flows, the shear-thinning behavior of blood and its relation to the blood cell structure and interactions, margination of white blood cells and platelets, and modeling hematologic diseases and disorders. Comparisons of the simulation predictions with existing experimental results are made whenever possible, and generally very satisfactory agreement is obtained.

  9. Open-access and multi-directional electroosmotic flow chip for positioning heterotypic cells.

    PubMed

    Terao, Kyohei; Kitazawa, Yuko; Yokokawa, Ryuji; Okonogi, Atsuhito; Kotera, Hidetoshi

    2011-04-21

    We propose a novel method of cell positioning using electroosmotic flow (EOF) to analyze cell-cell interactions. The EOF chip has an open-to-air configuration, is equipped with four electrodes to induce multi-directional EOF, and allows access of tools for liquid handling and of physical probes for cell measurements. Evaluation of the flow within this chip indicated that it controlled hydrodynamic transport of cells, in terms of both speed and direction. We also evaluated cell viability after EOF application and determined appropriate conditions for cell positioning. Two cells were successively positioned in pocket-like microstructures, one in each micropocket, by controlling the EOF direction. As an experimental demonstration, we observed contact interactions between two individual cells through gap junction channels. The EOF chip should provide ways to elucidate various cell-cell interactions between heterotypic cells. PMID:21350747

  10. Nuclear mgnetic resonance study of granular flows. Final report

    SciTech Connect

    1995-03-01

    We have demonstrated the efficacy of NMRI to granular flow studies of objects that yield NMR signals (seeds, pharmaceutical pills) in confined spaces (rotating cylinders, shaking boxes). Excellent results for velocity and concentration of flowing and colliding particles have been obtained non-invasively. We studied flows in geometries that are otherwise impossible to study because of the optical opacity of the materials. In addition, we obtained data for diffusion and collisional losses which must be related to granular temperature. We also measured the global collisional and frictional energy loss for a rotating cylinder containing granular material and compared it to that obtained from the NMRI velocity and acceleration data. We used the same NMR technique to study the interplay between radial and axial segregation of heterogeneous particles in a partially filled rotating cylinder. We also measured flow and diffusion of vibrating particles in the NMRI apparatus to ascertain that such experiments are, indeed, possible. Some theoretical studies were initiated to explain some of the velocity profiles and free surface shapes of rotating drum flow. A direct numerical simulation of the drum flow successfully predicted a number of features, including velocity, concentration, and dissipation profiles in rotating cylinders.

  11. Flow Liner Slot Edge Replication Feasibility Study

    NASA Technical Reports Server (NTRS)

    Newman, John A.; Willard, Scott A.; Smith, Stephen W.; Piascik, Robert S.

    2006-01-01

    Surface replication has been proposed as a method for crack detection in space shuttle main engine flowliner slots. The results of a feasibility study show that examination of surface replicas with a scanning electron microscope can result in the detection of cracks as small as 0.005 inch, and surface flaws as small as 0.001 inch, for the flowliner material.

  12. Simulation and study of stratified flows around finite bodies

    NASA Astrophysics Data System (ADS)

    Gushchin, V. A.; Matyushin, P. V.

    2016-06-01

    The flows past a sphere and a square cylinder of diameter d moving horizontally at the velocity U in a linearly density-stratified viscous incompressible fluid are studied. The flows are described by the Navier-Stokes equations in the Boussinesq approximation. Variations in the spatial vortex structure of the flows are analyzed in detail in a wide range of dimensionless parameters (such as the Reynolds number Re = Ud/ ν and the internal Froude number Fr = U/( Nd), where ν is the kinematic viscosity and N is the buoyancy frequency) by applying mathematical simulation (on supercomputers of Joint Supercomputer Center of the Russian Academy of Sciences) and three-dimensional flow visualization. At 0.005 < Fr < 100, the classification of flow regimes for the sphere (for 1 < Re < 500) and for the cylinder (for 1 < Re < 200) is improved. At Fr = 0 (i.e., at U = 0), the problem of diffusion-induced flow past a sphere leading to the formation of horizontal density layers near the sphere's upper and lower poles is considered. At Fr = 0.1 and Re = 50, the formation of a steady flow past a square cylinder with wavy hanging density layers in the wake is studied in detail.

