Phase-sensitive flow cytometer

DOEpatents

Steinkamp, J.A.

1993-12-14

A phase-sensitive flow cytometer (FCM) provides additional FCM capability to use the fluorescence lifetime of one or more fluorochromes bound to single cells to provide additional information regarding the cells. The resulting fluorescence emission can be resolved into individual fluorescence signals if two fluorochromes are present or can be converted directly to a decay lifetime from a single fluorochrome. The excitation light for the fluorochromes is modulated to produce an amplitude modulated fluorescence pulse as the fluorochrome is excited in the FCM. The modulation signal also forms a reference signal that is phase-shifted a selected amount for subsequent mixing with the output modulated fluorescence intensity signal in phase-sensitive detection circuitry. The output from the phase-sensitive circuitry is then an individual resolved fluorochrome signal or a single fluorochrome decay lifetime, depending on the applied phase shifts. 15 figures.

Design, simulation and characterisation of integrated optics for a microfabricated flow cytometer

NASA Astrophysics Data System (ADS)

Barat, David; Benazzi, Giuseppe; Mowlem, Matthew Charles; Ruano, Jesus Miguel; Morgan, Hywel

2010-05-01

Flow cytometry is widely used for analyzing micro-particles such as cells and bacteria. Microfabricated flow cytometers promise reduced instrument size and cost with increased robustness and have application in medicine, life sciences and environmental metrology. Further miniaturisation and robustness can be achieved if integrated optics are used instead of traditional free space optics. We present designs simulation and experimental characterisation of integrated optics for a microfabricated cytometer made from SU-8 resin on a glass substrate. The optics constructed from combinations of optical fibres (positioned with microgrooves), waveguides, and microlenses enable analysis of scattered light and fluorescence from particles positioned near the centre of a microchannel using one dimensional sheath flow. Four different methods for directing the incident light onto the particles are examined and the optimum design discussed.

Cytometer on a Chip

NASA Technical Reports Server (NTRS)

Fernandez, Salvador M.

2011-01-01

A cytometer now under development exploits spatial sorting of sampled cells on a microarray chip followed by use of grating-coupled surface-plasmon-resonance imaging (GCSPRI) to detect the sorted cells. This cytometer on a chip is a prototype of contemplated future miniature cytometers that would be suitable for rapidly identifying pathogens and other cells of interest in both field and laboratory applications and that would be attractive as alternatives to conventional flow cytometers. The basic principle of operation of a conventional flow cytometer requires fluorescent labeling of sampled cells, stringent optical alignment of a laser beam with a narrow orifice, and flow of the cells through the orifice, which is subject to clogging. In contrast, the principle of operation of the present cytometer on a chip does not require fluorescent labeling of cells, stringent optical alignment, or flow through a narrow orifice. The basic principle of operation of the cytometer on a chip also reduces the complexity, mass, and power of the associated laser and detection systems, relative to those needed in conventional flow cytometry. Instead of making cells flow in single file through a narrow flow orifice for sequential interrogation as in conventional flow cytometry, a liquid containing suspended sampled cells is made to flow over the front surface of a microarray chip on which there are many capture spots. Each capture spot is coated with a thin (approximately 50-nm) layer of gold that is, in turn, coated with antibodies that bind to cell-surface molecules characteristic of one the cell species of interest. The multiplicity of capture spots makes it possible to perform rapid, massively parallel analysis of a large cell population. The binding of cells to each capture spot gives rise to a minute change in the index of refraction at the surface of the chip. This change in the index of refraction is what is sensed in GCSPRI, as described briefly below. The identities of the

Cytometer on a Chip

NASA Technical Reports Server (NTRS)

Fernandez, Salvador M.

2011-01-01

A cytometer now under development exploits spatial sorting of sampled cells on a microarray chip followed by use of grating-coupled surface-plasmon-resonance imaging (GCSPRI) to detect the sorted cells. This cytometer on a chip is a prototype of contemplated future miniature cytometers that would be suitable for rapidly identifying pathogens and other cells of interest in both field and laboratory applications and that would be attractive as alternatives to conventional flow cytometers. The basic principle of operation of a conventional flow cytometer requires fluorescent labeling of sampled cells, stringent optical alignment of a laser beam with a narrow orifice, and flow of the cells through the orifice, which is subject to clogging. In contrast, the principle of operation of the present cytometer on a chip does not require fluorescent labeling of cells, stringent optical alignment, or flow through a narrow orifice. The basic principle of operation of the cytometer on a chip also reduces the complexity, mass, and power of the associated laser and detection systems, relative to those needed in conventional flow cytometry. Instead of making cells flow in single file through a narrow flow orifice for sequential interrogation as in conventional flow cytometry, a liquid containing suspended sampled cells is made to flow over the front surface of a microarray chip on which there are many capture spots. Each capture spot is coated with a thin (.50-nm) layer of gold that is, in turn, coated with antibodies that bind to cell-surface molecules characteristic of the cell species of interest. The multiplicity of capture spots makes it possible to perform rapid, massively parallel analysis of a large cell population. The binding of cells to each capture spot gives rise to a minute change in the index of refraction at the surface of the chip. This change in the index of refraction is what is sensed in GCSPRI, as described briefly below. The identities of the various species in

Flow cytometer measurement of binding assays

DOEpatents

Saunders, George C.

1987-01-01

A method of measuring the result of a binding assay that does not require separation of fluorescent smaller particles is disclosed. In a competitive binding assay the smaller fluorescent particles coated with antigen compete with antigen in the sample being analyzed for available binding sites on larger particles. In a sandwich assay, the smaller, fluorescent spheres coated with antibody attach themselves to molecules containing antigen that are attached to larger spheres coated with the same antibody. The separation of unattached, fluorescent smaller particles is made unnecessary by only counting the fluorescent events triggered by the laser of a flow cytometer when the event is caused by a particle with a light scatter measurement within a certain range corresponding to the presence of larger particles.

Flow cytometer jet monitor system

DOEpatents

Van den Engh, Ger

1997-01-01

A direct jet monitor illuminates the jet of a flow cytometer in a monitor wavelength band which is substantially separate from the substance wavelength band. When a laser is used to cause fluorescence of the substance, it may be appropriate to use an infrared source to illuminate the jet and thus optically monitor the conditions within the jet through a CCD camera or the like. This optical monitoring may be provided to some type of controller or feedback system which automatically changes either the horizontal location of the jet, the point at which droplet separation occurs, or some other condition within the jet in order to maintain optimum conditions. The direct jet monitor may be operated simultaneously with the substance property sensing and analysis system so that continuous monitoring may be achieved without interfering with the substance data gathering and may be configured so as to allow the front of the analysis or free fall area to be unobstructed during processing.

Monitoring tumor metastasis by in vivo imaging and flow cytometer

NASA Astrophysics Data System (ADS)

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

2009-08-01

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.

Graphical analysis of flow cytometer data for characterizing controlled fluorescent protein display on λ phage.

PubMed

Sokolenko, Stanislav; Nicastro, Jessica; Slavcev, Roderick; Aucoin, Marc G

2012-12-01

As native virus particles typically cannot be resolved using a flow cytometer, the general practice is to use fluorescent dyes to label the particles. In this work, an attempt was made to use a common commercial flow cytometer to characterize a phage display strategy that allows for controlled levels of protein display, in this case, eGFP. To achieve this characterization, a number of data processing steps were needed to ensure that the observed phenomena were indeed capturing differences in the phages produced. Phage display of eGFP resulted in altered side scatter and fluorescence profile, and sub-populations could be identified within what would otherwise be considered uniform populations. Surprisingly, this study has found that side scatter may be used in the future to characterize the display of nonfluorescent proteins. Copyright © 2012 International Society for Advancement of Cytometry.

Sample introduction apparatus for a flow cytometer

DOEpatents

Van den Engh, Ger

1998-01-01

A sample introduction system for a flow cytometer allows easy change of sample containers such as test tubes and facilitates use in high pressure environments. The sample container includes a cap having a pressure supply chamber and a sample container attachment cavity. A sample container may be automatically positioned into the attachment cavity so as to sealably engage the end of the sample container as its outer surface. This positioning may be accomplished through some sample introduction mechanism. To facilitate cleaning HPLC tubing and fittings may be used in a manner which facilitates removable of the entire tubing from both the nozzle container and other sample container cap to permit its replacement to avoid contamination. The sample container support may include horizontal stops which loosely limit the movement of the sample container and thus avoid further stresses upon it.

Sample introduction system for a flow cytometer

DOEpatents

Van den Engh, Ger

1997-01-01

A sample introduction system for a flow cytometer allows easy change of sample containers such as test tubes and facilitates use in high pressure environments. The sample container includes a cap having a pressure supply chamber and a sample container attachment cavity. A sample container may be automatically positioned into the attachment cavity so as to sealably engage the end of the sample container as its outer surface. This positioning may be accomplished through some sample introduction mechanism. To facilitate cleaning, HPLC tubing and fittings may be used in a manner which facilitates removing of the entire tubing from both the nozzle container and other sample container cap to permit its replacement to avoid contamination. The sample container support may include horizontal stops which loosely limit the movement of the sample container and thus avoid further stresses upon it.

Sample introduction apparatus for a flow cytometer

DOEpatents

Van den Engh, G.

1998-03-10

A sample introduction system for a flow cytometer allows easy change of sample containers such as test tubes and facilitates use in high pressure environments. The sample container includes a cap having a pressure supply chamber and a sample container attachment cavity. A sample container may be automatically positioned into the attachment cavity so as to sealably engage the end of the sample container as its outer surface. This positioning may be accomplished through some sample introduction mechanism. To facilitate cleaning HPLC tubing and fittings may be used in a manner which facilitates removable of the entire tubing from both the nozzle container and other sample container cap to permit its replacement to avoid contamination. The sample container support may include horizontal stops which loosely limit the movement of the sample container and thus avoid further stresses upon it. 3 figs.

Sample introduction system for a flow cytometer

DOEpatents

Engh, G. van den

1997-02-11

A sample introduction system for a flow cytometer allows easy change of sample containers such as test tubes and facilitates use in high pressure environments. The sample container includes a cap having a pressure supply chamber and a sample container attachment cavity. A sample container may be automatically positioned into the attachment cavity so as to sealably engage the end of the sample container as its outer surface. This positioning may be accomplished through some sample introduction mechanism. To facilitate cleaning, HPLC tubing and fittings may be used in a manner which facilitates removing of the entire tubing from both the nozzle container and other sample container cap to permit its replacement to avoid contamination. The sample container support may include horizontal stops which loosely limit the movement of the sample container and thus avoid further stresses upon it. 3 figs.

Studying liver cancer metastasis by in vivo imaging and flow cytometer

NASA Astrophysics Data System (ADS)

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

2009-11-01

Primary liver cancer (hepatocellular carcinoma, or HCC) is associated with liver cirrhosis 60-80% of the time. Liver cancer is one of the most common malignancies in the world, with approximately 1,000,000 cases reported every year. 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. 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 with 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.

UF6 Density and Mass Flow Measurements for Enrichment Plants using Acoustic Techniques

DOE Office of Scientific and Technical Information (OSTI.GOV)

Good, Morris S.; Smith, Leon E.; Warren, Glen A.

A key enabling capability for enrichment plant safeguards being considered by the International Atomic Energy Agency (IAEA) is high-accuracy, noninvasive, unattended measurement of UF6 gas density and mass flow rate. Acoustic techniques are currently used to noninvasively monitor gas flow in industrial applications; however, the operating pressures at gaseous centrifuge enrichment plants (GCEPs) are roughly two orders magnitude below the capabilities of commercial instrumentation. Pacific Northwest National Laboratory is refining acoustic techniques for estimating density and mass flow rate of UF6 gas in scenarios typical of GCEPs, with the goal of achieving 1% measurement accuracy. Proof-of-concept laboratory measurements using amore » surrogate gas for UF6 have demonstrated signatures sensitive to gas density at low operating pressures such as 10–50 Torr, which were observed over the background acoustic interference. Current efforts involve developing a test bed for conducting acoustic measurements on flowing SF6 gas at representative flow rates and pressures to ascertain the viability of conducting gas flow measurements under these conditions. Density and flow measurements will be conducted to support the evaluation. If successful, the approach could enable an unattended, noninvasive approach to measure mass flow in unit header pipes of GCEPs.« less

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.

A critical evaluation of a flow cytometer used for detecting enterococci in recreational waters.

PubMed

King, Dawn N; Brenner, Kristen P; Rodgers, Mark R

2007-06-01

The current U. S. Environmental Protection Agency-approved method for enterococci (Method 1600) in recreational water is a membrane filter (MF) method that takes 24 hours to obtain results. If the recreational water is not in compliance with the standard, the risk of exposure to enteric pathogens may occur before the water is identified as hazardous. Because flow cytometry combined with specific fluorescent antibodies has the potential to be used as a rapid detection method for microorganisms, this technology was evaluated as a rapid, same-day method to detect enterococci in bathing beach waters. The flow cytometer chosen for this study was a laser microbial detection system designed to detect labeled antibodies. A comparison of MF counts with flow cytometry counts of enterococci in phosphate buffer and sterile-filtered recreational water showed good agreement between the two methods. However, when flow cytometry was used, the counts were several orders of magnitude higher than the MF counts with no correlation to Enterococcus spike concentrations. The unspiked sample controls frequently had higher counts than the samples spiked with enterococci. Particles within the spiked water samples were probably counted as target cells by the flow cytometer because of autofluorescence or non-specific adsorption of antibody and carryover to subsequent samples. For these reasons, this technology may not be suitable for enterococci detection in recreational waters. Improvements in research and instrument design that will eliminate high background and carryover may make this a viable technology in the

Soluble Protein Analysis using a Compact Bench-top Flow Cytometer

NASA Technical Reports Server (NTRS)

Pappas, Dimitri; Kao, Shib-Hsin; Cyr, Johnathan

2004-01-01

Future space exploration missions will require analytical technology capable of providing both autonomous medical care to the crew and investigative capabilities to researchers. While several promising candidate technologies exist for further development, flow cytometry is an attractive technology as it offers both crew health (blood cell count, leukocyte differential, etc.) and a wide array of biochemistry and immunology assays. research settings, the application of this technique to soluble protein analysis is also possible. Proteomic beads using fluorescent dyes for optical encoding were used to monitor six cytokines simultaneously in cell medium of cell cultures in stationary and rotating cell culture systems. The results of this work demonstrate that a compact flow cytometer, such as a system proposed for space flight, can detect a variety of soluble proteins for crew health and biotechnology experiments during long-term missions.

Design and first results of CytoBuoy: a wireless flow cytometer for in situ analysis of marine and fresh waters.

PubMed

Dubelaar, G B; Gerritzen, P L; Beeker, A E; Jonker, R R; Tangen, K

1999-12-01

The high costs of microscopical determination and counting of phytoplankton often limit sampling frequencies below an acceptable level for the monitoring of dynamic ecosystems. Although having a limited discrimination power, flow cytometry allows the analysis of large numbers of samples to a level that is sufficient for many basic monitoring jobs. For this purpose, flow cytometers should not be restricted to research laboratories. We report here on the development of an in situ flow cytometer for autonomous operation inside a small moored buoy or on other platforms. Operational specifications served a wide range of applications in the aquatic field. Specific conditions had to be met with respect to the operation platform and autonomy. A small, battery-operated flow cytometer resulted, requiring no external sheath fluid supply. Because it was designed to operate in a buoy, we call it CytoBuoy. Sampling, analysis, and radio transmission of the data proceed automatically at user-defined intervals. A powerful feature is the acquisition and radio transmission of full detector pulse shapes of each particle. This provides valuable morphological information for particles larger than the 5-microm laser focus. CytoBuoy allows on-line in situ particle analysis, estimation of phytoplankton biomass, and discrimination between different phytoplankton groups. This will increase the applicability of flow cytometry in the field of environmental monitoring. Copyright 1999 Wiley-Liss, Inc.

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.

Analysis of particle in liquid using excitation-fluorescence spectral flow cytometer

NASA Astrophysics Data System (ADS)

Takenaka, Kei; Togashi, Shigenori

2018-01-01

We have developed a new flow cytometer that can measure the excitation-fluorescence spectra of a single particle. This system consists of a solution-transmitting unit and an optical unit. The solution-transmitting unit allows a sample containing particles to flow through the center of a flow cell by hydrodynamic focusing. The optical unit irradiates particles with dispersed white light (wavelength band: 400-650 nm) along the flow direction and measures their fluorescence spectra (wavelength band: 400-700 nm) using a spectroscopic photodetector array. The fluorescence spectrum of a particle changes with the shift of the wavelength of the excitation light. Using this system, the excitation-fluorescence spectra of a fluorescent particle were measured. Additionally, a homogenized tomato suspension and a homogenized spinach suspension were measured using the system. Measurement results show that it is possible to determine the components of vegetables by comparing measured fluorescence spectra of particles in a vegetable suspension.

Streak Imaging Flow Cytometer for Rare Cell Analysis.

PubMed

Balsam, Joshua; Bruck, Hugh Alan; Ossandon, Miguel; Prickril, Ben; Rasooly, Avraham

2017-01-01

There is a need for simple and affordable techniques for cytology for clinical applications, especially for point-of-care (POC) medical diagnostics in resource-poor settings. However, this often requires adapting expensive and complex laboratory-based techniques that often require significant power and are too massive to transport easily. One such technique is flow cytometry, which has great potential for modification due to the simplicity of the principle of optical tracking of cells. However, it is limited in that regard due to the flow focusing technique used to isolate cells for optical detection. This technique inherently reduces the flow rate and is therefore unsuitable for rapid detection of rare cells which require large volume for analysis.To address these limitations, we developed a low-cost, mobile flow cytometer based on streak imaging. In our new configuration we utilize a simple webcam for optical detection over a large area associated with a wide-field flow cell. The new flow cell is capable of larger volume and higher throughput fluorescence detection of rare cells than the flow cells with hydrodynamic focusing used in conventional flow cytometry. The webcam is an inexpensive, commercially available system, and for fluorescence analysis we use a 1 W 450 nm blue laser to excite Syto-9 stained cells with emission at 535 nm. We were able to detect low concentrations of stained cells at high flow rates of 10 mL/min, which is suitable for rapidly analyzing larger specimen volumes to detect rare cells at appropriate concentration levels. The new rapid detection capabilities, combined with the simplicity and low cost of this device, suggest a potential for clinical POC flow cytometry in resource-poor settings associated with global health.

A novel all-fiber optic flow cytometer technology for Point-of Care and Remote Environments

NASA Astrophysics Data System (ADS)

Mermut, Ozzy

Traditional flow cytometry designs tend to be bulky systems with a complex optical-fluidic sub-system and often require trained personnel for operation. This makes them difficult to readily translate to remote site testing applications. A new compact and portable fiber-optic flow cell (FOFC) technology has been developed at INO. We designed and engineered a specialty optical fiber through which a square hole is transversally bored by laser micromachining. A capillary is fitted into that hole to flow analyte within the fiber square cross-section for detection and counting. With demonstrated performance benchmarks potentially comparable to commercial flow cytometers, our FOFC provides several advantages compared to classic free-space con-figurations, e.g., sheathless flow, low cost, reduced number of optical components, no need for alignment (occurring in the fabrication process only), ease-of-use, miniaturization, portability, and robustness. This sheathless configuration, based on a fiber optic flow module, renders this cytometer amenable to space-grade microgravity environments. We present our recent results for an all-fiber approach to achieve a miniature FOFC to translate flow cytometry from bench to a portable, point-of-care device for deployment in remote settings. Our unique fiber approach provides the capability to illuminate a large surface with a uniform intensity distri-bution, independently of the initial shape originating from the light source, and without loss of optical power. The CVs and sensitivities are measured and compared to industry benchmarks. Finally, integration of LEDs enable several advantages in cost, compactness, and wavelength availability.

Evaluation and purchase of an analytical flow cytometer: some of the numerous factors to consider.

PubMed

Zucker, Robert M; Fisher, Nancy C

2013-01-01

When purchasing a flow cytometer, the decision of which brand, model, specifications, and accessories may be challenging. The decisions should initially be guided by the specific applications intended for the instrument. However, many other factors need to be considered, which include hardware, software, quality assurance, support, service, and price and recommendations from colleagues. These issues are discussed to help guide the purchasing process.

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.

Studying leukemia metastasis and therapy monitoring by in vivo imaging and flow cytometer

NASA Astrophysics Data System (ADS)

Wei, Xunbin; Li, Yan; Tan, Yuan; Zhang, Li; Chen, Yun; Liu, Guangda; Chen, Tong; Gu, Zhenqin; Wang, Guiying; Zhou, Zenghui; Wang, Li; Wang, Chen

2007-11-01

Cytotoxic chemotherapy agents are the foundation of current leukemia therapy. 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 leukemic cell growth and metastasis. A large body of elegant work in the field of immunology has demonstrated the mechanisms whereby leukocytes traffic to specific sites within the body. Vascular cell adhesion molecules and chemicalattractants combine to direct white blood cells to appropriate environments. Although it has been hypothesized that leukemic white blood cells home to hematopoietic organs using mechanisms similar to those of their benign leukocyte counterparts, detailed study of leukemic cell transit through bone marrow has yet to be undertaken. We develop the "in vivo microscopy" to study the mechanisms that govern leukemic cell spread through the bone marrow microenvironment in vivo in real-time confocal infrared fluorescence imaging. A recently developed "in vivo flow cytometer" and optical imaging are used to assess leukemic cell spreading and the circulation kinetics of leukemic cells. A real- time quantitative monitoring of circulating leukemic cells by the in vivo flow cytometer will be useful to assess the effectiveness of the potential therapeutic interventions.

High-speed cell recognition algorithm for ultrafast flow cytometer imaging system

NASA Astrophysics Data System (ADS)

Zhao, Wanyue; Wang, Chao; Chen, Hongwei; Chen, Minghua; Yang, Sigang

2018-04-01

An optical time-stretch flow imaging system enables high-throughput examination of cells/particles with unprecedented high speed and resolution. A significant amount of raw image data is produced. A high-speed cell recognition algorithm is, therefore, highly demanded to analyze large amounts of data efficiently. A high-speed cell recognition algorithm consisting of two-stage cascaded detection and Gaussian mixture model (GMM) classification is proposed. The first stage of detection extracts cell regions. The second stage integrates distance transform and the watershed algorithm to separate clustered cells. Finally, the cells detected are classified by GMM. We compared the performance of our algorithm with support vector machine. Results show that our algorithm increases the running speed by over 150% without sacrificing the recognition accuracy. This algorithm provides a promising solution for high-throughput and automated cell imaging and classification in the ultrafast flow cytometer imaging platform.

High-speed cell recognition algorithm for ultrafast flow cytometer imaging system.

PubMed

Zhao, Wanyue; Wang, Chao; Chen, Hongwei; Chen, Minghua; Yang, Sigang

2018-04-01

An optical time-stretch flow imaging system enables high-throughput examination of cells/particles with unprecedented high speed and resolution. A significant amount of raw image data is produced. A high-speed cell recognition algorithm is, therefore, highly demanded to analyze large amounts of data efficiently. A high-speed cell recognition algorithm consisting of two-stage cascaded detection and Gaussian mixture model (GMM) classification is proposed. The first stage of detection extracts cell regions. The second stage integrates distance transform and the watershed algorithm to separate clustered cells. Finally, the cells detected are classified by GMM. We compared the performance of our algorithm with support vector machine. Results show that our algorithm increases the running speed by over 150% without sacrificing the recognition accuracy. This algorithm provides a promising solution for high-throughput and automated cell imaging and classification in the ultrafast flow cytometer imaging platform. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Reconfigurable acquisition system with integrated optics for a portable flow cytometer.

PubMed

Kirleis, Matthew A; Mathews, Scott A; Verbarg, Jasenka; Erickson, Jeffrey S; Piqué, Alberto

2013-11-01

Portable and inexpensive scientific instruments that are capable of performing point of care diagnostics are needed for applications such as disease detection and diagnosis in resource-poor settings, for water quality and food supply monitoring, and for biosurveillance activities in autonomous vehicles. In this paper, we describe the development of a compact flow cytometer built from three separate, customizable, and interchangeable modules. The instrument as configured in this work is being developed specifically for the detection of selected Centers for Disease Control (CDC) category B biothreat agents through a bead-based assay: E. coli O157:H7, Salmonella, Listeria, and Shigella. It has two-color excitation, three-color fluorescence and light scattering detection, embedded electronics, and capillary based flow. However, these attributes can be easily modified for other applications such as cluster of differentiation 4 (CD4) counting. Proof of concept is demonstrated through a 6-plex bead assay with the results compared to a commercially available benchtop-sized instrument.

Reconfigurable acquisition system with integrated optics for a portable flow cytometer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kirleis, Matthew A., E-mail: matthew.kirleis@nrl.navy.mil; Mathews, Scott A.; Verbarg, Jasenka

2013-11-15

Portable and inexpensive scientific instruments that are capable of performing point of care diagnostics are needed for applications such as disease detection and diagnosis in resource-poor settings, for water quality and food supply monitoring, and for biosurveillance activities in autonomous vehicles. In this paper, we describe the development of a compact flow cytometer built from three separate, customizable, and interchangeable modules. The instrument as configured in this work is being developed specifically for the detection of selected Centers for Disease Control (CDC) category B biothreat agents through a bead-based assay: E. coli O157:H7, Salmonella, Listeria, and Shigella. It has two-colormore » excitation, three-color fluorescence and light scattering detection, embedded electronics, and capillary based flow. However, these attributes can be easily modified for other applications such as cluster of differentiation 4 (CD4) counting. Proof of concept is demonstrated through a 6-plex bead assay with the results compared to a commercially available benchtop-sized instrument.« less

Reconfigurable acquisition system with integrated optics for a portable flow cytometer

NASA Astrophysics Data System (ADS)

Kirleis, Matthew A.; Mathews, Scott A.; Verbarg, Jasenka; Erickson, Jeffrey S.; Piqué, Alberto

2013-11-01

Portable and inexpensive scientific instruments that are capable of performing point of care diagnostics are needed for applications such as disease detection and diagnosis in resource-poor settings, for water quality and food supply monitoring, and for biosurveillance activities in autonomous vehicles. In this paper, we describe the development of a compact flow cytometer built from three separate, customizable, and interchangeable modules. The instrument as configured in this work is being developed specifically for the detection of selected Centers for Disease Control (CDC) category B biothreat agents through a bead-based assay: E. coli O157:H7, Salmonella, Listeria, and Shigella. It has two-color excitation, three-color fluorescence and light scattering detection, embedded electronics, and capillary based flow. However, these attributes can be easily modified for other applications such as cluster of differentiation 4 (CD4) counting. Proof of concept is demonstrated through a 6-plex bead assay with the results compared to a commercially available benchtop-sized instrument.

Isomorphic red blood cells using automated urine flow cytometry is a reliable method in diagnosis of bladder cancer.

PubMed

Muto, Satoru; Sugiura, Syo-Ichiro; Nakajima, Akiko; Horiuchi, Akira; Inoue, Masahiro; Saito, Keisuke; Isotani, Shuji; Yamaguchi, Raizo; Ide, Hisamitsu; Horie, Shigeo

2014-10-01

We aimed to identify patients with a chief complaint of hematuria who could safely avoid unnecessary radiation and instrumentation in the diagnosis of bladder cancer (BC), using automated urine flow cytometry to detect isomorphic red blood cells (RBCs) in urine. We acquired urine samples from 134 patients over the age of 35 years with a chief complaint of hematuria and a positive urine occult blood test or microhematuria. The data were analyzed using the UF-1000i (®) (Sysmex Co., Ltd., Kobe, Japan) automated urine flow cytometer to determine RBC morphology, which was classified as isomorphic or dysmorphic. The patients were divided into two groups (BC versus non-BC) for statistical analysis. Multivariate logistic regression analysis was used to determine the predictive value of flow cytometry versus urine cytology, the bladder tumor antigen test, occult blood in urine test, and microhematuria test. BC was confirmed in 26 of 134 patients (19.4 %). The area under the curve for RBC count using the automated urine flow cytometer was 0.94, representing the highest reference value obtained in this study. Isomorphic RBCs were detected in all patients in the BC group. On multivariate logistic regression analysis, only isomorphic RBC morphology was significantly predictive for BC (p < 0.001). Analytical parameters such as sensitivity, specificity, positive predictive value, and negative predictive value of isomorphic RBCs in urine were 100.0, 91.7, 74.3, and 100.0 %, respectively. Detection of urinary isomorphic RBCs using automated urine flow cytometry is a reliable method in the diagnosis of BC with hematuria.

High speed flow cytometer droplet formation system and method

DOEpatents

Van den Engh, Ger

2000-01-01

A droplet forming flow cytometer system allows high speed processing without the need for high oscillator drive powers through the inclusion of an oscillator or piezoelectric crystal such as within the nozzle volume or otherwise unidirectionally coupled to the sheath fluid. The nozzle container continuously converges so as to amplify unidirectional oscillations which are transmitted as pressure waves through the nozzle volume to the nozzle exit so as to form droplets from the fluid jet. The oscillator is directionally isolated so as to avoid moving the entire nozzle container so as to create only pressure waves within the sheath fluid. A variation in substance concentration is achieved through a movable substance introduction port which is positioned within a convergence zone to vary the relative concentration of substance to sheath fluid while still maintaining optimal laminar flow conditions. This variation may be automatically controlled through a sensor and controller configuration. A replaceable tip design is also provided whereby the ceramic nozzle tip is positioned within an edge insert in the nozzle body so as to smoothly transition from nozzle body to nozzle tip. The nozzle tip is sealed against its outer surface to the nozzle body so it may be removable for cleaning or replacement.

Multi-parameter analysis using photovoltaic cell-based optofluidic cytometer

PubMed Central

Yan, Chien-Shun; Wang, Yao-Nan

2016-01-01

A multi-parameter optofluidic cytometer based on two low-cost commercial photovoltaic cells and an avalanche photodetector is proposed. The optofluidic cytometer is fabricated on a polydimethylsiloxane (PDMS) substrate and is capable of detecting side scattered (SSC), extinction (EXT) and fluorescence (FL) signals simultaneously using a free-space light transmission technique without the need for on-chip optical waveguides. The feasibility of the proposed device is demonstrated by detecting fluorescent-labeled polystyrene beads with sizes of 3 μm, 5 μm and 10 μm, respectively, and label-free beads with a size of 7.26 μm. The detection experiments are performed using both single-bead population samples and mixed-bead population samples. The detection results obtained using the SSC/EXT, EXT/FL and SSC/FL signals are compared with those obtained using a commercial flow cytometer. It is shown that the optofluidic cytometer achieves a high detection accuracy for both single-bead population samples and mixed-bead population samples. Consequently, the proposed device provides a versatile, straightforward and low-cost solution for a wide variety of point-of-care (PoC) cytometry applications. PMID:27699122

Utilization of microparticles in next-generation assays for microflow cytometers.

PubMed

Kim, Jason S; Ligler, Frances S

2010-11-01

Micron-sized particles have primarily been used in microfabricated flow cytometers for calibration purposes and proof-of-concept experiments. With increasing frequency, microparticles are serving as a platform for assays measured in these small analytical devices. Light scattering has been used to measure the agglomeration of antibody-coated particles in the presence of an antigen. Impedance detection is another technology being integrated into microflow cytometers for microparticle-based assays. Fluorescence is the most popular detection method in flow cytometry, enabling highly sensitive multiplexed assays. Finally, magnetic particles have also been used to measure antigen levels using a magnetophoretic micro-device. We review the progress of microparticle-based assays in microflow cytometry in terms of the advantages and limitations of each approach.

Development of a single-cell X-ray fluorescence flow cytometer

DOE PAGES

Crawford, Andrew M.; Kurecka, Patrick; Yim, Tsz Kwan; ...

2016-06-17

An X-ray fluorescence flow cytometer that can determine the total metal content of single cells has been developed. Capillary action or pressure was used to load cells into hydrophilic or hydrophobic capillaries, respectively. Once loaded, the cells were transported at a fixed vertical velocity past a focused X-ray beam. X-ray fluorescence was then used to determine the mass of metal in each cell. By making single-cell measurements, the population heterogeneity for metals in the µ M to m M concentration range on fL sample volumes can be directly measured, a measurement that is difficult using most analytical methods. This approachmore » has been used to determine the metal composition of 936 individual bovine red blood cells (bRBC), 31 individual 3T3 mouse fibroblasts (NIH3T3) and 18 Saccharomyces cerevisiae (yeast) cells with an average measurement frequency of ~4 cells min –1. These data show evidence for surprisingly broad metal distributions. Lastly, details of the device design, data analysis and opportunities for further sensitivity improvement are described.« less

Parametric analyses of planned flowing uranium hexafluoride critical experiments

NASA Technical Reports Server (NTRS)

Rodgers, R. J.; Latham, T. S.

1976-01-01

Analytical investigations were conducted to determine preliminary design and operating characteristics of flowing uranium hexafluoride (UF6) gaseous nuclear reactor experiments in which a hybrid core configuration comprised of UF6 gas and a region of solid fuel will be employed. The investigations are part of a planned program to perform a series of experiments of increasing performance, culminating in an approximately 5 MW fissioning uranium plasma experiment. A preliminary design is described for an argon buffer gas confined, UF6 flow loop system for future use in flowing critical experiments. Initial calculations to estimate the operating characteristics of the gaseous fissioning UF6 in a confined flow test at a pressure of 4 atm, indicate temperature increases of approximately 100 and 1000 K in the UF6 may be obtained for total test power levels of 100 kW and 1 MW for test times of 320 and 32 sec, respectively.

System for sensing droplet formation time delay in a flow cytometer

DOEpatents

Van den Engh, Ger; Esposito, Richard J.

1997-01-01

A droplet flow cytometer system which includes a system to optimize the droplet formation time delay based on conditions actually experienced includes an automatic droplet sampler which rapidly moves a plurality of containers stepwise through the droplet stream while simultaneously adjusting the droplet time delay. Through the system sampling of an actual substance to be processed can be used to minimize the effect of the substances variations or the determination of which time delay is optimal. Analysis such as cell counting and the like may be conducted manually or automatically and input to a time delay adjustment which may then act with analysis equipment to revise the time delay estimate actually applied during processing. The automatic sampler can be controlled through a microprocessor and appropriate programming to bracket an initial droplet formation time delay estimate. When maximization counts through volume, weight, or other types of analysis exists in the containers, the increment may then be reduced for a more accurate ultimate setting. This may be accomplished while actually processing the sample without interruption.

Development of a two-parameter slit-scan flow cytometer for screening of normal and aberrant chromosomes: application to a karyotype of Sus scrofa domestica (pig)

NASA Astrophysics Data System (ADS)

Hausmann, Michael; Doelle, Juergen; Arnold, Armin; Stepanow, Boris; Wickert, Burkhard; Boscher, Jeannine; Popescu, Paul C.; Cremer, Christoph

1992-07-01

Laser fluorescence activated slit-scan flow cytometry offers an approach to a fast, quantitative characterization of chromosomes due to morphological features. It can be applied for screening of chromosomal abnormalities. We give a preliminary report on the development of the Heidelberg slit-scan flow cytometer. Time-resolved measurement of the fluorescence intensity along the chromosome axis can be registered simultaneously for two parameters when the chromosome axis can be registered simultaneously for two parameters when the chromosome passes perpendicularly through a narrowly focused laser beam combined by a detection slit in the image plane. So far automated data analysis has been performed off-line on a PC. In its final performance, the Heidelberg slit-scan flow cytometer will achieve on-line data analysis that allows an electro-acoustical sorting of chromosomes of interest. Interest is high in the agriculture field to study chromosome aberrations that influence the size of litters in pig (Sus scrofa domestica) breeding. Slit-scan measurements have been performed to characterize chromosomes of pigs; we present results for chromosome 1 and a translocation chromosome 6/15.

A new hand-held microfluidic cytometer for evaluating irradiation damage by analysis of the damaged cells distribution.

PubMed

Wang, Junsheng; Fan, Zhiqiang; Zhao, Yile; Song, Younan; Chu, Hui; Song, Wendong; Song, Yongxin; Pan, Xinxiang; Sun, Yeqing; Li, Dongqing

2016-03-17

Space radiation brings uneven damages to cells. The detection of the distribution of cell damage plays a very important role in radiation medicine and the related research. In this paper, a new hand-held microfluidic flow cytometer was developed to evaluate the degree of radiation damage of cells. The device we propose overcomes the shortcomings (e.g., large volume and high cost) of commercial flow cytometers and can evaluate the radiation damage of cells accurately and quickly with potential for onsite applications. The distribution of radiation-damaged cells is analyzed by a simultaneous detection of immunofluorescence intensity of γ-H2AX and resistance pulse sensor (RPS) signal. The γ-H2AX fluorescence intensity provides information of the degree of radiation damage in cells. The ratio of the number of cells with γ-H2AX fluorescence signals to the total numbers of cells detected by RPS indicates the percentage of the cells that are damaged by radiation. The comparison experiment between the developed hand-held microfluidic flow cytometer and a commercial confocal microscope indicates a consistent and comparable detection performance.

A new hand-held microfluidic cytometer for evaluating irradiation damage by analysis of the damaged cells distribution

NASA Astrophysics Data System (ADS)

Wang, Junsheng; Fan, Zhiqiang; Zhao, Yile; Song, Younan; Chu, Hui; Song, Wendong; Song, Yongxin; Pan, Xinxiang; Sun, Yeqing; Li, Dongqing

2016-03-01

Space radiation brings uneven damages to cells. The detection of the distribution of cell damage plays a very important role in radiation medicine and the related research. In this paper, a new hand-held microfluidic flow cytometer was developed to evaluate the degree of radiation damage of cells. The device we propose overcomes the shortcomings (e.g., large volume and high cost) of commercial flow cytometers and can evaluate the radiation damage of cells accurately and quickly with potential for onsite applications. The distribution of radiation-damaged cells is analyzed by a simultaneous detection of immunofluorescence intensity of γ-H2AX and resistance pulse sensor (RPS) signal. The γ-H2AX fluorescence intensity provides information of the degree of radiation damage in cells. The ratio of the number of cells with γ-H2AX fluorescence signals to the total numbers of cells detected by RPS indicates the percentage of the cells that are damaged by radiation. The comparison experiment between the developed hand-held microfluidic flow cytometer and a commercial confocal microscope indicates a consistent and comparable detection performance.

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.

Classification of biological cells using a sound wave based flow cytometer

NASA Astrophysics Data System (ADS)

Strohm, Eric M.; Gnyawali, Vaskar; Van De Vondervoort, Mia; Daghighi, Yasaman; Tsai, Scott S. H.; Kolios, Michael C.

2016-03-01

A flow cytometer that uses sound waves to determine the size of biological cells is presented. In this system, a microfluidic device made of polydimethylsiloxane (PDMS) was developed to hydrodynamically flow focus cells in a single file through a target area. Integrated into the microfluidic device was an ultrasound transducer with a 375 MHz center frequency, aligned opposite the transducer was a pulsed 532 nm laser focused into the device by a 10x objective. Each passing cell was insonfied with a high frequency ultrasound pulse, and irradiated with the laser. The resulting ultrasound and photoacoustic waves from each cell were analyzed using signal processing methods, where features in the power spectra were compared to theoretical models to calculate the cell size. Two cell lines with different size distributions were used to test the system: acute myeloid leukemia cells (AML) and melanoma cells. Over 200 cells were measured using this system. The average calculated diameter of the AML cells was 10.4 +/- 2.5 μm using ultrasound, and 11.4 +/- 2.3 μm using photoacoustics. The average diameter of the melanoma cells was 16.2 +/- 2.9 μm using ultrasound, and 18.9 +/- 3.5 μm using photoacoustics. The cell sizes calculated using ultrasound and photoacoustic methods agreed with measurements using a Coulter Counter, where the AML cells were 9.8 +/- 1.8 μm and the melanoma cells were 16.0 +/- 2.5 μm. These results demonstrate a high speed method of assessing cell size using sound waves, which is an alternative method to traditional flow cytometry techniques.

CONTINUOUS PROCESS FOR THE CONVERSION OF UF$sub 6$ TO UF$sub 4$

DOEpatents

Smiley, S.H.; Brater, D.C.; Nimmo, R.H.

1959-10-01

A method is presented for reducing UF/sub 6/ to UF/sub 6/ with hydrogen. A preheated mixture of UF/sub 6/ and fluorine is contacted with a stoichiometric excess of preheated hydrogen in a reaction chamber thereby producing UF/sub 6/. The UF/sub 6/ reacts quantitatively and the UF/sub 6/ produced is of high purity and high density.

Numerical and experimental characterization of solid-state micropore-based cytometer for detection and enumeration of biological cells.

PubMed

Guo, Jinhong; Chen, Liang; Ai, Ye; Cheng, Yuanbing; Li, Chang Ming; Kang, Yuejun; Wang, Zhiming

2015-03-01

Portable diagnostic devices have emerged as important tools in various biomedical applications since they can provide an effective solution for low-cost and rapid clinical diagnosis. In this paper, we present a micropore-based resistive cytometer for the detection and enumeration of biological cells. The proposed device was fabricated on a silicon wafer by a standard microelectromechanical system processing technology, which enables a mass production of the proposed chip. The working principle of this cytometer is based upon a bias potential modulated pulse, originating from the biological particle's physical blockage of the micropore. Polystyrene particles of different sizes (7, 10, and 16 μm) were used to test and calibrate the proposed device. A finite element simulation was developed to predict the bias potential modulated pulse (peak amplitude vs. pulse bandwidth), which can provide critical insight into the design of this microfluidic flow cytometer. Furthermore, HeLa cells (a type of tumor cell lines) spiked in a suspension of blood cells, including red blood cells and white blood cells, were used to assess the performance for detecting and counting tumor cells. The proposed microfluidic flow cytometer is able to provide a promising platform to address the current unmet need for point-of-care clinical diagnosis. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A new concept and a comprehensive evaluation of SYSMEX UF-1000i  flow cytometer to identify culture-negative urine specimens in patients with UTI.

PubMed

Monsen, T; Ryden, P

2017-09-01

Urinary tract infections (UTIs) are among the most common bacterial infections in men and urine culture is gold standard for diagnosis. Considering the high prevalence of culture-negative specimens, any method that identifies such specimens is of interest. The aim was to evaluate a new screening concept for flow cytometry analysis (FCA). The outcomes were evaluated against urine culture, uropathogen species and three conventional screening methods. A prospective, consecutive study examined 1,312 urine specimens, collected during January and February 2012. The specimens were analyzed using the Sysmex UF1000i FCA. Based on the FCA data culture negative specimens were identified in a new model by use of linear discriminant analysis (FCA-LDA). In total 1,312 patients were included. In- and outpatients represented 19.6% and 79.4%, respectively; 68.3% of the specimens originated from women. Of the 610 culture-positive specimens, Escherichia coli represented 64%, enterococci 8% and Klebsiella spp. 7%. Screening with FCA-LDA at 95% sensitivity identified 42% (552/1312) as culture negative specimens when UTI was defined according to European guidelines. The proposed screening method was either superior or similar in comparison to the three conventional screening methods. In conclusion, the proposed/suggested and new FCA-LDA screening method was superior or similar to three conventional screening methods. We recommend the proposed screening method to be used in clinic to exclude culture negative specimens, to reduce workload, costs and the turnaround time. In addition, the FCA data may add information that enhance handling and support diagnosis of patients with suspected UTI pending urine culture [corrected].

Genotoxic effects of old landfill leachate on HepG2 cells after nitration/ultrafiltration/reverse osmosis membrane treatment process.

PubMed

Cheng, Rong; Zhao, Ling; Yin, Pinghe

2017-12-01

Toxicity assessment of nitration/ultrafiltration/reverse osmosis (nitration/UF/RO) project, which has recently been widely used as an efficient process with applications in practical leachate treatment, was very limited. In the present study, DNA damage of leachates was investigated before and after the nitration/UF/RO process by a battery of assays with human hepatoma cells. Methyletrazolium assay showed a high cytotoxicity of 97.1% after being exposed to the highest concentration of raw leachate for 24 h, and a cytotoxicity of 26% in effluent at a concentration of 30% (v/v). Both comet assay (24 h) and γH2AX flow cytometer assay (3 h) showed increased levels of DNA damage in cells exposed to raw leachate and after nitration/UF-treated leachate followed by a significant increase of 7-ethoxyresorufin-O-deethylase activity. However, the effluent after nitration/UF/RO treatment showed no significant difference compared to negative control for γH2AX flow cytometer assay but slight DNA damage at concentrations of 20% and 30% (v/v) as well as increase of 7-ethoxyresorufin-O-deethylase. Analysis showed that nitration/UF/RO process exhibited high removal of physicochemical indexes and significant reduction of toxic and genotoxic effects of leachate, but still demands an improvement to reduce all possible negative risks to the environment and humans. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

System and method for measuring particles in a sample stream of a flow cytometer or the like

DOEpatents

Graves, Steven W.; Habberset, Robert C.

2010-11-16

A system and method for analyzing a particle in a sample stream of a flow cytometer or the like. The system has a light source, such as a laser pointer module, for generating a low powered light beam and a fluidics apparatus which is configured to transport particles in the sample stream at substantially low velocity through the light beam for interrogation. Detectors, such as photomultiplier tubes, are configured to detect optical signals generated in response to the light beam impinging the particles. Signal conditioning circuitry is connected to each of the detectors to condition each detector output into electronic signals for processing and is designed to have a limited frequency response to filter high frequency noise from the detector output signals.

Argon/UF6 plasma exhaust gas reconstitution experiments using preheated fluorine and on-line diagnostics. [fissioning uranium plasma core reactor design

NASA Technical Reports Server (NTRS)

Roman, W. C.

1979-01-01

The feasibility of employing a flowing, high-temperature, pure fluorine/UF6 regeneration system to efficiently convert a large fraction of the effluent plasma exhaust back to pure UF6 was demonstrated. The custom built T.O.F. mass spectrometer sampling system permitted on-line measurements of the UF6 concentration at different locations in the exhaust system. Negligible amounts ( 100 ppm) of UF6 were detected in the axial bypass exhaust duct and the exhaust ducts downstream of the cryogenic trap system used to collect the UF6, thus verifying the overall system efficiency over a range of operating conditions. Use of a porous Monel duct as part of the exhaust duct system, including provision for injection of pure fluorine, provided a viable technique to eliminate uranium compound residue on the inside surface of the exhaust ducts. Typical uranium compound mass deposition per unit area of duct was 2 micron g/sq cm. This porous duct technique is directly applicable to future uranium compound transfer exhaust systems. Throughout these experiments, additional basic data on the corrosion aspects of hot, pressurized UF6/fluorine were also accumulated.

Comparison of MFI-UF constant pressure, MFI-UF constant flux and Crossflow Sampler-Modified Fouling Index Ultrafiltration (CFS-MFI UF).

PubMed

Sim, Lee Nuang; Ye, Yun; Chen, Vicki; Fane, Anthony G

2011-02-01

Understanding the foulant deposition mechanism during crossflow filtration is critical in developing indices to predict fouling propensity of feed water for reverse osmosis (RO). Factors affecting the performance on different fouling indices such as MFI-UF constant pressure, MFI-UF constant flux and newly proposed fouling index, CFS-MFI(UF) were investigated. Crossflow Sampler-Modified Fouling Index Ultrafiltration (CFS-MFI(UF)) utilises a typical crossflow unit to simulate the hydrodynamic conditions in the actual RO units followed by a dead-end unit to measure the fouling propensity of foulants. CFS-MFI(UF) was found sensitive to crossflow velocity. The crossflow velocity in the crossflow sampler unit influences the particle concentration and the particle size distribution in its permeate. CFS-MFI(UF) was also found sensitive to the permeate flux of both CFS and the dead-end cell. To closely simulate the hydrodynamic conditions of a crossflow RO unit, the flux used for CFS-MFI(UF) measurement was critical. The best option is to operate both the CFS and dead-end permeate flux at flux which is normally operated at industry RO units (∼20 L/m(2)h), but this would prolong the test duration excessively. In this study, the dead-end flux was accelerated by reducing the dead-end membrane area while maintaining the CFS permeate flux at 20 L/m(2)h. By doing so, a flux correction factor was investigated and applied to correlate the CFS-MFI(UF) measured at dead-end flux of 120 L/m(2)h to CFS-MFI(UF) measured at dead-end flux of 20 L/m(2)h for RO fouling rate prediction. Using this flux correction factor, the test duration of CFS-MFI(UF) can be shortened from 15 h to 2h. © 2010 Elsevier Ltd. All rights reserved.

System and method for measuring particles in a sample stream of a flow cytometer using low-power laser source

DOEpatents

Graves, Steven W.; Habbersett, Robert C.