  13. Study of two-phase flows in reduced gravity

    NASA Astrophysics Data System (ADS)

    Roy, Tirthankar

    Study of gas-liquid two-phase flows under reduced gravity conditions is extremely important. One of the major applications of gas-liquid two-phase flows under reduced gravity conditions is in the design of active thermal control systems for future space applications. Previous space crafts were characterized by low heat generation within the spacecraft which needed to be redistributed within the craft or rejected to space. This task could easily have been accomplished by pumped single-phase loops or passive systems such as heat pipes and so on. However with increase in heat generation within the space craft as predicted for future missions, pumped boiling two-phase flows are being considered. This is because of higher heat transfer co-efficients associated with boiling heat transfer among other advantages. Two-phase flows under reduced gravity conditions also find important applications in space propulsion as in space nuclear power reactors as well as in many other life support systems of space crafts. Two-fluid model along with Interfacial Area Transport Equation (IATE) is a useful tool available to predict the behavior of gas-liquid two-phase flows under reduced gravity conditions. It should be noted that considerable differences exist between two-phase flows under reduced and normal gravity conditions especially for low inertia flows. This is because due to suppression of the gravity field the gas-liquid two-phase flows take a considerable time to develop under reduced gravity conditions as compared to normal gravity conditions. Hence other common methods of analysis applicable for fully developed gas-liquid two-phase flows under normal gravity conditions, like flow regimes and flow regime transition criteria, will not be applicable to gas-liquid two-phase flows under reduced gravity conditions. However the two-fluid model and the IATE need to be evaluated first against detailed experimental data obtained under reduced gravity conditions. Although lot of studies

  14. Elucidation of flow-mediated tumour cell-induced platelet aggregation using an ultrasound standing wave trap

    PubMed Central

    Bazou, D; Santos-Martinez, MJ; Medina, C; Radomski, MW

    2011-01-01

    BACKGROUND AND PURPOSE Tumour cells activate and aggregate platelets [tumour cell-induced platelet aggregation (TCIPA)] and this process plays an important role in the successful metastasis of cancer cells. To date, most studies on TCIPA have been conducted under no-flow conditions. In this study, we have investigated TCIPA in real time under flow conditions, using an ultrasound standing wave trap that allows formation and levitation of cancer cell clusters in suspension, thus mimicking the conditions generated by flowing blood. EXPERIMENTAL APPROACH Using 59M adenocarcinoma and HT1080 fibrosarcoma cells and human platelets, cancer cell cluster–platelet aggregates were imaged in real time using epi-fluorescence microscopy (F-actin) and investigated in detail using confocal microscopy (matrix metalloproteinase-2-GPIIb/IIIa co-localization) and scanning electron and helium-ion microscopy (<1 nm resolution). The release of gelatinases from aggregates was studied using zymography. KEY RESULTS We found that platelet activation and aggregation takes place on the surface of cancer cells (TCIPA), leading to time-dependent disruption of cancer cell clusters. Pharmacological modulation of TCIPA revealed that EDTA, prostacyclin, o-phenanthroline and apyrase significantly down-regulated TCIPA and, in turn, delayed cell cluster disruption, However, EGTA and aspirin were ineffective. Pharmacological inhibition of TCIPA correlated with the down-regulation of platelet activation as shown by flow-cytometry assay of platelet P-selectin. CONCLUSION AND IMPLICATIONS Our results show for the first time, that during TCIPA, platelet activation disrupts cancer cell clusters and this can contribute to metastasis. Thus, selective targeting of platelet aggregate–cancer cell clusters may be an important strategy to control metastasis. PMID:21182493

  15. Novel Strategy for Phenotypic Characterization of Human B Lymphocytes from Precursors to Effector Cells by Flow Cytometry

    PubMed Central

    Vettier, Claire; Laurin, David; Pernollet, Martine; Raskovalova, Tatiana; Cesbron, Jean-Yves; Dumestre-Pérard, Chantal; Jacob, Marie-Christine