2014-07-01

A system and method for analyzing a particle in a sample stream of a flow cytometer or the like. The system has a light source, such as a laser pointer module, for generating a low powered light beam and a fluidics apparatus which is configured to transport particles in the sample stream at substantially low velocity through the light beam for interrogation. Detectors, such as photomultiplier tubes, are configured to detect optical signals generated in response to the light beam impinging the particles. Signal conditioning circuitry is connected to each of the detectors to condition each detector output into electronic signals for processing and is designed to have a limited frequency response to filter high frequency noise from the detector output signals.

System and method for measuring particles in a sample stream of a flow cytometer using low-power laser source

DOEpatents

Graves, Steven W.; Habbersett, Robert C.

2016-11-15

A system and method for analyzing a particle in a sample stream of a flow cytometer or the like. The system has a light source, such as a laser pointer module, for generating a low powered light beam and a fluidics apparatus which is configured to transport particles in the sample stream at substantially low velocity through the light beam for interrogation. Detectors, such as photomultiplier tubes, are configured to detect optical signals generated in response to the light beam impinging the particles. Signal conditioning circuitry is connected to each of the detectors to condition each detector output into electronic signals for processing and is designed to have a limited frequency response to filter high frequency noise from the detector output signals.

Urine flow cytometry can rule out urinary tract infection, but cannot identify bacterial morphologies correctly.

PubMed

Geerts, N; Jansz, A R; Boonen, K J M; Wijn, R P W F; Koldewijn, E L; Boer, A K; Scharnhorst, V

2015-08-25

The diagnosis of urinary tract infection (UTI) by urine culture is a time-consuming and costly procedure. Usage of a screening method, to identify negative samples, would therefore affect time-to-diagnosis and laboratory cost positively. Urine flow cytometers are able to identify particles in urine. Together with the introduction of a cut-off value, which determines if a urine sample is subsequently cultured or not, the number of cultures can be reduced, while maintaining a low level of false negatives and a high negative predictive value. Recently, Sysmex developed additional software for their urine flow cytometers. Besides measuring the number of bacteria present in urine, information is given on bacterial morphology, which may guide the physician in the choice of antibiotic. In this study, we evaluated this software update. The UF1000i classifies bacteria into two categories: 'rods' and 'cocci/mixed'. Compared to the actual morphology of the bacterial pathogen found, the 'rods' category scores reasonably well with 91% chance of classifying rod-shaped bacteria correctly. The 'cocci/mixed' category underperforms, with only 29% of spherical-shaped bacteria (cocci) classified as such. In its current version, the bacterial morphology software does not classify bacteria, according to their morphology, well enough to be of clinical use in this study population. Copyright © 2015 Elsevier B.V. All rights reserved.

Near infrared lasers in flow cytometry.

PubMed

Telford, William G

2015-07-01

Technology development in flow cytometry has closely tracked laser technology, the light source that flow cytometers almost exclusively use to excite fluorescent probes. The original flow cytometers from the 1970s and 1980s used large water-cooled lasers to produce only one or two laser lines at a time. Modern cytometers can take advantage of the revolution in solid state laser technology to use almost any laser wavelength ranging from the ultraviolet to the near infrared. Commercial cytometers can now be equipped with many small solid state lasers, providing almost any wavelength needed for cellular analysis. Flow cytometers are now equipped to analyze 20 or more fluorescent probes simultaneously, requiring multiple laser wavelengths. Instrument developers are now trying to increase this number by designing fluorescent probes that can be excited by laser wavelength at the "edges" of the visible light range, in the near ultraviolet and near-infrared region. A variety of fluorescent probes have been developed that excite with violet and long wavelength ultraviolet light; however, the near-infrared range (660-800 nm) has yet seen only exploitation in flow cytometry. Fortunately, near-infrared laser diodes and other solid state laser technologies appropriate for flow cytometry have been in existence for some time, and can be readily incorporated into flow cytometers to accelerate fluorescent probe development. The near infrared region represents one of the last "frontiers" to maximize the number of fluorescent probes that can be analyzed by flow cytometry. In addition, near infrared fluorescent probes used in biomedical tracking and imaging could also be employed for flow cytometry with the correct laser wavelengths. This review describes the available technology, including lasers, fluorescent probes and detector technology optimal for near infrared signal detection. Published by Elsevier Inc.

System and method for measuring particles in a sample stream of a flow cytometer using a low power laser source

DOEpatents

Graves, Steven W; Habbersett, Robert C

2013-10-22

A system and method for analyzing a particle in a sample stream of a flow cytometer or the like. The system has a light source, such as a laser pointer module, for generating a low powered light beam and a fluidics apparatus which is configured to transport particles in the sample stream at substantially low velocity through the light beam for interrogation. Detectors, such as photomultiplier tubes, are configured to detect optical signals generated in response to the light beam impinging the particles. Signal conditioning circuitry is connected to each of the detectors to condition each detector output into electronic signals for processing and is designed to have a limited frequency response to filter high frequency noise from the detector output signals.

Functional analysis and classification of phytoplankton based on data from an automated flow cytometer.

PubMed

Malkassian, Anthony; Nerini, David; van Dijk, Mark A; Thyssen, Melilotus; Mante, Claude; Gregori, Gerald

2011-04-01

Analytical flow cytometry (FCM) is well suited for the analysis of phytoplankton communities in fresh and sea waters. The measurement of light scatter and autofluorescence properties of particles by FCM provides optical fingerprints, which enables different phytoplankton groups to be separated. A submersible version of the CytoSense flow cytometer (the CytoSub) has been designed for in situ autonomous sampling and analysis, making it possible to monitor phytoplankton at a short temporal scale and obtain accurate information about its dynamics. For data analysis, a manual clustering is usually performed a posteriori: data are displayed on histograms and scatterplots, and group discrimination is made by drawing and combining regions (gating). The purpose of this study is to provide greater objectivity in the data analysis by applying a nonmanual and consistent method to automatically discriminate clusters of particles. In other words, we seek for partitioning methods based on the optical fingerprints of each particle. As the CytoSense is able to record the full pulse shape for each variable, it quickly generates a large and complex dataset to analyze. The shape, length, and area of each curve were chosen as descriptors for the analysis. To test the developed method, numerical experiments were performed on simulated curves. Then, the method was applied and validated on phytoplankton cultures data. Promising results have been obtained with a mixture of various species whose optical fingerprints overlapped considerably and could not be accurately separated using manual gating. Copyright © 2011 International Society for Advancement of Cytometry.

A microflow cytometer on a chip

NASA Astrophysics Data System (ADS)

Golden, Joel P.; Kim, Jason; Anderson, George P.; Hashemi, Nastaran; Howell, Peter J.; Ligler, Frances S.

2010-02-01

A rapid, automated, multi-analyte Microflow Cytometer is being developed as a portable, field-deployable sensor for onsite diagnosis of biothreat agent exposure and environmental monitoring. The technology relies on a unique method for ensheathing a sample stream in continuous flow past an interrogation region where optical fibers provide excitation and collect emission. This approach efficiently focuses particles in the interrogation region of the fluidic channel, avoids clogging and provides for subsequent separation of the core and sheath fluids in order to capture the target for confirmatory assays and recycling of the sheath fluid. Fluorescently coded microspheres provide the capability for highly multiplexed assays. Optical analysis at four different wavelengths identified six sets of the coded microspheres recognizing Escherichia coli, Listeria, and Salmonella as well as cholera toxin, staphylococcal enterotoxin B (SEB), and ricin, and assay results were compared with those of a commercial Luminex analysis system.

Argon/UF6 plasma experiments: UF6 regeneration and product analysis

NASA Technical Reports Server (NTRS)

Roman, W. C.

1980-01-01

An experimental and analytical investigation was conducted to aid in developing some of the technology necessary for designing a self-critical fissioning uranium plasma core reactors (PCR). This technology is applicable to gaseous uranium hexafluoride nuclear-pumped laser systems. The principal equipment used included 1.2 MW RF induction heater, a d.c. plasma torch, a uranium tetrafluoride feeder system, and batch-type fluorine/UF6 regeneration systems. Overall objectives were to continue to develop and test materials and handling techniques suitable for use with high-temperature, high-pressure, gaseous UF6; and to continue development of complementary diagnostic instrumentation and measurement techniques to characterize the effluent exhaust gases and residue deposited on the test chamber and exhaust system components. Specific objectives include: a development of a batch-type UF6 regeneration system employing pure high-temperature fluorine; development of a ruggedized time-of-flight mass spectrometer and associated data acquisition system capable of making on-line concentration measurements of the volatile effluent exhaust gas species in a high RF environment and corrosive environment of UF6 and related halide compounds.

Filling of high aspect ratio micro features of a microfluidic flow cytometer chip using micro injection moulding

NASA Astrophysics Data System (ADS)

Zhang, Haoyang; Fang, Fengzhou; Gilchrist, Michael D.; Zhang, Nan

2018-07-01

Micro injection moulding has been demonstrated as one of the most efficient mass production technologies for manufacturing polymeric microfluidic devices, which have been widely used in life sciences, environmental and analytical fields and agro-food industries. However, the filling of micro features for typical microfluidic devices is complicated and not yet fully understood, which consequently restricts the chip development. In the present work, a microfluidic flow cytometer chip with essential high aspect ratio micro features was used as a typical model to study their filling process. Short-shot experiments and single factor experiments were performed to examine the filling progress of such features during the injection and packing stages of the micro injection moulding process. The influence of process parameters such as shot size, packing pressure, packing time and mould temperature were systematically monitored, characterised and correlated with 3D measurements and real response of the machine such as screw velocity and screw position. A combined melt flow and creep deformation model was proposed to explain the complex influence of process on replication. An approach of over-shot micro injection moulding was proposed and was shown to be effective at improving the replication quality of high aspect ratio micro features.

Method for converting UF5 to UF4 in a molten fluoride salt

DOEpatents

Bennett, Melvin R.; Bamberger, Carlos E.; Kelmers, A. Donald

1977-01-01

The reduction of UF.sub.5 to UF.sub.4 in a molten fluoride salt by sparging with hydrogen is catalyzed by metallic platinum. The reaction is also catalyzed by platinum alloyed with gold reaction equipment.

Small lasers in flow cytometry.

PubMed

Telford, William G

2004-01-01

Laser technology has made tremendous advances in recent years, particularly in the area of diode and diode-pumped solid state sources. Flow cytometry has been a direct beneficiary of these advances, as these small, low-maintenance, inexpensive lasers with reasonable power outputs are integrated into flow cytometers. In this chapter we review the contribution and potential of solid-state lasers to flow cytometry, and show several examples of these novel sources integrated into production flow cytometers. Technical details and critical parameters for successful application of these lasers for biomedical analysis are reviewed.

METHOD FOR THE RECOVERY AND PURIFICATION OF GASEOUS UF$sub 6$ FROM GASEOUS MIXTURES AND UF$sub 7$NO AND UF$sub 7$NO$sub 2$ PRODUCTS PRODUCED THEREBY

DOEpatents

Ogle, P.R. Jr.

1962-06-16

A method is given for recovering uranium hexafluoride from a gaseous mixture containing said uranium hexafluoride and extraneous gaseous impurities. The method comprises reacting said mixture with a nitrogen oxyfluoride at a temperature in the range - 100 to 50 deg C to thereby form a solid compound having the empirical formula UF/sub 7/N(O)/sub x/ where x is a number from 1 to 2. (AEC)

Multinode acoustic focusing for parallel flow cytometry

PubMed Central

Piyasena, Menake E.; Suthanthiraraj, Pearlson P. Austin; Applegate, Robert W.; Goumas, Andrew M.; Woods, Travis A.; López, Gabriel P.; Graves, Steven W.

2012-01-01

Flow cytometry can simultaneously measure and analyze multiple properties of single cells or particles with high sensitivity and precision. Yet, conventional flow cytometers have fundamental limitations with regards to analyzing particles larger than about 70 microns, analyzing at flow rates greater than a few hundred microliters per minute, and providing analysis rates greater than 50,000 per second. To overcome these limits, we have developed multi-node acoustic focusing flow cells that can position particles (as small as a red blood cell and as large as 107 microns in diameter) into as many as 37 parallel flow streams. We demonstrate the potential of such flow cells for the development of high throughput, parallel flow cytometers by precision focusing of flow cytometry alignment microspheres, red blood cells, and the analysis of CD4+ cellular immunophenotyping assay. This approach will have significant impact towards the creation of high throughput flow cytometers for rare cell detection applications (e.g. circulating tumor cells), applications requiring large particle analysis, and high volume flow cytometry. PMID:22239072

UF.sub.6 -Recovery process utilizing desublimation

DOEpatents

Eby, Robert S.; Stephenson, Michael J.; Andrews, Deborah H.; Hamilton, Thomas H.

1985-01-01

The invention is a UF.sub.6 -recovery process of the kind in which a stream of substantially pure gaseous UF.sub.6 is directed through an externally chilled desublimer to convert the UF.sub.6 directly to an annular solid ring adhering to the interior wall of the desublimer. After accumulation of a desired amount of solid UF.sub.6, the desublimer is heated to liquefy the solid. Subsequently, the liquid is recovered from the desublimer. It has been found that during the heating operation the desublimer is subjected to excessive mechanical stresses. In addition, it has been found that the incorporation of a very small percentage of relatively noncondensable, nonreactive gas (e.g., nitrogen) in the UF.sub.6 input to the desublimer effects significant decreases in the stresses generated during the subsequent melting operation. This modification to the process provides valuable advantages in terms of reduced hazard, lower operating costs for the desublimer, and increased service life for the desublimer and its auxiliaries. The new process is especially suitable for the recovery of enriched UF.sub.6 from high-speed UF.sub.6 gas-centrifuge cascades.

UF/sub 6/-recovery process utilizing desublimation

DOEpatents

Eby, R.S.; Stephenson, M.J.; Andrews, D.H.; Hamilton, T.H.

1983-12-21

The invention is a UF/sub 6/-recovery process of the kind in which a stream of substantially pure gaseous UF/sub 6/ is directed through an externally chilled desublimer to convert the UF/sub 6/ directly to an annular solid ring adhering to the interior wall of the desublimer. After accumulation of a desired amount of solid UF/sub 6/, the desublimer is heated to liquefy the solid. Subsequently, the liquid is recovered from the desublimer. It has been found that during the heating operation the desublimer is subjected to excessive mechanical stresses. In addition, it has been found that the incorporation of a very small percentage of relatively noncondensable, nonreactive gas (e.g., nitrogen) in the UF/sub 6/ input to the desublimer effects significant decreases in the stresses generated during the subsequent melting operation. This modification to the process provides valuable advantages in terms of reduced hazard, lower operating costs for the desublimer, and increased service life for the desublimer and its auxiliaries. The new process is especially suitable for the recovery of enriched UF/sub 6/ from high-speed UF/sub 6/ gas-centrifuge cascades.

Circulation system for flowing uranium hexafluoride cavity reactor experiments

NASA Technical Reports Server (NTRS)

Jaminet, J. F.; Kendall, J. S.

1976-01-01

Research related to determining the feasibility of producing continuous power from fissile fuel in the gaseous state is presented. The development of three laboratory-scale flow systems for handling gaseous UF6 at temperatures up to 500 K, pressure up to approximately 40 atm, and continuous flow rates up to approximately 50g/s is presented. A UF6 handling system fabricated for static critical tests currently being conducted is described. The system was designed to supply UF6 to a double-walled aluminum core canister assembly at temperatures between 300 K and 400 K and pressure up to 4 atm. A second UF6 handling system designed to provide a circulating flow of up to 50g/s of gaseous UF6 in a closed-loop through a double-walled aluminum core canister with controlled temperature and pressure is described. Data from flow tests using UF6 and UF6/He mixtures with this system at flow rates up to approximately 12g/s and pressure up to 4 atm are presented. A third UF6 handling system fabricated to provide a continuous flow of UF6 at flow rates up to 5g/s and at pressures up to 40 atm for use in rf-heated, uranium plasma confinement experiments is described.

Thermal reactions of uranium metal, UO 2, U 3O 8, UF 4, and UO 2F 2 with NF 3 to produce UF 6

NASA Astrophysics Data System (ADS)

McNamara, Bruce; Scheele, Randall; Kozelisky, Anne; Edwards, Matthew

2009-11-01

This paper demonstrates that NF 3 fluorinates uranium metal, UO 2, UF 4, UO 3, U 3O 8, and UO 2F 2·2H 2O to produce the volatile UF 6 at temperatures between 100 and 550 °C. Thermogravimetric and differential thermal analysis reaction profiles are described that reflect changes in the uranium fluorination/oxidation state, physiochemical effects, and instances of discrete chemical speciation. Large differences in the onset temperatures for each system investigated implicate changes in mode of the NF 3 gas-solid surface interaction. These studies also demonstrate that NF 3 is a potential replacement fluorinating agent in the existing nuclear fuel cycle and in actinide volatility reprocessing.

Confocal Microscopy and Flow Cytometry System Performance: Assessment of QA Parameters that affect data Quanitification

EPA Science Inventory

Flow and image cytometers can provide useful quantitative fluorescence data. We have devised QA tests to be used on both a flow cytometer and a confocal microscope to assure that the data is accurate, reproducible and precise. Flow Cytometry: We have provided two simple perform...

Simultaneous ultrasound and photoacoustics based flow cytometry

NASA Astrophysics Data System (ADS)

Gnyawali, Vaskar; Strohm, Eric M.; Tsai, Scott S. H.; Kolios, Michael C.

2018-04-01

We have developed a flow cytometer based on simultaneous detection of ultrasound and photoacoustic waves from individual particles/cells flowing in a microfluidic channel. Our polydimethylsiloxane (PDMS) based hydrodynamic 3-dimensional (3D) flow-focusing microfluidic device contains a cross-junction channel, a micro-needle (ID 100 μm and OD 200 μm) insert, and a 3D printed frame to hold and align a high frequency (center frequency 375 MHz) ultrasound transducer. The focused flow passes through a narrow focal zone with lateral and axial focal lengths of 6-8 μm and 15-20 μm, respectively. Both the lateral and axial alignments are achieved by screwing the transducer to the frame onto the PDMS device. Individual particles pass through an interrogation zone in the microfluidic channel with a collinearly aligned ultrasound transducer and a focused 532 nm wavelength laser beam. The particles are simultaneously insonified by high-frequency ultrasound and irradiated by a laser beam. The ultrasound backscatter and laser generated photoacoustic waves are detected for each passing particle. The backscattered ultrasound and photoacoustic signal are strongly dependent on the size, morphology, mechanical properties, and material properties of the flowing particles; these parameters can be extracted by analyzing unique features in the power spectrum of the signals. Frequencies less than 100 MHz do not have these unique spectral signatures. We show that we can reliably distinguish between different particles in a sample using the acoustic-based flow cytometer. This technique, when extended to biomedical applications, allows us to rapidly analyze the spectral signatures from individual single cells of a large cell population, with applications towards label-free detection and characterization of healthy and diseased cells.

High throughput imaging cytometer with acoustic focussing.

PubMed

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

2015-10-31

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

Direct identification of bacteria causing urinary tract infections by combining matrix-assisted laser desorption ionization-time of flight mass spectrometry with UF-1000i urine flow cytometry.

PubMed

Wang, X-H; Zhang, G; Fan, Y-Y; Yang, X; Sui, W-J; Lu, X-X

2013-03-01

Rapid identification of bacterial pathogens from clinical specimens is essential to establish an adequate empirical antibiotic therapy to treat urinary tract infections (UTIs). We used matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) combined with UF-1000i urine flow cytometry of urine specimens to quickly and accurately identify bacteria causing UTIs. We divided each urine sample into three aliquots for conventional identification, UF-1000i, and MALDI-TOF MS, respectively. We compared the results of the conventional method with those of MALDI-TOF MS combined with UF-1000i, and discrepancies were resolved by 16S rRNA gene sequencing. We analyzed 1456 urine samples from patients with UTI symptoms, and 932 (64.0%) were negative using each of the three testing methods. The combined method used UF-1000i to eliminate negative specimens and then MALDI-TOF MS to identify the remaining positive samples. The combined method was consistent with the conventional method in 1373 of 1456 cases (94.3%), and gave the correct result in 1381 of 1456 cases (94.8%). Therefore, the combined method described here can directly provide a rapid, accurate, definitive bacterial identification for the vast majority of urine samples, though the MALDI-TOF MS software analysis capabilities should be improved, with regard to mixed bacterial infection. Copyright © 2012 Elsevier B.V. All rights reserved.

Assessment of Cumulative Trauma Disorder (CTD) Risk for 3 Different Tasks Constructing and Repairing Multi-Layer Insulation (MLI) Blankets, Preparing the Dough for a Pizza, and Operating the Becton-Dickinson FACSAria Flow Cytometer

NASA Technical Reports Server (NTRS)

Gentzler, Marc; Kline, Martin; Palmer, Andrew; Terrone, Mark

2007-01-01

The Cumulative Trauma Disorder (CTD) risks for three different tasks using McCauley-Bell and Badiru's (1993) formula based on task, personal, and organizational factors were examined. For the Multi-Layer Insulation (MLI) blanket task, the results showed that the task, personal, and organizational risks were at about the same level. The personal risk factors for this task were evaluated using a hypothetical female employee age 52. For the pizza dough task, it was shown that the organizational risk was particularly high, with task related factors also at quite dangerous levels. On the other hand, there was a very low level of personal risk factors, based on a female age 17. The flow cytometer task was assessed with three different participants, a11 of whom had quite disparate levels of personal risk, which slightly affected the overall CTD risk. This reveals how individual difference variables certainly need to be considered. The task and organizational risks for this task were rated at about the same moderate level. The overall CTD risk averaged across the three participants was .335, indicating some risk. Compruing across the tasks revealed that the pizza dough task created the greatest overall CTD risk by far (.568), with the MLI (.325) and flow cytometer task (.335) having some risk associated with them. Future research should look into different tasks for more of a comparison

Time-gated flow cytometry: an ultra-high selectivity method to recover ultra-rare-event μ-targets in high-background biosamples

NASA Astrophysics Data System (ADS)

Jin, Dayong; Piper, James A.; Leif, Robert C.; Yang, Sean; Ferrari, Belinda C.; Yuan, Jingli; Wang, Guilan; Vallarino, Lidia M.; Williams, John W.

2009-03-01

A fundamental problem for rare-event cell analysis is auto-fluorescence from nontarget particles and cells. Time-gated flow cytometry is based on the temporal-domain discrimination of long-lifetime (>1 μs) luminescence-stained cells and can render invisible all nontarget cell and particles. We aim to further evaluate the technique, focusing on detection of ultra-rare-event 5-μm calibration beads in environmental water dirt samples. Europium-labeled 5-μm calibration beads with improved luminescence homogeneity and reduced aggregation were evaluated using the prototype UV LED excited time-gated luminescence (TGL) flow cytometer (FCM). A BD FACSAria flow cytometer was used to sort accurately a very low number of beads (<100 events), which were then spiked into concentrated samples of environmental water. The use of europium-labeled beads permitted the demonstration of specific detection rates of 100%+/-30% and 91%+/-3% with 10 and 100 target beads, respectively, that were mixed with over one million nontarget autofluorescent background particles. Under the same conditions, a conventional FCM was unable to recover rare-event fluorescein isothiocyanate (FITC) calibration beads. Preliminary results on Giardia detection are also reported. We have demonstrated the scientific value of lanthanide-complex biolabels in flow cytometry. This approach may augment the current method that uses multifluorescence-channel flow cytometry gating.

Fluorescent Cell Barcoding for Multiplex Flow Cytometry

PubMed Central

Krutzik, Peter O.; Clutter, Matthew R.; Trejo, Angelica; Nolan, Garry P.

2011-01-01

Fluorescent Cell Barcoding (FCB) enables high throughput, i.e. high content flow cytometry by multiplexing samples prior to staining and acquisition on the cytometer. Individual cell samples are barcoded, or labeled, with unique signatures of fluorescent dyes so that they can be mixed together, stained, and analyzed as a single sample. By mixing samples prior to staining, antibody consumption is typically reduced 10 to 100-fold. In addition, data robustness is increased through the combination of control and treated samples, which minimizes pipetting error, staining variation, and the need for normalization. Finally, speed of acquisition is enhanced, enabling large profiling experiments to be run with standard cytometer hardware. In this unit, we outline the steps necessary to apply the FCB method to cell lines as well as primary peripheral blood samples. Important technical considerations such as choice of barcoding dyes, concentrations, labeling buffers, compensation, and software analysis are discussed. PMID:21207359

Removal of fluoride impurities from UF.sub.6 gas

DOEpatents

Beitz, James V.

1985-01-01

A method of purifying a UF.sub.6 gas stream containing one or more metal fluoride impurities composed of a transuranic metal, transition metal or mixtures thereof, is carried out by contacting the gas stream with a bed of UF.sub.5 in a reaction vessel under conditions where at least one impurity reacts with the UF.sub.5 to form a nongaseous product and a treated gas stream, and removing the treated gas stream from contact with the bed. The nongaseous products are subsequently removed in a reaction with an active fluorine affording agent to form a gaseous impurity which is removed from the reaction vessel. The bed of UF.sub.5 is formed by the reduction of UF.sub.6 in the presence of UV light. One embodiment of the reaction vessel includes a plurality of UV light sources as tubes on which UF.sub.5 is formed.

Purification of oily wastewater by hybrid UF/MD.

PubMed

Gryta, M; Karakulski, K; Morawski, A W

2001-10-01

Investigations on the treatment of oily wastewater by a combination of ultrafiltration (UF) and membrane distillation (MD) as a final purification method have been performed. A tubular UF module equipped with polyvinylidene fluoride (PVDF) membranes and a capillary MD module with polypropylene membranes were tested using a typical bilge water collected from a harbour without pretreatment. The permeate obtained from the UF process generally contains less than 5 ppm of oil. A further purification of the UF permeate by membrane distillation results in a complete removal of oil from wastewater and a very high reduction of the total organic carbon (99.5%) and total dissolved solids (99.9%).

Investigation of UF/sub 6/ behavior in a fire

DOE Office of Scientific and Technical Information (OSTI.GOV)

Williams, W.R.

Reactions between UF/sub 6/ and combustible gases and the potential for UF/sub 6/-filled cylinders to rupture when exposed to fire are addressed. Although the absence of kinetic data prevents specific identification and quantification of the chemical species formed, potential reaction products resulting from the release of UF/sub 6/ into a fire include UF/sub 4/, UO/sub 2/F/sub 2/, HF, C, CF/sub 4/,COF/sub 2/, and short chain, fluorinated or partially fluorinated hydrocarbons. Such a release adds energy to a fire relative to normal combustion reactions. Time intervals to an assumed point of rupture for UF/sub 6/-filled cylinders exposed to fire are estimatedmore » conservatively. Several related studies are also summarized, including a test series in which small UF/sub 6/-filled cylinders were immersed in fire resulting in valve failures and explosive ruptures. It is concluded that all sizes of UF/sub 6/ cylinders currently in use may rupture within 30 minutes when totally immersed in a fire. For cylinders adjacent to fires, rupture of the larger cylinders appears much less likely.« less

Removal of fluoride impurities from UF/sub 6/ gas

DOEpatents

Beitz, J.V.

1984-01-06

A method of purifying a UF/sub 6/ gas stream containing one or more metal fluoride impurities composed of a transuranic metal, transition metal or mixtures thereof, is carried out by contacting the gas stream with a bed of UF/sub 5/ in a reaction vessel under conditions where at least one impurity reacts with the UF/sub 5/ to form a nongaseous product and a treated gas stream, and removing the treated gas stream from contact with the bed. The nongaseous products are subsequently removed in a reaction with an active fluorine affording agent to form a gaseous impurity which is removed from the reaction vessel. The bed of UF/sub 5/ is formed by the reduction of UF/sub 6/ in the presence of uv light. One embodiment of the reaction vessel includes a plurality of uv light sources as tubes on which UF/sub 5/ is formed. 2 figures.

Characterization of uranium tetrafluoride (UF 4) with Raman spectroscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Villa-Aleman, Eliel; Wellons, Matthew S.

The Raman spectrum of uranium tetrafluoride (UF 4) is unambiguously characterized with multiple Raman excitation laser sources for the first time. Across different laser excitation wavelengths, UF 4 demonstrates 16 distinct Raman bands within the 50-400 cm -1 region. The observed Raman bands are representative of various F-F vibrational modes. UF 4 also shows intense fluorescent bands in the 325 – 750 nm spectral region. Comparison of the UF 4 spectrum with the ZrF 4 spectrum, its crystalline analog, demonstrates a similar Raman band structure consistent with group theory predictions for expected Raman bands. Additionally, a demonstration of combined scanningmore » electron microscopy (SEM) and in situ Raman spectroscopy microanalytical measurements of UF 4 particulates shows that despite the inherent weak intensity of Raman bands, identification and characterization are possible for micron-sized particulates with modern instrumentation. The published well characterized UF 4 spectrum is extremely relevant to nuclear materials and nuclear safeguard applications.« less

Characterization of uranium tetrafluoride (UF 4) with Raman spectroscopy

DOE PAGES

Villa-Aleman, Eliel; Wellons, Matthew S.

2016-03-22

The Raman spectrum of uranium tetrafluoride (UF 4) is unambiguously characterized with multiple Raman excitation laser sources for the first time. Across different laser excitation wavelengths, UF 4 demonstrates 16 distinct Raman bands within the 50-400 cm -1 region. The observed Raman bands are representative of various F-F vibrational modes. UF 4 also shows intense fluorescent bands in the 325 – 750 nm spectral region. Comparison of the UF 4 spectrum with the ZrF 4 spectrum, its crystalline analog, demonstrates a similar Raman band structure consistent with group theory predictions for expected Raman bands. Additionally, a demonstration of combined scanningmore » electron microscopy (SEM) and in situ Raman spectroscopy microanalytical measurements of UF 4 particulates shows that despite the inherent weak intensity of Raman bands, identification and characterization are possible for micron-sized particulates with modern instrumentation. The published well characterized UF 4 spectrum is extremely relevant to nuclear materials and nuclear safeguard applications.« less

Fundamentals of flow cytometry.

PubMed

Jaroszeski, M J; Radcliff, G

1999-02-01

Flow cytometers are instruments that are used primarily to measure the physical and biochemical characteristics of biological particles. This technology is used to perform measurements on whole cells as well as prepared cellular constituents, such as nuclei and organelles. Flow cytometers are investigative tools for a broad range of scientific disciplines because they make measurements on thousands of individual cells/particles in a matter of seconds. This is a unique advantage relative to other detection instruments that provide bulk particle measurements. Flow cytometry is a complex and highly technical field; therefore, a basic understanding of the technology is essential for all users. The purpose of this article is to provide fundamental information about the instrumentation used for flow cytometry as well as the methods used to analyze and interpret data. This information will provide a foundation to use flow cytometry effectively as a research tool.

Microbubble Sizing and Shell Characterization Using Flow Cytometry

PubMed Central

Tu, Juan; Swalwell, Jarred E.; Giraud, David; Cui, Weicheng; Chen, Weizhong; Matula, Thomas J.

2015-01-01

Experiments were performed to size, count, and obtain shell parameters for individual ultrasound contrast microbubbles using a modified flow cytometer. Light scattering was modeled using Mie theory, and applied to calibration beads to calibrate the system. The size distribution and population were measured directly from the flow cytometer. The shell parameters (shear modulus and shear viscosity) were quantified at different acoustic pressures (from 95 to 333 kPa) by fitting microbubble response data to a bubble dynamics model. The size distribution of the contrast agent microbubbles is consistent with manufacturer specifications. The shell shear viscosity increases with increasing equilibrium microbubble size, and decreases with increasing shear rate. The observed trends are independent of driving pressure amplitude. The shell elasticity does not vary with microbubble size. The results suggest that a modified flow cytometer can be an effective tool to characterize the physical properties of microbubbles, including size distribution, population, and shell parameters. PMID:21622051

SYSTEM FOR CONVERSION OF UF$sub 4$ TO UF$sub 6$

DOEpatents

Brater, D.G.; Pike, J.W.

1958-12-01

Method and apparatus are presented for rapid and complete conversion of solid, powdered uranium tetrafiuorlde to uranlum hexafluorlde by treating the UF/ sub 4/ with fluorine gas at a temperature of about 800 icient laborato C.

Reduced-gravity environment hardware demonstrations of a prototype miniaturized flow cytometer and companion microfluidic mixing technology.

PubMed

Phipps, William S; Yin, Zhizhong; Bae, Candice; Sharpe, Julia Z; Bishara, Andrew M; Nelson, Emily S; Weaver, Aaron S; Brown, Daniel; McKay, Terri L; Griffin, DeVon; Chan, Eugene Y

2014-11-13

Until recently, astronaut blood samples were collected in-flight, transported to earth on the Space Shuttle, and analyzed in terrestrial laboratories. If humans are to travel beyond low Earth orbit, a transition towards space-ready, point-of-care (POC) testing is required. Such testing needs to be comprehensive, easy to perform in a reduced-gravity environment, and unaffected by the stresses of launch and spaceflight. Countless POC devices have been developed to mimic laboratory scale counterparts, but most have narrow applications and few have demonstrable use in an in-flight, reduced-gravity environment. In fact, demonstrations of biomedical diagnostics in reduced gravity are limited altogether, making component choice and certain logistical challenges difficult to approach when seeking to test new technology. To help fill the void, we are presenting a modular method for the construction and operation of a prototype blood diagnostic device and its associated parabolic flight test rig that meet the standards for flight-testing onboard a parabolic flight, reduced-gravity aircraft. The method first focuses on rig assembly for in-flight, reduced-gravity testing of a flow cytometer and a companion microfluidic mixing chip. Components are adaptable to other designs and some custom components, such as a microvolume sample loader and the micromixer may be of particular interest. The method then shifts focus to flight preparation, by offering guidelines and suggestions to prepare for a successful flight test with regard to user training, development of a standard operating procedure (SOP), and other issues. Finally, in-flight experimental procedures specific to our demonstrations are described.

Reduced-gravity Environment Hardware Demonstrations of a Prototype Miniaturized Flow Cytometer and Companion Microfluidic Mixing Technology

PubMed Central

Bae, Candice; Sharpe, Julia Z.; Bishara, Andrew M.; Nelson, Emily S.; Weaver, Aaron S.; Brown, Daniel; McKay, Terri L.; Griffin, DeVon; Chan, Eugene Y.

2014-01-01

Until recently, astronaut blood samples were collected in-flight, transported to earth on the Space Shuttle, and analyzed in terrestrial laboratories. If humans are to travel beyond low Earth orbit, a transition towards space-ready, point-of-care (POC) testing is required. Such testing needs to be comprehensive, easy to perform in a reduced-gravity environment, and unaffected by the stresses of launch and spaceflight. Countless POC devices have been developed to mimic laboratory scale counterparts, but most have narrow applications and few have demonstrable use in an in-flight, reduced-gravity environment. In fact, demonstrations of biomedical diagnostics in reduced gravity are limited altogether, making component choice and certain logistical challenges difficult to approach when seeking to test new technology. To help fill the void, we are presenting a modular method for the construction and operation of a prototype blood diagnostic device and its associated parabolic flight test rig that meet the standards for flight-testing onboard a parabolic flight, reduced-gravity aircraft. The method first focuses on rig assembly for in-flight, reduced-gravity testing of a flow cytometer and a companion microfluidic mixing chip. Components are adaptable to other designs and some custom components, such as a microvolume sample loader and the micromixer may be of particular interest. The method then shifts focus to flight preparation, by offering guidelines and suggestions to prepare for a successful flight test with regard to user training, development of a standard operating procedure (SOP), and other issues. Finally, in-flight experimental procedures specific to our demonstrations are described. PMID:25490614

Scalable clustering algorithms for continuous environmental flow cytometry.

PubMed

Hyrkas, Jeremy; Clayton, Sophie; Ribalet, Francois; Halperin, Daniel; Armbrust, E Virginia; Howe, Bill

2016-02-01

Recent technological innovations in flow cytometry now allow oceanographers to collect high-frequency flow cytometry data from particles in aquatic environments on a scale far surpassing conventional flow cytometers. The SeaFlow cytometer continuously profiles microbial phytoplankton populations across thousands of kilometers of the surface ocean. The data streams produced by instruments such as SeaFlow challenge the traditional sample-by-sample approach in cytometric analysis and highlight the need for scalable clustering algorithms to extract population information from these large-scale, high-frequency flow cytometers. We explore how available algorithms commonly used for medical applications perform at classification of such a large-scale, environmental flow cytometry data. We apply large-scale Gaussian mixture models to massive datasets using Hadoop. This approach outperforms current state-of-the-art cytometry classification algorithms in accuracy and can be coupled with manual or automatic partitioning of data into homogeneous sections for further classification gains. We propose the Gaussian mixture model with partitioning approach for classification of large-scale, high-frequency flow cytometry data. Source code available for download at https://github.com/jhyrkas/seaflow_cluster, implemented in Java for use with Hadoop. hyrkas@cs.washington.edu Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Method and apparatus for measuring enrichment of UF6

DOEpatents

Hill, Thomas Roy [Santa Fe, NM; Ianakiev, Kiril Dimitrov [Los Alamos, NM

2011-06-07

A system and method are disclosed for determining the enrichment of .sup.235U in Uranium Hexafluoride (UF6) utilizing synthesized X-rays which are directed at a container test zone containing a sample of UF6. A detector placed behind the container test zone then detects and counts the X-rays which pass through the container and the UF6. In order to determine the portion of the attenuation due to the UF6 gas alone, this count rate may then be compared to a calibration count rate of X-rays passing through a calibration test zone which contains a vacuum, the test zone having experienced substantially similar environmental conditions as the actual test zone. Alternatively, X-rays of two differing energy levels may be alternately directed at the container, where either the container or the UF6 has a high sensitivity to the difference in the energy levels, and the other having a low sensitivity.

Uniform deposition of uranium hexafluoride (UF6): Standardized mass deposits and controlled isotopic ratios using a thermal fluorination method

DOE Office of Scientific and Technical Information (OSTI.GOV)

McNamara, Bruce K.; O’Hara, Matthew J.; Casella, Andrew M.

2016-07-01

Abstract: We report a convenient method for the generation of volatile uranium hexafluoride (UF6) from solid uranium oxides and other uranium compounds, followed by uniform deposition of low levels of UF6 onto sampling coupons. Under laminar flow conditions, UF6 is shown to interact with surfaces within the chamber to a highly predictable degree. We demonstrate the preparation of uranium deposits that range between ~0.01 and 470±34 ng∙cm-2. The data suggest the method can be extended to creating depositions at the sub-picogram∙cm-2 level. Additionally, the isotopic composition of the deposits can be customized by selection of the uranium source materials. Wemore » demonstrate a layering technique whereby two uranium solids, each with a different isotopic composition, are employed to form successive layers of UF6 on a surface. The result is an ultra-thin deposit of UF6 that bears an isotopic signature that is a composite of the two uranium sources. The reported deposition method has direct application to the development of unique analytical standards for nuclear safeguards and forensics.« less

Air sampling to assess potential generation of aerosolized viable bacteria during flow cytometric analysis of unfixed bacterial suspensions

PubMed Central

Carson, Christine F; Inglis, Timothy JJ

2018-01-01

This study investigated aerosolized viable bacteria in a university research laboratory during operation of an acoustic-assisted flow cytometer for antimicrobial susceptibility testing by sampling room air before, during and after flow cytometer use. The aim was to assess the risk associated with use of an acoustic-assisted flow cytometer analyzing unfixed bacterial suspensions. Air sampling in a nearby clinical laboratory was conducted during the same period to provide context for the existing background of microorganisms that would be detected in the air. The three species of bacteria undergoing analysis by flow cytometer in the research laboratory were Klebsiella pneumoniae, Burkholderia thailandensis and Streptococcus pneumoniae. None of these was detected from multiple 1000 L air samples acquired in the research laboratory environment. The main cultured bacteria in both locations were skin commensal and environmental bacteria, presumed to have been disturbed or dispersed in laboratory air by personnel movements during routine laboratory activities. The concentrations of bacteria detected in research laboratory air samples were reduced after interventional cleaning measures were introduced and were lower than those in the diagnostic clinical microbiology laboratory. We conclude that our flow cytometric analyses of unfixed suspensions of K. pneumoniae, B. thailandensis and S. pneumoniae do not pose a risk to cytometer operators or other personnel in the laboratory but caution against extrapolation of our results to other bacteria and/or different flow cytometric experimental procedures. PMID:29608197

Development of a diffraction imaging flow cytometer

PubMed Central

Jacobs, Kenneth M.; Lu, Jun Q.

2013-01-01

Diffraction images record angle-resolved distribution of scattered light from a particle excited by coherent light and can correlate highly with the 3D morphology of a particle. We present a jet-in-fluid design of flow chamber for acquisition of clear diffraction images in a laminar flow. Diffraction images of polystyrene spheres of different diameters were acquired and found to correlate highly with the calculated ones based on the Mie theory. Fast Fourier transform analysis indicated that the measured images can be used to extract sphere diameter values. These results demonstrate the significant potentials of high-throughput diffraction imaging flow cytometry for extracting 3D morphological features of cells. PMID:19794790

Copolymerization of UF Resins with Dimethylurea for Improving Storage Stability without Impairing Adhesive Performance.

PubMed

Pereira, Pedro; Pereira, João; Paiva, Nádia T; Ferra, João M; Martins, Jorge M; Carvalho, Luísa H; Magalhães, Fernão D

2018-06-19

Urea-formaldehyde (UF) resins are the most used resins in the wood industry due to high reactivity and low price. However, their reduced stability during storage is a drawback, imposing strict limits in terms of allowable shipping distances and storage times. This instability, manifested by viscosity increase that renders the resin unusable, occurs due to the progress of condensation reactions between the polymeric species present in the liquid medium. In order to achieve a stable resin formulation, dimethylurea (DMeU) was selected for being less reactive than urea. Dimethylurea is shown to co-polymerize with the UF polymer during the acidic synthesis condensation step. However, during storage it behaves like an end group blocker, due to its lower reactivity at basic pH. By adding 1.25% DMeU, it was possible to obtain a formulation that remained with stable viscosity during two-month storage at 40 °C. The reference UF resin remained stable only for eight days in these conditions. Wood particleboards produced with modified resins showed internal bond strengths of about 0.5 N·mm −2 , similar to the fresh reference UF resin, even when the resins were used after the two-month storage period. Formaldehyde content values were below the limit for E1 class, ≤8 mg/100 g oven dry board (EN 13986).

31 CFR 540.318 - Uranium Hexafluoride (UF6).

Code of Federal Regulations, 2012 CFR

2012-07-01

... 31 Money and Finance:Treasury 3 2012-07-01 2012-07-01 false Uranium Hexafluoride (UF6). 540.318... OF FOREIGN ASSETS CONTROL, DEPARTMENT OF THE TREASURY HIGHLY ENRICHED URANIUM (HEU) AGREEMENT ASSETS CONTROL REGULATIONS General Definitions § 540.318 Uranium Hexafluoride (UF6). The term uranium...

31 CFR 540.318 - Uranium Hexafluoride (UF6).

Code of Federal Regulations, 2011 CFR

2011-07-01

... 31 Money and Finance:Treasury 3 2011-07-01 2011-07-01 false Uranium Hexafluoride (UF6). 540.318... OF FOREIGN ASSETS CONTROL, DEPARTMENT OF THE TREASURY HIGHLY ENRICHED URANIUM (HEU) AGREEMENT ASSETS CONTROL REGULATIONS General Definitions § 540.318 Uranium Hexafluoride (UF6). The term uranium...

31 CFR 540.318 - Uranium Hexafluoride (UF6).

Code of Federal Regulations, 2013 CFR

2013-07-01

... 31 Money and Finance:Treasury 3 2013-07-01 2013-07-01 false Uranium Hexafluoride (UF6). 540.318... OF FOREIGN ASSETS CONTROL, DEPARTMENT OF THE TREASURY HIGHLY ENRICHED URANIUM (HEU) AGREEMENT ASSETS CONTROL REGULATIONS General Definitions § 540.318 Uranium Hexafluoride (UF6). The term uranium...

31 CFR 540.318 - Uranium Hexafluoride (UF6).

Code of Federal Regulations, 2014 CFR

2014-07-01

... 31 Money and Finance:Treasury 3 2014-07-01 2014-07-01 false Uranium Hexafluoride (UF6). 540.318... OF FOREIGN ASSETS CONTROL, DEPARTMENT OF THE TREASURY HIGHLY ENRICHED URANIUM (HEU) AGREEMENT ASSETS CONTROL REGULATIONS General Definitions § 540.318 Uranium Hexafluoride (UF6). The term uranium...

31 CFR 540.318 - Uranium Hexafluoride (UF6).

Code of Federal Regulations, 2010 CFR

2010-07-01

... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Uranium Hexafluoride (UF6). 540.318... OF FOREIGN ASSETS CONTROL, DEPARTMENT OF THE TREASURY HIGHLY ENRICHED URANIUM (HEU) AGREEMENT ASSETS CONTROL REGULATIONS General Definitions § 540.318 Uranium Hexafluoride (UF6). The term uranium...

Evaluating flow cytometer performance with weighted quadratic least squares analysis of LED and multi-level bead data

PubMed Central

Parks, David R.; Khettabi, Faysal El; Chase, Eric; Hoffman, Robert A.; Perfetto, Stephen P.; Spidlen, Josef; Wood, James C.S.; Moore, Wayne A.; Brinkman, Ryan R.

2017-01-01

We developed a fully automated procedure for analyzing data from LED pulses and multi-level bead sets to evaluate backgrounds and photoelectron scales of cytometer fluorescence channels. The method improves on previous formulations by fitting a full quadratic model with appropriate weighting and by providing standard errors and peak residuals as well as the fitted parameters themselves. Here we describe the details of the methods and procedures involved and present a set of illustrations and test cases that demonstrate the consistency and reliability of the results. The automated analysis and fitting procedure is generally quite successful in providing good estimates of the Spe (statistical photoelectron) scales and backgrounds for all of the fluorescence channels on instruments with good linearity. The precision of the results obtained from LED data is almost always better than for multi-level bead data, but the bead procedure is easy to carry out and provides results good enough for most purposes. Including standard errors on the fitted parameters is important for understanding the uncertainty in the values of interest. The weighted residuals give information about how well the data fits the model, and particularly high residuals indicate bad data points. Known photoelectron scales and measurement channel backgrounds make it possible to estimate the precision of measurements at different signal levels and the effects of compensated spectral overlap on measurement quality. Combining this information with measurements of standard samples carrying dyes of biological interest, we can make accurate comparisons of dye sensitivity among different instruments. Our method is freely available through the R/Bioconductor package flowQB. PMID:28160404

Estimated critical conditions for UF{sub 4}-oil systems in fully oil-reflected spherical geometry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Plaster, M.J.

Paraffinic oil has been exposed to UF{sub 6} gas in seal exhaust pumps and cascade equipment at the Portsmouth Gaseous Diffusion Plant. The resulting mixture is more nuclearly reactive than mixtures of UO{sub 2}F{sub 2} and H{sub 2}O and is not bounded by the subcritical mass limits presented in several nuclear criticality safety guides. The purpose of this analysis is to determine several critical parameters; specifically, (1) k{sub {infinity}} and the critical mass for several enrichments and moderation levels and (2) the mass limits for these mixtures. The estimated critical masses for the UF{sub 4}-oil systems are smaller than formore » the UO{sub 2}F{sub 2}-H{sub 2}O systems. The suggested mass limits for the UF{sub 4}-oil systems are 0.240, 0.280, 0.350, 0.430, and 0.670, and 1.170 kg {sup 235}U for enrichments of 100, 50, 20, 10, 5, and 3 wt.% {sup 235}U respectively.« less

Grafting of alginates on UF/NF ceramic membranes for wastewater treatment.

PubMed

Athanasekou, C P; Romanos, G E; Kordatos, K; Kasselouri-Rigopoulou, V; Kakizis, N K; Sapalidis, A A

2010-10-15

The mechanism of heavy metal ion removal in processes involving multi-layered tubular ultrafiltration and nanofiltration (UF/NF) membranes was investigated by conducting retention experiments in both flow-through and cross-flow modes. The prospect of the regeneration of the membranes through an acidic process was also examined and discussed. The UF/NF membranes were functionalised with alginates to develop hybrid inorganic/organic materials for continuous, single pass, wastewater treatment applications. The challenge laid in the induction of additional metal adsorption and improved regeneration capacity. This was accomplished by stabilizing alginates either into the pores or on the top-separating layer of the membrane. The preservation of efficient water fluxes at moderate trans-membrane pressures introduced an additional parameter that was pursued in parallel to the membrane modification process. The deposition and stabilization of alginates was carried out via physical (filtration/cross-linking) and chemical (grafting) procedures. The materials developed by means of the filtration process exhibited a 25-60% enhancement of their Cd(2+) binding capacity, depending on the amount of the filtered alginate solution. The grafting process led to the development of alginate layers with adequate stability under acidic regeneration conditions and metal retention enhancement of 25-180%, depending on the silane involved as grafting agent and the solvent of silanisation. 2010 Elsevier B.V. All rights reserved.

EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols

PubMed Central

Kalina, T; Flores-Montero, J; van der Velden, V H J; Martin-Ayuso, M; Böttcher, S; Ritgen, M; Almeida, J; Lhermitte, L; Asnafi, V; Mendonça, A; de Tute, R; Cullen, M; Sedek, L; Vidriales, M B; Pérez, J J; te Marvelde, J G; Mejstrikova, E; Hrusak, O; Szczepański, T; van Dongen, J J M; Orfao, A

2012-01-01

The EU-supported EuroFlow Consortium aimed at innovation and standardization of immunophenotyping for diagnosis and classification of hematological malignancies by introducing 8-color flow cytometry with fully standardized laboratory procedures and antibody panels in order to achieve maximally comparable results among different laboratories. This required the selection of optimal combinations of compatible fluorochromes and the design and evaluation of adequate standard operating procedures (SOPs) for instrument setup, fluorescence compensation and sample preparation. Additionally, we developed software tools for the evaluation of individual antibody reagents and antibody panels. Each section describes what has been evaluated experimentally versus adopted based on existing data and experience. Multicentric evaluation demonstrated high levels of reproducibility based on strict implementation of the EuroFlow SOPs and antibody panels. Overall, the 6 years of extensive collaborative experiments and the analysis of hundreds of cell samples of patients and healthy controls in the EuroFlow centers have provided for the first time laboratory protocols and software tools for fully standardized 8-color flow cytometric immunophenotyping of normal and malignant leukocytes in bone marrow and blood; this has yielded highly comparable data sets, which can be integrated in a single database. PMID:22948490

Uniform deposition of uranium hexafluoride (UF6): Standardized mass deposits and controlled isotopic ratios using a thermal fluorination method.

PubMed

McNamara, Bruce K; O'Hara, Matthew J; Casella, Andrew M; Carter, Jennifer C; Addleman, R Shane; MacFarlan, Paul J

2016-07-01

We report a convenient method for the generation of volatile uranium hexafluoride (UF6) from solid uranium oxides and other U compounds, followed by uniform deposition of low levels of UF6 onto sampling coupons. Under laminar flow conditions, UF6 is shown to interact with surfaces within a fixed reactor geometry to a highly predictable degree. We demonstrate the preparation of U deposits that range between approximately 0.01 and 500ngcm(-2). The data suggest the method can be extended to creating depositions at the sub-picogramcm(-2) level. The isotopic composition of the deposits can be customized by selection of the U source materials and we demonstrate a layering technique whereby two U solids, each with a different isotopic composition, are employed to form successive layers of UF6 on a surface. The result is an ultra-thin deposit that bears an isotopic signature that is a composite of the two U sources. The reported deposition method has direct application to the development of unique analytical standards for nuclear safeguards and forensics. Further, the method allows access to very low atomic or molecular coverages of surfaces. Copyright © 2016 Elsevier B.V. All rights reserved.

The Accuracy of the Sysmex UF-1000i in Urine Bacterial Detection Compared With the Standard Urine Analysis and Culture.

PubMed

Erdman, Patrick; Anderson, Brian; Zacko, J Christopher; Taylor, Kirk; Donaldson, Keri

2017-11-01

- Urinary tract infections are characterized by the presence of microbial pathogens within the urinary tract. They represent one of the most common infections in hospitalized and clinic patients. - To model the parameters of the Sysmex UF-1000i to the gold standard, urine culture, and to compare the detection of dipstick leukocyte esterase and nitrates to urine cultures and UF-1000i results. - Data were compared from urine samples collected in sterile containers for bacterial culture and microscopic analysis. One sample was used to inoculate a 5% sheep blood agar and MacConkey agar plate using a 0.001-mL calibrated loop. The second sample was analyzed by urinalysis-associated microscopy. The media plates were investigated for growth after 18 to 24 hours of aerobic incubation at 37°C. The second sample was analyzed for bacteria and leukocytes with the Sysmex UF-1000i according to the manufacturer's guidelines. Three definitions for culture results, sensitivity, and specificity at different cutoff values were calculated for the UF-1000i. - The negative predictive value for any positive culture in the adult population included in the study was 95.5%, and the negative predictive value for positive cultures containing growth of 100 000 or more colony-forming units was 99.3% using the Sysmex UF-1000i. - Sysmex UF-1000i showed 98% sensitivity and 93.7% specificity with a 95.5% negative predictive value. Thus, a negative screen with the UF-1000i using defined thresholds for white blood cell counts and bacteria was likely to be a true negative, decreasing the need for presumptive antibiotics.

Evaluating flow cytometer performance with weighted quadratic least squares analysis of LED and multi-level bead data.

PubMed

Parks, David R; El Khettabi, Faysal; Chase, Eric; Hoffman, Robert A; Perfetto, Stephen P; Spidlen, Josef; Wood, James C S; Moore, Wayne A; Brinkman, Ryan R

2017-03-01

We developed a fully automated procedure for analyzing data from LED pulses and multilevel bead sets to evaluate backgrounds and photoelectron scales of cytometer fluorescence channels. The method improves on previous formulations by fitting a full quadratic model with appropriate weighting and by providing standard errors and peak residuals as well as the fitted parameters themselves. Here we describe the details of the methods and procedures involved and present a set of illustrations and test cases that demonstrate the consistency and reliability of the results. The automated analysis and fitting procedure is generally quite successful in providing good estimates of the Spe (statistical photoelectron) scales and backgrounds for all the fluorescence channels on instruments with good linearity. The precision of the results obtained from LED data is almost always better than that from multilevel bead data, but the bead procedure is easy to carry out and provides results good enough for most purposes. Including standard errors on the fitted parameters is important for understanding the uncertainty in the values of interest. The weighted residuals give information about how well the data fits the model, and particularly high residuals indicate bad data points. Known photoelectron scales and measurement channel backgrounds make it possible to estimate the precision of measurements at different signal levels and the effects of compensated spectral overlap on measurement quality. Combining this information with measurements of standard samples carrying dyes of biological interest, we can make accurate comparisons of dye sensitivity among different instruments. Our method is freely available through the R/Bioconductor package flowQB. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

Neutron spectrometry for UF 6 enrichment verification in storage cylinders

DOE PAGES

Mengesha, Wondwosen; Kiff, Scott D.

2015-01-29

Verification of declared UF 6 enrichment and mass in storage cylinders is of great interest in nuclear material nonproliferation. Nondestructive assay (NDA) techniques are commonly used for safeguards inspections to ensure accountancy of declared nuclear materials. Common NDA techniques used include gamma-ray spectrometry and both passive and active neutron measurements. In the present study, neutron spectrometry was investigated for verification of UF 6 enrichment in 30B storage cylinders based on an unattended and passive measurement approach. MCNP5 and Geant4 simulated neutron spectra, for selected UF 6 enrichments and filling profiles, were used in the investigation. The simulated neutron spectra weremore » analyzed using principal component analysis (PCA). The PCA technique is a well-established technique and has a wide area of application including feature analysis, outlier detection, and gamma-ray spectral analysis. Results obtained demonstrate that neutron spectrometry supported by spectral feature analysis has potential for assaying UF 6 enrichment in storage cylinders. Thus the results from the present study also showed that difficulties associated with the UF 6 filling profile and observed in other unattended passive neutron measurements can possibly be overcome using the approach presented.« less

Capture of circulating tumor cells using photoacoustic flowmetry and two phase flow

NASA Astrophysics Data System (ADS)

O'Brien, Christine M.; Rood, Kyle D.; Bhattacharyya, Kiran; DeSouza, Thiago; Sengupta, Shramik; Gupta, Sagar K.; Mosley, Jeffrey D.; Goldschmidt, Benjamin S.; Sharma, Nikhilesh; Viator, John A.

2012-06-01

Melanoma is the deadliest form of skin cancer, yet current diagnostic methods are unable to detect early onset of metastatic disease. Patients must wait until macroscopic secondary tumors form before malignancy can be diagnosed and treatment prescribed. Detection of cells that have broken off the original tumor and travel through the blood or lymph system can provide data for diagnosing and monitoring metastatic disease. By irradiating enriched blood samples spiked with cultured melanoma cells with nanosecond duration laser light, we induced photoacoustic responses in the pigmented cells. Thus, we can detect and enumerate melanoma cells in blood samples to demonstrate a paradigm for a photoacoustic flow cytometer. Furthermore, we capture the melanoma cells using microfluidic two phase flow, a technique that separates a continuous flow into alternating microslugs of air and blood cell suspension. Each slug of blood cells is tested for the presence of melanoma. Slugs that are positive for melanoma, indicated by photoacoustic waves, are separated from the cytometer for further purification and isolation of the melanoma cell. In this paper, we evaluate the two phase photoacoustic flow cytometer for its ability to detect and capture metastastic melanoma cells in blood.

3D flow focusing for microfluidic flow cytometry with ultrasonics

NASA Astrophysics Data System (ADS)

Gnyawali, Vaskar; Strohm, Eric M.; Daghighi, Yasaman; van de Vondervoort, Mia; Kolios, Michael C.; Tsai, Scott S. H.

2015-11-01

We are developing a flow cytometer that detects unique acoustic signature waves generated from single cells due to interactions between the cells and ultrasound waves. The generated acoustic waves depend on the size and biomechanical properties of the cells and are sufficient for identifying cells in the medium. A microfluidic system capable of focusing cells through a 10 x 10 μm ultrasound beam cross section was developed to facilitate acoustic measurements of single cells. The cells are streamlined in a hydro-dynamically 3D focused flow in a 300 x 300 μm channel made using PDMS. 3D focusing is realized by lateral sheath flows and an inlet needle (inner diameter 100 μm). The accuracy of the 3D flow focusing is measured using a dye and detecting its localization using confocal microscopy. Each flowing cell would be probed by an ultrasound pulse, which has a center frequency of 375 MHz and bandwidth of 250 MHz. The same probe would also be used for recording the scattered waves from the cells, which would be processed to distinguish the physical and biomechanical characteristics of the cells, eventually identifying them. This technique has potential applications in detecting circulating tumor cells, blood cells and blood-related diseases.

Flow cytometry of mammalian sperm: progress in DNA and morphology measurement.

PubMed

Pinkel, D; Dean, P; Lake, S; Peters, D; Mendelsohn, M; Gray, J; Van Dilla, M; Gledhill, B

1979-01-01

Variability in DNA content and head shape of mammalian sperm are potentially useful markers for flow cytometric monitoring of genetic damage in spermatogenic cells. The high refractive index and extreme flatness of the sperm heads produce an optical effect which interferes with DNA measurements in flow cytometers which have dye excitation and fluorescence light collection normal to the axis of flow. Orientation of sperm in flow controls this effect and results in coefficients of variation of 2.5% and 4.2%, respectively, for DNA measurements of mouse and human sperm. Alternatively, the optical effect can be used to generate shape-related information. Measurements on randomly oriented sperm from three mammalian species using a pair of fluorescence detectors indicate that large shape differences are detectable. Acriflavine-Feulgen stained sperm nuclei are significantly bleached during flow cytometric measurements at power levels routinely used in many flow cytometers. Dual beam studies of this phenomenon indicate it may be useful in detecting abnormally shaped sperm.

Relationship between conventional culture and flow cytometry for the diagnosis of urinary tract infection.

PubMed

García-Coca, Marta; Gadea, Ignacio; Esteban, Jaime

2017-06-01

Urine culture is the gold standard for the diagnosis of urinary tract infections (UTI). The use of flow cytometry analyzers (FCA) prior to culture allows for the quantification and recognition of cell components in urine to be automated and makes it possible to relate these data to the urine pathogens subsequently identified in cultures. Urine samples were assessed with the Sysmex UF-1000i analyzer. Those that met the criteria for culture (> 25 leukocytes/μL or > 385 bacteria/μL) were subjected to quantitative urine culture on chromogenic agar. Counts of red blood cells (RBC), white blood cells (WBC), epithelial cells (EC), and the kind of microorganisms identified in cultures were evaluated. A total of 17,483 samples were processed by FCA. Of these, 9057 met the criteria for culture. Urine cultures were reduced by 48.2%. The most common urine pathogen was Escherichia coli (60.3%). Negative urine cultures were significantly (p < 0.001) associated with a lower WBC count than urine with E. coli, Klebsiella spp. and Proteus spp., but urine with Enterococcus spp. had a lower WBC than negative urine. Contaminated urine had a significantly (p < 0.001) lower WBC than urine with E. coli, Klebsiella spp. and Proteus spp., but no differences were found for Enterococcus spp. (p = 0.729). Negative urine cultures had significantly (p < 0.05) higher EC than all positive urine samples. Contaminated urine was associated (p < 0.001) with higher EC than cultures with E. coli and Klebsiella spp., in comparison with cultures with Enterococcus spp. (p = 0.091) and Proteus spp. (p = 0.251). The use of the Sysmex UF-1000i flow cytometer for screening urine samples allows for a reduction in the number of urine cultures. WBC values correlate well with the main urine pathogens related to UTI. The results observed for Enterococcus spp. suggest a low impact of these pathogens as a cause of UTI.

Clinical and laboratory applications of slide-based cytometry with the LSC, SFM, and the iCYTE imaging cytometer instruments

NASA Astrophysics Data System (ADS)

Bocsi, Jozsef; Luther, Ed; Mittag, Anja; Jensen, Ingo; Sack, Ulrich; Lenz, Dominik; Trezl, Lajos; Varga, Viktor S.; Molnar, Beea; Tarnok, Attila

2004-06-01

Background: Slide based cytometry (SBC) is a technology for the rapid stoichiometric analysis of cells fixed to surfaces. Its applications are highly versatile and ranges from the clinics to high throughput drug discovery. SBC is realized in different instruments such as the Laser Scanning Cytometer (LSC) and Scanning Fluorescent Microscope (SFM) and the novel inverted microscope based iCyte image cytometer (Compucyte Corp.). Methods: Fluorochrome labeled specimens were immobilized on microscopic slides. They were placed on a conventional fluorescence microscope and analyzed by photomultiplayers or digital camera. Data comparable to flow cytometry were generated. In addition, each individual event could be visualized. Applications: The major advantage of instruments is the combination of two features: a) the minimal sample volume needed, and b) the connection of fluorescence data and morphological information. Rare cells were detected, frequency of apoptosis by myricetin formaldehyde and H2O2 mixtures was determined;. Conclusion: LSC, SFM and the novel iCyte have a wide spectrum of applicability in SBC and can be introduced as a standard technology for multiple settings. In addition, the iCyte and SFM instrument is suited for high throughput screening by automation and may be in future adapted to telepathology due to their high quality images. (This study was supported by the IZKF-Leipzig, Germany and T 034245 OTKA, Hungary)

Analysis of UV-excited fluorochromes by flow cytometry using near-ultraviolet laser diodes.

PubMed

Telford, William G

2004-09-01

Violet laser diodes have become common and reliable laser sources for benchtop flow cytometers. While these lasers are very useful for a variety of violet and some ultraviolet-excited fluorochromes (e.g., DAPI), they do not efficiently excite most UV-stimulated probes. In this study, the next generation of InGaN near-UV laser diodes (NUVLDs) emitting in the 370-375-nm range have been evaluated as laser sources for cuvette-based flow cytometers. Several NUVLDs, ranging in wavelength from 370 to 374 nm and in power level from 1.5 to 10 mW, were mounted on a BD Biosciences LSR II and evaluated for their ability to excite cells labeled with the UV DNA binding dye DAPI, several UV phenotyping fluorochromes (including Alexa Fluor 350, Marina Blue, and quantum dots), and the fluorescent calcium chelator indo-1. NUVLDs at the 8-10-mW power range gave detection sensitivity levels comparable to more powerful solid-state and ion laser sources, using low-fluorescence microsphere beads as measurement standards. NUVLDs at all tested power levels allowed extremely high-resolution DAPI cell cycle analysis, and sources in the 8-10-mW power range excited Alexa Fluor 350, Marina Blue, and a variety of quantum dots at virtually the same signal-to-noise ratios as more powerful UV sources. These evaluations indicate that near-UV laser diodes installed on a cuvette-based flow cytometer performed nearly as well as more powerful solid-state UV lasers on the same instrumentation, and comparably to more powerful ion lasers on a jet-in-air system, and. Despite their limited power, integration of these small and inexpensive lasers into benchtop flow cytometers should allow the use of flow cytometric applications requiring UV excitation on a wide variety of instruments. Copyright 2004 Wiley-Liss, Inc.

Comparison of the Automated cobas u 701 Urine Microscopy and UF-1000i Flow Cytometry Systems and Manual Microscopy in the Examination of Urine Sediments.

PubMed

Lee, Wonmok; Ha, Jung-Sook; Ryoo, Nam-Hee

2016-09-01

The cobas u 701, a new automated image-based urine sediment analyzer, was introduced recently. In this study, we compared its performance with that of UF-1000i flow cytometry and manual microscopy in the examination of urine sediments. Precision, linearity, and carry-over were determined for the two urine sediment analyzers. For a comparison of the method, 300 urine samples were examined by the automated analyzers and by manual microscopy using a KOVA chamber. Within-run coefficients of variation (CVs) for the control materials were 7.0-8.8% and 1.7-5.7% for the cobas u 701 and UF-1000i systems, respectively. Between-run CVs were 8.5-9.8% and 2.7-5.4%, respectively. Both instruments showed good linearity and negligible carry-over. For red blood cells (RBC), white blood cells (WBC), and epithelial cells (EPI), the overall concordance rates within one grade of difference among the three methods were good (78.6-86.0%, 88.7-93.8%, and 81.3-90.7%, respectively). The concordance rate for casts was poor (66.5-68.9%). Compared with manual microscopy, the two automated sediment analyzers tested in this study showed satisfactory analytical performances for RBC, WBC, and EPI. However, for other urine sediment particles confirmation by visual microscopy is still required. © 2016 Wiley Periodicals, Inc.

UF insulation ban overturned by federal appeals court: but safety still in doubt

DOE Office of Scientific and Technical Information (OSTI.GOV)

Betts, M.

1983-05-02

Unless the federal appeals court honors a US Consumer Product Safety Commission (CPSC) request to reconsider, the ban on sales of urea-formaldehyde (UF) foam for insulation will be lifted for want of evidence that it constitutes a health threat. The CPSC defends its analysis of flu-like symptoms and a form of cancer in laboratory animals which led to the ban on sales for school and home use, a ban which effectively eliminated all UF markets. Some buildings have replaced the foam with fiber and cellulose products at considerable expense. In overturning the ban, the court agreed with the UF foammore » industry challenge of CPSC evidence, but made no claim about UF safety. (DCK)« less

miRNA detection at single-cell resolution using microfluidic LNA flow-FISH

DOE PAGES

Wu, Meiye; Piccini, Matthew Ernest; Koh, Chung -Yan; ...

2014-08-20

Flow cytometry in combination with fluorescent in situ hybridization (flow-FISH) is a powerful technique that can be utilized to rapidly detect nucleic acids at single-cell resolution without the need for homogenization or nucleic acid extraction. Here, we describe a microfluidic-based method which enables the detection of microRNAs or miRNAs in single intact cells by flow-FISH using locked nucleic acid (LNA)-containing probes. Our method can be applied to all RNA species including mRNA and small noncoding RNA and is suitable for multiplexing with protein immunostaining in the same cell. For demonstration of our method, this chapter details the detection of miR155more » and CD69 protein in PMA and ionomycin-stimulated Jurkat cells. Here, we also include instructions on how to set up a microfluidic chip sample preparation station to prepare cells for imaging and analysis on a commercial flow cytometer or a custom-built micro-flow cytometer.« less

Natural phenomena evaluations of the K-25 site UF{sub 6} cylinder storage yards

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fricke, K.E.

1996-09-15

The K-25 Site UF{sub 6} cylinder storage yards are used for the temporary storage of UF{sub 6} normal assay cylinders and long-term storage of other UF{sub 6} cylinders. The K-25 Site UF{sub 6} cylinder storage yards consist of six on-site areas: K-1066-B, K-1066-E, K-1066-F, K-1066-J, K-1066-K and K-1066-L. There are no permanent structures erected on the cylinder yards, except for five portable buildings. The operating contractor for the K-25 Site is preparing a Safety Analysis Report (SAR) to examine the safety related aspects of the K-25 Site UF{sub 6} cylinder storage yards. The SAR preparation encompasses many tasks terminating inmore » consequence analysis for the release of gaseous and liquid UF{sub 6}, one of which is the evaluation of natural phenomena threats, such as earthquakes, floods, and winds. In support of the SAR, the six active cylinder storage yards were evaluated for vulnerabilities to natural phenomena, earthquakes, high winds and tornados, tornado-generated missiles, floods (local and regional), and lightning. This report summarizes those studies. 30 refs.« less

Reliable and accurate CD4+ T cell count and percent by the portable flow cytometer CyFlow MiniPOC and "CD4 Easy Count Kit-Dry", as revealed by the comparison with the gold standard dual platform technology.

PubMed

Nasi, Milena; De Biasi, Sara; Bianchini, Elena; Gibellini, Lara; Pinti, Marcello; Scacchetti, Tiziana; Trenti, Tommaso; Borghi, Vanni; Mussini, Cristina; Cossarizza, Andrea

2015-01-01

An accurate and affordable CD4+ T cells count is an essential tool in the fight against HIV/AIDS. Flow cytometry (FCM) is the "gold standard" for counting such cells, but this technique is expensive and requires sophisticated equipment, temperature-sensitive monoclonal antibodies (mAbs) and trained personnel. The lack of access to technical support and quality assurance programs thus limits the use of FCM in resource-constrained countries. We have tested the accuracy, the precision and the carry-over contamination of Partec CyFlow MiniPOC, a portable and economically affordable flow cytometer designed for CD4+ count and percentage, used along with the "CD4% Count Kit-Dry". Venous blood from 59 adult HIV+ patients (age: 25-58 years; 43 males and 16 females) was collected and stained with the "MiniPOC CD4% Count Kit-Dry". CD4+ count and percentage were then determined in triplicate by the CyFlow MiniPOC. In parallel, CD4 count was performed using mAbs and a CyFlow Counter, or by a dual platform system (from Beckman Coulter) based upon Cytomic FC500 ("Cytostat tetrachrome kit" for mAbs) and Coulter HmX Hematology Analyzer (for absolute cell count). The accuracy of CyFlow MiniPOC against Cytomic FC500 showed a correlation coefficient (CC) of 0.98 and 0.97 for CD4+ count and percentage, respectively. The accuracy of CyFlow MiniPOC against CyFlow Counter showed a CC of 0.99 and 0.99 for CD4 T cell count and percentage, respectively. CyFlow MiniPOC showed an excellent repeatability: CD4+ cell count and percentage were analyzed on two instruments, with an intra-assay precision below ± 5% deviation. Finally, there was no carry-over contamination for samples at all CD4 values, regardless of their position in the sequence of analysis. The cost-effective CyFlow MiniPOC produces rapid, reliable and accurate results that are fully comparable with those from highly expensive dual platform systems.

Automated processing integrated with a microflow cytometer for pathogen detection in clinical matrices

PubMed Central

Golden, J.P.; Verbarg, J.; Howell, P.B.; Shriver-Lake, L.C.; Ligler, F.S.

2012-01-01

A spinning magnetic trap (MagTrap) for automated sample processing was integrated with a microflow cytometer capable of simultaneously detecting multiple targets to provide an automated sample-to-answer diagnosis in 40 min. After target capture on fluorescently coded magnetic microspheres, the magnetic trap automatically concentrated the fluorescently coded microspheres, separated the captured target from the sample matrix, and exposed the bound target sequentially to biotinylated tracer molecules and streptavidin-labeled phycoerythrin. The concentrated microspheres were then hydrodynamically focused in a microflow cytometer capable of 4-color analysis (two wavelengths for microsphere identification, one for light scatter to discriminate single microspheres and one for phycoerythrin bound to the target). A three-fold decrease in sample preparation time and an improved detection limit, independent of target preconcentration, was demonstrated for detection of Escherichia coli 0157:H7 using the MagTrap as compared to manual processing. Simultaneous analysis of positive and negative controls, along with the assay reagents specific for the target, was used to obtain dose–response curves, demonstrating the potential for quantification of pathogen load in buffer and serum. PMID:22960010

Method for selectively removing fluorine and fluorine-containing contaminants from gaseous UF.sub.6

DOEpatents

Jones, Robert L.; Otey, Milton G.; Perkins, Roy W.

1982-01-01

This invention is a method for effecting preferential removal and immobilization of certain gaseous contaminants from gaseous UF.sub.6. The contaminants include fluorine and fluorides which are more reactive with CaCO.sub.3 than is UF.sub.6. The method comprises contacting the contaminant-carrying UF.sub.6 with particulate CaCO.sub.3 at a temperature effecting reaction of the contaminant and the CaCO.sub.3.

Ultraviolet 320 nm laser excitation for flow cytometry.

PubMed

Telford, William; Stickland, Lynn; Koschorreck, Marco

2017-04-01

Although multiple lasers and high-dimensional analysis capability are now standard on advanced flow cytometers, ultraviolet (UV) lasers (usually 325-365 nm) remain an uncommon excitation source for cytometry. This is primarily due to their cost, and the small number of applications that require this wavelength. The development of the Brilliant Ultraviolet (BUV fluorochromes, however, has increased the importance of this formerly niche excitation wavelength. Historically, UV excitation was usually provided by water-cooled argon- and krypton-ion lasers. Modern flow cytometers primary rely on diode pumped solid state lasers emitting at 355 nm. While useful for all UV-excited applications, DPSS UV lasers are still large by modern solid state laser standards, and remain very expensive. Smaller and cheaper near UV laser diodes (NUVLDs) emitting at 375 nm make adequate substitutes for 355 nm sources in many situations, but do not work as well with very short wavelength probes like the fluorescent calcium chelator indo-1. In this study, we evaluate a newly available UV 320 nm laser for flow cytometry. While shorter in wavelength that conventional UV lasers, 320 is close to the 325 nm helium-cadmium wavelength used in the past on early benchtop cytometers. A UV 320 nm laser was found to excite almost all Brilliant Ultraviolet dyes to nearly the same level as 355 nm sources. Both 320 nm and 355 nm sources worked equally well for Hoechst and DyeCycle Violet side population analysis of stem cells in mouse hematopoetic tissue. The shorter wavelength UV source also showed excellent excitation of indo-1, a probe that is not compatible with NUVLD 375 nm sources. In summary, a 320 nm laser module made a suitable substitute for conventional 355 nm sources. This laser technology is available in a smaller form factor than current 355 nm units, making it useful for small cytometers with space constraints. © 2017 International Society for Advancement of Cytometry. © 2017 International

The Comparing Options for Management: Patient-centered Results for Uterine Fibroids (COMPARE-UF) Registry: Rationale and Design.

PubMed

Stewart, Elizabeth A; Lytle, Barbara L; Thomas, Laine; Wegienka, Ganesa R; Jacoby, Vanessa; Diamond, Michael P; Nicholson, Wanda K; Anchan, Raymond M; Venable, Sateria; Wallace, Kedra; Marsh, Erica E; Maxwell, George L; Borah, Bijan J; Catherino, William H; Myers, Evan R

2018-05-08

To design and establish a uterine fibroid (UF) registry based in the United States (US) to provide comparative effectiveness data regarding UF treatment. We report here the design and initial recruitment for the Comparing Options for Management: Patient-centered Results for Uterine Fibroids (COMPARE-UF) registry (Clinicaltrials.gov, NCT02260752), funded by the Agency for Healthcare Research and Quality (AHRQ) in collaboration with the-Patient-Centered Outcomes Research Institute (PCORI). COMPARE-UF is designed to help answer critical questions about treatment options for women with symptomatic UF. Women undergoing a procedure for UF (hysterectomy, myomectomy [abdominal, hysteroscopic, vaginal and laparoscopic/robotic], endometrial ablation, radiofrequency fibroid ablation, uterine artery embolization, magnetic resonance guided focused ultrasound or progestin-releasing intrauterine device insertion) at one of the COMPARE-UF sites are invited to participate in a prospective registry with three years follow-up for post-procedural outcomes. Enrolled participants provide annual follow-up through an online portal or through traditional phone contact. A central data abstraction center provides information obtained from imaging, operative or procedural notes and pathology reports. Women with uterine fibroids and other stakeholders are a key part of the COMPARE-UF registry and participate at all points from study design to dissemination of results. We built a network of nine clinical sites across the US with expertise in the care of women with UF to capture geographic, racial, ethnic and procedural diversity. Of the initial 2031 women enrolled in COMPARE-UF, 42% are self-identified as Black or African-American and 40% are age 40 years or younger with 16% of participants under age 35. Women undergoing myomectomy comprise the largest treatment group at 46% of all procedures with laparoscopic or robotic myomectomy comprising the largest subset of myomectomies at 19% of all

Integrating UF6 Cylinder RF Tracking With Continuous Load Cell Monitoring for Verifying Declared UF6 Feed and Withdrawal Operations Verifying Declared UF6 Feed and Withdrawal Operations

DOE Office of Scientific and Technical Information (OSTI.GOV)

Krichinsky, Alan M; Miller, Paul; Pickett, Chris A

2009-01-01

Oak Ridge National Laboratory is demonstrating the integration of UF6 cylinder tracking, using RF technology, with continuous load cell monitoring (CLCM) at mock UF6 feed and withdrawal (F&W) stations. CLCM and cylinder tracking are two of several continuous-monitoring technologies that show promise in providing integrated safeguards of F&W operations at enrichment plants. Integrating different monitoring technologies allows advanced, automated event processing to screen innocuous events thereby minimizing false alerts to independent inspectors. Traditionally, international inspectors rely on batch verification of material inputs and outputs derived from operator declarations and periodic on-site inspections at uranium enrichment plants or other nuclear processingmore » facilities. Continuously monitoring F&W activities between inspections while providing filtered alerts of significant operational events will substantially increase the amount of valuable information available to inspectors thereby promising to enhance the effectiveness of safeguards and to improve efficiency in conducting on-site inspections especially at large plants for ensuring that all operations are declared.« less

Line-Focused Optical Excitation of Parallel Acoustic Focused Sample Streams for High Volumetric and Analytical Rate Flow Cytometry.

PubMed

Kalb, Daniel M; Fencl, Frank A; Woods, Travis A; Swanson, August; Maestas, Gian C; Juárez, Jaime J; Edwards, Bruce S; Shreve, Andrew P; Graves, Steven W

2017-09-19

Flow cytometry provides highly sensitive multiparameter analysis of cells and particles but has been largely limited to the use of a single focused sample stream. This limits the analytical rate to ∼50K particles/s and the volumetric rate to ∼250 μL/min. Despite the analytical prowess of flow cytometry, there are applications where these rates are insufficient, such as rare cell analysis in high cellular backgrounds (e.g., circulating tumor cells and fetal cells in maternal blood), detection of cells/particles in large dilute samples (e.g., water quality, urine analysis), or high-throughput screening applications. Here we report a highly parallel acoustic flow cytometer that uses an acoustic standing wave to focus particles into 16 parallel analysis points across a 2.3 mm wide optical flow cell. A line-focused laser and wide-field collection optics are used to excite and collect the fluorescence emission of these parallel streams onto a high-speed camera for analysis. With this instrument format and fluorescent microsphere standards, we obtain analysis rates of 100K/s and flow rates of 10 mL/min, while maintaining optical performance comparable to that of a commercial flow cytometer. The results with our initial prototype instrument demonstrate that the integration of key parallelizable components, including the line-focused laser, particle focusing using multinode acoustic standing waves, and a spatially arrayed detector, can increase analytical and volumetric throughputs by orders of magnitude in a compact, simple, and cost-effective platform. Such instruments will be of great value to applications in need of high-throughput yet sensitive flow cytometry analysis.

Flow Cytometry Technician | Center for Cancer Research

Cancer.gov

PROGRAM DESCRIPTION The Basic Science Program (BSP) pursues independent, multidisciplinary research in basic and applied molecular biology, immunology, retrovirology, cancer biology, and human genetics. Research efforts and support are an integral part of the Center for Cancer Research (CCR) at the Frederick National Laboratory for Cancer Research (FNLCR). KEY ROLES/RESPONSIBILITIES The Flow Cytometry Core (Flow Core) of the Cancer and Inflammation Program (CIP) is a service core which supports the research efforts of the CCR by providing expertise in the field of flow cytometry (using analyzers and sorters) with the goal of gaining a more thorough understanding of the biology of cancer and cancer cells. The Flow Core provides service to 12-15 CIP laboratories and more than 22 non-CIP laboratories. Flow core staff provide technical advice on the experimental design of applications, which include immunological phenotyping, cell function assays, and cell cycle analysis. Work is performed per customer requirements, and no independent research is involved. The Flow Cytometry Technician will be responsible for: Monitor performance of and maintain high dimensional flow cytometer analyzers and cell sorters Operate high dimensional flow cytometer analyzers and cell sorters Monitoring lab supply levels and order lab supplies, perform various record keeping responsibilities Assist in the training of scientific end users on the use of flow cytometry in their research, as well as how to operate and troubleshoot the bench-top analyzer instruments Experience with sterile technique and tissue culture

Exergy analysis of an industrial-scale ultrafiltrated (UF) cheese production plant: a detailed survey

NASA Astrophysics Data System (ADS)

Nasiri, Farshid; Aghbashlo, Mortaza; Rafiee, Shahin

2017-02-01

In this study, a detailed exergy analysis of an industrial-scale ultrafiltrated (UF) cheese production plant was conducted based on actual operational data in order to provide more comprehensive insights into the performance of the whole plant and its main subcomponents. The plant included four main subsystems, i.e., steam generator (I), above-zero refrigeration system (II), Bactocatch-assisted pasteurization line (III), and UF cheese production line (IV). In addition, this analysis was aimed at quantifying the exergy destroyed in processing a known quantity of the UF cheese using the mass allocation method. The specific exergy destruction of the UF cheese production was determined at 2330.42 kJ/kg. The contributions of the subsystems I, II, III, and IV to the specific exergy destruction of the UF cheese production were computed as 1337.67, 386.18, 283.05, and 323.51 kJ/kg, respectively. Additionally, it was observed through the analysis that the steam generation system had the largest contribution to the thermodynamic inefficiency of the UF cheese production, accounting for 57.40 % of the specific exergy destruction. Generally, the outcomes of this survey further manifested the benefits of applying exergy analysis for design, analysis, and optimization of industrial-scale dairy processing plants to achieve the most cost-effective and environmentally-benign production strategies.

An experimental investigation of (UF-235)6 fission nuclear-pumped lasers

NASA Technical Reports Server (NTRS)

Miley, G. H.

1979-01-01

A UF6 handling system was designed for use in conjunction with the existing nuclear-pumped laser vacuum system at a nuclear reactor laboratory to perform the experiments described above. A modification to separate the gas fill system from the vacuum system and thus greatly reduce its volume is described as well as operating procedures for the first controlled nuclear pumping experiments with UF6 vapor contained in the laser cell.

Synthesis of phenol-urea-formaldehyde cocondensed resins from UF-concentrate and phenol

Treesearch

Bunchiro Tomita; Mashiko Ohyama; Chung-Yun Hse

1994-01-01

A new synthetic method to obtain phenol-urea-formaldehyde cocondensed resins was developed by reacting phenol with "UF-concentrate", which is a kind of urea-formaldehyde (UF) resin prepared with a high molar ratio of formaldehyde to urea (F/U) such as above 2.5. The products were analyzed with 13C-NMR spectroscopy and gel permeation...

Continuous-Wave Cavity Ring-Down Spectroscopy in a Pulsed Uniform Supersonic Flow

NASA Astrophysics Data System (ADS)

Thawoos, Shameemah; Suas-David, Nicolas; Suits, Arthur

2017-06-01

We introduce a new approach that couples a pulsed uniform supersonic flow with high sensitivity continuous wave cavity ringdown spectroscopy (UF-CRDS) operated in the near infrared (NIR). This combination is related to the CRESU technique developed in France and used for many years to study reaction kinetics at low temperature, and to the microwave based chirped-pulse uniform supersonic flow spectrometer (CPUF) developed in our group which has successfully demonstrated the use of pulsed uniform supersonic flow to probe reaction dynamics at temperatures as low as 22 K. CRDS operated with NIR permits access to the first overtones of C-H and O-H stretching/bending which, in combination with its extraordinary sensitivity opens new experiments complementary to the CPUF technique. The UF-CRDS apparatus (Figure) utilizes the pulsed uniform flow produced by means of a piezo-electric stack valve in combination with a Laval nozzle. At present, two machined aluminum Laval nozzles designed for carrier gases Ar and He generate flows with a temperature of approximately 25 K and pressure around 0.15 mbar. This flow is probed by an external cavity diode laser in the NIR (1280-1380 nm). Laval nozzles designed using a newly developed MATLAB-based program will be used in the future. A detailed illustration of the novel UF-CRDS instrumentation and its performance will be presented along with future directions and applications. I. Sims, J. L. Queffelec, A. Defrance, C. Rebrion-Rowe, D. Travers, P. Bocherel, B. Rowe, I. W. Smith, J. Chem. Phys. 100, 4229-4241, (1994). C. Abeysekera, B. Joalland, N. Ariyasingha, L. N. Zack, I. R. Sims, R. W. Field, A. G. Suits, J. Phys. Chem. Lett. 6, 1599-1604, (2015). N. Suas-David, T. Vanfleteren, T. Foldes, S. Kassi, R. Georges, M. Herman, J. Phys. Chem.A, 119, 10022-10034, (2015). C. Abeysekera, B. Joalland, Y. Shi, A. Kamasah, J. M. Oldham, A. G. Suits, Rev. Sci. Instrum. 85, 116107, (2014).

Thermodynamic assessment of the LiF-ThF4-PuF3-UF4 system

NASA Astrophysics Data System (ADS)

Capelli, E.; Beneš, O.; Konings, R. J. M.

2015-07-01

The LiF-ThF4-PuF3-UF4 system is the reference salt mixture considered for the Molten Salt Fast Reactor (MSFR) concept started with PuF3. In order to obtain the complete thermodynamic description of this quaternary system, two binary systems (ThF4-PuF3 and UF4-PuF3) and two ternary systems (LiF-ThF4-PuF3 and LiF-UF4-PuF3) have been assessed for the first time. The similarities between CeF3/PuF3 and ThF4/UF4 compounds have been taken into account for the presented optimization as well as in the experimental measurements performed, which have confirmed the temperatures predicted by the model. Moreover, the experimental results and the thermodynamic database developed have been used to identify potential compositions for the MSFR fuel and to evaluate the influence of partial substitution of ThF4 by UF4 in the salt.

Automated processing integrated with a microflow cytometer for pathogen detection in clinical matrices.

PubMed

Golden, J P; Verbarg, J; Howell, P B; Shriver-Lake, L C; Ligler, F S

2013-02-15

A spinning magnetic trap (MagTrap) for automated sample processing was integrated with a microflow cytometer capable of simultaneously detecting multiple targets to provide an automated sample-to-answer diagnosis in 40 min. After target capture on fluorescently coded magnetic microspheres, the magnetic trap automatically concentrated the fluorescently coded microspheres, separated the captured target from the sample matrix, and exposed the bound target sequentially to biotinylated tracer molecules and streptavidin-labeled phycoerythrin. The concentrated microspheres were then hydrodynamically focused in a microflow cytometer capable of 4-color analysis (two wavelengths for microsphere identification, one for light scatter to discriminate single microspheres and one for phycoerythrin bound to the target). A three-fold decrease in sample preparation time and an improved detection limit, independent of target preconcentration, was demonstrated for detection of Escherichia coli 0157:H7 using the MagTrap as compared to manual processing. Simultaneous analysis of positive and negative controls, along with the assay reagents specific for the target, was used to obtain dose-response curves, demonstrating the potential for quantification of pathogen load in buffer and serum. Published by Elsevier B.V.

ELECTROLYTIC PREPARATION OF UF$sub 4$

DOEpatents

Allen, A.L.; Anderson, R.W.; Powell, E.W.

1958-11-01

A method is presented for converting hexavalent aranium to uranium tetrafluoride. The method consists of electrolyzing a solution of uranyl fluoride in hydrofluoric acld at about 90 icient laborato C. The uranyl ions are reduced at the cathode and a hydrated uranium tetrafluoride precipitates. The precipitate is separated and subsequently dehydrated to UF/sub 4/.

Analysis of UF6 breeder reactor power plants

NASA Technical Reports Server (NTRS)

Clement, J. D.; Rust, J. H.

1976-01-01

Gaseous UF6 fueled breeder reactor design and technical applications of such concepts are summarized. Special attention was given to application in nuclear power plants and to reactor efficiency and safety factors.

A Burst Mode, Ultrahigh Temperature UF4 Vapor Core Reactor Rankine Cycle Space Power System Concept

NASA Technical Reports Server (NTRS)

Dugan, E. T.; Kahook, S. D.; Diaz, N. J.

1996-01-01

Static and dynamic neutronic analyses have been performed on an innovative burst mode (100's of MW output for a few thousand seconds) Ulvahigh Temperature Vapor Core Reactor (UTVR) space nuclear power system. The NVTR employs multiple, neutronically-coupled fissioning cores and operates on a direct, closed Rankine cycle using a disk Magnetohydrodynamic (MHD) generater for energy conversion. The UTVR includes two types of fissioning core regions: (1) the central Ultrahigh Temperature Vapor Core (UTVC) which contains a vapor mixture of highly enriched UF4 fuel and a metal fluoride working fluid and (2) the UF4 boiler column cores located in the BeO moderator/reflector region. The gaseous nature of the fuel the fact that the fuel is circulating, the multiple coupled fissioning cores, and the use of a two phase fissioning fuel lead to unique static and dynamic neutronic characteristics. Static neutronic analysis was conducted using two-dimensional S sub n, transport theory calculations and three-dimensional Monte Carlo transport theory calculations. Circulating-fuel, coupled-core point reactor kinetics equations were used for analyzing the dynamic behavior of the UTVR. In addition to including reactivity feedback phenomena associated with the individual fissioning cores, the effects of core-to-core neutronic and mass flow coupling between the UTVC and the surrounding boiler cores were also included in the dynamic model The dynamic analysis of the UTVR reveals the existence of some very effectlve inherent reactivity feedback effects that are capable of quickly stabilizing this system, within a few seconds, even when large positive reactivity insertions are imposed. If the UTVC vapor fuel density feedback is suppressed, the UTVR is still inherently stable because of the boiler core liquid-fuel volume feedback; in contrast, suppression of the vapor fuel density feedback in 'conventional" gas core cavity reactors causes them to become inherently unstable. Due to the

Features of free software packages in flow cytometry: a comparison between four non-commercial software sources.

PubMed

Sahraneshin Samani, Fazel; Moore, Jodene K; Khosravani, Pardis; Ebrahimi, Marzieh

2014-08-01

Flow cytometers designed to analyze large particles are enabling new applications in biology. Data analysis is a critical component of the process FCM. In this article we compare features of four free software packages including WinMDI, Cyflogic, Flowing software, and Cytobank.

Label-free high-throughput imaging flow cytometry

NASA Astrophysics Data System (ADS)

Mahjoubfar, A.; Chen, C.; Niazi, K. R.; Rabizadeh, S.; Jalali, B.

2014-03-01

Flow cytometry is an optical method for studying cells based on their individual physical and chemical characteristics. It is widely used in clinical diagnosis, medical research, and biotechnology for analysis of blood cells and other cells in suspension. Conventional flow cytometers aim a laser beam at a stream of cells and measure the elastic scattering of light at forward and side angles. They also perform single-point measurements of fluorescent emissions from labeled cells. However, many reagents used in cell labeling reduce cellular viability or change the behavior of the target cells through the activation of undesired cellular processes or inhibition of normal cellular activity. Therefore, labeled cells are not completely representative of their unaltered form nor are they fully reliable for downstream studies. To remove the requirement of cell labeling in flow cytometry, while still meeting the classification sensitivity and specificity goals, measurement of additional biophysical parameters is essential. Here, we introduce an interferometric imaging flow cytometer based on the world's fastest continuous-time camera. Our system simultaneously measures cellular size, scattering, and protein concentration as supplementary biophysical parameters for label-free cell classification. It exploits the wide bandwidth of ultrafast laser pulses to perform blur-free quantitative phase and intensity imaging at flow speeds as high as 10 meters per second and achieves nanometer-scale optical path length resolution for precise measurements of cellular protein concentration.

FlowCal: A user-friendly, open source software tool for automatically converting flow cytometry data from arbitrary to calibrated units

PubMed Central

Castillo-Hair, Sebastian M.; Sexton, John T.; Landry, Brian P.; Olson, Evan J.; Igoshin, Oleg A.; Tabor, Jeffrey J.

2017-01-01

Flow cytometry is widely used to measure gene expression and other molecular biological processes with single cell resolution via fluorescent probes. Flow cytometers output data in arbitrary units (a.u.) that vary with the probe, instrument, and settings. Arbitrary units can be converted to the calibrated unit molecules of equivalent fluorophore (MEF) using commercially available calibration particles. However, there is no convenient, non-proprietary tool available to perform this calibration. Consequently, most researchers report data in a.u., limiting interpretation. Here, we report a software tool named FlowCal to overcome current limitations. FlowCal can be run using an intuitive Microsoft Excel interface, or customizable Python scripts. The software accepts Flow Cytometry Standard (FCS) files as inputs and is compatible with different calibration particles, fluorescent probes, and cell types. Additionally, FlowCal automatically gates data, calculates common statistics, and produces publication quality plots. We validate FlowCal by calibrating a.u. measurements of E. coli expressing superfolder GFP (sfGFP) collected at 10 different detector sensitivity (gain) settings to a single MEF value. Additionally, we reduce day-to-day variability in replicate E. coli sfGFP expression measurements due to instrument drift by 33%, and calibrate S. cerevisiae mVenus expression data to MEF units. Finally, we demonstrate a simple method for using FlowCal to calibrate fluorescence units across different cytometers. FlowCal should ease the quantitative analysis of flow cytometry data within and across laboratories and facilitate the adoption of standard fluorescence units in synthetic biology and beyond. PMID:27110723

Validation of the Chemistry Module for the Euler Solver in Unified Flow Solver

DTIC Science & Technology

2012-03-01

traveling through the atmosphere there are three types of flow regimes that exist; the first is the continuum regime, second is the rarified regime and...The second method has been used in a program called Unified Flow Solver (UFS). UFS is currently being developed under collaborative efforts the Air...thermal non-equilibrium case and finally to a thermo-chemical non- equilibrium case. The data from the simulations will be compared to a second code

Reliable and Accurate CD4+ T Cell Count and Percent by the Portable Flow Cytometer CyFlow MiniPOC and “CD4 Easy Count Kit-Dry”, as Revealed by the Comparison with the Gold Standard Dual Platform Technology

PubMed Central

Nasi, Milena; De Biasi, Sara; Bianchini, Elena; Gibellini, Lara; Pinti, Marcello; Scacchetti, Tiziana; Trenti, Tommaso; Borghi, Vanni; Mussini, Cristina; Cossarizza, Andrea

2015-01-01

Background An accurate and affordable CD4+ T cells count is an essential tool in the fight against HIV/AIDS. Flow cytometry (FCM) is the “gold standard” for counting such cells, but this technique is expensive and requires sophisticated equipment, temperature-sensitive monoclonal antibodies (mAbs) and trained personnel. The lack of access to technical support and quality assurance programs thus limits the use of FCM in resource-constrained countries. We have tested the accuracy, the precision and the carry-over contamination of Partec CyFlow MiniPOC, a portable and economically affordable flow cytometer designed for CD4+ count and percentage, used along with the “CD4% Count Kit-Dry”. Materials and Methods Venous blood from 59 adult HIV+ patients (age: 25–58 years; 43 males and 16 females) was collected and stained with the “MiniPOC CD4% Count Kit-Dry”. CD4+ count and percentage were then determined in triplicate by the CyFlow MiniPOC. In parallel, CD4 count was performed using mAbs and a CyFlow Counter, or by a dual platform system (from Beckman Coulter) based upon Cytomic FC500 (“Cytostat tetrachrome kit” for mAbs) and Coulter HmX Hematology Analyzer (for absolute cell count). Results The accuracy of CyFlow MiniPOC against Cytomic FC500 showed a correlation coefficient (CC) of 0.98 and 0.97 for CD4+ count and percentage, respectively. The accuracy of CyFlow MiniPOC against CyFlow Counter showed a CC of 0.99 and 0.99 for CD4 T cell count and percentage, respectively. CyFlow MiniPOC showed an excellent repeatability: CD4+ cell count and percentage were analyzed on two instruments, with an intra-assay precision below ±5% deviation. Finally, there was no carry-over contamination for samples at all CD4 values, regardless of their position in the sequence of analysis. Conclusion The cost-effective CyFlow MiniPOC produces rapid, reliable and accurate results that are fully comparable with those from highly expensive dual platform systems. PMID:25622041

NASA/American Cancer Society High-Resolution Flow Cytometry Project-I

NASA Technical Reports Server (NTRS)

Thomas, R. A.; Krishan, A.; Robinson, D. M.; Sams, C.; Costa, F.