    2016-01-01

    A precise identification and phenotypic characterization of human B-cell subsets is of crucial importance in both basic research and medicine. In the literature, flow cytometry studies for the phenotypic characterization of B-lymphocytes are mainly focused on the description of a particular cell stage, or of specific cell stages observed in a single type of sample. In the present work, we propose a backbone of 6 antibodies (CD38, CD27, CD10, CD19, CD5 and CD45) and an efficient gating strategy to identify, in a single analysis tube, a large number of B-cell subsets covering the whole B-cell differentiation from precursors to memory and plasma cells. Furthermore, by adding two antibodies in an 8-color combination, our approach allows the analysis of the modulation of any cell surface marker of interest along B-cell differentiation. We thus developed a panel of seven 8-colour antibody combinations to phenotypically characterize B-cell subpopulations in bone marrow, peripheral blood, lymph node and cord blood samples. Beyond qualitative information provided by biparametric representations, we also quantified antigen expression on each of the identified B-cell subsets and we proposed a series of informative curves showing the modulation of seventeen cell surface markers along B-cell differentiation. Our approach by flow cytometry provides an efficient tool to obtain quantitative data on B-cell surface markers expression with a relative easy-to-handle technique that can be applied in routine explorations. PMID:27657694

  16. Orbital transfer vehicle launch operations study. Processing flows. Volume 3

    NASA Technical Reports Server (NTRS)

    1986-01-01

    The Orbit Transfer Vehicle (OTV) processing flow and Resource Identification Sheets (RISs) for the ground based orbit transfer vehicle and for the space based orbit transfer vehicle are the primary source of information for the rest of the Kennedy Space Center (KSC) OTV Launch Operations Study. Work is presented which identifies KSC facility requirements for the OTV Program, simplifies or automates either flow though the application technology, revises test practices and identifies crew sizes or skills used. These flows were used as the primary point of departure from current operations and practices. Analyses results were documented by revising the appropriate RIS page.

  17. A study of vapor-liquid flow in porous media

    SciTech Connect

    Satik, Cengiz; Yortsos, Yanis C.

    1994-01-20

    We study the heat transfer-driven liquid-to-vapor phase change in single-component systems in porous media by using pore network models and flow visualization experiments. Experiments using glass micromodels were conducted. The flow visualization allowed us to define the rules for the numerical pore network model. A numerical pore network model is developed for vapor-liquid displacement where fluid flow, heat transfer and capillarity are included at the pore level. We examine the growth process at two different boundary conditions.

  18. Concentrated Flow through a Riparian Buffer: A Case Study

    NASA Astrophysics Data System (ADS)

    Young, C. B.; Nogues, J. P.; Hutchinson, S. L.

    2004-05-01

    Riparian buffers are often used for in-situ treatment of agricultural runoff. Although the benefits of riparian buffers are well recongized, concentration of flow can restrict the efficiency of contaminant removal. This study evaluates flow concentration at a agricultural site near Manhattan, Kansas. Manual and automated GIS analyses of a high-resolution digital elevation model were used to determine the fraction of runoff contributing to each buffer segment. Subsequent simulation of the system in WEPP (Water Erosion and Prediction Project) demonstrates the extend to which flow concentration affects buffer efficiency.

  19. Concentrated Flow through a Riparian Buffer: A Case Study

    NASA Astrophysics Data System (ADS)

    Young, C. B.; Nogues, J. P.; Hutchinson, S. L.

    2005-05-01

    Riparian buffers are often used for in-situ treatment of agricultural runoff. Although the benefits of riparian buffers are well recongized, concentration of flow can restrict the efficiency of contaminant removal. This study evaluates flow concentration at a agricultural site near Manhattan, Kansas. Manual and automated GIS analyses of a high-resolution digital elevation model were used to determine the fraction of runoff contributing to each buffer segment. Subsequent simulation of the system in WEPP (Water Erosion and Prediction Project) demonstrates the extent to which flow concentration affects buffer efficiency. Recommendations are presented for the design of adaptive-width buffers.