2001-01-01

BACKGROUND: The NASA/American Cancer Society (ACS) flow cytometer can simultaneously analyze the electronic nuclear volume (ENV) and DNA content of cells. This study describes the schematics, resolution, reproducibility, and sensitivity of biological standards analyzed on this unit. METHODS: Calibrated beads and biological standards (lymphocytes, trout erythrocytes [TRBC], calf thymocytes, and tumor cells) were analyzed for ENV versus DNA content. Parallel data (forward scatter versus DNA) from a conventional flow cytometer were obtained. RESULTS: ENV linearity studies yielded an R value of 0.999. TRBC had a coefficient of variation (CV) of 1.18 +/- 0.13. DNA indexes as low as 1.02 were detectable. DNA content of lymphocytes from 42 females was 1.9% greater than that for 60 males, with a noninstrumental variability in total DNA content of 0.5%. The ENV/DNA ratio was constant in 15 normal human tissue samples, but differed in the four animal species tested. The ENV/DNA ratio for a hypodiploid breast carcinoma was 2.3 times greater than that for normal breast tissue. CONCLUSIONS: The high-resolution ENV versus DNA analyses are highly reliable, sensitive, and can be used for the detection of near-diploid tumor cells that are difficult to identify with conventional cytometers. ENV/DNA ratio may be a useful parameter for detection of aneuploid populations.

High sensitivity detection and sorting of infectious human immunodeficiency virus (HIV-1) particles by flow virometry

PubMed Central

Bonar, Micha M.; Tilton, John C.

2017-01-01

Detection of viruses by flow cytometry is complicated by their small size. Here, we characterized the ability of a standard (FACSAria II) and a sub-micron flow cytometer (A50 Micro) to resolve HIV-1 viruses. The A50 was superior at resolving small particles but did not reliably distinguish HIV-1, extracellular vesicles, and laser noise by light scatter properties alone. However, single fluorescent HIV-1 particles could readily be detected by both cytometers. Fluorescent particles were sorted and retained infectivity, permitting further exploration of the functional consequences of HIV-1 heterogeneity. Finally, flow cytometry had a limit of detection of 80 viruses/ml, nearly equal to PCR assays. These studies demonstrate the power of flow cytometry to detect and sort viral particles and provide a critical toolkit to validate methods to label wild-type HIV-1; quantitatively assess integrity and aggregation of viruses and virus-based therapeutics; and efficiently screen drugs inhibiting viral assembly and release. PMID:28235684

High sensitivity detection and sorting of infectious human immunodeficiency virus (HIV-1) particles by flow virometry.

PubMed

Bonar, Michał M; Tilton, John C

2017-05-01

Detection of viruses by flow cytometry is complicated by their small size. Here, we characterized the ability of a standard (FACSAria II) and a sub-micron flow cytometer (A50 Micro) to resolve HIV-1 viruses. The A50 was superior at resolving small particles but did not reliably distinguish HIV-1, extracellular vesicles, and laser noise by light scatter properties alone. However, single fluorescent HIV-1 particles could readily be detected by both cytometers. Fluorescent particles were sorted and retained infectivity, permitting further exploration of the functional consequences of HIV-1 heterogeneity. Finally, flow cytometry had a limit of detection of 80 viruses/ml, nearly equal to PCR assays. These studies demonstrate the power of flow cytometry to detect and sort viral particles and provide a critical toolkit to validate methods to label wild-type HIV-1; quantitatively assess integrity and aggregation of viruses and virus-based therapeutics; and efficiently screen drugs inhibiting viral assembly and release. Copyright © 2017 Elsevier Inc. All rights reserved.

Complete Genome Sequence of the Thermophilic Bacterium Geobacillus thermoleovorans CCB_US3_UF5

PubMed Central

Abdul Rahman, Ahmad Yamin; Saito, Jennifer A.; Hou, Shaobin

2012-01-01

Geobacillus thermoleovorans CCB_US3_UF5 is a thermophilic bacterium isolated from a hot spring in Malaysia. Here, we report the complete genome of G. thermoleovorans CCB_US3_UF5, which shows high similarity to the genome of Geobacillus kaustophilus HTA 426 in terms of synteny and orthologous genes. PMID:22328744

Scoping study to expedite development of a field deployable and portable instrument for UF6 enrichment assay

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chan, George; Valentine, John D.; Russo, Richard E.

The primary objective of the present study is to identity the most promising, viable technologies that are likely to culminate in an expedited development of the next-generation, field-deployable instrument for providing rapid, accurate, and precise enrichment assay of uranium hexafluoride (UF6). UF6 is typically involved, and is arguably the most important uranium compound, in uranium enrichment processes. As the first line of defense against proliferation, accurate analytical techniques to determine the uranium isotopic distribution in UF6 are critical for materials verification, accounting, and safeguards at enrichment plants. As nuclear fuel cycle technology becomes more prevalent around the world, international nuclearmore » safeguards and interest in UF6 enrichment assay has been growing. At present, laboratory-based mass spectrometry (MS), which offers the highest attainable analytical accuracy and precision, is the technique of choice for the analysis of stable and long-lived isotopes. Currently, the International Atomic Energy Agency (IAEA) monitors the production of enriched UF6 at declared facilities by collecting a small amount (between 1 to 10 g) of gaseous UF6 into a sample bottle, which is then shipped under chain of custody to a central laboratory (IAEA’s Nuclear Materials Analysis Laboratory) for high-precision isotopic assay by MS. The logistics are cumbersome and new shipping regulations are making it more difficult to transport UF6. Furthermore, the analysis is costly, and results are not available for some time after sample collection. Hence, the IAEA is challenged to develop effective safeguards approaches at enrichment plants. In-field isotopic analysis of UF6 has the potential to substantially reduce the time, logistics and expense of sample handling. However, current laboratory-based MS techniques require too much infrastructure and operator expertise for field deployment and operation. As outlined in the IAEA Department of Safeguards

Integrated aquitard-aquifer flow with a mixed-type well-face boundary and skin effect

NASA Astrophysics Data System (ADS)

Feng, Qinggao; Zhan, Hongbin

2016-03-01

A general analytical model describing groundwater flow to a partially penetrating well pumped at a constant rate in a leaky confined aquifer is developed. The model incorporates the effects of aquitard storage, aquifer anisotropy, wellbore storage and a finite well skin by treating the aquitard leakage as an aquitard-aquifer interface flow problem, and considers the well-face as a mixed-type or non-uniform flux (NUF) rather than a uniform flux (UF) boundary condition, which is novel. The solution is obtained using the Laplace transform coupled with separation of variables and discretization methods, followed by the numerical inverse Laplace transform. Moreover, the solution unifies some cases for flow to a partially penetrating well in a leaky confined aquifer including Perina and Lee (2006), Feng and Zhan (2015) and Hunt (2005) or confined aquifer including Chiu et al. (2007), Yang et al. (2006) and Hantush (1964). The newly developed NUF solution is compared with the UF solution. The NUF drawdown is larger than the UF drawdown at early time, while the NUF drawdown is smaller than the UF drawdown at intermediate and late times. The non-uniform flux along the well-face has significant impact on drawdown in the skin zone, while the UF solution can completely replace the NUF solution at a radial distance from the pumped well equaling to or greater than the aquifer thickness. The NUF and UF drawdowns for no skin case are remarkably smaller than that for the positive skin case and larger than that for the negative skin case. A thicker well skin results in a smaller drawdown in the skin zone.

On Line Enrichment Monitor (OLEM) UF 6 Tests for 1.5" Sch40 SS Pipe, Revision 1

DOE Office of Scientific and Technical Information (OSTI.GOV)

March-Leuba, José A.; Garner, Jim; Younkin, Jim

As global uranium enrichment capacity under international safeguards expands, the International Atomic Energy Agency (IAEA) is challenged to develop effective safeguards approaches at gaseous centrifuge enrichment plants while working within budgetary constraints. The “Model Safeguards Approach for Gas Centrifuge Enrichment Plants” (GCEPs) developed by the IAEA Division of Concepts and Planning in June 2006, defines the three primary Safeguards objectives to be the timely detection of: 1) diversion of significant quantities of natural (NU), depleted (DU) or low-enriched uranium (LEU) from declared plant flow, 2) facility misuse to produce undeclared LEU product from undeclared feed, and 3) facility misuse tomore » produce enrichments higher than the declared maximum, in particular, highly enriched uranium (HEU). The ability to continuously and independently (i.e. with a minimum of information from the facility operator) monitor not only the uranium mass balance but also the 235U mass balance in the facility could help support all three verification objectives described above. Two key capabilities required to achieve an independent and accurate material balance are 1) continuous, unattended monitoring of in-process UF 6 and 2) monitoring of cylinders entering and leaving the facility. The continuous monitoring of in-process UF 6 would rely on a combination of load-cell monitoring of the cylinders at the feed and withdrawal stations, online monitoring of gas enrichment, and a high-accuracy net weight measurement of the cylinder contents. The Online Enrichment Monitor (OLEM) is the instrument that would continuously measure the time-dependent relative uranium enrichment, E(t), in weight percent 235U, of the gas filling or being withdrawn from the cylinders. The OLEM design concept combines gamma-ray spectrometry using a collimated NaI(Tl) detector with gas pressure and temperature data to calculate the enrichment of the UF 6 gas within the unit header pipe as a

Whole-body voxel phantoms of paediatric patients—UF Series B

NASA Astrophysics Data System (ADS)

Lee, Choonik; Lee, Choonsik; Williams, Jonathan L.; Bolch, Wesley E.

2006-09-01

Following the previous development of the head and torso voxel phantoms of paediatric patients for use in medical radiation protection (UF Series A), a set of whole-body voxel phantoms of paediatric patients (9-month male, 4-year female, 8-year female, 11-year male and 14-year male) has been developed through the attachment of arms and legs from segmented CT images of a healthy Korean adult (UF Series B). Even though partial-body phantoms (head-torso) may be used in a variety of medical dose reconstruction studies where the extremities are out-of-field or receive only very low levels of scatter radiation, whole-body phantoms play important roles in general radiation protection and in nuclear medicine dosimetry. Inclusion of the arms and legs is critical for dosimetry studies of paediatric patients due to the presence of active bone marrow within the extremities of children. While the UF Series A phantoms preserved the body dimensions and organ masses as seen in the original patients who were scanned, comprehensive adjustments were made for the Series B phantoms to better match International Commission on Radiological Protection (ICRP) age-interpolated reference body masses, body heights, sitting heights and internal organ masses. The CT images of arms and legs of a Korean adult were digitally rescaled and attached to each phantom of the UF series. After completion, the resolutions of the phantoms for the 9-month, 4-year, 8-year, 11-year and 14-year were set at 0.86 mm × 0.86 mm × 3.0 mm, 0.90 mm × 0.90 mm × 5.0 mm, 1.16 mm × 1.16 mm × 6.0 mm, 0.94 mm × 0.94 mm × 6.00 mm and 1.18 mm × 1.18 mm × 6.72 mm, respectively.

Flow Cytometry Scientist | Center for Cancer Research

Cancer.gov

PROGRAM DESCRIPTION The Basic Science Program (BSP) pursues independent, multidisciplinary research in basic and applied molecular biology, immunology, retrovirology, cancer biology, and human genetics. Research efforts and support are an integral part of the Center for Cancer Research (CCR) at the Frederick National Laboratory for Cancer Research (FNLCR). KEY ROLES/RESPONSIBILITIES The Flow Cytometry Core (Flow Core) in the Cancer and Inflammation Program (CIP) is a service core which supports the research efforts of the CCR by providing expertise in the field of flow cytometry (using analyzers and sorters) with the goal of gaining a more thorough understanding of the biology of the immune system, cancer, and inflammation processes. The Flow Core provides service to 12-15 CIP laboratories and more than 22 non-CIP laboratories. Flow core staff provide technical advice on the experimental design of applications, which include immunological phenotyping, cell function assays, and cell cycle analysis. Work is performed per customer requirements, and no independent research is involved. The Flow Cytometry Scientist will be responsible for: Daily management of the Flow Cytometry Core, to include the supervision and guidance of technical staff members Monitor performance of and maintain high dimensional flow cytometer analyzers and cell sorters Operate high dimensional flow cytometer analyzers and cell sorters Provide scientific expertise to the user community and facilitate the development of cutting edge technologies Interact with Flow Core users and customers, and provide technical and scientific advice, and guidance regarding their experiments, including possible collaborations Train staff and scientific end users on the use of flow cytometry in their research, as well as teach them how to operate and troubleshoot the bench-top analyzer instruments Prepare and deliver lectures, as well as one-on-one training sessions, with customers/users Ensure that protocols are up

SINGLE-STEP CONVERSION OF UO$sub 3$ TO UF$sub 4$

DOEpatents

Moore, J.E.

1960-07-12

A description is given of the preparation of uranium tetrafluoride by reacting a hexavalent uranium compound with a pclysaccharide and gaseous hydrogen fluoride at an elevated temperature. Uranium trioxide and starch are combined with water to form a doughy mixture. which is extruded into pellets and dried. The pellets are then contacted with HF at a temperature from 500 to 700 deg C in a moving bed reactor to prcduce UF/sub 4/. Reduction of the hexavalent uranium to UO/sub 2/ and conversion of the UO/sub 2/ to UF/sub 4/ are accomplished simultaneously in this process.

Analysis of synthetic and biological microparticles on several flow cytometric platforms

EPA Science Inventory

Microparticles (MPs) are membrane vesicles (0.1 to 1 urn) released from cells upon activation. The limit of detection ofmost standard flow cytometers is just below 1 urn. Recent advances enable detection of particles lower than 0.5 urn, Synthetic. beads are used to define size ra...

Spaceflight Flow Cytometry: Design Challenges and Applications

NASA Technical Reports Server (NTRS)

Pappas, Dimitri; Kao, Shih-Hsin; Jeevarajan, Antony S.

2004-01-01

Future space exploration missions will require analytical technology capable of providing both autonomous medical care to the crew and investigative capabilities to researchers. While several promising candidate technologies exist for further development, flow cytometry is an attractive technology as it offers both crew health and a wide array of biochemistry and immunology assays. While flow cytometry has been widely used for cellular analysis in both clinical and research settings, the requirements for proper operation in spaceflight impose constraints on any instrument designs. The challenges of designing a spaceflight-ready flow cytometer are discussed, as well as some preliminary results using a prototype system.

Authors@UF Campus Conversation Series: A Case Study

ERIC Educational Resources Information Center

Silver, Isabel

2014-01-01

The Authors@UF program series emerged to showcase the scholarship and creativity of faculty, creating an intellectual forum within the academic library, and providing informal, extra-curriculum, academic engagement between students and faculty outside the classroom. This article identifies steps to launch an author program, and considerations in…

A New Submersible Imaging-in-flow Instrument to Monitor Nano- and Microplankton: Imaging FlowCytobot

NASA Astrophysics Data System (ADS)

Olson, R. J.; Sosik, H. M.; Shalapyonok, A.

2004-12-01

Understanding of how coastal plankton communities are regulated has traditionally been limited by undersampling, but cabled observatories now provide opportunities to deploy submersible sensors that have high power and data transmission requirements. We have developed an in situ instrument to carry out high-resolution, long term monitoring of phytoplankton and microzooplankton in the size range 10 to100 micrometers, to be deployed at cabled research facilities such as the Martha's Vineyard Coastal Observatory (MVCO). The new instrument is designed to complement FlowCytobot, a submersible flow cytometer currently deployed at MVCO that uses fluorescence and light scattering signals from a laser beam to characterize the smallest phytoplankton cells (less than 10 micrometers). Imaging FlowCytobot uses a combination of flow cytometric and video technology to capture images of organisms for identification and to measure chlorophyll fluorescence associated with each image. Images will be classified using neural net software, while the measurements of chlorophyll fluorescence will allow us to discriminate heterotrophic from phototrophic cells. The new instrument, like the original FlowCytobot is autonomous but remotely programmable. It utilizes a computer controlled syringe pump and distribution valve that allows periodic anti-fouling treatment and analysis of standard beads. Samples are analyzed continuously (0.25 to 2.5 ml per min) and data is sent over a fiber optic link to a remote computer for analysis. Preliminary results indicate that we can detect cells as small as 5 micrometers and discriminate several taxa of diatoms and dinoflagellates.

Method for selectively removing fluorine and fluorine-containing contaminants from gaseous UF/sub 6/. [ClF/sub 3/

DOEpatents

Jones, R.L.; Otey, M.G.; Perkins, R.W.

1980-11-24

This invention is a method for effecting preferential removal and immobilization of certain gaseous contaminants from gaseous UF/sub 6/. The contaminants include fluorine and fluorides which are more reactive with CaCO/sub 3/ than is UF/sub 6/. The method comprises contacting the contaminant-carrying UF/sub 6/ with particulate CaCO/sub 3/ at a temperature effecting reaction of the contaminant and the CaCO/sub 3/.

Microflow1, a sheathless fiber-optic flow cytometry biomedical platform: demonstration onboard the international space station.

PubMed

Dubeau-Laramée, Geneviève; Rivière, Christophe; Jean, Isabelle; Mermut, Ozzy; Cohen, Luchino Y

2014-04-01

A fiber-optic based flow cytometry platform was designed to build a portable and robust instrument for space applications. At the core of the Microflow1 is a unique fiber-optic flow cell fitted to a fluidic system and fiber coupled to the source and detection channels. A Microflow1 engineering unit was first tested and benchmarked against a commercial flow cytometer as a reference in a standard laboratory environment. Testing in parabolic flight campaigns was performed to establish Microflow1's performance in weightlessness, before operating the new platform on the International Space Station. Microflow1 had comparable performances to commercial systems, and operated remarkably and robustly in weightlessness (microgravity). Microflow1 supported immunophenotyping as well as microbead-based multiplexed cytokine assays in the space environment and independently of gravity levels. Results presented here provide evidence that this fiber-optic cytometer technology is inherently compatible with the space environment with negligible compromise to analytical performance. © 2013 International Society for Advancement of Cytometry.

Committee Report: Metrics & Methods for MF/UF System Optimization

EPA Science Inventory

After a membrane filtration (i.e., microfiltration (MF) and ultrafiltration (UF)) system is designed, installed, and commissioned, it is essential that the plant is well-maintained in order to proactively identify potential design or equipment problems and ensure its proper opera...

Depth resolved granular transport driven by shearing fluid flow

NASA Astrophysics Data System (ADS)

Allen, Benjamin; Kudrolli, Arshad

2017-02-01

We investigate granular transport by a fluid flow under steady-state driving conditions, from the bed-load regime to the suspension regime, with an experimental system based on a conical rheometer. The mean granular volume fraction ϕg, the mean granular velocity ug, and the fluid velocity uf are obtained as a function of depth inside the bed using refractive index matching and particle-tracking techniques. A torque sensor is utilized to measure the applied shear stress to complement estimates obtained from measured strain rates high above the bed where ϕg≈0 . The flow is found to be transitional at the onset of transport and the shear stress required to transport grains rises sharply as grains are increasingly entrained by the fluid flow. A significant slip velocity between the fluid and the granular phases is observed at the bed surface before the onset of transport as well as in the bed-load transport regime. We show that ug decays exponentially deep into the bed for ϕg>0.45 with a decay constant which is described by a nonlocal rheology model of granular flow that neglects fluid stress. Further, we show that uf and ug can be described using the applied shear stress and the Krieger-Dougherty model for the effective viscosity in the suspension regime, where 0 <ϕg<0.45 and where ug≈uf .

Separating the signal from the noise: Expanding flow cytometry into the sub-micron range.

EPA Science Inventory

Cytometry Part A Special Section: Separating the signal from the noise: Expanding flow cytometry into the sub-micron range. The current Cytometry Part A Special Section presents three studies that utilize cytometers to study sub-micron particles. The three studies involve the 1...

Onsite Gaseous Centrifuge Enrichment Plant UF6 Cylinder Destructive Analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Anheier, Norman C.; Cannon, Bret D.; Qiao, Hong

2012-07-17

The IAEA safeguards approach for gaseous centrifuge enrichment plants (GCEPs) includes measurements of gross, partial, and bias defects in a statistical sampling plan. These safeguard methods consist principally of mass and enrichment nondestructive assay (NDA) verification. Destructive assay (DA) samples are collected from a limited number of cylinders for high precision offsite mass spectrometer analysis. DA is typically used to quantify bias defects in the GCEP material balance. Under current safeguards measures, the operator collects a DA sample from a sample tap following homogenization. The sample is collected in a small UF6 sample bottle, then sealed and shipped under IAEAmore » chain of custody to an offsite analytical laboratory. Current practice is expensive and resource intensive. We propose a new and novel approach for performing onsite gaseous UF6 DA analysis that provides rapid and accurate assessment of enrichment bias defects. DA samples are collected using a custom sampling device attached to a conventional sample tap. A few micrograms of gaseous UF6 is chemically adsorbed onto a sampling coupon in a matter of minutes. The collected DA sample is then analyzed onsite using Laser Ablation Absorption Ratio Spectrometry-Destructive Assay (LAARS-DA). DA results are determined in a matter of minutes at sufficient accuracy to support reliable bias defect conclusions, while greatly reducing DA sample volume, analysis time, and cost.« less

Treatment of car wash wastewater by UF membranes

NASA Astrophysics Data System (ADS)

Istirokhatun, Titik; Destianti, Puti; Hargianintya, Adenira; Oktiawan, Wiharyanto; Susanto, Heru

2015-12-01

The existence of car wash service facilitates car owners to remove dirt and grime from their vehicles. However, the dirt washed off vehicles as well as the cleaning materials themselves may be harmful to the environment if they are not properly managed and discharged. Many technologies have been proposed to treat car wash wastewater such as coagulation flocculation, tricking filter and flocculation-flotation. Nevertheless, these technologies have low efficiency to eliminate oil and small organic compounds. Ultrafiltration (UF) membranes were used in this study to treat car wash wastewater. This study investigated the performance of UF membranes under various pressures to remove COD, oil and grease, and also turbidity from car wash waste water. The membrane performance was examined by investigation of permeate flux and membrane rejection. The results meet the standard of environmental regulation and it is possible to be reused. The highest rejection was shown by PES10 (polyethersulfone 10 kDa) in 1 bar operation with complete rejection for both turbidity and oil and grace and 95% rejection for COD.

UF/RO applications at the Browns Ferry Nuclear Power Station

DOE Office of Scientific and Technical Information (OSTI.GOV)

Palino, G.F.; Sailor, W.C.; Sawochka, S.G.

1981-04-01

In June 1979, NWT was contracted by TVA to review the applicability of reverse osmosis (RO) and ultrafiltration (UF) membrane treatment technology at the Browns Ferry Nuclear Power Station. Specific program tasks are described and results presented.

Multistate Evaluation of an Ultrafiltration-Based Procedure for Simultaneous Recovery of Enteric Microbes in 100-Liter Tap Water Samples▿

PubMed Central

Hill, Vincent R.; Kahler, Amy M.; Jothikumar, Narayanan; Johnson, Trisha B.; Hahn, Donghyun; Cromeans, Theresa L.

2007-01-01

Ultrafiltration (UF) is increasingly being recognized as a potentially effective procedure for concentrating and recovering microbes from large volumes of water and treated wastewater. Because of their very small pore sizes, UF membranes are capable of simultaneously concentrating viruses, bacteria, and parasites based on size exclusion. In this study, a UF-based water sampling procedure was used to simultaneously recover representatives of these three microbial classes seeded into 100-liter samples of tap water collected from eight cities covering six hydrologic areas of the United States. The UF-based procedure included hollow-fiber UF as the primary step for concentrating microbes and then used membrane filtration for bacterial culture assays, immunomagnetic separation for parasite recovery and quantification, and centrifugal UF for secondary concentration of viruses. Water samples were tested for nine water quality parameters to investigate whether water quality data correlated with measured recovery efficiencies and molecular detection levels. Average total method recovery efficiencies were 71, 97, 120, 110, and 91% for φX174 bacteriophage, MS2 bacteriophage, Enterococcus faecalis, Clostridium perfringens spores, and Cryptosporidium parvum oocysts, respectively. Real-time PCR and reverse transcription-PCR (RT-PCR) for seeded microbes and controls indicated that tap water quality could affect the analytical performance of molecular amplification assays, although no specific water quality parameter was found to correlate with reduced PCR or RT-PCR performance. PMID:17483281

Flow Cytometry and Solid Organ Transplantation: A Perfect Match

PubMed Central

Maguire, Orla; Tario, Joseph D.; Shanahan, Thomas C.; Wallace, Paul K.; Minderman, Hans

2015-01-01

In the field of transplantation, flow cytometry serves a well-established role in pre-transplant crossmatching and monitoring immune reconstitution following hematopoietic stem cell transplantation. The capabilities of flow cytometers have continuously expanded and this combined with more detailed knowledge of the constituents of the immune system, their function and interaction and newly developed reagents to study these parameters have led to additional utility of flow cytometry-based analyses, particularly in the post-transplant setting. This review discusses the impact of flow cytometry on managing alloantigen reactions, monitoring opportunistic infections and graft rejection and gauging immunosuppression in the context of solid organ transplantation. PMID:25296232

The cytometric future: it ain't necessarily flow!

PubMed

Shapiro, Howard M

2011-01-01

Initial approaches to cytometry for classifying and characterizing cells were based on microscopy; it was necessary to collect relatively high-resolution images of cells because only a few specific reagents usable for cell identification were available. Although flow cytometry, now the dominant cytometric technology, typically utilizes lenses similar to microscope lenses for light collection, improved, more quantitative reagents allow the necessary information to be acquired in the form of whole-cell measurements of the intensities of light transmission, scattering, and/or fluorescence.Much of the cost and complexity of both automated microscopes and flow cytometers arises from the necessity for them to measure one cell at a time. Recent developments in digital camera technology now offer an alternative in which one or more low-magnification, low-resolution images are made of a wide field containing many cells, using inexpensive light-emitting diodes (LEDs) for illumination. Minimalist widefield imaging cytometers can provide a smaller, less complex, and substantially less expensive alternative to flow cytometry, critical in systems intended for in resource-poor areas. Minimalism is, likewise, a good philosophy in developing instrumentation and methodology for both clinical and large-scale research use; it simplifies quality assurance and compliance with regulatory requirements, as well as reduces capital outlays, material costs, and personnel training requirements. Also, importantly, it yields "greener" technology.

Flow cytometric discrimination of seven lineage markers by using two fluorochromes

PubMed Central

Boin, Francesco; Giardino Torchia, Maria Letizia; Borrello, Ivan; Noonan, Kimberly A.; Neil, Matthew; Soloski, Mark J.

2017-01-01

Flow cytometry is the primary immunological technique used to analyze multiple parameters on complex cell populations. We present a staining method that identifies major human mononuclear lymphoid and myeloid populations (CD4+ and CD8+ T cells, γδ T cells, B cells, NK cells and monocytes), using only two fluorochromes and a minimal number of cells. Our approach increases the number of markers recordable on most flow cytometers allowing for a deeper and more comprehensive immunophenotyping. PMID:29190813

Detection of silver nanoparticles in cells by flow cytometry using light scatter and far-red fluorescence.

PubMed

Zucker, R M; Daniel, K M; Massaro, E J; Karafas, S J; Degn, L L; Boyes, W K

2013-10-01

The cellular uptake of different sized silver nanoparticles (AgNP) (10, 50, and 75 nm) coated with polyvinylpyrrolidone (PVP) or citrate on a human derived retinal pigment epithelial cell line (ARPE-19) was detected by flow cytometry following 24-h incubation of the cells with AgNP. A dose dependent increase of side scatter and far red fluorescence was observed with both PVP and citrate-coated 50 nm or 75 nm silver particles. Using five different flow cytometers, a far red fluorescence signal in the 700-800 nm range increased as much as 100 times background as a ratio comparing the intensity measurements of treated sample and controls. The citrate-coated silver nanoparticles (AgNP) revealed slightly more side scatter and far red fluorescence than did the PVP coated silver nanoparticles. This increased far red fluorescence signal was observed with 50 and 75 nm particles, but not with 10 nm particles. Morphological evaluation by dark field microscopy showed silver particles (50 and 75 nm) clumped and concentrated around the nucleus. One possible hypothesis to explain the emission of far red fluorescence from cells incubated with silver nanoparticles is that the silver nanoparticles inside cells agglomerate into small nano clusters that form surface plasmon resonance which interacts with laser light to emit a strong far red fluorescence signal. The results demonstrate that two different parameters (side scatter and far red fluorescence) on standard flow cytometers can be used to detect and observe metallic nanoparticles inside cells. The strength of the far red fluorescence suggests that it may be particularly useful for applications that require high sensitivity. © Published 2013 Wiley-Periodicals, Inc. Published 2013 Wiley‐Periodicals, Inc. This article is a US government work and, as such, is in the public domain in the United States of America.

Development of small and inexpensive digital data acquisition systems using a microcontroller-based approach†

PubMed Central

Naivar, Mark A.; Wilder, Mark E.; Habbersett, Robert C.; Woods, Travis A.; Sebba, David S.; Nolan, John P.; Graves, Steven W.

2014-01-01

Fully digital data acquisition systems for use in flow cytometry provide excellent flexibility and precision. Here, we demonstrate the development of a low cost, small, and low power digital flow cytometry data acquisition system using a single microcontroller chip with an integrated analog to digital converter (ADC). Our demonstration system uses a commercially available evaluation board making the system simple to integrate into a flow cytometer. We have evaluated this system using calibration microspheres analyzed on commercial, slow-flow, and CCD based flow cytometers. In our evaluations, our demonstration data system clearly resolves all eight peaks of a Rainbow microsphere set on both a slow-flow flow cytometer and a retrofitted BD FACScalibur, which indicates it has the sensitivity and resolution required for most flow cytometry applications. It is also capable of millisecond time resolution, full waveform collection, and selective triggering of data collection from a CCD camera. The capability of our demonstration system suggests that the use of microcontrollers for flow cytometry digital data-acquisition will be increasingly valuable for extending the life of older cytometers and provides a compelling data-system design approach for low-cost, portable flow cytometers. PMID:19852060

Development of small and inexpensive digital data acquisition systems using a microcontroller-based approach.

PubMed

Naivar, Mark A; Wilder, Mark E; Habbersett, Robert C; Woods, Travis A; Sebba, David S; Nolan, John P; Graves, Steven W

2009-12-01

Fully digital data acquisition systems for use in flow cytometry provide excellent flexibility and precision. Here, we demonstrate the development of a low cost, small, and low power digital flow cytometry data acquisition system using a single microcontroller chip with an integrated analog to digital converter (ADC). Our demonstration system uses a commercially available evaluation board making the system simple to integrate into a flow cytometer. We have evaluated this system using calibration microspheres analyzed on commercial, slow-flow, and CCD-based flow cytometers. In our evaluations, our demonstration data system clearly resolves all eight peaks of a Rainbow microsphere set on both a slow-flow flow cytometer and a retrofitted BD FACScalibur, which indicates it has the sensitivity and resolution required for most flow cytometry applications. It is also capable of millisecond time resolution, full waveform collection, and selective triggering of data collection from a CCD camera. The capability of our demonstration system suggests that the use of microcontrollers for flow cytometry digital data-acquisition will be increasingly valuable for extending the life of older cytometers and provides a compelling data-system design approach for low-cost, portable flow cytometers.

Analysis of an Indirect Neutron Signature for Enhanced UF6 Cylinder Verification

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kulisek, Jonathan A.; McDonald, Benjamin S.; Smith, Leon E.

2017-02-21

The International Atomic Energy Agency (IAEA) currently uses handheld gamma-ray spectrometers combined with ultrasonic wall-thickness gauges to verify the declared enrichment of uranium hexafluoride (UF6) cylinders. The current method provides relatively low accuracy for the assay of 235U enrichment, especially for natural and depleted UF6. Furthermore, the current method provides no capability to assay the absolute mass of 235U in the cylinder due to the localized instrument geometry and limited penetration of the 186-keV gamma-ray signature from 235U. Also, the current verification process is a time-consuming component of on-site inspections at uranium enrichment plants. Toward the goal of a more-capablemore » cylinder assay method, the Pacific Northwest National Laboratory has developed the hybrid enrichment verification array (HEVA). HEVA measures both the traditional 186-keV direct signature and a non-traditional, high-energy neutron-induced signature (HEVANT). HEVANT enables full-volume assay of UF6 cylinders by exploiting the relatively larger mean free paths of the neutrons emitted from the UF6. In this work, Monte Carlo modeling is used as the basis for characterizing HEVANT in terms of the individual contributions to HEVANT from nuclides and hardware components. Monte Carlo modeling is also used to quantify the intrinsic efficiency of HEVA for neutron detection in a cylinder-assay geometry. Modeling predictions are validated against neutron-induced gamma-ray spectra from laboratory measurements and a relatively large population of Type 30B cylinders spanning a range of enrichments. Implications of the analysis and findings on the viability of HEVA for cylinder verification are discussed, such as the resistance of the HEVANT signature to manipulation by the nearby placement of neutron-conversion materials.« less

Time-Domain Microfluidic Fluorescence Lifetime Flow Cytometry for High-Throughput Förster Resonance Energy Transfer Screening

PubMed Central

Nedbal, Jakub; Visitkul, Viput; Ortiz-Zapater, Elena; Weitsman, Gregory; Chana, Prabhjoat; Matthews, Daniel R; Ng, Tony; Ameer-Beg, Simon M

2015-01-01

Sensing ion or ligand concentrations, physico-chemical conditions, and molecular dimerization or conformation change is possible by assays involving fluorescent lifetime imaging. The inherent low throughput of imaging impedes rigorous statistical data analysis on large cell numbers. We address this limitation by developing a fluorescence lifetime-measuring flow cytometer for fast fluorescence lifetime quantification in living or fixed cell populations. The instrument combines a time-correlated single photon counting epifluorescent microscope with microfluidics cell-handling system. The associated computer software performs burst integrated fluorescence lifetime analysis to assign fluorescence lifetime, intensity, and burst duration to each passing cell. The maximum safe throughput of the instrument reaches 3,000 particles per minute. Living cells expressing spectroscopic rulers of varying peptide lengths were distinguishable by Förster resonant energy transfer measured by donor fluorescence lifetime. An epidermal growth factor (EGF)-stimulation assay demonstrated the technique's capacity to selectively quantify EGF receptor phosphorylation in cells, which was impossible by measuring sensitized emission on a standard flow cytometer. Dual-color fluorescence lifetime detection and cell-specific chemical environment sensing were exemplified using di-4-ANEPPDHQ, a lipophilic environmentally sensitive dye that exhibits changes in its fluorescence lifetime as a function of membrane lipid order. To our knowledge, this instrument opens new applications in flow cytometry which were unavailable due to technological limitations of previously reported fluorescent lifetime flow cytometers. The presented technique is sensitive to lifetimes of most popular fluorophores in the 0.5–5 ns range including fluorescent proteins and is capable of detecting multi-exponential fluorescence lifetime decays. This instrument vastly enhances the throughput of experiments involving

Multiplex immunoassay for persistent organic pollutants in tilapia: Comparison of imaging- and flow cytometry-based platforms using spectrally encoded paramagnetic microspheres

USDA-ARS?s Scientific Manuscript database

Recent developments in spectrally encoded microspheres (SEMs)-based technologies provide high multiplexing possibilities. Most SEMs-based assays required a flow cytometer with sophisticated fluidics and optics. The new imaging superparamagnetic SEMs-based platform transports SEMs with considerably ...

A Microfluidic Cytometer for Complete Blood Count With a 3.2-Megapixel, 1.1- μm-Pitch Super-Resolution Image Sensor in 65-nm BSI CMOS.

PubMed

Liu, Xu; Huang, Xiwei; Jiang, Yu; Xu, Hang; Guo, Jing; Hou, Han Wei; Yan, Mei; Yu, Hao

2017-08-01

Based on a 3.2-Megapixel 1.1- μm-pitch super-resolution (SR) CMOS image sensor in a 65-nm backside-illumination process, a lens-free microfluidic cytometer for complete blood count (CBC) is demonstrated in this paper. Backside-illumination improves resolution and contrast at the device level with elimination of surface treatment when integrated with microfluidic channels. A single-frame machine-learning-based SR processing is further realized at system level for resolution correction with minimum hardware resources. The demonstrated microfluidic cytometer can detect the platelet cells (< 2 μm) required in CBC, hence is promising for point-of-care diagnostics.

Capture of Fluorescence Decay Times by Flow Cytometry

PubMed Central

Naivar, Mark A.; Jenkins, Patrick; Freyer, James P.

2012-01-01

In flow cytometry, the fluorescence decay time of an excitable species has been largely underutilized and is not likely found as a standard parameter on any imaging cytometer, sorting, or analyzing system. Most cytometers lack fluorescence lifetime hardware mainly owing to two central issues. Foremost, research and development with lifetime techniques has lacked proper exploitation of modern laser systems, data acquisition boards, and signal processing techniques. Secondly, a lack of enthusiasm for fluorescence lifetime applications in cells and with bead-based assays has persisted among the greater cytometry community. In this unit, we describe new approaches that address these issues and demonstrate the simplicity of digitally acquiring fluorescence relaxation rates in flow. The unit is divided into protocol and commentary sections in order to provide a most comprehensive discourse on acquiring the fluorescence lifetime with frequency-domain methods. The unit covers (i) standard fluorescence lifetime acquisition (protocol-based) with frequency-modulated laser excitation, (ii) digital frequency-domain cytometry analyses, and (iii) interfacing fluorescence lifetime measurements onto sorting systems. Within the unit is also a discussion on how digital methods are used for aliasing in order to harness higher frequency ranges. Also, a final discussion is provided on heterodyning and processing of waveforms for multi-exponential decay extraction. PMID:25419263

Actual and Idealized Crystal Field Parameterizations for the Uranium Ions in UF 4

NASA Astrophysics Data System (ADS)

Gajek, Z.; Mulak, J.; Krupa, J. C.

1993-12-01

The crystal field parameters for the actual coordination symmetries of the uranium ions in UF 4, C2 and C1, and for their idealizations to D2, C2 v , D4, D4 d , and the Archimedean antiprism point symmetries are given. They have been calculated by means of both the perturbative ab initio model and the angular overlap model and are referenced to the recent results fitted by Carnall's group. The equivalency of some different sets of parameters has been verified with the standardization procedure. The adequacy of several idealized approaches has been tested by comparison of the corresponding splitting patterns of the 3H 4 ground state. Our results support the parameterization given by Carnall. Furthermore, the parameterization of the crystal field potential and the splitting diagram for the symmetryless uranium ion U( C1) are given. Having at our disposal the crystal field splittings for the two kinds of uranium ions in UF 4, U( C2) and U( C1), we calculate the model plots of the paramagnetic susceptibility χ( T) and the magnetic entropy associated with the Schottky anomaly Δ S( T) for UF 4.

Tight ceramic UF membrane as RO pre-treatment: the role of electrostatic interactions on phosphate rejection.

PubMed

Shang, Ran; Verliefde, Arne R D; Hu, Jingyi; Zeng, Zheyi; Lu, Jie; Kemperman, Antoine J B; Deng, Huiping; Nijmeijer, Kitty; Heijman, Sebastiaan G J; Rietveld, Luuk C

2014-01-01

Phosphate limitation has been reported as an effective approach to inhibit biofouling in reverse osmosis (RO) systems for water purification. The rejection of dissolved phosphate by negatively charged TiO2 tight ultrafiltration (UF) membranes (1 kDa and 3 kDa) was observed. These membranes can potentially be adopted as an effective process for RO pre-treatment in order to constrain biofouling by phosphate limitation. This paper focuses on electrostatic interactions during tight UF filtration. Despite the larger pore size, the 3 kDa ceramic membrane exhibited greater phosphate rejection than the 1 kDa membrane, because the 3 kDa membrane has a greater negative surface charge and thus greater electrostatic repulsion against phosphate. The increase of pH from 6 to 8.5 led to a substantial increase in phosphate rejection by both membranes due to increased electrostatic repulsion. At pH 8.5, the maximum phosphate rejections achieved by the 1 kDa and 3 kDa membrane were 75% and 86%, respectively. A Debye ratio (ratio of the Debye length to the pore radius) is introduced in order to evaluate double layer overlapping in tight UF membranes. Threshold Debye ratios were determined as 2 and 1 for the 1 kDa and 3 kDa membranes, respectively. A Debye ratio below the threshold Debye ratio leads to dramatically decreased phosphate rejection by tight UF membranes. The phosphate rejection by the tight UF, in combination with chemical phosphate removal by coagulation, might accomplish phosphate-limited conditions for biological growth and thus prevent biofouling in the RO systems. Copyright © 2013 Elsevier Ltd. All rights reserved.

Radiation-Triggered Surveillance for UF6 Monitoring

DOE Office of Scientific and Technical Information (OSTI.GOV)

Curtis, Michael M.

2015-12-01

This paper recommends the use of radiation detectors, singly or in sets, to trigger surveillance cameras. Ideally, the cameras will monitor cylinders transiting the process area as well as the process area itself. The general process area will be surveyed to record how many cylinders have been attached and detached to the process between inspections. Rad-triggered cameras can dramatically reduce the quantity of recorded images, because the movement of personnel and equipment not involving UF6 cylinders will not generate a surveillance review file.

Detection of Silver Nanoparticles in Cells by Flow Cytometry Using Light Scattering and Far-red Fluorescence

EPA Science Inventory

The cellular uptake of different sized silver nanoparticles (l0 nm, 50 nm, and 75nm) coated with polyvinylpyrrolidone (PVP) or citrate in ARPE-19 cells following 24 hour incubation was detected by side scatter through the use of a flow cytometer. A large far red fluorescence sign...

Immunophenotyping of posttraumatic neutrophils on a routine haematology analyser.

PubMed

Groeneveld, Kathelijne Maaike; Heeres, Marjolein; Leenen, Loek Petrus Hendrikus; Huisman, Albert; Koenderman, Leo

2012-01-01

Flow cytometry markers have been proposed as useful predictors for the occurrence of posttraumatic inflammatory complications. However, currently the need for a dedicated laboratory and the labour-intensive analytical procedures make these markers less suitable for clinical practice. We tested an approach to overcome these limitations. Neutrophils of healthy donors were incubated with antibodies commonly used in trauma research: CD11b (MAC-1), L-selectin (CD62L), FcγRIII (CD16), and FcγRII (CD32) in active form (MoPhab A27). Flow cytometric analysis was performed both on a FACSCalibur, a standard flow cytometer, and on a Cell-Dyn Sapphire, a routine haematology analyser. There was a high level of agreement between the two types of analysers, with 41% for FcγRIII, 80% for L-selectin, 98% for CD11b, and even a 100% agreement for active FcγRII. Moreover, analysis on the routine haematology analyser was possible in less than a quarter of the time in comparison to the flow cytometer. Analysis of neutrophil phenotype on the Cell-Dyn Sapphire leads to the same conclusion compared to a standard flow cytometer. The markedly reduced time necessary for analysis and reduced labour intensity constitutes a step forward in implementation of this type of analysis in clinical diagnostics in trauma research. Copyright © 2012 Kathelijne Maaike Groeneveld et al.

A Radiation-Triggered Surveillance System for UF6 Cylinder Monitoring

DOE Office of Scientific and Technical Information (OSTI.GOV)

Curtis, Michael M.; Myjak, Mitchell J.

This report provides background information and representative scenarios for testing a prototype radiation-triggered surveillance system at an operating facility that handles uranium hexafluoride (UF 6) cylinders. The safeguards objective is to trigger cameras using radiation, or radiation and motion, rather than motion alone, to reduce significantly the number of image files generated by a motion-triggered system. The authors recommend the use of radiation-triggered surveillance at all facilities where cylinder paths are heavily traversed by personnel. The International Atomic Energy Agency (IAEA) has begun using surveillance cameras in the feed and withdrawal areas of gas centrifuge enrichment plants (GCEPs). The camerasmore » generate imagery using elapsed time or motion, but this creates problems in areas occupied 24/7 by personnel. Either motion-or-interval-based triggering generates thousands of review files over the course of a month. Since inspectors must review the files to verify operator material-flow-declarations, a plethora of files significantly extends the review process. The primary advantage of radiation-triggered surveillance is the opportunity to obtain full-time cylinder throughput verification versus what presently amounts to part-time verification. Cost savings should be substantial, as the IAEA presently uses frequent unannounced inspections to verify cylinder-throughput declarations. The use of radiation-triggered surveillance allows the IAEA to implement less frequent unannounced inspections for the purpose of flow verification, but its principal advantage is significantly shorter and more effective inspector video reviews.« less

Relativistic scattered wave calculations on UF6

NASA Technical Reports Server (NTRS)

Case, D. A.; Yang, C. Y.

1980-01-01

Self-consistent Dirac-Slater multiple scattering calculations are presented for UF6. The results are compared critically to other relativistic calculations, showing that the results of all molecular orbital calculations are in qualitative agreement, as measured by energy levels, population analyses, and spin-orbit splittings. A detailed comparison is made to the relativistic X alpha(RX alpha) method of Wood and Boring, which also uses multiple scattering theory, but incorporates relativistic effects in a more approximate fashion. For the most part, the RX alpha results are in agreement with the present results.

Review of corrosion in 10- and 14-ton mild steel depleted UF{sub 6} storage cylinders

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lykins, M.L.

1995-08-01

A literature review was conducted to determine the type, extent and severity of corrosion found in the 10- and 14-ton mild steel depleted UF{sub 6} storage cylinders. Also discussed in this review is corrosion found in the valves and plugs used in the cylinders. Corrosion of the cylinders is a gradual process which occurs slowly over time. Understanding corrosion of the cylinders is an important concern for long term storage of the UF{sub 6} in the cylinder yards, as well as the final disposition of the depleted UF{sub 6} tails inventory in the future. The following conclusions are made frommore » the literature review: (1) The general external corrosion rate of the cylinders is about 1 to 2 mils per year (1 mil = 0.001{double_prime}). The highest general external corrosion rate was over 5 mpy on the 48G type cylinders. (2) General internal corrosion from the depleted UF{sub 6} is negligible under normal storage conditions. Crevice corrosion can occur at the cylinder/saddle interface from the retention of water in this area. Crevice corrosion can occur at the cylinder/skirt interface on the older skirted cylinders due to the lack of water drainage in this area. Crevice corrosion can occur on cylinders that have been in ground contact. Crevice corrosion and galvanic corrosion can occur where the stainless steel I.D. nameplates are attached to the cylinder. The packing nuts on the bronze one-inch valves used in the cylinders are susceptible to stress corrosion cracking (SCC). Mechanical damage from routine handling can lead to a breach in a cylinder with subsequent accelerated corrosion of the mild steel due to attack from HF and other UF{sub 6} hydrolysis by-products.« less

Masks in Imaging Flow Cytometry

PubMed Central

Dominical, Venina; Samsel, Leigh; McCoy, J. Philip

2016-01-01

Data analysis in imaging flow cytometry incorporates elements of flow cytometry together with other aspects of morphological analysis of images. A crucial early step in this analysis is the creation of a mask to distinguish the portion of the image upon which further examination of specified features can be performed. Default masks are provided by the manufacturer of the imaging flow cytometer but additional custom masks can be created by the individual user for specific applications. Flawed or inaccurate masks can have a substantial negative impact on the overall analysis of a sample, thus great care must be taken to ensure the accuracy of masks. Here we discuss various types of masks and cite examples of their use. Furthermore we provide our insight for how to approach selecting and assessing the optimal mask for a specific analysis. PMID:27461256

Separation Properties of Wastewater Containing O/W Emulsion Using Ceramic Microfiltration/Ultrafiltration (MF/UF) Membranes

PubMed Central

Nakamura, Kazuho; Matsumoto, Kanji

2013-01-01

Washing systems using water soluble detergent are used in electrical and mechanical industries and the wastewater containing O/W emulsion are discharged from these systems. Membrane filtration has large potential for the efficient separation of O/W emulsion for reuses of treated water and detergent. The separation properties of O/W emulsions by cross-flow microfiltration and ultrafiltration were studied with ceramic MF and UF membranes. The effects of pore size; applied pressure; cross-flow velocity; and detergent concentration on rejection of O/W emulsion and flux were systematically studied. At the condition achieving complete separation of O/W emulsion the pressure-independent flux was observed and this flux behavior was explained by gel-polarization model. The O/W emulsion tended to permeate through the membrane at the conditions of larger pore size; higher emulsion concentration; and higher pressure. The O/W emulsion could permeate the membrane pore structure by destruction or deformation. These results imply the stability of O/W emulsion in the gel-layer formed on membrane surface play an important role in the separation properties. The O/W emulsion was concentrated by batch cross-flow concentration filtration and the flux decline during the concentration filtration was explained by the gel- polarization model. PMID:24958621

Final Report Nucleic Acid System - Hybrid PCR and Multiplex Assay Project Phase 2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Koopman, R P; Langlois, R G; Nasarabadi, S

2002-04-17

This report covers phase 2 (year 2) of the Nucleic Acid System--Hybrid PCR and Multiplex Assay project. The objective of the project is to reduce to practice the detection and identification of biological warfare pathogens by the nucleic acid recognition technique of PCR (polymerase chain reaction) in a multiplex mode using flow cytometry. The Hybrid instrument consists of a flow-through PCR module capable of handling a multiplexed PCR assay, a hybridizing module capable of hybridizing multiplexed PCR amplicons and beads, and a flow cytometer module for bead-based identification, all controlled by a single computer. Multiplex immunoassay using bead-based Luminex flowmore » cytometry is available, allowing rapid screening for many agents. PCR is highly specific and complements and verifies immunoassay. It can also be multiplexed and detection provided using the bead-based Luminex flow cytometer. This approach allows full access to the speed and 100-fold multiplex capability of flow cytometry for rapid screening as well as the accuracy and specificity of PCR. This project has two principal activities: (1) Design, build and test a prototype hybrid PCR/flow cytometer with the basic capabilities for rapid, broad spectrum detection and identification, and (2) Develop and evaluate multiplex flow analysis assay protocols and reagents for the simultaneous detection of PCR products. This project requires not only building operationally functional instrumentation but also developing the chemical assays for detection of priority pathogens. This involves development and evaluation of multiplex flow analysis assay protocols and reagents for the simultaneous detection of PCR products.« less

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.