  20. Circulating tumor cell detection using a parallel flow micro-aperture chip system.

    PubMed

    Chang, Chun-Li; Huang, Wanfeng; Jalal, Shadia I; Chan, Bin-Da; Mahmood, Aamer; Shahda, Safi; O'Neil, Bert H; Matei, Daniela E; Savran, Cagri A

    2015-04-01

    We report on-chip isolation and detection of circulating tumor cells (CTCs) from blood samples using a system that integrates a microchip with immunomagnetics, high-throughput fluidics and size-based filtration. CTCs in a sample are targeted via their surface antigens using magnetic beads functionalized with antibodies. The mixture is then run through a fluidic chamber that contains a micro-fabricated chip with arrays of 8 μm diameter apertures. The fluid runs parallel to the microchip while a magnetic field is generated underneath to draw the beads and cells bound to them toward the chip surface for detection of CTCs that are larger than the apertures and clear out free beads and other smaller particles bound to them. The parallel flow configuration allows high volumetric flow rates, which reduces nonspecific binding to the chip surface and enables multiple circulations of the sample fluid through the system in a short period of time. In this study we first present models of the magnetic and fluidic forces in the system using a finite element method. We then verify the simulation results experimentally to determine an optimal flow rate. Next, we characterize the system by detecting cancer cell lines spiked into healthy human blood and show that on average 89% of the spiked MCF-7 breast cancer cells were detected. We finally demonstrate detection of CTCs in 49 out of 50 blood samples obtained from non-small cell lung cancer (NSCLC) patients and pancreatic cancer (PANC) patients. The number of CTCs detected ranges from 2 to 122 per 8 mL s of blood. We also demonstrate a statistically significant difference between the CTC counts of NSCLC patients who have received therapy and those who have not. PMID:25687986

  1. Circulating tumor cell detection using a parallel flow micro-aperture chip system.

    PubMed

    Chang, Chun-Li; Huang, Wanfeng; Jalal, Shadia I; Chan, Bin-Da; Mahmood, Aamer; Shahda, Safi; O'Neil, Bert H; Matei, Daniela E; Savran, Cagri A

    2015-04-01

    We report on-chip isolation and detection of circulating tumor cells (CTCs) from blood samples using a system that integrates a microchip with immunomagnetics, high-throughput fluidics and size-based filtration. CTCs in a sample are targeted via their surface antigens using magnetic beads functionalized with antibodies. The mixture is then run through a fluidic chamber that contains a micro-fabricated chip with arrays of 8 μm diameter apertures. The fluid runs parallel to the microchip while a magnetic field is generated underneath to draw the beads and cells bound to them toward the chip surface for detection of CTCs that are larger than the apertures and clear out free beads and other smaller particles bound to them. The parallel flow configuration allows high volumetric flow rates, which reduces nonspecific binding to the chip surface and enables multiple circulations of the sample fluid through the system in a short period of time. In this study we first present models of the magnetic and fluidic forces in the system using a finite element method. We then verify the simulation results experimentally to determine an optimal flow rate. Next, we characterize the system by detecting cancer cell lines spiked into healthy human blood and show that on average 89% of the spiked MCF-7 breast cancer cells were detected. We finally demonstrate detection of CTCs in 49 out of 50 blood samples obtained from non-small cell lung cancer (NSCLC) patients and pancreatic cancer (PANC) patients. The number of CTCs detected ranges from 2 to 122 per 8 mL s of blood. We also demonstrate a statistically significant difference between the CTC counts of NSCLC patients who have received therapy and those who have not.

  2. Computer simulation of flow through a lattice flow-cell model

    SciTech Connect

    Mazaheri, A.R.; Zerai, B.; Ahmadi, Goodarz; Kadambi, J.R.; Saylor, B.Z.; Oliver, M.; Bromhal, G.S.; Smith, D.H.