Very High Density of Chinese Hamster Ovary Cells in Perfusion by Alternating Tangential Flow or Tangential Flow Filtration in WAVE Bioreactor™—Part II: Applications for Antibody Production and Cryopreservation

PubMed Central

Clincke, Marie-Françoise; Mölleryd, Carin; Samani, Puneeth K; Lindskog, Eva; Fäldt, Eric; Walsh, Kieron; Chotteau, Véronique

2013-01-01

A high cell density perfusion process of monoclonal antibody (MAb) producing Chinese hamster ovary (CHO) cells was developed in disposable WAVE Bioreactor™ using external hollow fiber (HF) filter as cell separation device. Tangential flow filtration (TFF) and alternating tangential flow (ATF) systems were compared and process applications of high cell density perfusion were studied here: MAb production and cryopreservation. Operations by perfusion using microfiltration (MF) or ultrafiltration (UF) with ATF or TFF and by fed-batch were compared. Cell densities higher than 108 cells/mL were obtained using UF TFF or UF ATF. The cells produced comparable amounts of MAb in perfusion by ATF or TFF, MF or UF. MAbs were partially retained by the MF using ATF or TFF but more severely using TFF. Consequently, MAbs were lost when cell broth was discarded from the bioreactor in the daily bleeds. The MAb cell-specific productivity was comparable at cell densities up to 1.3 × 108 cells/mL in perfusion and was comparable or lower in fed-batch. After 12 days, six times more MAbs were harvested using perfusion by ATF or TFF with MF or UF, compared to fed-batch and 28× more in a 1-month perfusion at 108 cells/mL density. Pumping at a recirculation rate up to 2.75 L/min did not damage the cells with the present TFF settings with HF short circuited. Cell cryopreservation at 0.5 × 108 and 108 cells/mL was performed using cells from a perfusion run at 108 cells/mL density. Cell resuscitation was very successful, showing that this system was a reliable process for cell bank manufacturing. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:768–777, 2013 PMID:23436783

Thermodynamic assessment of the LiF-NaF-BeF2-ThF4-UF4 system

NASA Astrophysics Data System (ADS)

Capelli, E.; Beneš, O.; Konings, R. J. M.

2014-06-01

The present study describes the full thermodynamic assessment of the LiF-NaF-BeF2-ThF4-UF4 system which is one of the key systems considered for a molten salt reactor fuel. The work is an extension of the previously assessed LiF-NaF-ThF4-UF4 system with addition of BeF2 which is characterized by very low neutron capture cross section and a relatively low melting point. To extend the database the binary BeF2-ThF4 and BeF2-UF4 systems were optimized and the novel data were used for the thermodynamic assessment of BeF2 containing ternary systems for which experimental data exist in the literature. The obtained database is used to optimize the molten salt reactor fuel composition and to assess its properties with the emphasis on the melting behaviour.

Spectroscopic properties of K 5Li 2UF 10

NASA Astrophysics Data System (ADS)

Karbowiak, M.; Gajek, Z.; Drożdżyński, J.

2005-04-01

A new uranium (III) fluoro-complex of the formula K 5Li 2UF 10 has been synthesised and characterised by X-ray powder diffraction and electronic absorption spectra measurements. The compound crystallises in the orthorhombic system, space group Pnma, with a = 20.723, b = 7.809, c = 6.932 Å, V = 1121.89 Å 3, Z = 4 and is isostructural with its K 5Li 2NdF 10 and K 5Li 2LaF 10 analogous. The absorption spectrum of a polycrystalline sample of K 5Li 2UF 10 was recorded at 4.2 K in the 3500-45,000 cm -1 range and is discussed. The observed crystal-field levels were assigned and fitted to parameters of the simplified angular overlap model (AOM) and next to those of a semi-empirical Hamiltonian, which was representing the combined atomic and one-electron crystal-field interactions. The starting values of the AOM parameters were obtained from ab initio calculations. The analysis of the spectra enabled the assignment of 71 crystal-field levels of U 3+ with a relatively small r.m.s. deviation of 37 cm -1. The total splitting of 714 cm -1 was calculated for the 4I 9/2 ground multiplet.

Styrene process condensate treatment with a combination process of UF and NF for reuse.

PubMed

Wang, Aijun; Liu, Guangmin; Huang, Jin; Wang, Lijuan; Li, Guangbin; Su, Xudong; Qi, Hong

2013-01-15

Aiming at reusing the SPC to save water resource and heat energy, a combination treatment process of UF/NF was applied to remove inorganic irons, suspended particles and little amount of organic contaminants in this article. To achieve the indexes of CODM≤5.00 mg L(-1), oil≤2.00 mg L(-1), conductivity≤10.00 μs cm(-1), pH of 6.0-8.0, the NF membrane process was adopted. It was necessary to employ a pretreatment process to reduce NF membrane fouling. Hence UF membrane as an efficient pretreatment unit was proposed to remove the inorganic particles, such as iron oxide catalyst, to meet the influent demands of NF. The effluent of UF, which was less than 0.02 mg L(-1) of total iron, went into a security filter and then was pumped into the NF process unit. High removal efficiencies of CODM, oil and conductivity were achieved by using NF process. The ABS grafting copolymerization experiment showed that the effluent of the combination process met the criteria of ABS production process, meanwhile the process could alleviate the environment pollution. It was shown that this combination process concept was feasible and successful in treating the SPC. Copyright © 2012 Elsevier B.V. All rights reserved.

PDB-UF: database of predicted enzymatic functions for unannotated protein structures from structural genomics.

PubMed

von Grotthuss, Marcin; Plewczynski, Dariusz; Ginalski, Krzysztof; Rychlewski, Leszek; Shakhnovich, Eugene I

2006-02-06

The number of protein structures from structural genomics centers dramatically increases in the Protein Data Bank (PDB). Many of these structures are functionally unannotated because they have no sequence similarity to proteins of known function. However, it is possible to successfully infer function using only structural similarity. Here we present the PDB-UF database, a web-accessible collection of predictions of enzymatic properties using structure-function relationship. The assignments were conducted for three-dimensional protein structures of unknown function that come from structural genomics initiatives. We show that 4 hypothetical proteins (with PDB accession codes: 1VH0, 1NS5, 1O6D, and 1TO0), for which standard BLAST tools such as PSI-BLAST or RPS-BLAST failed to assign any function, are probably methyltransferase enzymes. We suggest that the structure-based prediction of an EC number should be conducted having the different similarity score cutoff for different protein folds. Moreover, performing the annotation using two different algorithms can reduce the rate of false positive assignments. We believe, that the presented web-based repository will help to decrease the number of protein structures that have functions marked as "unknown" in the PDB file. http://paradox.harvard.edu/PDB-UF and http://bioinfo.pl/PDB-UF.

Performance of a commercial industrial-scale UF-based process for treatment of oily wastewaters.

PubMed

Karhu, M; Kuokkanen, T; Rämö, J; Mikola, M; Tanskanen, J

2013-10-15

An evaluation was made of the performance of a commercial industrial-scale ultrafiltration (UF)-based process for treatment of highly concentrated oily wastewaters. Wastewater samples were gathered from two plants treating industrial wastewaters in 2008, and in 2011 (only from one of the plants), from three points of a UF-based treatment train. The wastewater samples were analyzed by measuring the BOD7, COD, TOC and total surface charge (TSC). The inorganic content and zeta potentials of the samples were analyzed and GC-FID/MS analyses were performed. The removal performances of BOD7, COD, TOC and TSC in 2008 and 2011 for both plants were very high. Initial concentrations of contaminants in 2011 were lower than in 2008, therefore the COD and TSC reductions were also lower in 2011 than three years before. Regardless of the high performance of UF-based processes in both plants, at times the residual concentrations were considerable. This could be explained by the high initial concentrations and also by the presence of the dissolved compounds that were characterized. Linear correlation was observed between COD and TOC, and between COD and TSC. The correlation between COD and TSC could be utilized for process control purposes. Copyright © 2013 Elsevier Ltd. All rights reserved.

Quasi-3D Modeling and Efficient Simulation of Laminar Flows in Microfluidic Devices.

PubMed

Islam, Md Zahurul; Tsui, Ying Yin

2016-10-03

A quasi-3D model has been developed to simulate the flow in planar microfluidic systems with low Reynolds numbers. The model was developed by decomposing the flow profile along the height of a microfluidic system into a Fourier series. It was validated against the analytical solution for flow in a straight rectangular channel and the full 3D numerical COMSOL Navier-Stokes solver for flow in a T-channel. Comparable accuracy to the full 3D numerical solution was achieved by using only three Fourier terms with a significant decrease in computation time. The quasi-3D model was used to model flows in a micro-flow cytometer chip on a desktop computer and good agreement between the simulation and the experimental results was found.

Imaging flow cytometry for phytoplankton analysis.

PubMed

Dashkova, Veronika; Malashenkov, Dmitry; Poulton, Nicole; Vorobjev, Ivan; Barteneva, Natasha S

2017-01-01

This review highlights the concepts and instrumentation of imaging flow cytometry technology and in particular its use for phytoplankton analysis. Imaging flow cytometry, a hybrid technology combining speed and statistical capabilities of flow cytometry with imaging features of microscopy, is rapidly advancing as a cell imaging platform that overcomes many of the limitations of current techniques and contributed significantly to the advancement of phytoplankton analysis in recent years. This review presents the various instrumentation relevant to the field and currently used for assessment of complex phytoplankton communities' composition and abundance, size structure determination, biovolume estimation, detection of harmful algal bloom species, evaluation of viability and metabolic activity and other applications. Also we present our data on viability and metabolic assessment of Aphanizomenon sp. cyanobacteria using Imagestream X Mark II imaging cytometer. Herein, we highlight the immense potential of imaging flow cytometry for microalgal research, but also discuss limitations and future developments. Copyright © 2016 Elsevier Inc. All rights reserved.

Dynamic thermoregulation of the sample in flow cytometry.

PubMed

Graves, Steven W; Habbersett, Robert C; Nolan, John P

2002-05-01

Fine control of temperature is an important capability for any analytical platform. A circulating water bath has been the traditional means of maintaining constant temperature in the sample chamber of a flow cytometer, but this approach does not permit rapid changes in sample temperature. This unit explains the use of Peltier modules for regulation of sample temperature. The heat pumping generated by the passage of current through properly matched semiconductors, known as the Peltier effect, makes it possible for these thermoelectric modules to both heat and cool. The authors describe the construction of a Peltier module based thermoregulation unit in step-by-step detail and present a demonstration of flow cytometry measurements as a function of temperature.

A novel flow cytometry-based method of analyzing Heinz bodies.

PubMed

Palasuwan, D; Palasuwan, A; Charoensappakit, A; Noulsri, E

2017-02-01

Heinz bodies are important to diagnosing and managing patients. However, microscopic examination of Heinz bodies has several disadvantages, demonstrating the need for a better method. We explored the potential use of flow cytometry to examine Heinz bodies. Whole-blood samples were collected from patients deficient in G6PD and healthy volunteers. Acetylphenylhydrazine was used to induce formation of Heinz bodies in red blood cells (RBCs). Then, RBCs positive for Heinz bodies were examined using a FACSCanto II cytometer. RBCs treated with acetylphenylhydrazine formed Heinz bodies and emitted a broad spectrum of fluorescence that could be detected by flow cytometry. The maximum emission of fluorescence was observed at 45 min after the incubation with acetylphenylhydrazine. In addition, the fluorescence emitted was stable for at least 72 h. The flow cytometer could detect the RBCs positive for Heinz bodies even if they made up as little as 0.1% of the total RBC population. Furthermore, the percentage and number, respectively, of RBCs positive for Heinz bodies in G6PD-deficient patients and normal donors exhibited a mean ± standard deviation (SD) of 68.9 ± 27.5 vs. 50.9 ± 28.6 and 96 014 ±35 732 cells/μL vs. 74 688 ± 36 514 cells/μL. Heinz bodies induced by acetylphenylhydrazine emit fluorescence, and this fluorescence could be examined using flow cytometry. Our study suggests the potential use of the developed method to investigate the formation of Heinz bodies in clinical samples. © 2016 John Wiley & Sons Ltd.

Imaging flow cytometry for the screening of compounds that disrupt the Plasmodium falciparum digestive vacuole.

PubMed

Chia, Wan Ni; Lee, Yan Quan; Tan, Kevin Shyong-Wei

2017-01-01

Malaria, despite being one of the world's oldest infectious diseases, remains difficult to eradicate because the parasite is rapidly developing resistance to frontline chemotherapies. Previous studies have shown that the parasite exhibits features resembling programmed cell death upon treatment with drugs that disrupt its digestive vacuole (DV), providing a phenotypic readout that can be detected using the imaging flow cytometer. Large compound collections can thus be screened to identify drugs that are able to disrupt the DV of the malaria parasite using this high-content high-throughput screening platform. As a proof-of-concept, 4440 compounds were screened using this platform in 4months and 254 hits (5.7% hit rate) were obtained. Additionally, 25 compounds (0.6% top hit rate) were observed to retain potent DV disruption activity that was comparable to the canonical DV disruptive drug chloroquine when tested at a ten-fold lower concentration from the original screen. This pilot study demonstrates the robustness and high-throughput capability of the imaging flow cytometer and we report herein the methodology of this screening assay. Copyright © 2016 Elsevier Inc. All rights reserved.

Performance evaluation of the new fully automated urine particle analyser UF-5000 compared to the reference method of the Fuchs-Rosenthal chamber.

PubMed

Previtali, Giulia; Ravasio, Rudi; Seghezzi, Michela; Buoro, Sabrina; Alessio, Maria Grazia

2017-09-01

UF-5000 is the new fully automated urine particle analyser. We validated its performance. 736 urines were analysed and results were compared by two pathologists on uncentrifuged samples, using Fuchs-Rosenthal chamber. AUC of ROC curve ranged between 0.86 and 0.99. Sensitivity was >0.90 for all the elements and similar for RBC and yeasts. Specificity ranged between 0.74 and 0.89 for total cast, epithelial/non-squamous/renal-tubular cells and RBC. For all the other parameters specificity was >0.90. Comparison with Fuchs-Rosenthal chamber was very good for all the parameters; r ranged between 0.52 and 0.99 except for pathological cast because of the lack of the pathological samples in medium and higher ranges. Linearity performance (R 2 ) was 1.00, 1.00 and 0.99 respectively for RBC, WBC and epithelial cells (EC). No carry-over was observed. The within-run imprecision was 25.42%,13.81%,1.36% for RBC; 37.50%,10.16%,1.41% for WBC and 35.25%, 17.85%,6.30% for EC at low, near the cut off level and high concentrations, respectively. The between-run imprecision was 6.90%,1.60% for RBC, 4.10%,1.90% for WBC and 7.60%,7.30% for EC, using low and high positive quality controls, respectively. UF-5000 is an analyser of great interest to detect urine particle related to pathological process of kidney and urinary tract. Copyright © 2017 Elsevier B.V. All rights reserved.

Quasi-3D Modeling and Efficient Simulation of Laminar Flows in Microfluidic Devices

PubMed Central

Islam, Md. Zahurul; Tsui, Ying Yin

2016-01-01

A quasi-3D model has been developed to simulate the flow in planar microfluidic systems with low Reynolds numbers. The model was developed by decomposing the flow profile along the height of a microfluidic system into a Fourier series. It was validated against the analytical solution for flow in a straight rectangular channel and the full 3D numerical COMSOL Navier-Stokes solver for flow in a T-channel. Comparable accuracy to the full 3D numerical solution was achieved by using only three Fourier terms with a significant decrease in computation time. The quasi-3D model was used to model flows in a micro-flow cytometer chip on a desktop computer and good agreement between the simulation and the experimental results was found. PMID:27706104

Developments of solid materials for UF 6 sampling

DOE Office of Scientific and Technical Information (OSTI.GOV)

Smith, Nicholas; Hebden, Andrew; Savina, Joseph

2017-11-15

This project demonstrated that a device using majority Commercial Off the Shelf (COTS) components could be used to collect uranium hexafluoride samples safely from gaseous or solid sources. The device was based on the successful Cristallini method developed by ABACC over the past 10 years. The system was designed to capture and store the UF 6 as an inert fluoride salt to ease transportation regulations. In addition, the method was considerably faster than traditional cryogenic methods, collected enough material to perform analyses without undue waste, and could be used either inside a facility or in the storage yard.

Depleted UF6 Management Information Network - A resource for the public,

Science.gov Websites

Depleted UF6 Management Information Network Web Site is an online repository of information about the U.S ) and DUF6, research and development efforts for beneficial uses of DU, DOE's program for management of line DUF6 Guide DUF6 Guide line Introductory information about depleted uranium: how it is created

Design of portable ultraminiature flow cytometers for medical diagnostics

NASA Astrophysics Data System (ADS)

Leary, James F.

2018-02-01

Design of portable microfluidic flow/image cytometry devices for measurements in the field (e.g. initial medical diagnostics) requires careful design in terms of power requirements and weight to allow for realistic portability. True portability with high-throughput microfluidic systems also requires sampling systems without the need for sheath hydrodynamic focusing both to avoid the need for sheath fluid and to enable higher volumes of actual sample, rather than sheath/sample combinations. Weight/power requirements dictate use of super-bright LEDs with top-hat excitation beam architectures and very small silicon photodiodes or nanophotonic sensors that can both be powered by small batteries. Signal-to-noise characteristics can be greatly improved by appropriately pulsing the LED excitation sources and sampling and subtracting noise in between excitation pulses. Microfluidic cytometry also requires judicious use of small sample volumes and appropriate statistical sampling by microfluidic cytometry or imaging for adequate statistical significance to permit real-time (typically in less than 15 minutes) initial medical decisions for patients in the field. This is not something conventional cytometry traditionally worries about, but is very important for development of small, portable microfluidic devices with small-volume throughputs. It also provides a more reasonable alternative to conventional tubes of blood when sampling geriatric and newborn patients for whom a conventional peripheral blood draw can be problematical. Instead one or two drops of blood obtained by pin-prick should be able to provide statistically meaningful results for use in making real-time medical decisions without the need for blood fractionation, which is not realistic in the doctor's office or field.

Using digital flow cytometry to assess the degradation of three cyanobacteria species after oxidation processes.

PubMed

Wert, Eric C; Dong, Mei Mei; Rosario-Ortiz, Fernando L

2013-07-01

Depending on drinking water treatment conditions, oxidation processes may result in the degradation of cyanobacteria cells causing the release of toxic metabolites (microcystin), odorous metabolites (MIB, geosmin), or disinfection byproduct precursors. In this study, a digital flow cytometer (FlowCAM(®)) in combination with chlorophyll-a analysis was used to evaluate the ability of ozone, chlorine, chlorine dioxide, and chloramine to damage or lyse cyanobacteria cells added to Colorado River water. Microcystis aeruginosa (MA), Oscillatoria sp. (OSC) and Lyngbya sp. (LYN) were selected for the study due to their occurrence in surface water supplies, metabolite production, and morphology. Results showed that cell damage was observed without complete lysis or fragmentation of the cell membrane under many of the conditions tested. During ozone and chlorine experiments, the unicellular MA was more susceptible to oxidation than the filamentous OSC and LYN. Rate constants were developed based on the loss of chlorophyll-a and oxidant exposure, which showed the oxidants degraded MA, OSC, and LYN according to the order of ozone > chlorine ~ chlorine dioxide > chloramine. Digital and binary images taken by the digital flow cytometer provided qualitative insight regarding cell damage. When applying this information, drinking water utilities can better understand the risk of cell damage or lysis during oxidation processes. Copyright © 2013 Elsevier Ltd. All rights reserved.

Laboratory Tests of Multiplex Detection of PCR Amplicons Using the Luminex 100 Flow Analyzer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Venkateswaran, K.S.; Nasarabadi, S.; Langlois, R.G.

2000-05-05

Lawrence Livermore National Laboratory (LLNL) demonstrated the power of flow cytometry in detecting the biological agents simulants at JFT III. LLNL pioneered in the development of advanced nucleic acid analyzer (ANM) for portable real time identification. Recent advances in flow cytometry provide a means for multiplexed nucleic acid detection and immunoassay of pathogenic microorganisms. We are presently developing multiplexed immunoassays for the simultaneous detection of different simulants. Our goal is to build an integrated instrument for both nucleic acid analysis and immuno detection. In this study we evaluated the Luminex LX 100 for concurrent identification of more than one PCRmore » amplified product. ANAA has real-time Taqman fluorescent detection capability for rapid identification of field samples. However, its multiplexing ability is limited by the combination of available fluorescent labels. Hence integration of ANAA with flow cytometry can give the rapidity of ANAA amplification and the multiplex capability of flow cytometry. Multiplexed flow cytometric analysis is made possible using a set of fluorescent latex microsphere that are individually identified by their red and infrared fluorescence. A green fluorochrome is used as the assay signal. Methods were developed for the identification of specific nucleic acid sequences from Bacillus globigii (Bg), Bacillus thuringensis (Bt) and Erwinia herbicola (Eh). Detection sensitivity using different reporter fluorochromes was tested with the LX 100, and also different assay formats were evaluated for their suitability for rapid testing. A blind laboratory trial was carried out December 22-27, 1999 to evaluate bead assays for multiplex identification of Bg and Bt PCR products. This report summarizes the assay development, fluorochrome comparisons, and the results of the blind trial conducted at LLNL for the laboratory evaluation of the LX 100 flow analyzer.« less

77 FR 48130 - Prospective Grant of Exclusive Patent License

Federal Register 2010, 2011, 2012, 2013, 2014

2012-08-13

.... Department of Commerce, is contemplating the grant of an exclusive license in the United States of America... Provisional Application for Patent Application No. 61,638,362 titled ``Flow Cytometer Systems and Associated... Secretary of Commerce and the Regents of the University of Colorado. The invention is a flow cytometer...

Low-Intensity Sprint Training With Blood Flow Restriction Improves 100-m Dash.

PubMed

Behringer, Michael; Behlau, Daniel; Montag, Johannes C K; McCourt, Molly L; Mester, Joachim

2017-09-01

Behringer, M, Behlau, D, Montag, JCK, McCourt, ML, and Mester, J. Low-intensity sprint training with blood flow restriction improves 100-m dash. J Strength Cond Res 31(9): 2462-2472, 2017-We investigated the effects of practical blood flow restriction (pBFR) of leg muscles during sprint training on the 100-m dash time in well-trained sport students. Participants performed 6 × 100-m sprints at 60-70% of their maximal 100-m sprinting speed twice a week for 6 weeks, either with (intervention group [IG]; n = 12) or without pBFR (control group [CG]; n = 12). The 100-m dash time significantly decreased more in the IG (-0.38 ± 0.24 seconds) than in the CG (-0.16 ± 0.17 seconds). The muscle thickness of the rectus femoris increased only in the IG, whereas no group-by-time interactions were found for the muscle thickness of the biceps femoris and the biceps brachii. The maximal isometric force, measured using a leg press, did not change in either group. However, the rate of force development improved in the IG. Growth hormone, testosterone, insulin-like growth factor 1, and cortisol concentrations did not significantly differ between both groups at any measurement time point (pre, 1 minute, 20 minutes, 120 minutes, and 24 hours after the 6 all-out sprints of the first training session). The muscle damage marker h-FABP increased significantly more in the CG than in the IG. The pBFR improved the 100-m dash time significantly more than low-intensity sprint interval training alone. Other noted benefits of training with pBFR were a decreased level of muscle damage, a greater increase of the rectus femoris muscle thickness, and a higher rate of force development. However, the tested hormones were unable to explain the additional beneficial effects.

A portable battery powered microfluidic impedance cytometer with smartphone readout: towards personal health monitoring.

PubMed

Talukder, Niloy; Furniturewalla, Abbas; Le, Tuan; Chan, Matthew; Hirday, Shreyas; Cao, Xinnan; Xie, Pengfei; Lin, Zhongtian; Gholizadeh, Azam; Orbine, Steve; Javanmard, Mehdi

2017-06-01

We present a portable system for personalized blood cell counting consisting of a microfluidic impedance cytometer and portable analog readout electronics, feeding into an analog-to-digital converter (ADC), and being transmitted via Bluetooth to a user-accessible mobile application. We fabricated a microfluidic impedance cytometer with a novel portable analog readout. The novel design of the analog readout, which consists of a lock-in-amplifier followed by a high-pass filter stage for subtraction of drift and DC offset, and a post-subtraction high gain stage, enables detection of particles and cells as small as 1 μm in diameter, despite using a low-end 8-bit ADC. The lock-in-amplifier and the ADC were set up to receive and transmit data from a Bluetooth module. In order to initiate the system, as well as to transmit all of the data, a user friendly mobile application was developed, and a proof-of-concept trial was run on a blood sample. Applications such as personalized health monitoring require robust device operation and resilience to clogging. It is desirable to avoid using channels comparable in size to the particles being detected thus requiring high levels of sensitivity. Despite using low-end off-the-shelf hardware, our sensing platform was capable of detecting changes in impedance as small as 0.032%, allowing detection of 3 μm diameter particles in a 300 μm wide channel. The sensitivity of our system is comparable to that of a high-end bench-top impedance spectrometer when tested using the same sensors. The novel analog design allowed for an instrument with a footprint of less than 80 cm 2 . The aim of this work is to demonstrate the potential of using microfluidic impedance spectroscopy for low cost health monitoring. We demonstrated the utility of the platform technology towards cell counting, however, our platform is broadly applicable to assaying wide panels of biomarkers including proteins, nucleic acids, and various cell types.

Hollow-fiber ultrafiltration for simultaneous recovery of viruses, bacteria and parasites from reclaimed water.

PubMed

Liu, Pengbo; Hill, Vincent R; Hahn, Donghyun; Johnson, Trisha B; Pan, Yi; Jothikumar, Narayanan; Moe, Christine L

2012-01-01

Hollow-fiber ultrafiltration (UF) is a technique that has been reported to be effective for recovering a diverse array of microbes from water, and may also be potentially useful for microbial monitoring of effluent from water reclamation facilities. However, few data are available to indicate the potential limitations and efficacy of the UF technique for treated wastewater. In this study, recovery efficiencies were determined for various options available for performing the tangential-flow UF technique, including hollow-fiber ultrafilter (i.e., dialyzer) type, ultrafilter pre-treatment (i.e., blocking), and elution. MS2 and ΦX174 bacteriophages, Clostridium perfringens spores, Escherichia coli, and Cryptosporidium parvum oocysts were seeded into 10-L reclaimed water samples to evaluate UF options. Then a single UF protocol was established and studied using seeded and non-seeded 100-L samples from two water reclamation facilities in Georgia, USA. Baxter Exeltra Plus 210 and Fresenius F200NR dialyzers were found to provide significantly higher microbial recovery than Minntech HPH 1400 hemoconcentrators. The selected final UF method incorporated use of a non-blocked ultrafilter for UF followed by elution using a surfactant-based solution. For 10-L samples, this method achieved recovery efficiencies of greater than 50% recovery of seeded viruses, bacteria, and parasites. There was no significant difference in overall microbial recovery efficiency when the method was applied to 10- and 100-L samples. In addition, detection levels for pathogens in seeded 100-L reclaimed water samples were 1000 PFU HAV, 10,000 GI norovirus particles, <500 Salmonella and <200 Cryptosporidium oocysts. These data demonstrate that UF can be an effective technique for recovering diverse microbes in reclaimed water to monitor and improve effluent water quality in wastewater treatment plants. Published by Elsevier B.V.

Nutrient recovery from the dry grind process using sequential micro and ultrafiltration of thin stillage.

PubMed

Arora, Amit; Dien, Bruce S; Belyea, Ronald L; Singh, Vijay; Tumbleson, M E; Rausch, Kent D

2010-06-01

The effectiveness of microfiltration (MF) and ultrafiltration (UF) for nutrient recovery from a thin stillage stream was determined. When a stainless steel MF membrane (0.1microm pore size) was used, the content of solids increased from 7.0% to 22.8% with a mean permeate flux rate of 45L/m(2)/h (LMH), fat increased and ash content decreased. UF experiments were conducted in batch mode under constant temperature and flow rate conditions. Permeate flux profiles were evaluated for regenerated cellulose membranes (YM1, YM10 and YM100) with molecular weight cut offs of 1, 10 and 100kDa. UF increased total solids, protein and fat and decreased ash in retentate stream. When permeate streams from MF were subjected to UF, retentate total solids concentrations similar to those of commercial syrup (23-28.8%) were obtained. YM100 had the highest percent permeate flux decline (70% of initial flux) followed by YM10 and YM1 membranes. Sequential filtration improved permeate flux rates of the YM100 membrane (32.6-73.4LMH) but the percent decline was also highest in a sequential MF+YM100 system. Protein recovery was the highest in YM1 retentate. Removal of solids, protein and fat from thin stillage may generate a permeate stream that may improve water removal efficiency and increase water recycling. Copyright 2010 Elsevier Ltd. All rights reserved.

Development of a bead-based multiplexed assay for simultaneous quantification of five bovine cytokines by flow cytometry.

PubMed

Rodrigues, Valérie; Baudier, Jean Baptiste; Chantal, Isabelle

2017-09-01

Quantifying cytokines is extremely important in studies of host-pathogen interactions. Multiplex assays are commercially available but only for human and mouse cytokines. Here a method for the simultaneous quantification of five important bovine cytokines IFNγ, IL-4, IL-10, IL-12, and TNFα in cell culture supernatants, using flow cytometry was reported. Functional beads from BD Biosciences expressing specific APC intensity were used. Commercially available antibodies against bovine cytokines were covalently coupled to beads as capture antibodies. Fixed recombinant cytokines were revealed with a second monoclonal antibody coupled with biotin, then revealed with streptavidin-PE. This complex was analyzed using a standard flow cytometer. Experiments were performed to check no cross reactions had occurred. The limits of detection ranged between 0.08 and 0.4 ng/ml depending on the cytokine, and the linearity between the lower and higher limits was remarkable (R 2  > 99.8%). Finally, native cytokines from cell culture supernatants were tested. Results were compared using the standard ELISA test and showed that concentrations of native cytokine in cell culture supernatants were comparable with the two methods, with a wider dynamic range using beads and flow cytometry than with ELISA assays. Bovine IFNγ, IL-4, IL-10, IL-12, and TNFα in culture supernatants can be now simultaneously detected in a single assay, using a standard flow cytometer for both basic and high-throughput analyses. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

Fate of antibiotics in activated sludge followed by ultrafiltration (CAS-UF) and in a membrane bioreactor (MBR).

PubMed

Sahar, Eyal; Messalem, Rami; Cikurel, Haim; Aharoni, Avi; Brenner, Asher; Godehardt, Manuel; Jekel, Martin; Ernst, Mathias

2011-10-15

The fates of several macrolide, sulphonamide, and trimethoprim antibiotics contained in the raw sewage of the Tel-Aviv wastewater treatment plant (WWTP) were investigated after the sewage was treated using either a full-scale conventional activated sludge (CAS) system coupled with a subsequent ultrafiltration (UF) step or a pilot membrane bioreactor (MBR) system. Antibiotics removal in the MBR system, once it achieved stable operation, was 15-42% higher than that of the CAS system. This advantage was reduced to a maximum of 20% when a UF was added to the CAS. It was hypothesized that the contribution of membrane separation (in both systems) to antibiotics removal was due either to sorption to biomass (rather than improvement in biodegradation) or to enmeshment in the membrane biofilm (since UF membrane pores are significantly larger than the contaminant molecules). Batch experiments with MBR biomass showed a markedly high potential for sorption of the tested antibiotics onto the biomass. Moreover, methanol extraction of MBR biomass released significant amounts of sorbed antibiotics. This finding implies that more attention must be devoted to the management of excess sludge. Copyright © 2011 Elsevier Ltd. All rights reserved.

Thermodynamic properties of a high pressure subcritical UF6/He gas volume (irradiated by an external source)

NASA Technical Reports Server (NTRS)

Sterritt, D. E.; Lalos, G. T.; Schneider, R. T.

1976-01-01

A computer simulation study concerning a compressed fissioning UF6 gas is presented. The compression is to be achieved by a ballistic piston compressor. Data on UF6 obtained with this compressor were incorporated in the simulation study. As a neutron source to create the fission events in the compressed gas, a fast burst reactor was considered. The conclusion is that it takes a neutron flux in excess of 10 to the 15th power n/sec sq cm to produce measurable increases in pressure and temperature, while a flux in excess of 10 to 19th power n/sq cm sec would probably damage the compressor.

Treatment of poultry slaughterhouse wastewater using a static granular bed reactor (SGBR) coupled with ultrafiltration (UF) membrane system.

PubMed

Basitere, M; Rinquest, Z; Njoya, M; Sheldon, M S; Ntwampe, S K O

2017-07-01

The South African poultry industry has grown exponentially in recent years due to an increased demand for their products. As a result, poultry plants consume large volumes of high quality water to ensure that hygienically safe poultry products are produced. Furthermore, poultry industries generate high strength wastewater, which can be treated successfully at low cost using anaerobic digesters. In this study, the performance of a bench-scale mesophilic static granular bed reactor (SGBR) containing fully anaerobic granules coupled with an ultrafiltration (UF) membrane system, as a post-treatment system, was investigated. The poultry slaughterhouse wastewater was characterized by a chemical oxygen demand (COD) range between 1,223 and 9,695mg/L, average biological oxygen demand of 2,375mg/L and average fats, oil and grease (FOG) of 554mg/L. The SGBR anaerobic reactor was operated for 9 weeks at different hydraulic retention times (HRTs), i.e. 55 and 40 h, with an average organic loading rate (OLR) of 1.01 and 3.14g COD/L.day. The SGBR results showed an average COD, total suspended solids (TSS) and FOG removal of 93%, 95% and 90% respectively, for both OLR. The UF post-treatment results showed an average of COD, TSS and FOG removal of 64%, 88% and 48%, respectively. The overall COD, TSS and FOG removal of the system (SGBR and UF membrane) was 98%, 99.8%, and 92.4%, respectively. The results of the combined SGBR reactor coupled with the UF membrane showed a potential to ensure environmentally friendly treatment of poultry slaughterhouse wastewater.

The role of flow cytometry in companion animal diagnostic medicine.

PubMed

Tarrant, Jacqueline M

2005-11-01

Flow cytometry is a powerful tool for characterising the composition of complex cell populations. The accuracy and precision of this technology for describing and enumerating cells exceeds traditional methods. The number of diagnostic veterinary laboratories with access to a dedicated machine is increasing, and there is the potential to offer a clinical flow cytometry service. The improved availability of monoclonal antibodies (mAb) to cell markers expressed by the leukocytes of companion animals, permits the implementation of comprehensive mAb panels suitable for diagnosis of lympho- and myeloproliferative disease. Reticulated erythrocyte and platelet quantification, antiglobulin assays for immune-mediated cytopenias, lymphocyte subset analysis, and immunophenotyping of lymphoma and leukemia, have been validated for companion animal samples on the flow cytometer. It is now timely to consider the role of flow cytometry in diagnostic practice, and the requirement for quality assurance and standardization of testing procedures.

High temperature UF6 RF plasma experiments applicable to uranium plasma core reactors

NASA Technical Reports Server (NTRS)

Roman, W. C.

1979-01-01

An investigation was conducted using a 1.2 MW RF induction heater facility to aid in developing the technology necessary for designing a self critical fissioning uranium plasma core reactor. Pure, high temperature uranium hexafluoride (UF6) was injected into an argon fluid mechanically confined, steady state, RF heated plasma while employing different exhaust systems and diagnostic techniques to simulate and investigate some potential characteristics of uranium plasma core nuclear reactors. The development of techniques and equipment for fluid mechanical confinement of RF heated uranium plasmas with a high density of uranium vapor within the plasma, while simultaneously minimizing deposition of uranium and uranium compounds on the test chamber peripheral wall, endwall surfaces, and primary exhaust ducts, is discussed. The material tests and handling techniques suitable for use with high temperature, high pressure, gaseous UF6 are described and the development of complementary diagnostic instrumentation and measurement techniques to characterize the uranium plasma, effluent exhaust gases, and residue deposited on the test chamber and exhaust system components is reported.

Evaluation of the automated urine particle analyzer UF-1000i screening for urinary tract infection in nonpregnant women.

PubMed

Dai, Qingkai; Jiang, Yongmei; Shi, Hua; Zhou, Wei; Zhou, Shengjie; Yang, Hui

2014-01-01

Urinary tract infection (UTI) is a widespread disease in women. Urine culture is still the "gold standard" diagnostic test for UTI, but most of them are negative. To reduce unnecessary culture, we evaluated the automated urine particle analyzer UF-1000i screening for UTI in nonpregnant women. The urine specimens submitted to our laboratory were submitted for culture and tested by the Sysmex UF-1000i. Bacteria and white blood cell (WBC) counts were compared to standard urine culture results to assess the best cutoff values. In this study, 272 urine samples were included, of which 98 (36.0%) were culture positive with a bacterial cutoff value of 10 x 10(5) CFU/mL. A combination of bacterial (> 95/microL) and/or WBC count (> 24/microL) provided the best screening for UTI, with a sensitivity of 0.99 and a specificity of 0.82 compared with the urine culture. Sysmex UF-1000i could be used as a screening test for UTI in nonpregnant women. According to the distribution and range of the bacterial scattergram, we could primarily identify and differentiate between Gram-negative and Gram-positive bacteria.

Thermodynamic properties of a high pressure subcritical UF/sub 6/He gas volume (irradiated by an external source)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sterritt, D.E.; Lalos, G.T.; Schneider, R.T.

1976-12-01

A computer simulation study concerning a compressed fissioning UF/sub 6/ gas is presented. The compression is to be achieved by a ballistic piston compressor. Data on UF/sub 6/ obtained with this compressor were incorporated in the simulation study. As a neutron source to create the fission events in the compressed gas, a fast burst reactor was considered. The conclusion is that it takes a neutron flux in excess of 10/sup 15/ n/cm/sup 2/-s to produce measurable increases in pressure and temperature, while a flux in excess of 10/sup 19/ n/cm/sup 2/-s would probably damage the compressor.

Novel full-spectral flow cytometry with multiple spectrally-adjacent fluorescent proteins and fluorochromes and visualization of in vivo cellular movement.

PubMed

Futamura, Koji; Sekino, Masashi; Hata, Akihiro; Ikebuchi, Ryoyo; Nakanishi, Yasutaka; Egawa, Gyohei; Kabashima, Kenji; Watanabe, Takeshi; Furuki, Motohiro; Tomura, Michio

2015-09-01

Flow cytometric analysis with multicolor fluoroprobes is an essential method for detecting biological signatures of cells. Here, we present a new full-spectral flow cytometer (spectral-FCM). Unlike conventional flow cytometer, this spectral-FCM acquires the emitted fluorescence for all probes across the full-spectrum from each cell with 32 channels sequential PMT unit after dispersion with prism, and extracts the signals of each fluoroprobe based on the spectral shape of each fluoroprobe using unique algorithm in high speed, high sensitive, accurate, automatic and real-time. The spectral-FCM detects the continuous changes in emission spectra from green to red of the photoconvertible protein, KikGR with high-spectral resolution and separates spectrally-adjacent fluoroprobes, such as FITC (Emission peak (Em) 519 nm) and EGFP (Em 507 nm). Moreover, the spectral-FCM can measure and subtract autofluorescence of each cell providing increased signal-to-noise ratios and improved resolution of dim samples, which leads to a transformative technology for investigation of single cell state and function. These advances make it possible to perform 11-color fluorescence analysis to visualize movement of multilinage immune cells by using KikGR-expressing mice. Thus, the novel spectral flow cytometry improves the combinational use of spectrally-adjacent various FPs and multicolor fluorochromes in metabolically active cell for the investigation of not only the immune system but also other research and clinical fields of use. © 2015 International Society for Advancement of Cytometry.

An experimental investigation of 235 sub UF sub 6 fission produced plasmas. [gas handling system for use with nuclear pumped laser experiments

NASA Technical Reports Server (NTRS)

Miley, G. H.

1981-01-01

A gas handling system capable of use with uranium fluoride was designed and constructed for use with nuclear pumped laser experiments using the TRIGA research reactor. By employing careful design and temperature controls, the UF6 can be first transported into the irradiation chamber, and then, at the conclusion of the experiment, returned to gas cylinders. The design of the system is described. Operating procedures for the UF6 and gas handling systems are included.

Electromigration Critical Product to Measure Effect of Underfill Material in Suppressing Bi Segregation in Sn-58Bi Solder

NASA Astrophysics Data System (ADS)

Zhao, Xu; Takaya, Satoshi; Muraoka, Mikio

2017-08-01

Recently, we detected length-dependent electromigration (EM) behavior in Sn-58Bi (SB) solder and revealed the existence of Bi back-flow, which retards EM-induced Bi segregation and is dependent on solder length. The cause of the back-flow is attributed to an oxide layer formed on the SB solder. At present, underfill (UF) material is commonly used in flip-chip packaging as filler between chip and substrate to surround solder bumps. In this study, we quantitatively investigated the effect of UF material as a passivation layer on EM in SB solder strips. EM tests on SB solder strips with length of 50 μm, 100 μm, and 150 μm were conducted simultaneously. Some samples were coated with commercial thermosetting epoxy UF material, which acted as a passivation layer on the Cu-SB-Cu interconnections. The value of the critical product for SB solder was estimated to be 38 A/cm to 43 A/cm at 353 K to 373 K without UF coating and 59 A/cm at 373 K with UF coating. The UF material acting as a passivation layer suppressed EM-induced Bi segregation and increased the threshold current density by 37% to 55%. However, at very high current density, this effect became very slight. In addition, Bi atoms can diffuse to the anode side through the Sn phase, hence addition of microelements to the Sn phase to form obstacles, such as intermetallic compounds, may retard Bi segregation in SB solder.

Multiparametric Flow Cytometry Using Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes

PubMed Central

Telford, William G.; Shcherbakova, Daria M.; Buschke, David; Hawley, Teresa S.; Verkhusha, Vladislav V.

2015-01-01

Engineering of fluorescent proteins (FPs) has followed a trend of achieving longer fluorescence wavelengths, with the ultimate goal of producing proteins with both excitation and emission in the near-infrared (NIR) region of the spectrum. Flow cytometers are now almost universally equipped with red lasers, and can now be equipped with NIR lasers as well. Most red-shifted FPs of the GFP-like family are maximally excited by orange lasers (590 to 610 nm) not commonly found on cytometers. This has changed with the development of the iRFP series of NIR FPs from the protein family of bacterial phytochromes. The shortest wavelength variants of this series, iRFP670 and iRFP682 showed maximal excitation with visible red lasers. The longer wavelength variants iRFP702, iRFP713 and iRFP720 could be optimally excited by NIR lasers ranging from 685 to 730 nm. Pairs of iRFPs could be detected simultaneously by using red and NIR lasers. Moreover, a novel spectral cytometry technique, which relies on spectral deconvolution rather than optical filters, allowed spectra of all five iRFPs to be analyzed simultaneously with no spectral overlap. Together, the combination of iRFPs with the advanced flow cytometry will allow to first image tissues expressing iRFPs deep in live animals and then quantify individual cell intensities and sort out the distinct primary cell subpopulations ex vivo. PMID:25811854

Multiparametric flow cytometry using near-infrared fluorescent proteins engineered from bacterial phytochromes.

PubMed

Telford, William G; Shcherbakova, Daria M; Buschke, David; Hawley, Teresa S; Verkhusha, Vladislav V

2015-01-01

Engineering of fluorescent proteins (FPs) has followed a trend of achieving longer fluorescence wavelengths, with the ultimate goal of producing proteins with both excitation and emission in the near-infrared (NIR) region of the spectrum. Flow cytometers are now almost universally equipped with red lasers, and can now be equipped with NIR lasers as well. Most red-shifted FPs of the GFP-like family are maximally excited by orange lasers (590 to 610 nm) not commonly found on cytometers. This has changed with the development of the iRFP series of NIR FPs from the protein family of bacterial phytochromes. The shortest wavelength variants of this series, iRFP670 and iRFP682 showed maximal excitation with visible red lasers. The longer wavelength variants iRFP702, iRFP713 and iRFP720 could be optimally excited by NIR lasers ranging from 685 to 730 nm. Pairs of iRFPs could be detected simultaneously by using red and NIR lasers. Moreover, a novel spectral cytometry technique, which relies on spectral deconvolution rather than optical filters, allowed spectra of all five iRFPs to be analyzed simultaneously with no spectral overlap. Together, the combination of iRFPs with the advanced flow cytometry will allow to first image tissues expressing iRFPs deep in live animals and then quantify individual cell intensities and sort out the distinct primary cell subpopulations ex vivo.

Definition of hydraulic stability of KVGM-100 hot-water boiler and minimum water flow rate

NASA Astrophysics Data System (ADS)

Belov, A. A.; Ozerov, A. N.; Usikov, N. V.; Shkondin, I. A.

2016-08-01

In domestic power engineering, the methods of quantitative and qualitative-quantitative adjusting the load of the heat supply systems are widely distributed; furthermore, during the greater part of the heating period, the actual discharge of network water is less than estimated values when changing to quantitative adjustment. Hence, the hydraulic circuits of hot-water boilers should ensure the water velocities, minimizing the scale formation and excluding the formation of stagnant zones. The results of the calculations of hot-water KVGM-100 boiler and minimum water flow rate for the basic and peak modes at the fulfillment of condition of the lack of surface boil are presented in the article. The minimal flow rates of water at its underheating to the saturation state and the thermal flows in the furnace chamber were defined. The boiler hydraulic calculation was performed using the "Hydraulic" program, and the analysis of permissible and actual velocities of the water movement in the pipes of the heating surfaces was carried out. Based on the thermal calculations of furnace chamber and thermal- hydraulic calculations of heating surfaces, the following conclusions were drawn: the minimum velocity of water movement (by condition of boiling surface) at lifting movement of environment increases from 0.64 to 0.79 m/s; it increases from 1.14 to 1.38 m/s at down movement of environmental; the minimum water flow rate by the boiler in the basic mode (by condition of the surface boiling) increased from 887 t/h at the load of 20% up to 1074 t/h at the load of 100%. The minimum flow rate is 1074 t/h at nominal load and is achieved at the pressure at the boiler outlet equal to 1.1 MPa; the minimum water flow rate by the boiler in the peak mode by condition of surface boiling increases from 1669 t/h at the load of 20% up to 2021 t/h at the load of 100%.

Flow cytometry and sorting

NASA Astrophysics Data System (ADS)

Yentsch, Clarice M.