    2005-12-01

    For single-phase flow through a network model of a porous medium, we report (1) solutions of the Navier–Stokes equation for the flow, (2) micro-particle imaging velocimetry (PIV) measurements of local flow velocity vectors in the “pores throats” and “pore bodies,” and (3) comparisons of the computed and measured velocity vectors. A “two-dimensional” network of cylindrical pores and parallelepiped connecting throats was constructed and used for the measurements. All pore bodies had the same dimensions, but three-different (square cross-section) pore-throat sizes were randomly distributed throughout the network. An unstructured computational grid for flow through an identical network was developed and used to compute the local pressure gradients and flow vectors for several different (macroscopic) flow rates. Numerical solution results were compared with the experimental data, and good agreement was found. Cross-over from Darcy flow to inertial flow was observed in the computational results, and the permeability and inertia coefficients of the network were estimated. The development of inertial flow was seen as a “two-step” process: (1) recirculation zones appeared in more and more pore bodies as the flow rate was increased, and (2) the strengths of individual recirculation zones increased with flow rate. Because each pore-throat and pore-body dimension is known, in this approach an experimental (and/or computed) local Reynolds number is known for every location in the porous medium at which the velocity has been measured (and/or computed).

  3. Separation of platelets from other blood cells in continuous-flow by dielectrophoresis field-flow-fractionation

    PubMed Central

    Piacentini, Niccolò; Mernier, Guillaume; Tornay, Raphaël; Renaud, Philippe

    2011-01-01

    We present a microfluidic device capable of separating platelets from other blood cells in continuous flow using dielectrophoresis field-flow-fractionation. The use of hydrodynamic focusing in combination with the application of a dielectrophoretic force allows the separation of platelets from red blood cells due to their size difference. The theoretical cell trajectory has been calculated by numerical simulations of the electrical field and flow speed, and is in agreement with the experimental results. The proposed device uses the so-called “liquid electrodes” design and can be used with low applied voltages, as low as 10 Vpp. The obtained separation is very efficient, the device being able to achieve a very high purity of platelets of 98.8% with less than 2% cell loss. Its low-voltage operation makes it particularly suitable for point-of-care applications. It could further be used for the separation of other cell types based on their size difference, as well as in combination with other sorting techniques to separate multiple cell populations from each other. PMID:22662047

  4. Inertia-dependent dynamics of three-dimensional vesicles and red blood cells in shear flow.

    PubMed

    Luo, Zheng Yuan; Wang, Shu Qi; He, Long; Xu, Feng; Bai, Bo Feng

    2013-10-28

    A three-dimensional (3D) simulation study of the effect of inertia on the dynamics of vesicles and red blood cells (RBCs) has not been reported. Here, we developed a 3D model based on the front tracking method to investigate how inertia affects the dynamics of spherical/non-spherical vesicles and biconcave-shaped RBCs with the Reynolds number ranging from 0.1 to 10. The results showed that inertia induced non-spherical vesicles transitioned from tumbling to swinging, which was not observed in previous 2D models. The critical viscosity ratio of inner/outer fluids for the tumbling–swinging transition remarkably increased with an increasing Reynolds number. The deformation of vesicles was greatly enhanced by inertia, and the frequency of tumbling and tank-treading was significantly decreased by inertia. We also found that RBCs can transit from tumbling to steady tank-treading through the swinging regime when the Reynolds number increased from 0.1 to 10. These results indicate that inertia needs to be considered at moderate Reynolds number (Re ~ 1) in the study of blood flow in the human body and the flow of deformable particle suspension in inertial microfluidic devices. The developed 3D model provided new insights into the dynamics of RBCs under shear flow, thus holding great potential to better understand blood flow behaviors under normal/disease conditions.

  5. Influence of airway wall compliance on epithelial cell injury and adhesion during interfacial flows

    PubMed Central

    Higuita-Castro, Natalia; Mihai, Cosmin; Hansford, Derek J.

    2014-01-01

    Interfacial flows during cyclic airway reopening are an important source of ventilator-induced lung injury. However, it is not known how changes in airway wall compliance influence cell injury during airway reopening. We used an in vitro model of airway reopening in a compliant microchannel to investigate how airway wall stiffness influences epithelial cell injury. Epithelial cells were grown on gel substrates with different rigidities, and cellular responses to substrate stiffness were evaluated in terms of metabolic activity, mechanics, morphology, and adhesion. Repeated microbubble propagations were used to simulate cyclic airway reopening, and cell injury and detachment were quantified via live/dead staining. Although cells cultured on softer gels exhibited a reduced elastic modulus, these cells experienced less plasma membrane rupture/necrosis. Cells on rigid gels exhibited a minor, but statistically significant, increase in the power law exponent and also exhibited a significantly larger height-to-length aspect ratio. Previous studies indicate that this change in morphology amplifies interfacial stresses and, therefore, correlates with the increased necrosis observed during airway reopening. Although cells cultured on stiff substrates exhibited more plasma membrane rupture, these cells experienced significantly less detachment and monolayer disruption during airway reopening. Western blotting and immunofluorescence indicate that this protection from detachment and monolayer disruption correlates with increased focal adhesion kinase and phosphorylated paxillin expression. Therefore, changes in cell morphology and focal adhesion structure may govern injury responses during compliant airway reopening. In addition, these results indicate that changes in airway compliance, as occurs during fibrosis or emphysema, may significantly influence cell injury during mechanical ventilation. PMID:25213636

  6. Experimental Studies of Unsteady Flow through Compliant Vessels

    NASA Astrophysics Data System (ADS)

    Sturgeon, Victoria; Tsai, William; Saloner, David; Savas, Omer

    2004-11-01

    Hemodynamic forces are a significant cause of device failure when stent-grafts are used to treat abdominal aortic aneurysms and even have a causative relationship with the formation and rupture of atherosclerosis. A better comprehension of the forces at play in this environment would help further the understanding and treatment of aneurysmal diseases. In this experimental study, we characterize the behavior of physiologically correct pulsatile input flow in an straight compliant vessel as an analog for the hemodynamic behavior in an abdominal aorta. Flow visualization and particle image velocimetry are used to study the flow in simplified geometries simulating segments of human abdominal aorta in various stages of disease progression. The effects of external pressure are examined to shed light on the interactions between pressure differential across the vessel wall, blood flow, and vessel deformation.

  7. Experimental studies of unsteady flow through compliant vessels

    NASA Astrophysics Data System (ADS)

    Sturgeon, Victoria; Saloner, David; Savas, Omer

    2003-11-01

    Hemodynamic forces are a significant cause of device failure when stent-grafts are used to treat abdominal aortic aneurysms and even have a strong causative relationship with the very formation and rupture of atherosclerosis. A better comprehension of the forces at play in this environment is highly desirable in furthering the understanding and treatment of aneurysmal diseases. The purpose of this experimental study is to characterize the behavior of physiologically correct pulsatile input flow in a straight compliant vessel as an analog for the behavior in an abdominal aorta. Flow visualization and particle image velocimetry are used to study the flow in simplified geometries replicating healthy and diseased segments of human abdominal aorta. The effects of external pressure are examined to shed light on the interactions between pressure differential across the vessel wall, blood flow, and vessel deformation.

  8. Micro/macroscopic fluid flow in open cell fibrous structures and porous media

    NASA Astrophysics Data System (ADS)

    Tamayol, Ali

    Fibrous porous materials are involved in a wide range of applications including composite fabrication, filtration, compact heat exchangers, fuel cell technology, and tissue engineering to name a few. Fibrous structures, such as metalfoams, have unique characteristics such as low weight, high porosity, high mechanical strength, and high surface to volume ratio. More importantly, in many applications the fibrous microstructures can be tailored to meet a range of requirements. Therefore, fibrous materials have the potential to be used in emerging sustainable energy conversion applications. The first step for analyzing transport phenomena in porous materials is to determine the micro/macroscopic flow-field inside the medium. In applications where the porous media is confined in a channel, the system performance is tightly related to the flow properties of the porous medium and its interaction with the channel walls, i.e., macroscopic velocity distribution. Therefore, the focus of the study has been on: developing new mechanistic model(s) for determining permeability and inertial coefficient of fibrous porous materials; investigating the effects of microstructural and mechanical parameters such as porosity, fiber orientation, mechanical compression, and fiber distribution on the flow properties and pressure drop of fibrous structures; determining the macroscopic flow-field in confined porous media where the porous structure fills the channel cross-section totally or partially. A systematic approach has been followed to study different aspects of the flow through fibrous materials. The complex microstructure of real materials has been modelled using unit cells that have been assumed to be repeated throughout the media. Implementing various exact and approximate analytical techniques such as integral technique, point matching, blending rules, and scale analysis the flow properties of such media have been modelled; the targeted properties include permeability and inertial