For the first time oceanographers have a tool, known as a flow cytometer and sorter, which is useful for simultaneous measurement of multiple parameters of individual cells and particles at rapid rates. We are now able to exploit the fluorescent capability of pigments and stains as signals to quantify and separate subpopulations of cells and particles in the 1.0 to 150 μm size range. Analysis rates exceed 1000 cells per second and high sensitivity is attained using laser excitation.The addition of this new technology to the ocean sciences will enable researchers to address problems which were previously intractable. The first unit, funded by the Office of Naval Research and the National Science Foundation, will be at Bigelow Laboratory for Ocean Sciences in West Boothbay Harbor, Maine, in the laboratory of Clarice M. Yentsch and David A. Phinney. In anticipation of this award, a workshop course on flow cytometry (FCM) and sorting techniques was held from October 24 through November 1, 1982, at the Bermuda Biological Station.

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM).

PubMed

Tang, Anson H L; Lai, Queenie T K; Chung, Bob M F; Lee, Kelvin C M; Mok, Aaron T Y; Yip, G K; Shum, Anderson H C; Wong, Kenneth K Y; Tsia, Kevin K

2017-06-28

Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the main trend in the advanced development of flow cytometry. Notably, adding high-resolution imaging capabilities allows for the complex morphological analysis of cellular/sub-cellular structures. This is not possible with standard flow cytometers. However, it is valuable for advancing our knowledge of cellular functions and can benefit life science research, clinical diagnostics, and environmental monitoring. Incorporating imaging capabilities into flow cytometry compromises the assay throughput, primarily due to the limitations on speed and sensitivity in the camera technologies. To overcome this speed or throughput challenge facing imaging flow cytometry while preserving the image quality, asymmetric-detection time-stretch optical microscopy (ATOM) has been demonstrated to enable high-contrast, single-cell imaging with sub-cellular resolution, at an imaging throughput as high as 100,000 cells/s. Based on the imaging concept of conventional time-stretch imaging, which relies on all-optical image encoding and retrieval through the use of ultrafast broadband laser pulses, ATOM further advances imaging performance by enhancing the image contrast of unlabeled/unstained cells. This is achieved by accessing the phase-gradient information of the cells, which is spectrally encoded into single-shot broadband pulses. Hence, ATOM is particularly advantageous in high-throughput measurements of single-cell morphology and texture - information indicative of cell types, states, and even functions. Ultimately, this could become a powerful imaging flow cytometry platform for the biophysical phenotyping of cells, complementing the current state-of-the-art biochemical-marker-based cellular assay. This work describes a protocol to establish the key modules of an ATOM system (from optical frontend to data processing and visualization

The UF family of reference hybrid phantoms for computational radiation dosimetry

NASA Astrophysics Data System (ADS)

Lee, Choonsik; Lodwick, Daniel; Hurtado, Jorge; Pafundi, Deanna; Williams, Jonathan L.; Bolch, Wesley E.

2010-01-01

organ masses from ICRP Publication 89, (3) reference elemental compositions provided in ICRP 89 as well as ICRU Report 46, and (4) reference data on the alimentary tract organs given in ICRP Publications 89 and 100. Various adjustments and refinements to the organ systems of the previously described newborn, 15 year and adult phantoms are also presented. The UF series of hybrid phantoms retain the non-uniform scalability of stylized phantoms while maintaining the anatomical realism of patient-specific voxel phantoms with respect to organ shape, depth and inter-organ distance. While the final versions of these phantoms are in a voxelized format for radiation transport simulation, their primary format is given as NURBS and polygon mesh surfaces, thus permitting one to sculpt non-reference phantoms using the reference phantoms as an anatomic template.

Simulation and performance analysis of a novel high-accuracy sheathless microfluidic impedance cytometer with coplanar electrode layout.

PubMed

Caselli, Federica; Bisegna, Paolo

2017-10-01

The performance of a novel microfluidic impedance cytometer (MIC) with coplanar configuration is investigated in silico. The main feature of the device is the ability to provide accurate particle-sizing despite the well-known measurement sensitivity to particle trajectory. The working principle of the device is presented and validated by means of an original virtual laboratory providing close-to-experimental synthetic data streams. It is shown that a metric correlating with particle trajectory can be extracted from the signal traces and used to compensate the trajectory-induced error in the estimated particle size, thus reaching high-accuracy. An analysis of relevant parameters of the experimental setup is also presented. Copyright © 2017 IPEM. Published by Elsevier Ltd. All rights reserved.

Parallel pulse processing and data acquisition for high speed, low error flow cytometry

DOEpatents

Engh, G.J. van den; Stokdijk, W.

1992-09-22

A digitally synchronized parallel pulse processing and data acquisition system for a flow cytometer has multiple parallel input channels with independent pulse digitization and FIFO storage buffer. A trigger circuit controls the pulse digitization on all channels. After an event has been stored in each FIFO, a bus controller moves the oldest entry from each FIFO buffer onto a common data bus. The trigger circuit generates an ID number for each FIFO entry, which is checked by an error detection circuit. The system has high speed and low error rate. 17 figs.

Parallel pulse processing and data acquisition for high speed, low error flow cytometry

DOEpatents

van den Engh, Gerrit J.; Stokdijk, Willem

1992-01-01

A digitally synchronized parallel pulse processing and data acquisition system for a flow cytometer has multiple parallel input channels with independent pulse digitization and FIFO storage buffer. A trigger circuit controls the pulse digitization on all channels. After an event has been stored in each FIFO, a bus controller moves the oldest entry from each FIFO buffer onto a common data bus. The trigger circuit generates an ID number for each FIFO entry, which is checked by an error detection circuit. The system has high speed and low error rate.

Novel full‐spectral flow cytometry with multiple spectrally‐adjacent fluorescent proteins and fluorochromes and visualization of in vivo cellular movement

PubMed Central

Futamura, Koji; Sekino, Masashi; Hata, Akihiro; Ikebuchi, Ryoyo; Nakanishi, Yasutaka; Egawa, Gyohei; Kabashima, Kenji; Watanabe, Takeshi; Furuki, Motohiro

2015-01-01

Abstract Flow cytometric analysis with multicolor fluoroprobes is an essential method for detecting biological signatures of cells. Here, we present a new full‐spectral flow cytometer (spectral‐FCM). Unlike conventional flow cytometer, this spectral‐FCM acquires the emitted fluorescence for all probes across the full‐spectrum from each cell with 32 channels sequential PMT unit after dispersion with prism, and extracts the signals of each fluoroprobe based on the spectral shape of each fluoroprobe using unique algorithm in high speed, high sensitive, accurate, automatic and real‐time. The spectral‐FCM detects the continuous changes in emission spectra from green to red of the photoconvertible protein, KikGR with high‐spectral resolution and separates spectrally‐adjacent fluoroprobes, such as FITC (Emission peak (Em) 519 nm) and EGFP (Em 507 nm). Moreover, the spectral‐FCM can measure and subtract autofluorescence of each cell providing increased signal‐to‐noise ratios and improved resolution of dim samples, which leads to a transformative technology for investigation of single cell state and function. These advances make it possible to perform 11‐color fluorescence analysis to visualize movement of multilinage immune cells by using KikGR‐expressing mice. Thus, the novel spectral flow cytometry improves the combinational use of spectrally‐adjacent various FPs and multicolor fluorochromes in metabolically active cell for the investigation of not only the immune system but also other research and clinical fields of use. © 2015 The Authors. Cytometry Part A Published by Wiley Periodicals, Inc. on behalf of ISAC PMID:26217952

Aged spontaneously hypertensive rats (SHRs) are predisposed to ultrafine carbon particle (UfCP) mediated cardiovascular impairment.

EPA Science Inventory

Background: Elderly individuals are considered to be more susceptible to particulate matter (PM) related cardiovascular (CV) health effects. In this study we investigated UfCP mediated CV effects on aged SHRs and compared the findings to that of adult SHRs to identify age related...

Technology development for the LISA using the UF Torsion Pendulu

NASA Astrophysics Data System (ADS)

Conklin, John W.; Chilton, Andrew; Olatunde, Taiwo; Apple, Stephen; Ciani, Giacomo; Mueller, Guido

2015-08-01

Space-based gravitational wave observatories like LISA measure picometer changes in the distances between free falling test masses separated by millions of kilometers caused by gravitational waves. A test mass and its associated sensing, actuation, charge control and caging subsystems are referred to as a gravitational reference sensor (GRS). LISA will observe gravitational wave sources ranging from super-massive black hole mergers to compact galactic binaries in the millihertz region, and LISA science has consistently been ranked in the top two for future large space missions in the last two NASA astrophysics decadal reviews. With the 2015 launch of LISA Pathfinder (LPF) and the expected detection of gravitational waves by aLIGO and/or Pulsar Timing Arrays within in the next several years, this can arguably be called the decade of gravitational waves. Following a successful demonstration of the baseline LISA GRS by LPF, the measurement principle will be carried forward, but improvements in several GRS components are possible over the next ten years that will lead to cost savings and potential noise reductions. The UF LISA group has constructed the UF Torsion Pendulum to increase U.S. competency in this critical area and to have a facility where new technologies can be developed and evaluated. This experimental facility is based on the design of a similar facility at the University of Trento, and consists of a vacuum enclosed torsion pendulum that suspends mock-ups of the LISA test masses, surrounded by electrode housings. This presentation will describe this facility, focusing on its mechanical design, capacitive sensing and electrostatic actuation systems, and overall acceleration noise performance

Magnetic fingerprints of rolling cells for quantitative flow cytometry in whole blood

NASA Astrophysics Data System (ADS)

Reisbeck, Mathias; Helou, Michael Johannes; Richter, Lukas; Kappes, Barbara; Friedrich, Oliver; Hayden, Oliver

2016-09-01

Over the past 50 years, flow cytometry has had a profound impact on preclinical and clinical applications requiring single cell function information for counting, sub-typing and quantification of epitope expression. At the same time, the workflow complexity and high costs of such optical systems still limit flow cytometry applications to specialized laboratories. Here, we present a quantitative magnetic flow cytometer that incorporates in situ magnetophoretic cell focusing for highly accurate and reproducible rolling of the cellular targets over giant magnetoresistance sensing elements. Time-of-flight analysis is used to unveil quantitative single cell information contained in its magnetic fingerprint. Furthermore, we used erythrocytes as a biological model to validate our methodology with respect to precise analysis of the hydrodynamic cell diameter, quantification of binding capacity of immunomagnetic labels, and discrimination of cell morphology. The extracted time-of-flight information should enable point-of-care quantitative flow cytometry in whole blood for clinical applications, such as immunology and primary hemostasis.

Microfabrication and Test of a Three-Dimensional Polymer Hydro-focusing Unit for Flow Cytometry Applications

NASA Technical Reports Server (NTRS)

Yang, Ren; Feeback, Daniel L.; Wang, Wan-Jun

2005-01-01

This paper details a novel three-dimensional (3D) hydro-focusing micro cell sorter for micro flow cytometry applications. The unit was microfabricated by means of SU-8 3D lithography. The 3D microstructure for coaxial sheathing was designed, microfabricated, and tested. Three-dimensional hydrofocusing capability was demonstrated with an experiment to sort labeled tanned sheep erythrocytes (red blood cells). This polymer hydro-focusing microstructure is easily microfabricated and integrated with other polymer microfluidic structures. Keywords: SU-8, three-dimensional hydro-focusing, microfluidic, microchannel, cytometer

Spectrophotometer properties of vein blood plasma in UF-region patients with sharp surgical pathology of abdominal region organs

NASA Astrophysics Data System (ADS)

Guminestskij, S. G.; Polianski, I. J.; Motrich, A. V.; Grunchuk, F. W.

2006-05-01

It is set that there are two maximums in UF- region absorption of vein blood plasma of a man: at λ = 235 nm and at λ = 280 nm. It is shown that there are the substantial changes of values of the optical density D comparative with controls (for donors) exactly in a maximum at development of sharp surgical diseases of organs of abdominal region λ = 280 nm, in that time as maximum at λ = 235 nm in this plan is not informing. Resulted results of researches of dynamics of changes of optical properties of vein blood plasma in UF- region of patients with pathology of abdominal region organs in after operating period (sharp appendicitis, sharp pancreatitis, intestinal impassability and others like that), which can have the diagnostic value.

The substitution of sand filtration by immersed-UF for surface water treatment: pilot-scale studies.

PubMed

Lihua, Sun; Xing, Li; Guoyu, Zhang; Jie, Chen; Zhe, Xu; Guibai, Li

2009-01-01

The newly issued National Drinking Water Standard required that turbidity should be lower than 1 NTU, and the substitution of sand filtration by immersed ultrafiltration (immersed-UF) is feasible to achieve the standard. This study aimed to optimise the operational processes (i.e. aeration, backwashing) through pilot scale studies, to control membrane fouling while treating the sedimentation effluent. Results indicated that the immersed-UF was promising to treat the sedimentation effluent. The turbidity was below 0.10 NTU, bacteria and E. coli were not detected in the permeate water. The intermittent filtration with aeration is beneficial to inhibit membrane fouling. The critical aeration intensity is observed to be 60.0 m(3) m(-2) h(-1). At this aeration intensity, the decline rate of permeate flux in one period of backwashing was 1.94% and 7.03% for intermittent filtration and sustained filtration respectively. The different membrane backwashing methods (i.e. aeration 1.5 min, synchronous aeration and water backwashing 2 min, water backwashing 1.5 min; synchronous aeration and water backwashing 3 min, water backwashing 2 min; aeration 3 min, single water backwashing 2 min; synchronous aeration and water backwashing 5 min; single water backwashing 5 min) on the recovery of permeate flux were compared, indicating that the synchronous aeration and water backwashing exhibited best potential for permeate flux recovery. The optimal intensity of water backwashing is shown to be 90.0 L m(-2) h(-1). When the actual water intensity was below or exceeded the value, the recovery rate of permeate flux would be reduced. Additionally, the average operating cost for the immersed UF membrane, including the power, the chemical cleaning reagents, and membrane modules replacement, was about 0.31 RMB/m(3).

Evaluation of a flow cytometry method to determine size and real refractive index distributions in natural marine particle populations.

PubMed

Agagliate, Jacopo; Röttgers, Rüdiger; Twardowski, Michael S; McKee, David

2018-03-01

A flow cytometric (FC) method was developed to retrieve particle size distributions (PSDs) and real refractive index (n r ) information in natural waters. Geometry and signal response of the sensors within the flow cytometer (CytoSense, CytoBuoy b.v., Netherlands) were characterized to form a scattering inversion model based on Mie theory. The procedure produced a mesh of diameter and n r isolines where each particle is assigned the diameter and n r values of the closest node, producing PSDs and particle real refractive index distributions. The method was validated using polystyrene bead standards of known diameter and polydisperse suspensions of oil with known n r , and subsequently applied to natural samples collected across a broad range of UK shelf seas. FC PSDs were compared with independent PSDs produced from data of two LISST-100X instruments (type B and type C). PSD slopes and features were found to be consistent between the FC and the two LISST-100X instruments, but LISST concentrations were found in disagreement with FC concentrations and with each other. FC n r values were found to agree with expected refractive index values of typical marine particle components across all samples considered. The determination of particle size and refractive index distributions enabled by the FC method has potential to facilitate identification of the contribution of individual subpopulations to the bulk inherent optical properties and biogeochemical properties of the particle population.

Accurate live and dead bacterial cell enumeration using flow cytometry (Conference Presentation)

NASA Astrophysics Data System (ADS)

Ou, Fang; McGoverin, Cushla; Swift, Simon; Vanholsbeeck, Frédérique

2017-03-01

Flow cytometry (FCM) is based on the detection of scattered light and fluorescence to identify cells with particular characteristics of interest. However most FCM cannot precisely control the flow through its interrogation point and hence the volume and concentration of the sample cannot be immediately obtained. The easiest, most reliable and inexpensive way of obtaining absolute counts with FCM is by using reference beads. We investigated a method of using FCM with reference beads to measure live and dead bacterial concentration over the range of 106 to 108 cells/mL and ratio varying from 0 to 100%. We believe we are the first to use this method for such a large cell concentration range while also establishing the effect of varying the live/dead bacteria ratios. Escherichia coli solutions with differing ratios of live:dead cells were stained with fluorescent dyes SYTO 9 and propidium iodide (PI), which label live and dead cells, respectively. Samples were measured using a LSR II Flow Cytometer (BD Biosciences); using 488 nm excitation with 20 mW power. Both SYTO 9 and PI fluorescence were collected and threshold was set to side scatter. Traditional culture-based plate count was done in parallel to the FCM analysis. The concentration of live bacteria from FCM was compared to that obtained by plate counts. Preliminary results show that the concentration of live bacteria obtained by FCM and plate counts correlate well with each other and indicates this may be extended to a wider concentration range or for studying other cell characteristics.

How mole ratio of UF resin affects formaldehyde emission and other properties : a literature critique

Treesearch

George E. Myers

1984-01-01

A critical review was made of the literature concerned with how the formaldehyde to urea mole ratio (F/U) affects formaldehyde emission from particleboard and plywood bonded with urea-formaldehyde (UF) adhesives, and how this ratio affects certain other adhesive and board properties. It is difficult to quantify the dependence of various properties on mole ratio or...

Non-linear optical flow cytometry using a scanned, Bessel beam light-sheet.

PubMed

Collier, Bradley B; Awasthi, Samir; Lieu, Deborah K; Chan, James W

2015-05-29

Modern flow cytometry instruments have become vital tools for high-throughput analysis of single cells. However, as issues with the cellular labeling techniques often used in flow cytometry have become more of a concern, the development of label-free modalities for cellular analysis is increasingly desired. Non-linear optical phenomena (NLO) are of growing interest for label-free analysis because of the ability to measure the intrinsic optical response of biomolecules found in cells. We demonstrate that a light-sheet consisting of a scanned Bessel beam is an optimal excitation geometry for efficiently generating NLO signals in a microfluidic environment. The balance of photon density and cross-sectional area provided by the light-sheet allowed significantly larger two-photon fluorescence intensities to be measured in a model polystyrene microparticle system compared to measurements made using other excitation focal geometries, including a relaxed Gaussian excitation beam often used in conventional flow cytometers.

Visualization of Pulmonary Clearance Mechanisms via Noninvasive Optical Imaging Validated by Near-Infrared Flow Cytometry

PubMed Central

Zhou, Haiying; Gunsten, Sean P.; Zhegalova, Natalia G.; Bloch, Sharon; Achilefu, Samuel; Holley, J. Christopher; Schweppe, Daniel; Akers, Walter; Brody, Steven L.; Eades, William; Berezin, Mikhail Y.

2016-01-01

In vivo optical imaging with near-infrared (NIR) probes is an established method of diagnostics in preclinical and clinical studies. However, the specificities of these probes are difficult to validate ex vivo due to the lack of NIR flow cytometry. To address this limitation, we modified a flow cytometer to include an additional NIR channel using a 752 nm laser line. The flow cytometry system was tested using NIR microspheres and cell lines labeled with a combination of visible range and NIR fluorescent dyes. The approach was verified in vivo in mice evaluated for immune response in lungs after intratracheal delivery of the NIR contrast agent. Flow cytometry of cells obtained from the lung bronchoalveolar lavage demonstrated that the NIR dye was taken up by pulmonary macrophages as early as four-hours post-injection. This combination of optical imaging with NIR flow cytometry extends the capability of imaging and enables complementation of in vivo imaging with cell-specific studies. PMID:25808737

URANIUM RECOVERY FROM COMPOSITE UF$sub 4$ REDUCTION BOMB WASTES

DOE Office of Scientific and Technical Information (OSTI.GOV)

Johnson, E R; Doyle, R L; Coleman, J R

1954-01-28

A number of techniques have been investigated on a laboratory-scale for separating uranium from fluorides during the recovery of uranium from UF4 reduction bomb wastes (C-oxide) by an HCl leach - NH4OH precipitation process. Among these are included adsorption of fluorides from filtered leach liquors, fractional precipitation of fluorides and uranium, complexing of fluorides into forms soluble in slightly acid solutions, and fluoride volatilization from the uranium concentrate. Solubility studies of CaF2 and MgF2 in aqueous hydrochloric acid at various acidities and temperatures were also conducted. A description of the production-scale processing of C-oxide in the FMPC scrap plant hasmore » been included.« less

Hybrid coagulation-UF processes for spent filter backwash water treatment: a comparison studies for PAFCl and FeCl3 as a pre-treatment.

PubMed

Ebrahimi, Afshin; Amin, Mohammad Mehdi; Pourzamani, Hamidreza; Hajizadeh, Yaghoub; Mahvi, Amir Hossein; Mahdavi, Mokhtar; Rad, Mohammad Hassan Rabie

2017-08-01

In this study, the reclamation of clean water from spent filter backwash water (SFBW) was investigated through pilot-scale experiments. The pilot plant consisted of pre-sedimentation, coagulation, flocculation, clarification, and ultrafiltration (UF). Two coagulants of PAFCl and FeCl 3 were investigated with respect to their performance on treated SFBW quality and UF membrane fouling. At the optimum dose of PAFCl and FeCl 3 turbidity removal of 99.6 and 99.4% was attained, respectively. PAFCl resulted in an optimum UV 254 , TOC, and DOC removal of 80, 83.6, and 72.7%, respectively, and FeCl 3 caused the removal of those parameters by 76.7, 80.9, and 65.9%, respectively. PAFCl removed hydrophilic and transphilic constituent better than FeCl 3 , but FeCl 3 had, to some extent, higher affinities to a hydrophobic fraction. It was concluded that PAFCl showed a better coagulation performance in most cases and caused a lower membrane fouling rate compared to FeCl 3 . Finally, the treated SFBW with both coagulant-UF systems met the drinking water standards.

Particle analysis in an acoustic cytometer

DOEpatents

Kaduchak, Gregory; Ward, Michael D

2012-09-18

The present invention is a method and apparatus for acoustically manipulating one or more particles. Acoustically manipulated particles may be separated by size. The particles may be flowed in a flow stream and acoustic radiation pressure, which may be radial, may be applied to the flow stream. This application of acoustic radiation pressure may separate the particles. In one embodiment, the particles may be separated by size, and as a further example, the larger particles may be transported to a central axis.

Scattering matrix elements of biological particles measured in a flow through system: theory and practice.

PubMed

Sloot, P M; Hoekstra, A G; van der Liet, H; Figdor, C G

1989-05-15

Light scattering techniques (including depolarization experiments) applied to biological cells provide a fast nondestructive probe that is very sensitive to small morphological differences. Until now quantitative measurement of these scatter phenomena were only described for particles in suspension. In this paper we discuss the symmetry conditions applicable to the scattering matrices of monodisperse biological cells in a flow cytometer and provide evidence that quantitative measurement of the elements of these scattering matrices is possible in flow through systems. Two fundamental extensions to the theoretical description of conventional scattering experiments are introduced: large cone integration of scattering signals and simultaneous implementation of the localization principle to account for scattering by a sharply focused laser beam. In addition, a specific calibration technique is proposed to account for depolarization effects of the highly specialized optics applied in flow through equipment.

Preliminary Studies on Membrane Filtration for the Production of Potable Water: A Case of Tshaanda Rural Village in South Africa

PubMed Central

Molelekwa, Gomotsegang F.; Mukhola, Murembiwa S.; Van der Bruggen, Bart; Luis, Patricia

2014-01-01

Ultrafiltration (UF) systems have been used globally for treating water from resources including rivers, reservoirs, and lakes for the production of potable water in the past decade. UF membranes with a pore size of between 0.1 and 0.01 micrometres provide an effective barrier for bacteria, viruses, suspended particles, and colloids. The use of UF membrane technology in treating groundwater for the supply of potable water in the impoverished and rural village, Tshaanda (i.e., the study area) is demonstrated. The technical and administrative processes that are critical for the successful installation of the pilot plant were developed. Given the rural nature of Tshaanda, the cultural and traditional protocols were observed. Preliminary results of the water quality of untreated water and the permeate are presented. Escherichia coli in the untreated water during the dry season (i.e., June and July) was 2 cfu/100 ml and was <1 cfu/100 ml (undetected) following UF, which complied with the WHO and South African National Standards and Guidelines of <1 cfu/100 ml. During the wet/rainy season (February) total coliform was unacceptably high (>2419.2 cfu/100 ml) before UF. Following UF, it dramatically reduced to acceptable level (7 cfu/100 ml) which is within the WHO recommended level of <10 cfu/100 ml. Additionally, during the wet/rainy season E. coli and enterococci were unacceptably high (40.4 cfu/100 ml and 73.3 cfu/100 ml, respectively) before UF but were completely removed following UF, which are within the WHO and SANS recommended limit. The values for electrical conductivity (EC) and turbidity were constantly within the WHO recommended limits of 300 µS/cm corrected at 25°C and <5 NTU, respectively, before and after UF, during dry season and wet season. This suggests that there is no need for pre-treatment of the water for suspended particles and colloids. Considering these data, it can be concluded that the water is suitable for human consumption, following UF. PMID

Preliminary studies on membrane filtration for the production of potable water: a case of Tshaanda rural village in South Africa.

PubMed

Molelekwa, Gomotsegang F; Mukhola, Murembiwa S; Van der Bruggen, Bart; Luis, Patricia

2014-01-01

Ultrafiltration (UF) systems have been used globally for treating water from resources including rivers, reservoirs, and lakes for the production of potable water in the past decade. UF membranes with a pore size of between 0.1 and 0.01 micrometres provide an effective barrier for bacteria, viruses, suspended particles, and colloids. The use of UF membrane technology in treating groundwater for the supply of potable water in the impoverished and rural village, Tshaanda (i.e., the study area) is demonstrated. The technical and administrative processes that are critical for the successful installation of the pilot plant were developed. Given the rural nature of Tshaanda, the cultural and traditional protocols were observed. Preliminary results of the water quality of untreated water and the permeate are presented. Escherichia coli in the untreated water during the dry season (i.e., June and July) was 2 cfu/100 ml and was <1 cfu/100 ml (undetected) following UF, which complied with the WHO and South African National Standards and Guidelines of <1 cfu/100 ml. During the wet/rainy season (February) total coliform was unacceptably high (>2419.2 cfu/100 ml) before UF. Following UF, it dramatically reduced to acceptable level (7 cfu/100 ml) which is within the WHO recommended level of <10 cfu/100 ml. Additionally, during the wet/rainy season E. coli and enterococci were unacceptably high (40.4 cfu/100 ml and 73.3 cfu/100 ml, respectively) before UF but were completely removed following UF, which are within the WHO and SANS recommended limit. The values for electrical conductivity (EC) and turbidity were constantly within the WHO recommended limits of 300 µS/cm corrected at 25°C and <5 NTU, respectively, before and after UF, during dry season and wet season. This suggests that there is no need for pre-treatment of the water for suspended particles and colloids. Considering these data, it can be concluded that the water is suitable for human consumption, following UF.

235U enrichment determination on UF6 cylinders with CZT detectors

NASA Astrophysics Data System (ADS)

Berndt, Reinhard; Mortreau, Patricia

2018-04-01

Measurements of uranium enrichment in UF6 transit cylinders are an important nuclear safeguards verification task, which is performed using a non-destructive assay method, the traditional enrichment meter, which involves measuring the count rate of the 186 keV gamma ray. This provides a direct measure of the 235U enrichment. Measurements are typically performed using either high-resolution detectors (Germanium) with e-cooling and battery operation, or portable devices equipped with low resolution detectors (NaI). Despite good results being achieved when measuring Low Enriched Uranium in 30B type cylinders and natural uranium in 48Y type containers using both detector systems, there are situations, which preclude the use of one or both of these systems. The focus of this work is to address some of the recognized limitations in relation to the current use of the above detector systems by considering the feasibility of an inspection instrument for 235U enrichment measurements on UF6 cylinders using the compact and light Cadmium Zinc Telluride (CZT) detectors. In the present work, test measurements were carried out, under field conditions and on full-size objects, with different CZT detectors, in particular for situations where existing systems cannot be used e.g. for stacks of 48Y type containers with depleted uranium. The main result of this study shows that the CZT detectors, actually a cluster of four μCZT1500 micro spectrometers provide as good results as the germanium detector in the ORTEC Micro-trans SPEC HPGe Portable spectrometer, and most importantly in particular for natural and depleted uranium in 48Y cylinders.

Electrochemical separation of uranium in the molten system LiF-NaF-KF-UF4

NASA Astrophysics Data System (ADS)

Korenko, M.; Straka, M.; Szatmáry, L.; Ambrová, M.; Uhlíř, J.

2013-09-01

This article is focused on the electrochemical investigation (cyclic voltammetry and related studies) of possible reduction of U4+ ions to metal uranium in the molten system LiF-NaF-KF(eut.)-UF4 that can provide basis for the electrochemical extraction of uranium from molten salts. Two-step reduction mechanism for U4+ ions involving one electron exchange in soluble/soluble U4+/U3+ system and three electrons exchange in the second step were found on the nickel working electrode. Both steps were found to be reversible and diffusion controlled. Based on cyclic voltammetry, the diffusion coefficients of uranium ions at 530 °C were found to be D(U4+) = 1.64 × 10-5 cm2 s-1 and D(U3+) 1.76 × 10-5 cm2 s-1. Usage of the nickel spiral electrode for electrorefining of uranium showed fairly good feasibility of its extraction. However some oxidant present during the process of electrorefining caused that the solid deposits contained different uranium species such as UF3, UO2 and K3UO2F5.

Study on the removal efficiency of UF membranes using bacteriophages in bench-scale and semi-technical scale.

PubMed

Kreissel, K; Bösl, M; Lipp, P; Franzreb, M; Hambsch, B

2012-01-01

To determine the removal efficiency of ultrafiltration (UF) membranes for nano-particles in the size range of viruses the state of the art uses challenge tests with virus-spiked water. This work focuses on bench-scale and semi-technical scale experiments. Different experimental parameters influencing the removal efficiency of the tested UF membrane modules were analyzed and evaluated for bench- and semi-technical scale experiments. Organic matter in the water matrix highly influenced the removal of the tested bacteriophages MS2 and phiX174. Less membrane fouling (low ΔTMP) led to a reduced phage reduction. Increased flux positively affected phage removal in natural waters. The tested bacteriophages MS2 and phiX174 revealed different removal properties. MS2, which is widely used as a model organism to determine virus removal efficiencies of membranes, mostly showed a better removal than phiX174 for the natural water qualities tested. It seems that MS2 is possibly a less conservative surrogate for human enteric virus removal than phiX174. In bench-scale experiments log removal values (LRV) for MS2 of 2.5-6.0 and of 2.5-4.5 for phiX174 were obtained for the examined range of parameters. Phage removal obtained with differently fabricated semi-technical modules was quite variable for comparable parameter settings, indicating that module fabrication can lead to differing results. Potting temperature and module size were identified as influencing factors. In conclusion, careful attention has to be paid to the choice of experimental settings and module potting when using bench-scale or semi-technical scale experiments for UF membrane challenge tests.

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.

Thermodynamic properties of UF sub 6 measured with a ballistic piston compressor

NASA Technical Reports Server (NTRS)

Sterritt, D. E.; Lalos, G. T.; Schneider, R. T.

1973-01-01

From experiments performed with a ballistic piston compressor, certain thermodynamic properties of uranium hexafluoride were investigated. Difficulties presented by the nonideal processes encountered in ballistic compressors are discussed and a computer code BCCC (Ballistic Compressor Computer Code) is developed to analyze the experimental data. The BCCC unfolds the thermodynamic properties of uranium hexafluoride from the helium-uranium hexafluoride mixture used as the test gas in the ballistic compressor. The thermodynamic properties deduced include the specific heat at constant volume, the ratio of specific heats for UF6, and the viscous coupling constant of helium-uranium hexafluoride mixtures.

In vivo plant flow cytometry: A first proof-of-concept

PubMed Central

Nedosekin, Dmitry A.; Khodakovskaya, Mariya V.; Biris, Alexandru S.; Wang, Daoyuan; Xu, Yang; Villagarcia, Hector; Galanzha, Ekaterina I.; Zharov, Vladimir P.

2011-01-01

In vivo flow cytometry has facilitated advances in the ultrasensitive detection of tumor cells, bacteria, nanoparticles, dyes, and other normal and abnormal objects directly in blood and lymph circulatory systems. Here, we propose in vivo plant flow cytometry for the real-time noninvasive study of nanomaterial transport in xylem and phloem plant vascular systems. As a proof of this concept, we demonstrate in vivo real-time photoacoustic monitoring of quantum dot-carbon nanotube conjugate uptake and uptake by roots and spreading through stem to leaves in a tomato plant. In addition, in vivo scanning cytometry using multimodal photoacoustic, photothermal, and fluorescent detection schematics provided multiplex detection and identification of nanoparticles accumulated in plant leaves in the presence of intensive absorption, scattering, and autofluorescent backgrounds. The use of a portable fiber-based photoacoustic flow cytometer for studies of plant vasculature was demonstrated. These integrated cytometry modalities using both endogenous and exogenous contrast agents have a potential to open new avenues of in vivo study of the nutrients, products of photosynthesis and metabolism, nanoparticles, infectious agents, and other objects transported through plant vasculature. PMID:21905208

A general method for bead-enhanced quantitation by flow cytometry

PubMed Central

Montes, Martin; Jaensson, Elin A.; Orozco, Aaron F.; Lewis, Dorothy E.; Corry, David B.

2009-01-01

Flow cytometry provides accurate relative cellular quantitation (percent abundance) of cells from diverse samples, but technical limitations of most flow cytometers preclude accurate absolute quantitation. Several quantitation standards are now commercially available which, when added to samples, permit absolute quantitation of CD4+ T cells. However, these reagents are limited by their cost, technical complexity, requirement for additional software and/or limited applicability. Moreover, few studies have validated the use of such reagents in complex biological samples, especially for quantitation of non-T cells. Here we show that addition to samples of known quantities of polystyrene fluorescence standardization beads permits accurate quantitation of CD4+ T cells from complex cell samples. This procedure, here termed single bead-enhanced cytofluorimetry (SBEC), was equally capable of enumerating eosinophils as well as subcellular fragments of apoptotic cells, moieties with very different optical and fluorescent characteristics. Relative to other proprietary products, SBEC is simple, inexpensive and requires no special software, suggesting that the method is suitable for the routine quantitation of most cells and other particles by flow cytometry. PMID:17067632

A vectorization of the Hess McDonnell Douglas potential flow program NUED for the STAR-100 computer

NASA Technical Reports Server (NTRS)

Boney, L. R.; Smith, R. E., Jr.

1979-01-01

The computer program NUED for analyzing potential flow about arbitrary three dimensional lifting bodies using the panel method was modified to use vector operations and run on the STAR-100 computer. A high speed of computation and ability to approximate the body surface with a large number of panels are characteristics of NUEDV. The new program shows that vector operations can be readily implemented in programs of this type to increase the computational speed on the STAR-100 computer. The virtual memory architecture of the STAR-100 facilitates the use of large numbers of panels to approximate the body surface.

Portable dual field gradient force multichannel flow cytometer device with a dual wavelength low noise detection scheme

DOEpatents

James, Conrad D; Galambos, Paul C; Derzon, Mark S; Graf, Darin C; Pohl, Kenneth R; Bourdon, Chris J

2012-10-23

Systems and methods for combining dielectrophoresis, magnetic forces, and hydrodynamic forces to manipulate particles in channels formed on top of an electrode substrate are discussed. A magnet placed in contact under the electrode substrate while particles are flowing within the channel above the electrode substrate allows these three forces to be balanced when the system is in operation. An optical detection scheme using near-confocal microscopy for simultaneously detecting two wavelengths of light emitted from the flowing particles is also discussed.

Guide to red fluorescent proteins and biosensors for flow cytometry.

PubMed

Piatkevich, Kiryl D; Verkhusha, Vladislav V

2011-01-01

Since the discovery of the first red fluorescent protein (RFP), named DsRed, 12 years ago, a wide pallet of red-shifted fluorescent proteins has been cloned and biotechnologically developed into monomeric fluorescent probes for optical microscopy. Several new types of monomeric RFPs that change the emission wavelength either with time, called fluorescent timers, or after a brief irradiation with violet light, known as photoactivatable proteins, have been also engineered. Moreover, RFPs with a large Stokes shift of fluorescence emission have been recently designed. Because of their distinctive excitation and fluorescence detection conditions developed specifically for microscopy, these fluorescent probes can be suboptimal for flow cytometry. Here, we have selected and summarized the advanced orange, red, and far-red fluorescent proteins with the properties specifically required for the flow cytometry applications. Their effective brightness was calculated for the laser sources available for the commercial flow cytometers and sorters. Compatibility of the fluorescent proteins of different colors in a multiparameter flow cytometry was determined. Novel FRET pairs, utilizing RFPs, RFP-based intracellular biosensors, and their application to a high-throughput screening, are also discussed. Copyright © 2011 Elsevier Inc. All rights reserved.

Technical feasibility study of a low-cost hybrid PAC-UF system for wastewater reclamation and reuse: a focus on feedwater production for low-pressure boilers.

PubMed

Amosa, Mutiu Kolade; Jami, Mohammed Saedi; Alkhatib, Ma'an Fahmi R; Majozi, Thokozani

2016-11-01

This study has applied the concept of the hybrid PAC-UF process in the treatment of the final effluent of the palm oil industry for reuse as feedwater for low-pressure boilers. In a bench-scale set-up, a low-cost empty fruit bunch-based powdered activated carbon (PAC) was employed for upstream adsorption of biotreated palm oil mill effluent (BPOME) with the process conditions: 60 g/L dose of PAC, 68 min of mixing time and 200 rpm of mixing speed, to reduce the feedwater strength, alleviate probable fouling of the membranes and thus improve the process flux (productivity). Three polyethersulfone ultrafiltration membranes of molecular weight cut-off (MWCO) of 1, 5 and 10 kDa were investigated in a cross-flow filtration mode, and under constant transmembrane pressures of 40, 80, and 120 kPa. The permeate qualities of the hybrid processes were evaluated, and it was found that the integrated process with the 1 kDa MWCO UF membrane yielded the best water quality that falls within the US EPA reuse standard for boiler-feed and cooling water. It was also observed that the permeate quality is fit for extended reuse as process water in the cement, petroleum and coal industries. In addition, the hybrid system's operation consumed 37.13 Wh m -3 of energy at the highest applied pressure of 120 kPa, which is far lesser than the typical energy requirement range (0.8-1.0 kWh m -3 ) for such wastewater reclamation.

A Monte Carlo Analysis of Weight Data from UF 6 Cylinder Feed and Withdrawal Stations

DOE Office of Scientific and Technical Information (OSTI.GOV)

Garner, James R; Whitaker, J Michael

2015-01-01

As the number of nuclear facilities handling uranium hexafluoride (UF 6) cylinders (e.g., UF 6 production, enrichment, and fuel fabrication) increase in number and throughput, more automated safeguards measures will likely be needed to enable the International Atomic Energy Agency (IAEA) to achieve its safeguards objectives in a fiscally constrained environment. Monitoring the process data from the load cells built into the cylinder feed and withdrawal (F/W) stations (i.e., cylinder weight data) can significantly increase the IAEA’s ability to efficiently achieve the fundamental safeguards task of confirming operations as declared (i.e., no undeclared activities). Researchers at the Oak Ridge Nationalmore » Laboratory, Los Alamos National Laboratory, the Joint Research Center (in Ispra, Italy), and University of Glasgow are investigating how this weight data can be used for IAEA safeguards purposes while fully protecting the operator’s proprietary and sensitive information related to operations. A key question that must be resolved is, what is the necessary frequency of recording data from the process F/W stations to achieve safeguards objectives? This paper summarizes Monte Carlo simulations of typical feed, product, and tails withdrawal cycles and evaluates longer sampling frequencies to determine the expected errors caused by low-frequency sampling and its impact on material balance calculations.« less

Development of wide-angle 2D light scattering static cytometry

NASA Astrophysics Data System (ADS)

Xie, Linyan; Liu, Qiao; Shao, Changshun; Su, Xuantao

2016-10-01

We have recently developed a 2D light scattering static cytometer for cellular analysis in a label-free manner, which measures side scatter (SSC) light in the polar angular range from 79 to 101 degrees. Compared with conventional flow cytometry, our cytometric technique requires no fluorescent labeling of the cells, and static cytometry measurements can be performed without flow control. In this paper we present an improved label-free static cytometer that can obtain 2D light scattering patterns in a wider angular range. By illuminating the static microspheres on chip with a scanning optical fiber, wide-angle 2D light scattering patterns of single standard microspheres with a mean diameter of 3.87 μm are obtained. The 2D patterns of 3.87 μm microspheres contain both large-angle forward scatter (FSC) and SSC light in the polar angular range from 40 to 100 degrees, approximately. Experimental 2D patterns of 3.87 μm microspheres are in good agreement with Mie theory simulated ones. The wide-angle light scattering measurements may provide a better resolution for particle analysis as compared with the SSC measurements. Two dimensional light scattering patterns of HL-60 human acute leukemia cells are obtained by using our static cytometer. Compared with SSC 2D light scattering patterns, wide-angle 2D patterns contain richer information of the HL-60 cells. The obtaining of 2D light scattering patterns in a wide angular range could help to enhance the capabilities of our label-free static cytometry for cell analysis.

Novel Quantitative Autophagy Analysis by Organelle Flow Cytometry after Cell Sonication

PubMed Central

Degtyarev, Michael; Reichelt, Mike; Lin, Kui

2014-01-01

Autophagy is a dynamic process of bulk degradation of cellular proteins and organelles in lysosomes. Current methods of autophagy measurement include microscopy-based counting of autophagic vacuoles (AVs) in cells. We have developed a novel method to quantitatively analyze individual AVs using flow cytometry. This method, OFACS (organelle flow after cell sonication), takes advantage of efficient cell disruption with a brief sonication, generating cell homogenates with fluorescently labeled AVs that retain their integrity as confirmed with light and electron microscopy analysis. These AVs could be detected directly in the sonicated cell homogenates on a flow cytometer as a distinct population of expected organelle size on a cytometry plot. Treatment of cells with inhibitors of autophagic flux, such as chloroquine or lysosomal protease inhibitors, increased the number of particles in this population under autophagy inducing conditions, while inhibition of autophagy induction with 3-methyladenine or knockdown of ATG proteins prevented this accumulation. This assay can be easily performed in a high-throughput format and opens up previously unexplored avenues for autophagy analysis. PMID:24489953

Lab-on-Chip Cytometry Based on Magnetoresistive Sensors for Bacteria Detection in Milk

PubMed Central

Fernandes, Ana C.; Duarte, Carla M.; Cardoso, Filipe A.; Bexiga, Ricardo.; Cardoso, Susana.; Freitas, Paulo P.

2014-01-01

Flow cytometers have been optimized for use in portable platforms, where cell separation, identification and counting can be achieved in a compact and modular format. This feature can be combined with magnetic detection, where magnetoresistive sensors can be integrated within microfluidic channels to detect magnetically labelled cells. This work describes a platform for in-flow detection of magnetically labelled cells with a magneto-resistive based cell cytometer. In particular, we present an example for the validation of the platform as a magnetic counter that identifies and quantifies Streptococcus agalactiae in milk. PMID:25196163

Lab-on-chip cytometry based on magnetoresistive sensors for bacteria detection in milk.

PubMed

Fernandes, Ana C; Duarte, Carla M; Cardoso, Filipe A; Bexiga, Ricardo; Cardoso, Susana; Freitas, Paulo P

2014-08-21

Flow cytometers have been optimized for use in portable platforms, where cell separation, identification and counting can be achieved in a compact and modular format. This feature can be combined with magnetic detection, where magnetoresistive sensors can be integrated within microfluidic channels to detect magnetically labelled cells. This work describes a platform for in-flow detection of magnetically labelled cells with a magneto-resistive based cell cytometer. In particular, we present an example for the validation of the platform as a magnetic counter that identifies and quantifies Streptococcus agalactiae in milk.

Flow Cytometry: Impact On Early Drug Discovery

PubMed Central

Edwards, Bruce S.; Sklar, Larry A.

2015-01-01

Summary Modern flow cytometers can make optical measurements of 10 or more parameters per cell at tens-of-thousands of cells per second and over five orders of magnitude dynamic range. Although flow cytometry is used in most drug discovery stages, “sip-and-spit” sampling technology has restricted it to low sample throughput applications. The advent of HyperCyt sampling technology has recently made possible primary screening applications in which tens-of-thousands of compounds are analyzed per day. Target-multiplexing methodologies in combination with extended multi-parameter analyses enable profiling of lead candidates early in the discovery process, when the greatest numbers of candidates are available for evaluation. The ability to sample small volumes with negligible waste reduces reagent costs, compound usage and consumption of cells. Improved compound library formatting strategies can further extend primary screening opportunities when samples are scarce. Dozens of targets have been screened in 384- and 1536-well assay formats, predominantly in academic screening lab settings. In concert with commercial platform evolution and trending drug discovery strategies, HyperCyt-based systems are now finding their way into mainstream screening labs. Recent advances in flow-based imaging, mass spectrometry and parallel sample processing promise dramatically expanded single cell profiling capabilities to bolster systems level approaches to drug discovery. PMID:25805180

Validation of a single-platform, volumetric, flow cytometry for CD4 T cell count monitoring in therapeutic mobile unit

PubMed Central

2012-01-01

Background A mobile health unit may be useful to follow up adult and pediatric patients on antiretroviral treatment and living in remote areas devoid of laboratory facilities. The study evaluated the use of the simplified, robust, single-plateform, volumetric, pan-leucogating Auto40 flow cytometer (Apogee Flow Systems Ltd, Hemel Hempstead, UK) for CD4 T cell numeration in a mobile unit, compared against a reference flow cytometry method. Methods The therapeutic mobile unit of the Laboratoire National de Santé Hygiène Mobile, Yaoundé, Cameroon, was equipped with the Auto40. A FACSCalibur flow cytometer (Becton Dickinson Immuno-cytometry System, San Jose, CA, USA) was used as reference method. EDTA-blood samples from volunteers were first subjected to CD4 T cell count in the mobile unit, and an aliquot was sent within 4 hours to Centre International de Référence Chantal Biya, Yaoundé, for FACSCalibur assay. Results Two HIV screening campaigns with the mobile unit were organised in December 2009 and January 2010. The campaign in the suburb of Yaoundé which was 20 km from the reference laboratory included 188 volunteers comprising 93 children less than 5 years old. The campaign in Ambang Bikok (53 km far from Yaoundé) included 69 adult volunteers. In Yaoundé suburb, mean ± standard deviation (SD) CD4 T cell count was 996 ± 874 cells/μl by Auto40, and 989 ± 883 cells/μl by FACSCalibur; in Ambang Bikok, mean ± SD CD4 T cell count was 1041 ± 317 cells/μl by Auto40, and 1032 ± 294 cells/μl by FACSCalibur. Results by Auto40 and FACSCalibur were highly correlated in Yaoundé (r2 = 0.982) as in Ambang Bikok (r2 = 0.921). Bland-Altman analysis showed a close agreement between Auto40 and FACSCalibur results expressed in absolute count as in percentage in Yaoundé and Ambang Bikok. When pooling the 257 CD4 T cell count measurements, the Auto40 yielded a mean difference of +7.6 CD4 T cells/μl higher than by reference flow cytometry; and the sensitivity and

RESULTS FROM A DEMONSTRATION OF RF-BASED UF6 CYLINDER ACCOUNTING AND TRACKING SYSTEM INSTALLED AT A USEC FACILITY

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pickett, Chris A; Kovacic, Donald N; Morgan, Jim

Approved industry-standard cylinders are used globally for storing and transporting uranium hexafluoride (UF{sub 6}) at uranium enrichment plants and processing facilities. To verify that no diversion or undeclared production of nuclear material involving UF{sub 6} cylinders at the facility has occurred, the International Atomic Energy Agency (IAEA) conducts periodic, labor-intensive physical inspections to validate facility records, cylinder identities, and cylinder weights. A reliable cylinder monitoring system that would improve overall inspector effectiveness would be a significant improvement to the current international safeguards inspection regime. Such a system could include real-time unattended monitoring of cylinder movements, situation-specific rules-based event detection algorithms,more » and the capability to integrate with other types of safeguards technologies. This type of system could provide timely detection of abnormal operational activities that may be used to ensure more appropriate and efficient responses by the IAEA. A system of this type can reduce the reliance on paper records and have the additional benefit of facilitating domestic safeguards at the facilities at which it is installed. A radio-frequency (RF)-based system designed to track uranium hexafluoride (UF{sub 6}) cylinders during processing operations was designed, assembled, and tested at the United States Enrichment Corporation (USEC) facility in Portsmouth, Ohio, to determine the operational feasibility and durability of RF technology. The overall objective of the effort was to validate the robustness of RF technology for potential use as a future international safeguards tool for tracking UF6 cylinders at uranium-processing facilities. The results to date indicate that RF tags represent a feasible technique for tracking UF{sub 6} cylinders in operating facilities. Additional work will be needed to improve the operational robustness of the tags for repeated autoclave

Nanoparticle-based flow virometry for the analysis of individual virions

PubMed Central

Arakelyan, Anush; Fitzgerald, Wendy; Margolis, Leonid; Grivel, Jean-Charles

2013-01-01

While flow cytometry has been used to analyze the antigenic composition of individual cells, the antigenic makeup of viral particles is still characterized predominantly in bulk. Here, we describe a technology, “flow virometry,” that can be used for antigen detection on individual virions. The technology is based on binding magnetic nanoparticles to virions, staining the virions with monoclonal antibodies, separating the formed complexes with magnetic columns, and characterizing them with flow cytometers. We used this technology to study the distribution of two antigens (HLA-DR and LFA-1) that HIV-1 acquires from infected cells among individual HIV-1 virions. Flow virometry revealed that the antigenic makeup of virions from a single preparation is heterogeneous. This heterogeneity could not be detected with bulk analysis of viruses. Moreover, in two preparations of the same HIV-1 produced by different cells, the distribution of antigens among virions was different. In contrast, HIV-1 of two different HIV-1 genotypes replicating in the same cells became somewhat antigenically similar. This nanotechnology allows the study of virions in bodily fluids without virus propagation and in principle is not restricted to the analysis of HIV, but can be applied to the analysis of the individual surface antigenic makeup of any virus. PMID:23925291

Evaluation of the platelet counting by Abbott CELL-DYN SAPPHIRE haematology analyser compared with flow cytometry.

PubMed

Grimaldi, E; Del Vecchio, L; Scopacasa, F; Lo Pardo, C; Capone, F; Pariante, S; Scalia, G; De Caterina, M

2009-04-01

The Abbot Cell-Dyn Sapphire is a new generation haematology analyser. The system uses optical/fluorescence flow cytometry in combination with electronic impedance to produce a full blood count. Optical and impedance are the default methods for platelet counting while automated CD61-immunoplatelet analysis can be run as selectable test. The aim of this study was to determine the platelet count performance of the three counting methods available on the instrument and to compare the results with those provided by Becton Dickinson FACSCalibur flow cytometer used as reference method. A lipid interference experiment was also performed. Linearity, carryover and precision were good, and satisfactory agreement with reference method was found for the impedance, optical and CD61-immunoplatelet analysis, although this latter provided the closest results in comparison with flow cytometry. In the lipid interference experiment, a moderate inaccuracy of optical and immunoplatelet counts was observed starting from a very high lipid value.

Real-time viability and apoptosis kinetic detection method of 3D multicellular tumor spheroids using the Celigo Image Cytometer.

PubMed

Kessel, Sarah; Cribbes, Scott; Bonasu, Surekha; Rice, William; Qiu, Jean; Chan, Leo Li-Ying

2017-09-01

The development of three-dimensional (3D) multicellular tumor spheroid models for cancer drug discovery research has increased in the recent years. The use of 3D tumor spheroid models may be more representative of the complex in vivo tumor microenvironments in comparison to two-dimensional (2D) assays. Currently, viability of 3D multicellular tumor spheroids has been commonly measured on standard plate-readers using metabolic reagents such as CellTiter-Glo® for end point analysis. Alternatively, high content image cytometers have been used to measure drug effects on spheroid size and viability. Previously, we have demonstrated a novel end point drug screening method for 3D multicellular tumor spheroids using the Celigo Image Cytometer. To better characterize the cancer drug effects, it is important to also measure the kinetic cytotoxic and apoptotic effects on 3D multicellular tumor spheroids. In this work, we demonstrate the use of PI and caspase 3/7 stains to measure viability and apoptosis for 3D multicellular tumor spheroids in real-time. The method was first validated by staining different types of tumor spheroids with PI and caspase 3/7 and monitoring the fluorescent intensities for 16 and 21 days. Next, PI-stained and nonstained control tumor spheroids were digested into single cell suspension to directly measure viability in a 2D assay to determine the potential toxicity of PI. Finally, extensive data analysis was performed on correlating the time-dependent PI and caspase 3/7 fluorescent intensities to the spheroid size and necrotic core formation to determine an optimal starting time point for cancer drug testing. The ability to measure real-time viability and apoptosis is highly important for developing a proper 3D model for screening tumor spheroids, which can allow researchers to determine time-dependent drug effects that usually are not captured by end point assays. This would improve the current tumor spheroid analysis method to potentially better

An enzyme-free flow cytometric bead assay for the sensitive detection of microRNAs based on click nucleic acid ligation-mediated signal amplification.

PubMed

Qi, Yan; Qiu, Liying; Fan, Wenjiao; Liu, Chenghui; Li, Zhengping

2017-08-07

A versatile flow cytometric bead assay (FCBA) coupled with a completely enzyme-free signal amplification mechanism is developed for the sensitive detection of microRNAs (miRNAs). This new strategy integrates click chemistry-mediated ligation chain reaction (CLCR) with hybridization chain reaction (HCR) for enzyme-free signal amplification on magnetic beads (MBs), and a flow cytometer for the robust fluorescence readout of the MBs. Firstly, target miRNA can initiate CLCR on the surface of MBs based on the click chemical ligation between dibenzocyclooctyne (DBCO)- and azide-modified single-stranded DNA (ssDNA) probes, and the amount of ligated ssDNA sequences on the MBs will be proportional to the dosage of target miRNA. Afterward, each of the ligated ssDNA products can trigger a cascade chain reaction of hybridization events between two alternating fluorophore-tagged hairpin probes, resulting in another signal amplification pathway with an amplified accumulation of fluorophores on the MBs. Finally, the fluorophore-anchored MBs are directly and rapidly analyzed by using a flow cytometer without any separation or elution processes. Herein, the click nucleic acid ligation only occurs on the surface of MBs, so the nonspecific ligations are greatly inhibited compared with that of ligation reaction performed in homogeneous solution. Furthermore, the signal amplification by CLCR-HCR is highly efficient but totally enzyme-free, which may overcome the potential drawbacks of conventional enzyme-catalyzed signal amplification protocols and lead to a high sensitivity. The CLCR-HCR-based FCBA has pushed the detection limit of let-7a miRNA down to the femtomolar (fM) level, showing great potential in miRNA-related biological studies and disease diagnosis.

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

PubMed Central

Hossain, Md. Sharoare; Johannisson, Anders; Wallgren, Margareta; Nagy, Szabolcs; Siqueira, Amanda Pimenta; Rodriguez-Martinez, Heriberto

2011-01-01

Flow cytometry is now a recognized methodology within animal spermatology, and has moved from being a research tool to become routine in the assessment of animal semen destined to breeding. The availability of ‘bench-top' flow cytometers and of newer and versatile markers for cell structure and function had allowed the instrumentation to measure more sperm parameters, from viability to reactiveness when exposed to exogenous stimuli, and to increase our capabilities to sort spermatozoa for potential fertilizing capacity, or chromosomal sex. The present review summarizes the state of the art regarding flow cytometry applied to animal andrology, albeit keeping an open comparative intent. It critically evaluates the present and future capabilities of flow cytometry for the diagnostics of potential fertility and for the development of current reproductive technologies such as sperm freezing, sperm selection and sperm sorting. The flow cytometry methods will probably further revolutionize our understanding of the sperm physiology and their functionality, and will undoubtedly extend its application in isolating many uncharacterized features of spermatozoa. However, continuous follow-up of the methods is a necessity owing to technical developments and the complexity of mapping spermatozoa. PMID:21478895

High throughput imaging cytometer with acoustic focussing† †Electronic supplementary information (ESI) available: High throughput imaging cytometer with acoustic focussing. See DOI: 10.1039/c5ra19497k Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file.

PubMed Central

Zmijan, Robert; Jonnalagadda, Umesh S.; Carugo, Dario; Kochi, Yu; Lemm, Elizabeth; Packham, Graham; Hill, Martyn

2015-01-01

We demonstrate an imaging flow cytometer that uses acoustic levitation to assemble cells and other particles into a sheet structure. This technique enables a high resolution, low noise CMOS camera to capture images of thousands of cells with each frame. While ultrasonic focussing has previously been demonstrated for 1D cytometry systems, extending the technology to a planar, much higher throughput format and integrating imaging is non-trivial, and represents a significant jump forward in capability, leading to diagnostic possibilities not achievable with current systems. A galvo mirror is used to track the images of the moving cells permitting exposure times of 10 ms at frame rates of 50 fps with motion blur of only a few pixels. At 80 fps, we demonstrate a throughput of 208 000 beads per second. We investigate the factors affecting motion blur and throughput, and demonstrate the system with fluorescent beads, leukaemia cells and a chondrocyte cell line. Cells require more time to reach the acoustic focus than beads, resulting in lower throughputs; however a longer device would remove this constraint. PMID:29456838

Functional analysis of unfermented and fermented citrus peels and physical properties of citrus peel-added doughs for bread making.

PubMed

Shyu, Yung-Shin; Lu, Tzu-Chi; Lin, Chuan-Chuan

2014-12-01

Several studies have indicated citrus peels (CP) contain specific methoxy flavones, e.g. nobiletin and tangeretin, which have been shown to prevent numerous diseases. However, research reports regarding their application as food additive in healthy baked products is scarce. In our study, both unfermented (UF) and fermented (F) citrus peels were processed under different dry hot-air temperatures to make four citrus peel powders , UF-100 °C,UF-150 °C, F-100 °C, F-150 °C, respectively. The analysis of the basic components and nutraceuticals as well as antioxidant activity were conducted. Various percentages of CP were added to dough and toast bread for physical property and sensory evaluations. The results indicated the contents of crude proteins (3.3-4.3 mg/g) and fibers (10.9-14.9 %) among the four samples were similar. The UF extracts showed better antioxidant activities than F extracts. HPLC analysis indicated the contents of hesperidine, nobiletin and tangeretin in CP extracts were UF-150 °C > UF-100 °C. Farinograph analysis indicated a linear relation between CP powder content and the parameters of the physical properties of dough. A high percentage of fibrous CP powder in dough increases the water adsorption capacity of the dough, resulting in a decrease in its stability The sensory evaluation results indicated a greater acceptability of UF-added toast bread relative to the F-added one. Among these, according to the statistical anaylsis, the UF-150 °C 4 % and UF-100 °C 6 % groups were the best and F-150 °C 2 % group was the poorest in overall acceptability.

Flow Cytometry: Impact on Early Drug Discovery.

PubMed

Edwards, Bruce S; Sklar, Larry A

2015-07-01

Modern flow cytometers can make optical measurements of 10 or more parameters per cell at tens of thousands of cells per second and more than five orders of magnitude dynamic range. Although flow cytometry is used in most drug discovery stages, "sip-and-spit" sampling technology has restricted it to low-sample-throughput applications. The advent of HyperCyt sampling technology has recently made possible primary screening applications in which tens of thousands of compounds are analyzed per day. Target-multiplexing methodologies in combination with extended multiparameter analyses enable profiling of lead candidates early in the discovery process, when the greatest numbers of candidates are available for evaluation. The ability to sample small volumes with negligible waste reduces reagent costs, compound usage, and consumption of cells. Improved compound library formatting strategies can further extend primary screening opportunities when samples are scarce. Dozens of targets have been screened in 384- and 1536-well assay formats, predominantly in academic screening lab settings. In concert with commercial platform evolution and trending drug discovery strategies, HyperCyt-based systems are now finding their way into mainstream screening labs. Recent advances in flow-based imaging, mass spectrometry, and parallel sample processing promise dramatically expanded single-cell profiling capabilities to bolster systems-level approaches to drug discovery. © 2015 Society for Laboratory Automation and Screening.

Monitoring circulating prostate tumor cells after tumor resection by in vivo flow cytometry

NASA Astrophysics Data System (ADS)

Ding, Nan; Zhu, Xi; Xie, Chengying; Wei, Dan; Yang, Zhangru; Suo, Yuanzhen; Wei, Xunbin

2018-02-01

Prostate cancer has already become the biggest threat among all cancer types for male people and many people died because of its bone metastases. Circulating tumor cells (CTCs) can be used as early metastasis marker so that the detection of CTCs in blood is meaningful for early diagnosis and treatment. However, the relationship between these therapies and metastasis has not been fully clarified yet. Hence, we built PC3 subcutaneous tumor model and developed in vivo flow cytometer (IVFC) platform to record the dynamics of CTC before and after tumor resection. We found out that tumor resection can reduce CTC quantities instantaneously while having a good control of metastasis. CTC re-occurred 7 days after surgery, which might be correlated with early disseminated and deposited tumors. In conclusion, in vivo flow cytometry (IVFC) is capable of detecting CTC dynamics in prostate subcutaneous tumor model and this method could facilitate further research about relationship between other cancer therapies and circulating tumor cells.

Geomorphic Change Induced by 100 years of Flow Alteration on the Diamond Fork River, Central Utah

NASA Astrophysics Data System (ADS)

Jones, J.; Belmont, P.; Wilcock, P. R.

2017-12-01

Changes in hydrology and sediment supply affect the form of rivers. The rate of change of fluvial form is controlled by a variety of factors, including valley confinement, sediment size, and antecedent condition. The Diamond Fork River in central Utah has been altered by trans-basin flows delivered from the Colorado River system for over a century. Beginning in 1915, water used for irrigation was delivered through a tributary, Sixth Water Creek, with daily summer flows regularly exceeding the 50 - 100 year flood. Elevated flows caused drastic geomorphic change - resulting in incision and widening of the channel, and the destruction of riparian vegetation. Beginning in 1997, the outlet for the trans-basin diversion was moved downstream on Sixth Water, bypassing a large landslide, and flows were drastically reduced in 2004 through management actions. We delineated eight distinct process domains for the Sixth Water-Diamond Fork system and examined the response of each process domain to the altered flow and sediment regimes through the analysis of aerial photographs and repeat cross-sections. We measured a variety of channel metrics, including channel width, areal extent of bars and islands, and sinuosity in ArcGIS. Results indicate that unconfined reaches that were wide and braided during the period of elevated flows have narrowed to become single threaded and meandering in response to the reduced flows. Confined reaches have experienced minor changes since the reduction in flows, suggesting that confinement is a primary control on the degree of channel response. These findings and complimentary studies will provide managers of Sixth Water and Diamond Fork with a greater understanding of the physical response of the streams, and the resulting effects on ecological communities.

Flexible planar microfluidic chip employing a light emitting diode and a PIN-photodiode for portable flow cytometers.

PubMed

Kettlitz, Siegfried W; Valouch, Sebastian; Sittel, Wiebke; Lemmer, Uli

2012-01-07

Detection of fluorescence particles is a key method of flow cytometry. We evaluate the performance of a design for a microfluidic fluorescence particle detection device. Due to the planar design with low layer thicknesses, we avoid optical components such as lenses or dichroic mirrors and substitute them with a shadow mask and colored film filters. A commercially available LED is used as the light source and a PIN-photodiode as detector. This design approach reduces component cost and power consumption and enables supplying the device with power from a standard USB port. From evaluation of this design, we obtain a maximum particle detection frequency of up to 600 particles per second at a sensitivity of better than 4.7 × 10(5) MESF (molecules of equivalent soluble fluorochrome) measured with particles for FITC sensitivity calibration. Lowering the flow rate increases the instrument sensitivity by an order of magnitude enabling the detection of particles with 4.5 × 10(4) MESF.

Simulation of water-surface elevations for a hypothetical 100-year peak flow in Birch Creek at the Idaho National Engineering and Environmental Laboratory, Idaho

DOE Office of Scientific and Technical Information (OSTI.GOV)

Berenbrock, C.; Kjelstrom, L.C.

1997-10-01

Delineation of areas at the Idaho National Engineering and Environmental Laboratory that would be inundated by a 100-year peak flow in Birch Creek is needed by the US Department of Energy to fulfill flood-plain regulatory requirements. Birch Creek flows southward about 40 miles through an alluvium-filled valley onto the northern part of the Idaho National Engineering and Environmental laboratory site on the eastern Snake River Plain. The lower 10-mile reach of Birch Creek that ends in Birch Creek Playa near several Idaho National Engineering and Environmental Laboratory facilities is of particular concern. Twenty-six channel cross sections were surveyed to developmore » and apply a hydraulic model to simulate water-surface elevations for a hypothetical 100-year peak flow in Birch Creek. Model simulation of the 100-year peak flow (700 cubic feet per second) in reaches upstream from State Highway 22 indicated that flow was confined within channels even when all flow was routed to one channel. Where the highway crosses Birch Creek, about 315 cubic feet per second of water was estimated to move downstream--115 cubic feet per second through a culvert and 200 cubic feet per second over the highway. Simulated water-surface elevation at this crossing was 0.8 foot higher than the elevation of the highway. The remaining 385 cubic feet per second flowed southwestward in a trench along the north side of the highway. Flow also was simulated with the culvert removed. The exact location of flood boundaries on Birch Creek could not be determined because of the highly braided channel and the many anthropogenic features (such as the trench, highway, and diversion channels) in the study area that affect flood hydraulics and flow. Because flood boundaries could not be located exactly, only a generalized flood-prone map was developed.« less

Enhancing the Detection of Dysmorphic Red Blood Cells and Renal Tubular Epithelial Cells with a Modified Urinalysis Protocol.

PubMed

Chu-Su, Yu; Shukuya, Kenichi; Yokoyama, Takashi; Lin, Wei-Chou; Chiang, Chih-Kang; Lin, Chii-Wann

2017-01-11

Urinary sediment is used to evaluate patients with possible urinary tract diseases. Currently, numerous protocols are applied to detect dysmorphic red blood cells (RBCs) and renal tubular epithelial cells (RTECs) in urinary sediment. However, distinct protocols are used by nephrologists and medical technologists for specimen concentration and observation, which leads to major discrepancies in the differential counts of formed elements such as dysmorphic RBCs and RTECs and might interfere with an accurate clinical diagnosis. To resolve these problems, we first tested a modified urinalysis protocol with an increased relative centrifuge force and concentration factor in 20 biopsy-confirmed glomerulonephritis patients with haematuria. We successfully improved the recovery ratio of dysmorphic RBCs in clinical specimens from 34.7% to 42.0% (P < 0.001). Furthermore, we confirmed the correlation between counts by the modified urinary protocol and Sysmex UF-1000i urinary flow cytometer (r ≥ 0.898, P < 0.001). A total of 28 types of isomorphic and dysmorphic RBCs were detected using a bright field microscope, with results comparable to those using a standard phase contrast microscope. Finally, we applied Sternheimer stain to enhance the contrast of RTECs in the urinary sediments. We concluded that this modified urinalysis protocol significantly enhanced the quality of urinalysis.

Enhancing the Detection of Dysmorphic Red Blood Cells and Renal Tubular Epithelial Cells with a Modified Urinalysis Protocol

PubMed Central

Chu-Su, Yu; Shukuya, Kenichi; Yokoyama, Takashi; Lin, Wei-Chou; Chiang, Chih-Kang; Lin, Chii-Wann

2017-01-01

Urinary sediment is used to evaluate patients with possible urinary tract diseases. Currently, numerous protocols are applied to detect dysmorphic red blood cells (RBCs) and renal tubular epithelial cells (RTECs) in urinary sediment. However, distinct protocols are used by nephrologists and medical technologists for specimen concentration and observation, which leads to major discrepancies in the differential counts of formed elements such as dysmorphic RBCs and RTECs and might interfere with an accurate clinical diagnosis. To resolve these problems, we first tested a modified urinalysis protocol with an increased relative centrifuge force and concentration factor in 20 biopsy-confirmed glomerulonephritis patients with haematuria. We successfully improved the recovery ratio of dysmorphic RBCs in clinical specimens from 34.7% to 42.0% (P < 0.001). Furthermore, we confirmed the correlation between counts by the modified urinary protocol and Sysmex UF-1000i urinary flow cytometer (r ≥ 0.898, P < 0.001). A total of 28 types of isomorphic and dysmorphic RBCs were detected using a bright field microscope, with results comparable to those using a standard phase contrast microscope. Finally, we applied Sternheimer stain to enhance the contrast of RTECs in the urinary sediments. We concluded that this modified urinalysis protocol significantly enhanced the quality of urinalysis. PMID:28074941

Schneefernerhaus as a mountain research station for clouds and turbulence - Part 1: Flow conditions and large-scale turbulence

NASA Astrophysics Data System (ADS)

Risius, S.; Xu, H.; Di Lorenzo, F.; Xi, H.; Siebert, H.; Shaw, R. A.; Bodenschatz, E.

2015-01-01

Cloud measurements are usually carried out with airborne campaigns, which are expensive and are limited by temporal duration and weather conditions. Ground based measurements at high-altitude research stations therefore play a complementary role in cloud study. Using the meteorological data (wind speed, direction, temperature, humidity, visibility, etc.) collected by the German Weather Service (DWD) from 2000 to 2012 and turbulence measurements recorded by multiple ultrasonic sensors (sampled at 10 Hz) in 2010, we show that the Umweltforschungsstation Schneefernerhaus (UFS) located just below the peak of Zugspitze in the German Alps, at a height of 2650 m, is a well-suited station for cloud-turbulence research. The wind at UFS is dominantly in the east-west direction and nearly horizontal. During the summer time (July and August) the UFS is immersed in warm clouds about 25% of the time. The clouds are either from convection originating in the valley in the east, or associated with synoptic-scale weather systems typically advected from the west. Air turbulence, as measured from the second and third order velocity structure functions that exhibit well-developed inertial ranges, possesses Taylor microscale Reynolds numbers up to 104, with the most probable value at ~ 3000. In spite of the complex topography, the turbulence appears to be nearly as isotropic as many laboratory flows when evaluated on the so called "Lumley-triangle".

ENVIRONMENTAL TECHNOLOGY VERIFICATION REPORT - PHYSICAL REMOVAL OF PARTICULATE CONTAMINANTS IN DRINKING WATER: POLYMEM UF 120 S2 ULTRAFILTRATION MEMBRANE MODULE, LUXENBURG, WISCONSIN

EPA Science Inventory

Verification testing of the Polymem UF120 S2 Ultrafiltration Membrane Module was conducted over a 46-day period at the Green Bay Water Utility Filtration Plant, Luxemburg, Wisconsin. The ETV testing described herein was funded in conjunction with a 12-month membrane pilot study f...

A polishing hybrid AER/UF membrane process for the treatment of a high DOC content surface water.

PubMed

Humbert, H; Gallard, H; Croué, J-P

2012-03-15

The efficacy of a combined AER/UF (Anion Exchange Resin/Ultrafiltration) process for the polishing treatment of a high DOC (Dissolved Organic Carbon) content (>8 mgC/L) surface water was investigated at lab-scale using a strong base AER. Both resin dose and bead size had a significant impact on the kinetic removal of DOC for short contact times (i.e. <15 min). For resin doses higher than 700 mg/L and median bead sizes below 250 μm DOC removal remained constant after 30 min of contact time with very high removal rates (80%). Optimum AER treatment conditions were applied in combination with UF membrane filtration on water previously treated by coagulation-flocculation (i.e. 3 mgC/L). A more severe fouling was observed for each filtration run in the presence of AER. This fouling was shown to be mainly reversible and caused by the progressive attrition of the AER through the centrifugal pump leading to the production of resin particles below 50 μm in diameter. More important, the presence of AER significantly lowered the irreversible fouling (loss of permeability recorded after backwash) and reduced the DOC content of the clarified water to l.8 mgC/L (40% removal rate), concentration that remained almost constant throughout the experiment. Copyright © 2011 Elsevier Ltd. All rights reserved.

Impedance Flow Cytometry as a Tool to Analyze Microspore and Pollen Quality.

PubMed

Heidmann, Iris; Di Berardino, Marco

2017-01-01

Analyzing pollen quality in an efficient and reliable manner is of great importance to the industries involved in seed and fruit production, plant breeding, and plant research. Pollen quality parameters, viability and germination capacity, are analyzed by various staining methods or by in vitro germination assays, respectively. These methods are time-consuming, species-dependent, and require a lab environment. Furthermore, the obtained viability data are often poorly related to in vivo pollen germination and seed set. Here, we describe a quick, label-free method to analyze pollen using microfluidic chips inserted into an impedance flow cytometer (IFC). Using this approach, pollen quality parameters are determined by a single measurement in a species-independent manner. The advantage of this protocol is that pollen viability and germination can be analyzed quickly by a reliable and standardized method.

Sunspot Cycle 24: Smallest Cycle in 100 Years?

DTIC Science & Technology

2005-01-11

and power systems Wilcox Solar Observatories (WSO [Svalgaard et al., [National Oceanic and Atmospheric Administration, 2004; 1978], since 1976). The...are _W taken to be a measure of how well the procedure works (the oun* Wilson Solor Obs. Wico ,*Uf ts,. only real measure as far as we are concerned

Evaluation of human enteric viruses in surface water and drinking water resources in southern Ghana.

PubMed

Gibson, Kristen E; Opryszko, Melissa C; Schissler, James T; Guo, Yayi; Schwab, Kellogg J

2011-01-01

An estimated 884 million people worldwide do not have access to an improved drinking water source, and the microbial quality of these sources is often unknown. In this study, a combined tangential flow, hollow fiber ultrafiltration (UF), and real-time PCR method was applied to large volume (100 L) groundwater (N = 4), surface water (N = 9), and finished (i.e., receiving treatment) drinking water (N = 6) samples for the evaluation of human enteric viruses and bacterial indicators. Human enteric viruses including norovirus GI and GII, adenovirus, and polyomavirus were detected in five different samples including one groundwater, three surface water, and one drinking water sample. Total coliforms and Escherichia coli assessed for each sample before and after UF revealed a lack of correlation between bacterial indicators and the presence of human enteric viruses.

Evaluation of Human Enteric Viruses in Surface Water and Drinking Water Resources in Southern Ghana

PubMed Central

Gibson, Kristen E.; Opryszko, Melissa C.; Schissler, James T.; Guo, Yayi; Schwab, Kellogg J.

2011-01-01

An estimated 884 million people worldwide do not have access to an improved drinking water source, and the microbial quality of these sources is often unknown. In this study, a combined tangential flow, hollow fiber ultrafiltration (UF), and real-time PCR method was applied to large volume (100 L) groundwater (N = 4), surface water (N = 9), and finished (i.e., receiving treatment) drinking water (N = 6) samples for the evaluation of human enteric viruses and bacterial indicators. Human enteric viruses including norovirus GI and GII, adenovirus, and polyomavirus were detected in five different samples including one groundwater, three surface water, and one drinking water sample. Total coliforms and Escherichia coli assessed for each sample before and after UF revealed a lack of correlation between bacterial indicators and the presence of human enteric viruses. PMID:21212196

Cellular vacuolation and mitochondrial-associated factors induced by Clostridium perfringens epsilon toxin detected using acoustic flow cytometry.

PubMed

Ferrarezi, Marina C; Curci, Vera C L M; Cardoso, Tereza C

2013-12-01

Epsilon toxin (ETX) produced by Clostridium perfringens types B and D is a potent toxin that is responsible for fatal enterotoxaemia. In vitro, ETX, which is considered as a pore-forming toxin, forms a heptamer in Madin-Darby canine kidney (MDCK) cell membranes, which is considered to be a pre-pore stage. After binding of the ETX, vacuoles inside cell cytoplasm are produced. ETX causes decreased levels of essential coenzymes required for host cell energy. Here, we optimized and applied acoustic flow cytometry analysis in order to gain further insight into ETX-pathogenesis. Using acoustic flow cytometer analysis, which considered highly sensitive, ETX-exposed MDCK cells revealed mitochondrial membrane decreases followed by 25.48% and 45.45% of the exposed cells expressing the Bax and BCL-2 proteins at a pre-pore stage, respectively. These results together with high cytotoxicity and visualization of cell vacuoles, demonstrates that acoustic flow cytometry analysis potentially represents an effective tool to study ETX pathogenesis. Copyright © 2013. Published by Elsevier Ltd.

Evaluation of the UF/NCI hybrid computational phantoms for use in organ dosimetry of pediatric patients undergoing fluoroscopically guided cardiac procedures

NASA Astrophysics Data System (ADS)

Marshall, Emily L.; Borrego, David; Tran, Trung; Fudge, James C.; Bolch, Wesley E.

2018-03-01

Epidemiologic data demonstrate that pediatric patients face a higher relative risk of radiation induced cancers than their adult counterparts at equivalent exposures. Infants and children with congenital heart defects are a critical patient population exposed to ionizing radiation during life-saving procedures. These patients will likely incur numerous procedures throughout their lifespan, each time increasing their cumulative radiation absorbed dose. As continued improvements in long-term prognosis of congenital heart defect patients is achieved, a better understanding of organ radiation dose following treatment becomes increasingly vital. Dosimetry of these patients can be accomplished using Monte Carlo radiation transport simulations, coupled with modern anatomical patient models. The aim of this study was to evaluate the performance of the University of Florida/National Cancer Institute (UF/NCI) pediatric hybrid computational phantom library for organ dose assessment of patients that have undergone fluoroscopically guided cardiac catheterizations. In this study, two types of simulations were modeled. A dose assessment was performed on 29 patient-specific voxel phantoms (taken as representing the patient’s true anatomy), height/weight-matched hybrid library phantoms, and age-matched reference phantoms. Two exposure studies were conducted for each phantom type. First, a parametric study was constructed by the attending pediatric interventional cardiologist at the University of Florida to model the range of parameters seen clinically. Second, four clinical cardiac procedures were simulated based upon internal logfiles captured by a Toshiba Infinix-i Cardiac Bi-Plane fluoroscopic unit. Performance of the phantom library was quantified by computing both the percent difference in individual organ doses, as well as the organ dose root mean square values for overall phantom assessment between the matched phantoms (UF/NCI library or reference) and the patient

An Operator Perspective from a Facility Evaluation of an RFID-Based UF6 Cylinder Accounting and Tracking System

DOE Office of Scientific and Technical Information (OSTI.GOV)

Martyn, Rose; Fitzgerald, Peter; Stehle, Nicholas D

An operational field test of a Radio-Frequency Identification (RFID) system for tracking and accounting UF6 cylinders was conducted at the Global Nuclear Fuel Americas (GNF) fuel fabrication plant in 2009. The Cylinder Accountability and Tracking System (CATS) was designed and deployed by Oak Ridge National Laboratory (ORNL) and evaluated in cooperation with GNF. The system required that passive RFID be attached to several UF6 30B cylinders as they were received at the site; then the cylinders were tracked as they proceeded to interim storage, to processing in an autoclave, and eventually to disposition from the site. This CATS deployment alsomore » provided a direct integration of scale data from the site accountability scales. The integration of this information into the tracking data provided an attribute for additional safeguards for evaluation. The field test provided insight into the advantages and challenges of using RFID at an operating nuclear facility. The RFID system allowed operators to interact with the technology and demonstrated the survivability of the tags and reader equipment in the process environment. This paper will provide the operator perspective on utilizing RFID technology for locating cylinders within the facility, thereby tracking the cylinders for process and for Material Control & Accounting functions. The paper also will present the operator viewpoint on RFID implemented as an independent safeguards system.« less

Flow cytometric estimation on cytotoxic activity of leaf extracts from seashore plants in subtropical Japan: isolation, quantification and cytotoxic action of (-)-deoxypodophyllotoxin.

PubMed

Masuda, Toshiya; Oyama, Yasuo; Yonemori, Shigetomo; Takeda, Yoshio; Yamazaki, Yuko; Mizuguchi, Shinichi; Nakata, Mami; Tanaka, Tomochika; Chikahisa, Lumi; Inaba, Yuzuru; Okada, Yoshihiko

2002-06-01

The cytotoxic activity of methanol extracts of leaves collected from 39 seashore plants in Iriomote Island, subtropical Japan was examined on human leukaemia cells (K562 cells) using a flow cytometer with two fluorescent probes, ethidium bromide and annexin V-FITC. Five extracts (10 microg/mL) from Hernandia nymphaeaefolia, Cerbera manghas, Pongamia pinnata, Morus australis var. glabra and Thespesia populnea greatly inhibited the growth of K562 cells. When the concentration was decreased to 1 microg/mL, only one extract from H. nymphaeaefolia still inhibited the cell growth. A cytotoxic compound was isolated from the leaves by bioassay-guided fractionation and was identified as (-)-deoxypodophyllotoxin (DPT). The fresh leaves of H. nymphaeaefolia contained a remarkably high amount of DPT (0.21 +/- 0.07% of fresh leaf weight), being clarified by a quantitative HPLC analysis. DPT at 70-80 pM started to inhibit the growth of K562 cells in an all-or-none fashion and at 100 pM or more it produced complete inhibition in all cases. Therefore, the slope of the dose-response curve was very steep. DPT at 100 pM or more decreased the cell viability to 50%-60% and increased the number of cells undergoing apoptosis (annexin V-positive cells). The results indicate that DPT contributes to the cytotoxic action of the extract from the leaves of H. nymphaeaefolia on K562 cells. Copyright 2002 John Wiley & Sons, Ltd.

Combination of CD157 and FLAER to Detect Peripheral Blood Eosinophils by Multiparameter Flow Cytometry.

PubMed

Carulli, Giovanni; Marini, Alessandra; Sammuri, Paola; Domenichini, Cristiana; Ottaviano, Virginia; Pacini, Simone; Petrini, Mario

2015-01-01

The identification of eosinophils by flow cytometry is difficult because most of the surface antigens expressed by eosinophils are shared with neutrophils. Some methods have been proposed, generally based on differential light scatter properties, enhanced autofluorescence, lack of CD16 or selective positivity of CD52. Such methods, however, show several limitations. In the present study we report a novel method based on the analysis of glycosylphosphatidylinositol (GPI)-linked molecules. The combination of CD157 and FLAER was used, since FLAER recognizes all GPI-linked molecules, while CD157 is absent on the membrane of eosinophils and expressed by neutrophils. Peripheral blood samples from normal subjects and patients with variable percentages of eosinophils (n = 31), and without any evidence for circulating immature myeloid cells, were stained with the combination of FLAER-Alexa Fluor and CD157-PE. A FascCanto II cytometer was used. Granulocytes were gated after CD33 staining and eosinophils were identified as CD157(-)/FLAER(+) events. Neutrophils were identified as CD157(+)/FLAER(+) events. The percentages of eosinophils detected by this method showed a very significant correlation both with automated counting and with manual counting (r = 0.981 and 0.989, respectively). Sorting assays were carried out by a S3 Cell Sorter: cytospins obtained from CD157(-)/FLAER(+) events consisted of 100% eosinophils, while samples from CD157(+)/FLAER(+) events were represented only by neutrophils. In conclusion, this method shows high sensitivity and specificity in order to distinguish eosinophils from neutrophils by flow cytometry. However, since CD157 is gradually up-regulated throughout bone marrow myeloid maturation, our method cannot be applied to cases characterized by immature myeloid cells.

A Mock UF6 Feed and Withdrawal System for Testing Safeguards Monitoring Systems and Strategies Intended for Nuclear Fuel Enrichment and Processing Plants

DOE Office of Scientific and Technical Information (OSTI.GOV)

Krichinsky, Alan M; Bates, Bruce E; Chesser, Joel B

2009-12-01

This report describes an engineering-scale, mock UF6 feed and withdrawal (F&W) system, its operation, and its intended uses. This system has been assembled to provide a test bed for evaluating and demonstrating new methodologies that can be used in remote, unattended, continuous monitoring of nuclear material process operations. These measures are being investigated to provide independent inspectors improved assurance that operations are being conducted within declared parameters, and to increase the overall effectiveness of safeguarding nuclear material. Testing applicable technologies on a mock F&W system, which uses water as a surrogate for UF6, enables thorough and cost-effective investigation of hardware,more » software, and operational strategies before their direct installation in an industrial nuclear material processing environment. Electronic scales used for continuous load-cell monitoring also are described as part of the basic mock F&W system description. Continuous monitoring components on the mock F&W system are linked to a data aggregation computer by a local network, which also is depicted. Data collection and storage systems are described only briefly in this report. The mock UF{sub 6} F&W system is economical to operate. It uses a simple process involving only a surge tank between feed tanks and product and withdrawal (or waste) tanks. The system uses water as the transfer fluid, thereby avoiding the use of hazardous UF{sub 6}. The system is not tethered to an operating industrial process involving nuclear materials, thereby allowing scenarios (e.g., material diversion) that cannot be conducted otherwise. These features facilitate conducting experiments that yield meaningful results with a minimum of expenditure and quick turnaround time. Technologies demonstrated on the engineering-scale system lead to field trials (described briefly in this report) for determining implementation issues and performance of the monitoring technologies under plant

Cutting-edge analysis of extracellular microparticles using ImageStream(X) imaging flow cytometry.

PubMed

Headland, Sarah E; Jones, Hefin R; D'Sa, Adelina S V; Perretti, Mauro; Norling, Lucy V

2014-06-10

Interest in extracellular vesicle biology has exploded in the past decade, since these microstructures seem endowed with multiple roles, from blood coagulation to inter-cellular communication in pathophysiology. In order for microparticle research to evolve as a preclinical and clinical tool, accurate quantification of microparticle levels is a fundamental requirement, but their size and the complexity of sample fluids present major technical challenges. Flow cytometry is commonly used, but suffers from low sensitivity and accuracy. Use of Amnis ImageStream(X) Mk II imaging flow cytometer afforded accurate analysis of calibration beads ranging from 1 μm to 20 nm; and microparticles, which could be observed and quantified in whole blood, platelet-rich and platelet-free plasma and in leukocyte supernatants. Another advantage was the minimal sample preparation and volume required. Use of this high throughput analyzer allowed simultaneous phenotypic definition of the parent cells and offspring microparticles along with real time microparticle generation kinetics. With the current paucity of reliable techniques for the analysis of microparticles, we propose that the ImageStream(X) could be used effectively to advance this scientific field.

One-dimensional acoustic standing waves in rectangular channels for flow cytometry.

PubMed

Austin Suthanthiraraj, Pearlson P; Piyasena, Menake E; Woods, Travis A; Naivar, Mark A; Lόpez, Gabriel P; Graves, Steven W

2012-07-01

Flow cytometry has become a powerful analytical tool for applications ranging from blood diagnostics to high throughput screening of molecular assemblies on microsphere arrays. However, instrument size, expense, throughput, and consumable use limit its use in resource poor areas of the world, as a component in environmental monitoring, and for detection of very rare cell populations. For these reasons, new technologies to improve the size and cost-to-performance ratio of flow cytometry are required. One such technology is the use of acoustic standing waves that efficiently concentrate cells and particles to the center of flow channels for analysis. The simplest form of this method uses one-dimensional acoustic standing waves to focus particles in rectangular channels. We have developed one-dimensional acoustic focusing flow channels that can be fabricated in simple capillary devices or easily microfabricated using photolithography and deep reactive ion etching. Image and video analysis demonstrates that these channels precisely focus single flowing streams of particles and cells for traditional flow cytometry analysis. Additionally, use of standing waves with increasing harmonics and in parallel microfabricated channels is shown to effectively create many parallel focused streams. Furthermore, we present the fabrication of an inexpensive optical platform for flow cytometry in rectangular channels and use of the system to provide precise analysis. The simplicity and low-cost of the acoustic focusing devices developed here promise to be effective for flow cytometers that have reduced size, cost, and consumable use. Finally, the straightforward path to parallel flow streams using one-dimensional multinode acoustic focusing, indicates that simple acoustic focusing in rectangular channels may also have a prominent role in high-throughput flow cytometry. Copyright © 2012 Elsevier Inc. All rights reserved.

The Spatial and Temporal Variability of Meltwater Flow Paths: Insights From a Grid of Over 100 Snow Lysimeters

NASA Astrophysics Data System (ADS)

Webb, R. W.; Williams, M. W.; Erickson, T. A.

2018-02-01

Snowmelt is an important part of the hydrologic cycle and ecosystem dynamics for headwater systems. However, the physical process of water flow through snow is a poorly understood aspect of snow hydrology as meltwater flow paths tend to be highly complex. Meltwater flow paths diverge and converge as percolating meltwater reaches stratigraphic layer interfaces creating high spatial variability. Additionally, a snowpack is temporally heterogeneous due to rapid localized metamorphism that occurs during melt. This study uses a snowmelt lysimeter array at tree line in the Niwot Ridge study area of northern Colorado. The array is designed to address the issue of spatial and temporal variability of basal discharge at 105 locations over an area of 1,300 m2. Observed coefficients of variation ranged from 0 to almost 10 indicating more variability than previously observed, though this variability decreased throughout each melt season. Snowmelt basal discharge also significantly increases as snow depth decreases displaying a cluster pattern that peaks during weeks 3-5 of the snowmelt season. These results are explained by the flow of meltwater along snow layer interfaces. As the snowpack becomes less stratified through the melt season, the pattern transforms from preferential flow paths to uniform matrix flow. Correlation ranges of the observed basal discharge correspond to a mean representative elementary area of 100 m2, or a characteristic length of 10 m. Snowmelt models representing processes at scales less than this will need to explicitly incorporate the spatial variability of snowmelt discharge and meltwater flow paths through snow between model pixels.

Changes in CSF flow after one-stage posterior vertebral column resection in scoliosis patients with syringomyelia and Chiari malformation type I.

PubMed

Wang, Yingsong; Xie, Jingming; Zhao, Zhi; Zhang, Ying; Li, Tao; Si, Yongyu

2013-05-01

Phase contrast-cine MRI (PC-cine MRI) studies in patients with syringomyelia and Chiari malformation Type I (CM-I) have demonstrated abnormal CSF flow across the foramen magnum, which can revert to normal after craniocervical decompression with syrinx shrinkage. In order to investigate the mechanisms leading to postoperative syringomyelia shrinkage, the authors studied the hydrodynamic changes of CSF flow in the craniocervical junction and spinal canal in patients with scoliosis associated with syringomyelia after one-stage deformity correction by posterior vertebral column resection. Preoperative and postoperative CSF flow dynamics at the levels of the foramen magnum, C-7, T-7 (or apex), and L-1 were assessed by electrocardiogram-synchronized cardiac-gated PC-cine MRI in 8 adolescent patients suffering from severe scoliosis with syringomyelia and CM-I (scoliosis group) and undergoing posterior vertebral column resection. An additional 8 patients with syringomyelia and CM-I without spinal deformity (syrinx group) and 8 healthy volunteers (control group) were also enrolled. Mean values were obtained for the following parameters: the duration of a CSF cycle, the duration of caudad CSF flow (CSF downflow [DF]) and cephalad CSF flow (CSF upflow [UF]), the ratio of DF duration to CSF cycle duration (DF%), and the ratio of UF duration to CSF cycle duration (UF%). The ratio of the stationary phase (SP) duration to CSF cycle duration was calculated (SP%). The maximum downflow velocities (VD max) and maximum upflow velocities (VU max) were measured. SPSS (version 14.0) was used for all statistical analysis. Patients in the scoliosis group underwent one-stage posterior vertebral column resection for deformity correction without suboccipital decompression. The mean preoperative coronal Cobb angle was 102.4° (range 76°-138°). The mean postoperative Cobb angle was 41.7° (range 12°-75°), with an average correction rate of 59.3%. During the follow-up, 1 patient with

Flow Cytometry of Spinach Chloroplasts 1

PubMed Central

Schröder, Wolfgang P.; Petit, Patrice X.

1992-01-01

Intact spinach (Spinacia oleracea) chloroplasts, thylakoid membranes, and inside-out or right-side-out thylakoid vesicles have been characterized by flow cytometry with respect to forward angle light scatter, right angle light scatter, and chlorophyll fluorescence. Analysis of intact chloroplasts with respect to forward light scatter and the chlorophyll fluorescence parameter revealed the presence of truly “intact” and “disrupted” chloroplasts. The forward light scatter parameter, normally considered to reflect object size, was instead found to reflect the particle density. One essential advantage of flow cytometry is that additional parameters such as Ricinus communis agglutinin (linked to fluorescein isothiocyanate) fluorescence can be determined through logical conditions placed on bit-maps, amounting to an analytical purification procedure. In the present case, chloroplast subpopulations with fully preserved envelopes, thylakoid membrane, and inside-out or right-side-out thylakoid membranes vesicles can be distinguished. Flow cytometry is also a useful tool to address the question of availability of glycosyl moities on the membrane surfaces if one keeps in mind that organelle-to-organelle interactions could be partially mediated through a recognition process. A high specific binding of R. communis agglutinin and peanut lectin to the chloroplast envelope was detected. This showed that galactose residues were exposed and accessible to specific lectins on the chloroplast surface. No exposed glucose, fucose, or mannose residues could be detected by the appropriate lectins. Ricin binding to the intact chloroplasts caused a strong aggregation. Disruption of these aggregates by resuspension or during passage in the flow cytometer induced partial breakage of the chloroplasts. Only minor binding of R. communis agglutinin and peanut lectin to the purified thylakoid membranes was detected; the binding was found to be low for both inside-out and right

Performance evaluation of cross-flow membrane system for wastewater reuse from the wood-panels industry.

PubMed

Dizge, Nadir

2014-01-01

The objectives of this investigation were to perform a series of lab-scale membrane separation experiments under various operating conditions to investigate the performance behaviour of nanofiltration membrane (NF 270) for wastewater reuse from the wood-panels industry. The operating condition effects, e.g. cross-flow velocity (CFV), trans membrane pressure (TMP) and temperature, on the permeate flux and contaminant rejection efficiency were investigated. Moreover, three different samples: (1) raw wastewater collected from the wood-panels industry; (2) ultrafiltration pre-treated wastewater (UF-NF); and (3) coagulation/flocculation pre-treated wastewater (CF-NF) were employed in this study. The UF-NF was proposed as a pre-treatment process because it could reduce the chemical oxygen demand (COD) effectively with lower energy consumption than CF-NF. The performance of NF 270 membrane was assessed by measurements of the many parameters (pH, conductivity, total dissolved solids, COD, suspended solids, total nitrogen, nitrite, nitrate, and total phosphate) under various operating conditions. It was noted that the contaminant rejection was affected by changing TMP and CFV. It was concluded that the purified water stream can be recycled into the process for water reuse or safely disposed to the river.

Detection of dilute sperm samples using photoacoustic flowmetry

NASA Astrophysics Data System (ADS)

Viator, J. A.; Sutovsky, P.; Weight, R. M.

2008-02-01

Detection of sperm cells in dilute samples may have application in forensic testing and diagnosis of male reproductive health. Due to the optically dense subcellular structures in sperm cells, irradiation by nanosecond laser pulses induces a photoacoustic response detectable using a custom flow cytometer. We determined the detection threshold of bull sperm using various concentrations, from 200 to 1,000,000 sperm cells per milliliter. Using a tunable laser system set to 450nm with a 5 ns pulse duration and 11-12 mJ/pulse, we obtained a detection threshold of 3 sperm cells. The flow rate was 4 ml/minute through the flow chamber. The acoustic sensor was a 100 μm PVDF film attached to the glass flow chamber. The acoustic signal was preamplified and sent to an oscilloscope. The threshold signal indicated a signal to noise ratio of approximately 6 to 1. Improved system design may decrease the threshold to single sperm cells.

Status Report on the Passive Neutron Enrichment Meter (PNEM) for UF6 Cylinder Assay

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, Karen A.; Swinhoe, Martyn T.; Menlove, Howard O.