  9. Spaceflight bioreactor studies of cells and tissues.

    PubMed

    Freed, Lisa E; Vunjak-Novakovic, Gordana

    2002-01-01

    Studies of the fundamental role of gravity in the development and function of biological organisms are a central component of the human exploration of space. Microgravity affects numerous physical phenomena relevant to biological research, including the hydrostatic pressure in fluid filled vesicles, sedimentation of organelles, and buoyancy-driven convection of flow and heat. These physical phenomena can in turn directly and indirectly affect cellular morphology, metabolism, locomotion, secretion of extracellular matrix and soluble signals, and assembly into functional tissues. Studies aimed at distinguishing specific effects of gravity on biological systems require the ability to: (i) control and systematically vary gravity, e.g. by utilizing the microgravity environment of space in conjunction with an in-flight centrifuge; and (ii) maintain constant all other factors in the immediate environment, including in particular concentrations and exchange rates of biochemical species and hydrodynamic shear. The latter criteria imply the need for gravity-independent mechanisms to provide for mass transport between the cells and their environment. Available flight hardware has largely determined the experimental design and scientific objectives of spaceflight cell and tissue culture studies carried out to date. Simple culture vessels have yielded important quantitative data, and helped establish in vitro models of cell locomotion, growth and differentiation in various mammalian cell types including embryonic lung cells [6], lymphocytes [2,8], and renal cells [7,31]. Studies done using bacterial cells established the first correlations between gravity-dependent factors such as cell settling velocity and diffusional distance and the respective cell responses [12]. The development of advanced bioreactors for microgravity cell and tissue culture and for tissue engineering has benefited both research areas and provided relevant in vitro model systems for studies of astronaut

  10. CFD study on flow characteristics of pump sump and performance analysis of the mixed flow pump

    NASA Astrophysics Data System (ADS)

    Zhao, Y. X.; Kim, C. G.; Lee, Y. H.

    2013-12-01

    Head-capacity curves provided by the pump manufacturer are obtained on the condition of no vortices flowing into the pump intake. The efficiency and performance of pumping stations depend not only on the performance of the selected pumps but also on the proper design of the intake sumps. A faulty design of pump sump can lead to the occurrence of swirl and vortices, which reduce the pump performance. Therefore, sump model test is necessary in order to check the flow condition around intake structure. Numerical simulation is a good facility for reducing the time and cost involved throughout the design process. In this study, the commercial software ANSYS CFX-13.0 has been used for the CFD analysis of the pump sump. The effect of an anti-vortex device (AVD) for the submerged vortex has been examined. Hydraulic performances for the head rise, shaft power, pump efficiencies versus flow rate are studied by the performance curves. In addition, numerical simulation of cavitation phenomenon in a mixed flow pump has been performed by calculating the full cavitation model with k-ε turbulence model. According to the result, the efficacy of the AVD to ensure the uniform flow conditions around the pump intake is confirmed. From the numerical analysis, the inception of cavitation is observed on the suction surface where the leading edges meet the tip, and then the cavitation zone expands.

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

    PubMed Central

    Jagannadh, Veerendra Kalyan; Adhikari, Jayesh Vasudeva; Gorthi, Sai Siva

    2015-01-01

    In this work, we report a system-level integration of portable microscopy and microfluidics for the realization of optofluidic imaging flow analyzer with a throughput of 450 cells/s. With the use of a cellphone augmented with off-the-shelf optical components and custom designed microfluidics, we demonstrate a portable optofluidic imaging flow analyzer. A multiple microfluidic channel geometry was employed to demonstrate the enhancement of throughput in the context of low frame-rate imaging systems. Using the cell-phone based digital imaging flow analyzer, we have imaged yeast cells present in a suspension. By digitally processing the recorded videos of the flow stream on the cellphone, we demonstrated an automated cell viability assessment of the yeast cell population. In addition, we also demonstrate the suitability of the system for blood cell counting. PMID:26015835

  12. Non-Flow-Through Fuel Cell System Test Results and Demonstration on the SCARAB Rover

    NASA Technical Reports Server (NTRS)

    Scheidegger, Brianne; Burke, Kenneth; Jakupca, Ian

    2012-01-01

    This presentation describes the results of the demonstration of a non-flow-through PEM fuel cell as part of a power system on the SCARAB rover at the NASA Glenn Research Center. A 16-cell non-flow-through fuel cell stack from Infinity Fuel Cell and Hydrogen, Inc. was incorporated into a power system designed to act as a range extender by providing power to the SCARAB rover s hotel loads. The power system, including the non-flow-through fuel cell technology, successfully demonstrated its goal as a range extender by powering hotel loads on the SCARAB rover, making this demonstration the first to use the non-flow-through fuel cell technology on a mobile platform.

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

    PubMed

    Jagannadh, Veerendra Kalyan; Adhikari, Jayesh Vasudeva; Gorthi, Sai Siva

    2015-03-01

    In this work, we report a system-level integration of portable microscopy and microfluidics for the realization of optofluidic imaging flow analyzer with a throughput of 450 cells/s. With the use of a cellphone augmented with off-the-shelf optical components and custom designed microfluidics, we demonstrate a portable optofluidic imaging flow analyzer. A multiple microfluidic channel geometry was employed to demonstrate the enhancement of throughput in the context of low frame-rate imaging systems. Using the cell-phone based digital imaging flow analyzer, we have imaged yeast cells present in a suspension. By digitally processing the recorded videos of the flow stream on the cellphone, we demonstrated an automated cell viability assessment of the yeast cell population. In addition, we also demonstrate the suitability of the system for blood cell counting. PMID:26015835

  14. Performance Characteristics of a PEM Fuel Cell with Parallel Flow Channels at Different Cathode Relative Humidity Levels.

    PubMed

    Lee, Pil Hyong; Hwang, Sang Soon

    2009-01-01

    In fuel cells flow configuration and operating conditions such as cell temperature, humidity at each electrode and stoichiometric number are very crucial for improving performance. Too many flow channels could enhance the performance but result in high parasite loss. Therefore a trade-off between pressure drop and efficiency of a fuel cell should be considered for optimum design. This work focused on numerical simulation of the effects of operating conditions, especially cathode humidity, with simple micro parallel flow channels. It is known that the humidity at the cathode flow channel becomes very important for enhancing the ion conductivity of polymer membrane because fully humidified condition was normally set at anode. To investigate the effect of humidity on the performance of a fuel cell, in this study humidification was set to 100% at the anode flow channel and was changed by 0-100% at the cathode flow channel. Results showed that the maximum power density could be obtained under 60% humidified condition at the cathode where oxygen concentration was moderately high while maintaining high ion conductivity at a membrane.

  15. Monoclonal antibodies to proliferating cell nuclear antigen (PCNA)/cyclin as probes for proliferating cells by immunofluorescence microscopy and flow cytometry.

    PubMed

    Kurki, P; Ogata, K; Tan, E M

    1988-04-22

    Proliferating cell nuclear antigen (PCNA)/cyclin is an intranuclear polypeptide antigen that is found in both normal and transformed proliferating cells. We have recently described two mouse monoclonal antibodies reacting with PCNA. In this report we describe the application of these antibodies to the study of proliferating human cells by indirect immunofluorescence microscopy and by flow cytometry. A fixation/permeation procedure was developed in order to obtain satisfactory binding of monoclonal PCNA-specific antibodies to proliferating cells. This method involved fixation with 1% paraformaldehyde followed by methanol treatment. For the staining of cells in suspension with the IgM type monoclonal antibodies lysolecithin was added to the paraformaldehyde solution to achieve a better permeation by the antibody molecules. This procedure gave a good ratio of specific staining relative to the background staining. It also preserved the shape and normal architecture of the cells as judged by visual microscopic observation and by light scatter measurements using a flow cytometer. Furthermore, this fixation technique permits simultaneous labeling of DNA by propidium iodide and PCNA by monoclonal antibodies. PCNA was detected in various types of normal and transformed proliferating cells by indirect immunofluorescence. Quiescent peripheral blood mononuclear cells were PCNA-negative whereas a fraction of lectin-stimulated lymphocytes became PCN