2012-05-02

The Passive Neutron Enrichment Meter (PNEM) is a nondestructive assay (NDA) system being developed at Los Alamos National Laboratory (LANL). It was designed to determine {sup 235}U mass and enrichment of uranium hexafluoride (UF{sub 6}) in product, feed, and tails cylinders (i.e., 30B and 48Y cylinders). These cylinders are found in the nuclear fuel cycle at uranium conversion, enrichment, and fuel fabrication facilities. The PNEM is a {sup 3}He-based neutron detection system that consists of two briefcase-sized detector pods. A photograph of the system during characterization at LANL is shown in Fig. 1. Several signatures are currently being studied tomore » determine the most effective measurement and data reduction technique for unfolding {sup 235}U mass and enrichment. The system collects total neutron and coincidence data for both bare and cadmium-covered detector pods. The measurement concept grew out of the success of the Uranium Cylinder Assay System (UCAS), which is an operator system at Rokkasho Enrichment Plant (REP) that uses total neutron counting to determine {sup 235}U mass in UF{sub 6} cylinders. The PNEM system was designed with higher efficiency than the UCAS in order to add coincidence counting functionality for the enrichment determination. A photograph of the UCAS with a 48Y cylinder at REP is shown in Fig. 2, and the calibration measurement data for 30B product and 48Y feed and tails cylinders is shown in Fig. 3. The data was collected in a low-background environment, meaning there is very little scatter in the data. The PNEM measurement concept was first presented at the 2010 Institute of Nuclear Materials Management (INMM) Annual Meeting. The physics design and uncertainty analysis were presented at the 2010 International Atomic Energy Agency (IAEA) Safeguards Symposium, and the mechanical and electrical designs and characterization measurements were published in the ESARDA Bulletin in 2011.« less

Recovery of Whey Proteins and Enzymatic Hydrolysis of Lactose Derived from Casein Whey Using a Tangential Flow Ultrafiltration Module

NASA Astrophysics Data System (ADS)

Das, Bipasha; Bhattacharjee, Sangita; Bhattacharjee, Chiranjib

2013-09-01

In this study, ultrafiltration (UF) of pretreated casein whey was carried out in a cross-flow module fitted with 5 kDa molecular weight cut-off polyethersulfone membrane to recover whey proteins in the retentate and lactose in the permeate. Effects of processing conditions, like transmembrane pressure and pH on permeate flux and rejection were investigated and reported. The polarised layer resistance was found to increase with time during UF even in this high shear device. The lactose concentration in the permeate was measured using dinitro salicylic acid method. Enzymatic kinetic study for lactose hydrolysis was carried out at three different temperatures ranging from 30 to 50 °C using β-galactosidase enzyme. The glucose formed during lactose hydrolysis was analyzed using glucose oxidase-peroxidase method. Kinetics of enzymatic hydrolysis of lactose solution was found to follow Michaelis-Menten model and the model parameters were estimated by Lineweaver-Burk plot. The hydrolysis rate was found to be maximum (with Vmax = 5.5091 mmol/L/min) at 30 °C.

Highly multiparametric analysis by mass cytometry.

PubMed

Ornatsky, Olga; Bandura, Dmitry; Baranov, Vladimir; Nitz, Mark; Winnik, Mitchell A; Tanner, Scott

2010-09-30

This review paper describes a new technology, mass cytometry, that addresses applications typically run by flow cytometer analyzers, but extends the capability to highly multiparametric analysis. The detection technology is based on atomic mass spectrometry. It offers quantitation, specificity and dynamic range of mass spectrometry in a format that is familiar to flow cytometry practitioners. The mass cytometer does not require compensation, allowing the application of statistical techniques; this has been impossible given the constraints of fluorescence noise with traditional cytometry instruments. Instead of "colors" the mass cytometer "reads" the stable isotope tags attached to antibodies using metal-chelating labeling reagents. Because there are many available stable isotopes, and the mass spectrometer provides exquisite resolution between detection channels, many parameters can be measured as easily as one. For example, in a single tube the technique allows for the ready detection and characterization of the major cell subsets in blood or bone marrow. Here we describe mass cytometric immunophenotyping of human leukemia cell lines and leukemia patient samples, differential cell analysis of normal peripheral and umbilical cord blood; intracellular protein identification and metal-encoded bead arrays. Copyright © 2010 Elsevier B.V. All rights reserved.

A Miniature Couette to Generate Shear for Flow Cytometry: Studying Real-Time Modulation of Intracellular Calcium in Monocytic Cells

PubMed Central

Zwartz, Gordon J.; Chigaev, Alexandre; Foutz, Terry D.; Edwards, Bruce; Sklar, Larry A.

2013-01-01

Extracellular hydrodynamic forces may be transmitted to the interior of cells through the alteration of integrin conformation and affinity. Integrin activation regulates leukocyte recruitment, cell activation, and transmigration. The cellular and molecular mechanisms for integrin activation are not precisely known, although intracellular calcium signaling is involved. Flow cytometry offers a versatile way to study intracellular calcium signaling in real-time. We report a novel method to generate defined shear by using a miniature Couette. Testing involved measuring shear induced intracellular calcium signals of human monoblastoid U937 cells in suspension. The Couette was connected externally to a flow cytometer and pressurized at 6 PSI (4.1 N/m2). Cells were subjected to well-defined shear between 0 and 1000 s−1 and delivered continuously within 10 s to a FACScan at 1 μl/s. Intracellular calcium levels and the percentage of cells activated increased as shear increased in duration and intensity. PMID:22045643

A simple fabrication of plasmonic surface-enhanced Raman scattering (SERS) substrate for pesticide analysis via the immobilization of gold nanoparticles on UF membrane

NASA Astrophysics Data System (ADS)

Hong, Jangho; Kawashima, Ayato; Hamada, Noriaki

2017-06-01

In this study, we developed a facile fabrication method to access a highly reproducible plasmonic surface enhanced Raman scattering substrate via the immobilization of gold nanoparticles on an Ultrafiltration (UF) membrane using a suction technique. This was combined with a simple and rapid analyte concentration and detection method utilizing portable Raman spectroscopy. The minimum detectable concentrations for aqueous thiabendazole standard solution and thiabendazole in orange extract are 0.01 μg/mL and 0.125 μg/g, respectively. The partial least squares (PLS) regression plot shows a good linear relationship between 0.001 and 100 μg/mL of analyte, with a root mean square error of prediction (RMSEP) of 0.294 and a correlation coefficient (R2) of 0.976 for the thiabendazole standard solution. Meanwhile, the PLS plot also shows a good linear relationship between 0.0 and 2.5 μg/g of analyte, with an RMSEP value of 0.298 and an R2 value of 0.993 for the orange peel extract. In addition to the detection of other types of pesticides in agricultural products, this highly uniform plasmonic substrate has great potential for application in various environmentally-related areas.

Rise of the micromachines: microfluidics and the future of cytometry.

PubMed

Wlodkowic, Donald; Darzynkiewicz, Zbigniew

2011-01-01

The past decade has brought many innovations to the field of flow and image-based cytometry. These advancements can be seen in the current miniaturization trends and simplification of analytical components found in the conventional flow cytometers. On the other hand, the maturation of multispectral imaging cytometry in flow imaging and the slide-based laser scanning cytometers offers great hopes for improved data quality and throughput while proving new vistas for the multiparameter, real-time analysis of cells and tissues. Importantly, however, cytometry remains a viable and very dynamic field of modern engineering. Technological milestones and innovations made over the last couple of years are bringing the next generation of cytometers out of centralized core facilities while making it much more affordable and user friendly. In this context, the development of microfluidic, lab-on-a-chip (LOC) technologies is one of the most innovative and cost-effective approaches toward the advancement of cytometry. LOC devices promise new functionalities that can overcome current limitations while at the same time promise greatly reduced costs, increased sensitivity, and ultra high throughputs. We can expect that the current pace in the development of novel microfabricated cytometric systems will open up groundbreaking vistas for the field of cytometry, lead to the renaissance of cytometric techniques and most importantly greatly support the wider availability of these enabling bioanalytical technologies. Copyright © 2011 Elsevier Inc. All rights reserved.

Rise of the Micromachines: Microfluidics and the Future of Cytometry

PubMed Central

Wlodkowic, Donald; Darzynkiewicz, Zbigniew

2011-01-01

The past decade has brought many innovations to the field of flow and image-based cytometry. These advancements can be seen in the current miniaturization trends and simplification of analytical components found in the conventional flow cytometers. On the other hand, the maturation of multispectral imaging cytometry in flow imaging and the slide-based laser scanning cytometers offers great hopes for improved data quality and throughput while proving new vistas for the multiparameter, real-time analysis of cells and tissues. Importantly, however, cytometry remains a viable and very dynamic field of modern engineering. Technological milestones and innovations made over the last couple of years are bringing the next generation of cytometers out of centralized core facilities while making it much more affordable and user friendly. In this context, the development of microfluidic, lab-on-a-chip (LOC) technologies is one of the most innovative and cost-effective approaches toward the advancement of cytometry. LOC devices promise new functionalities that can overcome current limitations while at the same time promise greatly reduced costs, increased sensitivity, and ultra high throughputs. We can expect that the current pace in the development of novel microfabricated cytometric systems will open up groundbreaking vistas for the field of cytometry, lead to the renaissance of cytometric techniques and most importantly greatly support the wider availability of these enabling bioanalytical technologies. PMID:21704837

Optimization of air plasma reconversion of UF6 to UO2 based on thermodynamic calculations

NASA Astrophysics Data System (ADS)

Tundeshev, Nikolay; Karengin, Alexander; Shamanin, Igor

2018-03-01

The possibility of plasma-chemical conversion of depleted uranium-235 hexafluoride (DUHF) in air plasma in the form of gas-air mixtures with hydrogen is considered in the paper. Calculation of burning parameters of gas-air mixtures is carried out and the compositions of mixtures obtained via energy-efficient conversion of DUHF in air plasma are determined. With the help of plasma-chemical conversion, thermodynamic modeling optimal composition of UF6-H2-Air mixtures and its burning parameters, the modes for production of uranium dioxide in the condensed phase are determined. The results of the conducted researches can be used for creation of technology for plasma-chemical conversion of DUHF in the form of air-gas mixtures with hydrogen.

NUCLEAR FISSION CHAIN REACTING SYSTEM

DOEpatents

Anderson, H.L.; Brown, H.S.

1961-06-27

The patent describes a reactor consisting of a plurality of tubes passing through a body of heavy water or graphite, a heat exchanger, means for flowing UF/sub 6/ through the tubes and the heat exchangar, and means for bleeding off some of the UF/sub 6/ and separating plutonium therefrom. A specific suggestion contained is that the amount of the UF/sub 6/ outside the reaction unit be a multiple of that within it.

High-lift flow-physics flight experiments on a subsonic civil transport aircraft (B737-100)

NASA Technical Reports Server (NTRS)

Vandam, Cornelis P.

1994-01-01

As part of the subsonic transport high-lift program, flight experiments are being conducted using NASA Langley's B737-100 to measure the flow characteristics of the multi-element high-lift system at full-scale high-Reynolds-number conditions. The instrumentation consists of hot-film anemometers to measure boundary-layer states, an infra-red camera to detect transition from laminar to turbulent flow, Preston tubes to measure wall shear stress, boundary-layer rakes to measure off-surface velocity profiles, and pressure orifices to measure surface pressure distributions. The initial phase of this research project was recently concluded with two flights on July 14. This phase consisted of a total of twenty flights over a period of about ten weeks. In the coming months the data obtained in this initial set of flight experiments will be analyzed and the results will be used to finalize the instrumentation layout for the next set of flight experiments scheduled for Winter and Spring of 1995. The main goal of these upcoming flights will be: (1) to measure more detailed surface pressure distributions across the wing for a range of flight conditions and flap settings; (2) to visualize the surface flows across the multi-element wing at high-lift conditions using fluorescent mini tufts; and (3) to measure in more detail the changes in boundary-layer state on the various flap elements as a result of changes in flight condition and flap deflection. These flight measured results are being correlated with experimental data measured in ground-based facilities as well as with computational data calculated with methods based on the Navier-Stokes equations or a reduced set of these equations. Also these results provide insight into the extent of laminar flow that exists on actual multi-element lifting surfaces at full-scale high-life conditions. Preliminary results indicate that depending on the deflection angle, the slat and flap elements have significant regions of laminar flow over

Effective factors in the treatment of kerosene-water emulsion by using UF membranes.

PubMed

Rezvanpour, Alireza; Roostaazad, Reza; Hesampour, Mehrdad; Nyström, Marianne; Ghotbi, Cyrus

2009-01-30

The effects of different parameters including membrane type (regenerated cellulose and polysulphone), transmembrane pressure (TMP), the content of oil in the feed, the flow velocity of the feed and pH on the ultrafiltration of an emulsion of kerosene in water were studied. It was found that the important factors affecting ultrafiltration were, in order, membrane type, pressure and oil concentration. The greatest flux at the optimum conditions here of 3 bar, an oil content of 3% (v/v) and with membrane type C30F was predicted as 108 L/(m(2)h) that was within the range of the confidence limit of the measured value of 106 L/(m(2)h). The normalised FTIR results of the virgin cellulosic membranes C30F and C100F showed more abundant OH groups. The bigger number of OH groups implies a greater hydrophilicity. The larger observed flux in the C30F is related to a higher number of pores as well (surface porosity) compared with the C100F membrane. In the "polarised regime" from 3 bar upwards, flux was independent of pressure for all membranes and was assumed to be determined by the back diffusion transport. Despite the fact that both the PS100H and C100F membranes had the same cut-off (100 kg/mol), the hydrophilic C100F showed a superior permeate flux. The strongest drop of flux with time due to oil fouling was observed for the C100F although it was hydrophilic. In the case of the PS100H, both FTIR and SEM showed that cake layer formation was not the cause of fouling. Meanwhile the SEM and FTIR results of fouled C100F provided evidence of adsorptive and gel formation fouling.

Light-scattering flow cytometry for identification and characterization of blood microparticles

NASA Astrophysics Data System (ADS)

Konokhova, Anastasiya I.; Yurkin, Maxim A.; Moskalensky, Alexander E.; Chernyshev, Andrei V.; Tsvetovskaya, Galina A.; Chikova, Elena D.; Maltsev, Valeri P.

2012-05-01

We describe a novel approach to study blood microparticles using the scanning flow cytometer, which measures light scattering patterns (LSPs) of individual particles. Starting from platelet-rich plasma, we separated spherical microparticles from non-spherical plasma constituents, such as platelets and cell debris, based on similarity of their LSP to that of sphere. This provides a label-free method for identification (detection) of microparticles, including those larger than 1 μm. Next, we rigorously characterized each measured particle, determining its size and refractive index including errors of these estimates. Finally, we employed a deconvolution algorithm to determine size and refractive index distributions of the whole population of microparticles, accounting for largely different reliability of individual measurements. Developed methods were tested on a blood sample of a healthy donor, resulting in good agreement with literature data. The only limitation of this approach is size detection limit, which is currently about 0.5 μm due to used laser wavelength of 0.66 μm.

Smart catheter flow sensor for real-time continuous regional cerebral blood flow monitoring

NASA Astrophysics Data System (ADS)

Li, Chunyan; Wu, Pei-Ming; Hartings, Jed A.; Wu, Zhizhen; Ahn, Chong H.; LeDoux, David; Shutter, Lori A.; Narayan, Raj K.

2011-12-01

We present a smart catheter flow sensor for real-time, continuous, and quantitative measurement of regional cerebral blood flow using in situ temperature and thermal conductivity compensation. The flow sensor operates in a constant-temperature mode and employs a periodic heating and cooling technique. This approach ensures zero drift and provides highly reliable data with microelectromechanical system-based thin film sensors. The developed flow sensor has a sensitivity of 0.973 mV/ml/100 g/min in the range from 0 to 160 ml/100 g/min with a linear correlation coefficient of R2 = 0.9953. It achieves a resolution of 0.25 ml/100 g/min and an accuracy better than 5 ml/100 g/min.

Synthesis of a combined system for precise stabilization of the Spektr-UF observatory: II

NASA Astrophysics Data System (ADS)

Bychkov, I. V.; Voronov, V. A.; Druzhinin, E. I.; Kozlov, R. I.; Ul'yanov, S. A.; Belyaev, B. B.; Telepnev, P. P.; Ul'yashin, A. I.

2014-03-01

The paper presents the second part of the results of search studies for the development of a combined system of high-precision stabilization of the optical telescope for the designed Spectr-UF international observatory [1]. A new modification of the strict method of the synthesis of nonlinear discrete-continuous stabilization systems with uncertainties is described, which is based on the minimization of the guaranteed accuracy estimate calculated using vector Lyapunov functions. Using this method, the synthesis of the feedback parameters in the mode of precise inertial stabilization of the optical telescope axis is performed taking the design nonrigidity, quantization of signals over time and level, and errors of orientation meters, as well as the errors and limitation of control moments of executive engine-flywheels into account. The results of numerical experiments that demonstrate the quality of the synthesized system are presented.

MANUFACTURE OF UF$sub 4$

DOEpatents

Calcott, W.S.

1959-10-13

The manufacture of uranium tetrafluoride from urarium dioxide is described. Uranium dioxide is heated to about 500 deg C in a reactor. Anhydrous hydrogen fluoride is passed through the reactor in contact with uranium dioxide for several hours, the flow of hydrogen fluoride is discontinued, and hydrogen passed through the reactor for less than an hour. The flow of hydrogen fluoride is resumed for several hours, and then nitrogen is passed for a few minutes to expel unreacted hydrogen fluoride as water vapor. The reactor is cooled to room temperature and the uranium tetrafluoride removed.

Electronic, magnetic, transport, and thermal properties of single-crystalline UF e2A l10

NASA Astrophysics Data System (ADS)

Troć, R.; Samsel-Czekała, M.; Talik, E.; Wawryk, R.; Gajek, Z.; Pasturel, M.

2015-09-01

The valence and core-level x-ray photoemission spectra (XPS), performed on an UF e2A l10 single crystal, were measured using the Al Kα radiation. The results of valence XPS show practically two separate regions of spectral intensity, one just at the Fermi level (EF) and the other one being a wide content with its maximum at about 0.8 eV below EF. These give rise to two electronic configurations of the 5 f states in the studied aluminide, itinerant and localized ones, i.e., their dual character. In such a situation the corresponding valence spectra, calculated within the local density approximation (LDA), well explain the former configuration, being responsible for a metallic behavior of the studied compound. Moreover, this behavior is confirmed clearly also by our results of magnetotransport measurements. On the other hand, the obtained magnetic susceptibility, specific heat, electrical resistivity, and thermoelectric power data support very well the local character of the 5 f2 -electron configuration of the U4 + ion in UF e2A l10 having the orthorhombic and cage-type crystal structure. Based on that configuration, the magnetic and thermal characteristics of the compound were modeled by the effective crystal field (CF) potential in the intermediate coupling scheme using initial parameters obtained by the angular overlap model (AOM). The obtained final CF parameters yielded the CF level scheme, composed of only singlets, proper for orthorhombic symmetry. Such a set of singlets reproduces in a satisfactory way both the strongly anisotropic temperature variations of the magnetic susceptibility, measured along the three main crystallographic directions, as well as the Schottky anomaly, evaluated using specific heat results of isomorphic ThF e2A l10 as a phonon reference. Also, the strongly anisotropic behavior of the Seebeck coefficient and its low temperature maxima observed for the compound studied here have been explained roughly by the CF effect.

Flow cytometric analysis of microbial contamination in food industry technological lines--initial study.

PubMed

Józwa, Wojciech; Czaczyk, Katarzyna

2012-04-02

Flow cytometry constitutes an alternative for traditional methods of microorganisms identification and analysis, including methods requiring cultivation step. It enables the detection of pathogens and other microorganisms contaminants without the need to culture microbial cells meaning that the sample (water, waste or food e.g. milk, wine, beer) may be analysed directly. This leads to a significant reduction of time required for analysis allowing monitoring of production processes and immediate reaction in case of contamination or any disruption occurs. Apart from the analysis of raw materials or products on different stages of manufacturing process, the flow cytometry seems to constitute an ideal tool for the assessment of microbial contamination on the surface of technological lines. In the present work samples comprising smears from 3 different surfaces of technological lines from fruit and vegetable processing company from Greater Poland were analysed directly with flow cytometer. The measured parameters were forward and side scatter of laser light signals allowing the estimation of microbial cell contents in each sample. Flow cytometric analysis of the surface of food industry production lines enable the preliminary evaluation of microbial contamination within few minutes from the moment of sample arrival without the need of sample pretreatment. The presented method of fl ow cytometric initial evaluation of microbial state of food industry technological lines demonstrated its potential for developing a robust, routine method for the rapid and labor-saving detection of microbial contamination in food industry.

From the Lab to the real world : sources of error in UF {sub 6} gas enrichment monitoring

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lombardi, Marcie L.

2012-03-01

Safeguarding uranium enrichment facilities is a serious concern for the International Atomic Energy Agency (IAEA). Safeguards methods have changed over the years, most recently switching to an improved safeguards model that calls for new technologies to help keep up with the increasing size and complexity of today’s gas centrifuge enrichment plants (GCEPs). One of the primary goals of the IAEA is to detect the production of uranium at levels greater than those an enrichment facility may have declared. In order to accomplish this goal, new enrichment monitors need to be as accurate as possible. This dissertation will look at themore » Advanced Enrichment Monitor (AEM), a new enrichment monitor designed at Los Alamos National Laboratory. Specifically explored are various factors that could potentially contribute to errors in a final enrichment determination delivered by the AEM. There are many factors that can cause errors in the determination of uranium hexafluoride (UF{sub 6}) gas enrichment, especially during the period when the enrichment is being measured in an operating GCEP. To measure enrichment using the AEM, a passive 186-keV (kiloelectronvolt) measurement is used to determine the {sup 235}U content in the gas, and a transmission measurement or a gas pressure reading is used to determine the total uranium content. A transmission spectrum is generated using an x-ray tube and a “notch” filter. In this dissertation, changes that could occur in the detection efficiency and the transmission errors that could result from variations in pipe-wall thickness will be explored. Additional factors that could contribute to errors in enrichment measurement will also be examined, including changes in the gas pressure, ambient and UF{sub 6} temperature, instrumental errors, and the effects of uranium deposits on the inside of the pipe walls will be considered. The sensitivity of the enrichment calculation to these various parameters will then be evaluated

Moxa-stick suffumigation for disinfecting air in hematology and hematopoietic stem cell transplantation wards with class 100 laminar flow.

PubMed

He, Jing-song; Yang, Qing; Huang, Wei-jia; Hu, Xiao-rong

2014-04-01

To evaluate the effect of moxa-stick suffumigation in the hematology and hematopoietic stem cell transplantation (HSCT) wards with luminar flow. The plate exposure method was used to measure the effect of air-disinfection of moxa-stick suffumigation in hematology and HSCT wards. The yearly average qualified rates of air sampling in HSCT wards were evaluated from 2007 to 2010. To further investigate the disinfecting effect of moxa-stick suffumigation, the colony counts of common pathogens (including Staphylcoccus aureus and Pseudomonas aeruginosa) before and after moxa-stick suffumigation were compared. The mean air quality rates of the HSCT wards with class 100 laminar flow were all above 90.0% (91.2%-96.2%) from 2007 to 2010. Moxa-stick suffumigation effectively decreased the presence of bacteria in the hematology ward's air (P<0.01). The most notable effect was the drastic reduction in the colony counts of Staphylococcus aureus and Pseudomonas aeruginosa on the blood plates exposed to air treated with moxa-stick suffumigation (77.1±52.9 cfu/m(2) vs 196.1±87.5 cfu/m(2), P<0.01; and 100.2±35.3 cfu/m(2) vs 371.5±35.3 cfu/m(2), P<0.01). Moxa-stick suffumigation proved to be a reliable and effective airdisinfection method for hematology and HSCT wards, and hence, it should be employed extensively.

Detection of fumonisin b1 and ochratoxin a in grain products using microsphere-based fluid array immunoassays.

PubMed

Anderson, George P; Kowtha, Vasudha A; Taitt, Chris R

2010-02-01

Grain products are a staple of diets worldwide and therefore, the ability to accurately and efficiently detect foodborne contaminants such as mycotoxins is of importance to everyone. Here we describe an indirect competitive fluid array fluoroimmunoassay to quantify the mycotoxins, fumonisin B1 and ochratoxin A. Both toxins were immobilized to the surface of microspheres using a variety of intermediate molecules and binding of biotinylated "tracer" antibody tracers determined through flow cytometry using streptavidin-phycoerythrin conjugates and the Luminex100 flow cytometer. Competitive assays were developed where the binding of biotinylated monoclonal antibodies to fumonisin B and ochratoxin A was competitively inhibited by different concentrations of those toxins in solution. Concentrations of fumonisin giving 50% inhibition were 300 pg/mL in buffer, 100 ng/g in spiked oats, and 1 μg/g in spiked cornmeal; analogous concentrations for ochratoxin A were 30 ng/mL in buffer, 30 ng/g in spiked oats, and 10 ng/g in spiked corn. The future challenge will be to expand the number of mycotoxins tested both individually and in multiplexed format using this platform.

Detection and capture of breast cancer cells with photoacoustic flow cytometry

NASA Astrophysics Data System (ADS)

Bhattacharyya, Kiran; Goldschmidt, Benjamin S.; Viator, John A.

2016-08-01

According to the Centers for Disease Control and Prevention, breast cancer is the most common cancer and the second leading cause of cancer related deaths among women. Metastasis-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. A technique is developed to detect circulating breast cancer cells in human blood using a photoacoustic flow cytometry method. A Q-switched laser is used to interrogate thousands of blood cells with one pulse as they flow through the beam path. Cells that are optically absorbing, 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 enhance optical absorption. After which, the PA cytometry device is calibrated to demonstrate the ability to detect single cells. Cultured breast cancer cells are added to whole blood to reach a biologically relevant concentration of about 25 to 45 breast cancer cells per 1 mL of blood. An in vitro PA 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 but also it can be used for genetic testing and in vitro drug trials since the circulating cell can be captured and studied.

Experimental and Numerical Characterization of a Pulsed Supersonic Uniform Flow for Kinetics and Spectroscopy

NASA Astrophysics Data System (ADS)

Suas-David, Nicolas; Thawoos, Shameemah; Broderick, Bernadette M.; Suits, Arthur

2017-06-01

The current CPUF (Chirped Pulse Uniform Flow) and the new UF-CRDS (Uniform Flow Cavity Ring-Down Spectroscopy) setups relie mostly on the production of a good quality supersonic uniform flow. A supersonic uniform flow is produced by expanding a gas through a Laval nozzle - similar to the nozzles used in aeronautics - linked to a vacuum chamber. The expansion is characterized by an isentropic core where constant very low kinetic temperature (down to 20K) and constant density are observed. The relatively large diameter of the isentropic core associated with homogeneous thermodynamic conditions makes it a relevant tool for low temperature spectroscopy. On the other hand, the length along the axis of the flow of this core (could be longer than 50cm) allows kinetic studies which is one of the main interest of this setup (CRESU technique. The formation of a uniform flow requires an extreme accuracy in the design of the shape of the nozzle for a set of defined temperature/density. The design is based on a Matlab program which retrieves the shape of the isentropic core according to the method of characteristics prior to calculate the thickness of the boundary layer. Two different approaches are used to test the viability of a new nozzle derived from the program. First, a computational fluid dynamic software (OpenFOAM) models the distribution of the thermodynamic properties of the expansion. Then, fabricated nozzles using 3-D printing are tested based on Pitot measurements and spectroscopic analyses. I will present comparisons of simulation and measured performance for a range of nozzles. We will see how the high level of accuracy of numerical simulations provides a deeper knowledge of the experimental conditions. J. M. Oldham, C. Abeysekera, J. Joalland, L. N. Zack, K. Prozument, I. R. Sims, G. Barrat Park, R. W. Filed and A. G. Suits, J. Chem. Phys. 141, 154202, (2014). I. Sims, J. L. Queffelec, A. Defrance, C. Rebrion-Rowe, D. Travers, P. Bocherel, B. Rowe, I. W. Smith

Four-Channel Threshold Detector With Optical Isolation

DTIC Science & Technology

2009-02-01

1uF tantalum 35 V Capacitor 6 75ohm 1l4 W Carbon or Metal Film Resistor I .1 uF tantalum 35V Capacitor 37 HCC / HCF 40688 CMOS 8 input NAND / AND 1...100uF aluminum 35 V Capacitor 3 1M340T15 1 5 V, 1 A TO –220 Regulator 1 MC7915ACT neg15 V, 1 A TO –220 Regulator 1 35952503 50 Kohm, 10-turn

Comparison of flow cytometry, fluorescence microscopy and spectrofluorometry for analysis of gene electrotransfer efficiency.

PubMed

Marjanovič, Igor; Kandušer, Maša; Miklavčič, Damijan; Keber, Mateja Manček; Pavlin, Mojca

2014-12-01

In this study, we compared three different methods used for quantification of gene electrotransfer efficiency: fluorescence microscopy, flow cytometry and spectrofluorometry. We used CHO and B16 cells in a suspension and plasmid coding for GFP. The aim of this study was to compare and analyse the results obtained by fluorescence microscopy, flow cytometry and spectrofluorometry and in addition to analyse the applicability of spectrofluorometry for quantifying gene electrotransfer on cells in a suspension. Our results show that all the three methods detected similar critical electric field strength, around 0.55 kV/cm for both cell lines. Moreover, results obtained on CHO cells showed that the total fluorescence intensity and percentage of transfection exhibit similar increase in response to increase electric field strength for all the three methods. For B16 cells, there was a good correlation at low electric field strengths, but at high field strengths, flow cytometer results deviated from results obtained by fluorescence microscope and spectrofluorometer. Our study showed that all the three methods detected similar critical electric field strengths and high correlations of results were obtained except for B16 cells at high electric field strengths. The results also demonstrated that flow cytometry measures higher values of percentage transfection compared to microscopy. Furthermore, we have demonstrated that spectrofluorometry can be used as a simple and consistent method to determine gene electrotransfer efficiency on cells in a suspension.

An improved method for differentiating cell-bound from internalized particles by imaging flow cytometry.

PubMed

Smirnov, Asya; Solga, Michael D; Lannigan, Joanne; Criss, Alison K

2015-08-01

Recognition, binding, internalization, and elimination of pathogens and cell debris are important functions of professional as well as non-professional phagocytes. However, high-throughput methods for quantifying cell-associated particles and discriminating bound from internalized particles have been lacking. Here we describe a protocol for using imaging flow cytometry to quantify the attached and phagocytosed particles that are associated with a population of cells. Cells were exposed to fluorescent particles, fixed, and exposed to an antibody of a different fluorophore that recognizes the particles. The antibody is added without cell permeabilization, such that the antibody only binds extracellular particles. Cells with and without associated particles were identified by imaging flow cytometry. For each cell with associated particles, a spot count algorithm was employed to quantify the number of extracellular (double fluorescent) and intracellular (single fluorescent) particles per cell, from which the percent particle internalization was determined. The spot count algorithm was empirically validated by examining the fluorescence and phase contrast images acquired by the flow cytometer. We used this protocol to measure binding and internalization of the bacterium Neisseria gonorrhoeae by primary human neutrophils, using different bacterial variants and under different cellular conditions. The results acquired using imaging flow cytometry agreed with findings that were previously obtained using conventional immunofluorescence microscopy. This protocol provides a rapid, powerful method for measuring the association and internalization of any particle by any cell type. Copyright © 2015 Elsevier B.V. All rights reserved.

CROSS-DISCIPLINARY PHYSICS AND RELATED AREAS OF SCIENCE AND TECHNOLOGY: Surface Micromachined Adjustable Micro-Concave Mirror for Bio-Detection Applications

NASA Astrophysics Data System (ADS)

Kuo, Ju-Nan; Chen, Wei-Lun; Jywe, Wen-Yuh

2009-08-01

We present a bio-detection system integrated with an adjustable micro-concave mirror. The bio-detection system consists of an adjustable micro-concave mirror, micro flow cytometer chip and optical detection module. The adjustable micro-concave mirror can be fabricated with ease using commercially available MEMS foundry services (such as multiuser MEMS processes, MUMPs) and its curvature can be controlled utilizing thermal or electrical effects. Experimental results show that focal lengths of the micro-concave mirror ranging from 313.5 to 2275.0 μm are achieved. The adjustable micro-concave mirror can be used to increase the efficiency of optical detection and provide a high signal-to-noise ratio. The developed micro-concave mirror is integrated with a micro flow cytometer for cell counting applications. Successful counting of fluorescent-labeled beads is demonstrated using the developed method.

Systematic review and meta-analysis of flow cytometry in urinary tract infection screening.

PubMed

Shang, Yan-Jun; Wang, Qian-Qian; Zhang, Jian-Rong; Xu, Yu-Lian; Zhang, Wei-Wei; Chen, Yan; Gu, Ming-Li; Hu, Zhi-De; Deng, An-Mei

2013-09-23

Automated urine sediment analysis of white blood cells (WBCs) and bacteria is a promising approach for urinary tract infections (UTIs) screening. However, available data on their screening efficacy is inconsistent. English articles from Pubmed, EMBASE, and Web of Science published before December 1, 2012 were analyzed. The Quality Assessment for Studies of Diagnostic Accuracy (QUADAS) tool was used to evaluate the quality of eligible studies. Performance characteristics of WBCs and bacteria (sensitivity, specificity, and other measures of accuracy) were pooled and examined by random-effects models. Nineteen studies containing 22,305 samples were included. Pooled sensitivities were 0.87 (95% confidence interval [CI], 0.86-0.89) for WBCs and 0.92 (95% CI, 0.91-0.93) for bacteria. Corresponding pooled specificities were 0.67 (95% CI, 0.66-0.68) for WBCs and 0.60 (95% CI, 0.59-0.61) for bacteria. Areas under the summary receiver operating characteristics curves were 0.87 and 0.93 for WBCs and bacteria, respectively. The major limitation of eligible studies was that enrolled subjects were often not representative of clinical patient populations in which UTI would be suspected. WBC and bacterial measurements by the UF-100 and UF-1000i are useful indicators in UTI screening; however, the performances of these systems should be rigorously evaluated by additional studies. Copyright © 2013 Elsevier B.V. All rights reserved.

Removal of Trace Pharmaceuticals from Water using coagulation and powdered activated carbon as pretreatment to ultrafiltration membrane system.

PubMed

Sheng, Chenguang; Nnanna, A G Agwu; Liu, Yanghe; Vargo, John D

2016-04-15

In this study, the efficacy of water treatment technologies: ultra-filtration (UF), powdered activated carbon (PAC), coagulation (COA) and a combination of these technologies (PAC/UF and COA/UF) to remove target pharmaceuticals (Acetaminophen, Bezafibrate, Caffeine, Carbamazepine, Cotinine, Diclofenac, Gemfibrozil, Ibuprofen, Metoprolol, Naproxen, Sulfadimethoxine, Sulfamethazine, Sulfamethoxazole, Sulfathiazole, Triclosan and Trimethoprim) was investigated. Samples of wastewater from municipal WWTPs were analyzed using direct aqueous injection High Performance Liquid Chromatography with Tandem Quadrupole Mass Spectrometric (LC/MS/MS) detection. On concentration basis, results showed an average removal efficiency of 29%, 50%, and 7%, respectively, for the UF, PAC dosage of 50ppm, and COA dosage of 10ppm. When PAC dosage of 100ppm was used as pretreatment to the combined PAC and UF in-line membrane system, a 90.3% removal efficiency was achieved. The removal efficiency of UF in tandem with COA was 33%, an increase of 4% compared with the single UF treatment. The adsorption effect of PAC combined with the physical separation process of UF revealed the best treatment strategy for removing pharmaceutical contaminant from water. Copyright © 2016 Elsevier B.V. All rights reserved.

Regression equations for estimating flood flows for the 2-, 10-, 25-, 50-, 100-, and 500-Year recurrence intervals in Connecticut

USGS Publications Warehouse

Ahearn, Elizabeth A.

2004-01-01

Multiple linear-regression equations were developed to estimate the magnitudes of floods in Connecticut for recurrence intervals ranging from 2 to 500 years. The equations can be used for nonurban, unregulated stream sites in Connecticut with drainage areas ranging from about 2 to 715 square miles. Flood-frequency data and hydrologic characteristics from 70 streamflow-gaging stations and the upstream drainage basins were used to develop the equations. The hydrologic characteristics?drainage area, mean basin elevation, and 24-hour rainfall?are used in the equations to estimate the magnitude of floods. Average standard errors of prediction for the equations are 31.8, 32.7, 34.4, 35.9, 37.6 and 45.0 percent for the 2-, 10-, 25-, 50-, 100-, and 500-year recurrence intervals, respectively. Simplified equations using only one hydrologic characteristic?drainage area?also were developed. The regression analysis is based on generalized least-squares regression techniques. Observed flows (log-Pearson Type III analysis of the annual maximum flows) from five streamflow-gaging stations in urban basins in Connecticut were compared to flows estimated from national three-parameter and seven-parameter urban regression equations. The comparison shows that the three- and seven- parameter equations used in conjunction with the new statewide equations generally provide reasonable estimates of flood flows for urban sites in Connecticut, although a national urban flood-frequency study indicated that the three-parameter equations significantly underestimated flood flows in many regions of the country. Verification of the accuracy of the three-parameter or seven-parameter national regression equations using new data from Connecticut stations was beyond the scope of this study. A technique for calculating flood flows at streamflow-gaging stations using a weighted average also is described. Two estimates of flood flows?one estimate based on the log-Pearson Type III analyses of the annual

Nine color eleven parameter immunophenotyping using three laser flow cytometry.

PubMed

Bigos, M; Baumgarth, N; Jager, G C; Herman, O C; Nozaki, T; Stovel, R T; Parks, D R; Herzenberg, L A

1999-05-01

This study describes a three laser flow cytometer, reagents, and software used to simultaneously evaluate nine distinct fluorescent parameters on one cell sample. We compare the quality of data obtained with (1) full software compensation and (2) the use of partial spectral compensation of selected pairs of parameters in analog hardware, in combination with final software compensation. An application characterizing low frequency murine B cell subpopulations is given. The fluorochromes used are: fluorescein (FITC), phycoerythrin (PE), Cy5PE and Cy7PE, excited at 488 nm by an argon laser; Texas Red (TR), allophycocyanin (APC), and Cy7APC excited at 595 nm by a pumped dye laser; and cascade blue (CB) and cascade yellow (CY) excited at 407 nm by a violet-enhanced krypton laser. Custom additions to commercial electronics and an extended optical bench allow the measurement of these nine parameters plus forward and side scatter light signals. We find the use of partial analog compensation reduces the variation in the background staining levels introduced by the compensation process. Novel B cell populations with frequencies below 1% are characterized. Nine color flow cytometry is capable of providing measurements with high information content. The choice of reagent-dye combinations and the ability to compensate in multi-parameter measurement space are crucial to obtaining satisfactory results.

Dose-Dependent Thresholds of 10-ns Electric Pulse Induced Plasma Membrane Disruption and Cytotoxicity in Multiple Cell Lines

DTIC Science & Technology

2011-01-01

normalized to parallel controls. Flow Cytometry and Confocal Microscopy Upon exposure to 10-ns EP, aliquots of the cellular suspension were added to a tube...Survival data was processed and plotted using GrapherH software (Golden Software, Golden, Colorado). Flow cytometry results were processed in C6 software...Accuri Cytometers, Inc., Ann Arbor, MI) and FCSExpress software (DeNovo Software, Los Angeles, CA). Final analysis and presentation of flow cytometry

Effect of perforation on flow past a conic cylinder at Re = 100: vortex-shedding pattern and force history

NASA Astrophysics Data System (ADS)

Lin, L. M.; Zhong, X. F.; Wu, Y. X.

2017-09-01

The flow past a circular-section cylinder with a conic shroud perforated with four holes at the peak was simulated numerically at Re=100 , considering two factors, viz. the angle of attack and the diameter of the holes. The effects of the perforated conic shroud on the vortex shedding pattern in the near wake was mainly investigated, as well as the time history of the drag and lift forces. In the investigated parameter space, three flow regimes were generally identified, corresponding to weak, moderate, and strong disturbance effects. In regime I, the wake can mainly be described by alternately shedding Kármán or Kármán-like vortices. In regime II, the spanwise vortices are obviously disturbed along the span due to the appearance of additional vorticity components and their interactions with the spanwise vortices, but still shed in synchronization along the spanwise direction. In regime III, the typical Kármán vortices partially or totally disappear, and some new vortex shedding patterns appear, such as Ω -type, obliquely shedding, and crossed spanwise vortices with opposite sign. Corresponding to these complex vortex shedding patterns in the near wake, the fluid forces no longer oscillate regularly at a single vortex shedding frequency, but rather with a lower modulation frequency and multiple amplitudes. An overview of these flow regimes is presented.

HoloMonitor M4: holographic imaging cytometer for real-time kinetic label-free live-cell analysis of adherent cells

NASA Astrophysics Data System (ADS)

Sebesta, Mikael; Egelberg, Peter J.; Langberg, Anders; Lindskov, Jens-Henrik; Alm, Kersti; Janicke, Birgit

2016-03-01

Live-cell imaging enables studying dynamic cellular processes that cannot be visualized in fixed-cell assays. An increasing number of scientists in academia and the pharmaceutical industry are choosing live-cell analysis over or in addition to traditional fixed-cell assays. We have developed a time-lapse label-free imaging cytometer HoloMonitorM4. HoloMonitor M4 assists researchers to overcome inherent disadvantages of fluorescent analysis, specifically effects of chemical labels or genetic modifications which can alter cellular behavior. Additionally, label-free analysis is simple and eliminates the costs associated with staining procedures. The underlying technology principle is based on digital off-axis holography. While multiple alternatives exist for this type of analysis, we prioritized our developments to achieve the following: a) All-inclusive system - hardware and sophisticated cytometric analysis software; b) Ease of use enabling utilization of instrumentation by expert- and entrylevel researchers alike; c) Validated quantitative assay end-points tracked over time such as optical path length shift, optical volume and multiple derived imaging parameters; d) Reliable digital autofocus; e) Robust long-term operation in the incubator environment; f) High throughput and walk-away capability; and finally g) Data management suitable for single- and multi-user networks. We provide examples of HoloMonitor applications of label-free cell viability measurements and monitoring of cell cycle phase distribution.

Applications of Flow Cytometry to Clinical Microbiology†

PubMed Central

Álvarez-Barrientos, Alberto; Arroyo, Javier; Cantón, Rafael; Nombela, César; Sánchez-Pérez, Miguel

2000-01-01

Classical microbiology techniques are relatively slow in comparison to other analytical techniques, in many cases due to the need to culture the microorganisms. Furthermore, classical approaches are difficult with unculturable microorganisms. More recently, the emergence of molecular biology techniques, particularly those on antibodies and nucleic acid probes combined with amplification techniques, has provided speediness and specificity to microbiological diagnosis. Flow cytometry (FCM) allows single- or multiple-microbe detection in clinical samples in an easy, reliable, and fast way. Microbes can be identified on the basis of their peculiar cytometric parameters or by means of certain fluorochromes that can be used either independently or bound to specific antibodies or oligonucleotides. FCM has permitted the development of quantitative procedures to assess antimicrobial susceptibility and drug cytotoxicity in a rapid, accurate, and highly reproducible way. Furthermore, this technique allows the monitoring of in vitro antimicrobial activity and of antimicrobial treatments ex vivo. The most outstanding contribution of FCM is the possibility of detecting the presence of heterogeneous populations with different responses to antimicrobial treatments. Despite these advantages, the application of FCM in clinical microbiology is not yet widespread, probably due to the lack of access to flow cytometers or the lack of knowledge about the potential of this technique. One of the goals of this review is to attempt to mitigate this latter circumstance. We are convinced that in the near future, the availability of commercial kits should increase the use of this technique in the clinical microbiology laboratory. PMID:10755996

LOW PRESSURE ULTRAVEIOLET STUDIES FOR INACTIVATION OF GIARDIA MURIS CYSTS

EPA Science Inventory

Cysts of Giardia muris were inactivated using a low pressure ultravolet (UV) light source. Cyst viability was detemined by both in vitro excystation and animal infectivity. Cyst doeses were counted using a flow cytometer for the animal infectivity experiments. Using in vitro excy...

LOW PRESSURE ULTRAVIOLET STUDIES FOR INACTIVATION OF GIARDIA MURIS CYSTS

EPA Science Inventory

Cysts of Giardia muris were inactivated using a low pressure ultravolet (UV) light source. Cyst viability was detemined by both in vitro excystation and animal infectivity. Cyst doeses were counted using a flow cytometer for the animal infectivity experiments. Using in vitro excy...

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.

Tangential flow ultrafiltration for detection of white spot syndrome virus (WSSV) in shrimp pond water.

PubMed

Alavandi, S V; Ananda Bharathi, R; Satheesh Kumar, S; Dineshkumar, N; Saravanakumar, C; Joseph Sahaya Rajan, J

2015-06-15

Water represents the most important component in the white spot syndrome virus (WSSV) transmission pathway in aquaculture, yet there is very little information. Detection of viruses in water is a challenge, since their counts will often be too low to be detected by available methods such as polymerase chain reaction (PCR). In order to overcome this difficulty, viruses in water have to be concentrated from large volumes of water prior to detection. In this study, a total of 19 water samples from aquaculture ecosystem comprising 3 creeks, 10 shrimp culture ponds, 3 shrimp broodstock tanks and 2 larval rearing tanks of shrimp hatcheries and a sample from a hatchery effluent treatment tank were subjected to concentration of viruses by ultrafiltration (UF) using tangential flow filtration (TFF). Twenty to 100l of water from these sources was concentrated to a final volume of 100mL (200-1000 fold). The efficiency of recovery of WSSV by TFF ranged from 7.5 to 89.61%. WSSV could be successfully detected by PCR in the viral concentrates obtained from water samples of three shrimp culture ponds, one each of the shrimp broodstock tank, larval rearing tank, and the shrimp hatchery effluent treatment tank with WSSV copy numbers ranging from 6 to 157mL(-1) by quantitative real time PCR. The ultrafiltration virus concentration technique enables efficient detection of shrimp viral pathogens in water from aquaculture facilities. It could be used as an important tool to understand the efficacy of biosecurity protocols adopted in the aquaculture facility and to carry out epidemiological investigations of aquatic viral pathogens. Copyright © 2015 Elsevier B.V. All rights reserved.

Utility of light scatter in the morphological analysis of sperm

EPA Science Inventory

We were able to differentiate the morphologically diverse sperm nuclei of four animal species by using an Ortho flow cytometer to detect the forward light scatter from a red (helium-neon) laser. Cytograms depicting the axial light loss and forward red scatter signals revealed uni...

An Investigation of Coccolithophore Optical Properties under Bloom Conditions: A Continuation

DTIC Science & Technology

1990-01-01

particles are produced annually throughout much of the temperate oceans of the world; mes-scale vatches of the coccolithophore, Emiliania huxleyi have been...cruise, our experiments with the EPICS flow cytometer at Biaelow Laboratory demonstrated that the optical properties of the coccolithophore, E. huxleyi

Analysis of fluid fuel flow to the neutron kinetics on molten salt reactor FUJI-12

NASA Astrophysics Data System (ADS)

Aji, Indarta Kuncoro; Waris, Abdul; Permana, Sidik

2015-09-01

Molten Salt Reactor is a reactor are operating with molten salt fuel flowing. This condition interpret that the neutron kinetics of this reactor is affected by the flow rate of the fuel. This research analyze effect by the alteration velocity of the fuel by MSR type Fuji-12, with fuel composition LiF-BeF2-ThF4-233UF4 respectively 71.78%-16%-11.86%-0.36%. Calculation process in this study is performed numerically by SOR and finite difference method use C programming language. Data of reactivity, neutron flux, and the macroscopic fission cross section for calculation process obtain from SRAC-CITATION (Standard thermal Reactor Analysis Code) and JENDL-4.0 data library. SRAC system designed and developed by JAEA (Japan Atomic Energy Agency). This study aims to observe the effect of the velocity of fuel salt to the power generated from neutron precursors at fourth year of reactor operate (last critical condition) with number of multiplication effective; 1.0155.

Evaluation of the treatability of a winery distillery (vinasse) wastewater by UASB, anoxic-aerobic UF-MBR and chemical precipitation/adsorption.

PubMed

Petta, Luigi; De Gisi, Sabino; Casella, Patrizia; Farina, Roberto; Notarnicola, Michele

2017-10-01

A multi-stage pilot-scale treatment cycle consisting of an Upflow Anaerobic Sludge Blanket reactor (UASB) followed by an anoxic-aerobic Ultra Filtration Membrane Bio Reactor (UF-MBR) and a post treatment based on chemical precipitation with lime or adsorption on Granular Activated Carbons (GAC), was applied in order to evaluate the treatment feasibility of a real winery distillery wastewater at laboratory and bench scale. The wastewater was classified as high strength with acidic pH (3.8), and concentrations of 44,600, 254, 604 and 660 mg/l for COD tot , total nitrogen, total phosphorous and phenols, respectively. The UASB reactor was operated at Organic Loading Rates (OLR) in the range 3.0-11.5 kgCOD tot /m 3 /d achieving treatment efficiency up to 97%, with an observed methane production of 340 L of CH 4 /kgCOD. The MBR system was operated with an organic load in the range 0.070-0.185 kgCOD/kgVSS/d, achieving a removal up to 48%, 67% and 65% of the influent COD, total nitrogen and phenols, respectively. The combination of UASB and UF-MBR treatment units was not effective in phosphate and colour removal assigning to further chemical precipitation and adsorption processes, respectively, their complete removal in order to comply with legal standards for wastewater discharge. Subsequently, the optimization of the investigated treatment chain was assessed by applying a chemical precipitation step upstream and downstream the UASB reactor, and a related treatment unit cost assessment is presented in view of a further technological scale-up. Copyright © 2017 Elsevier Ltd. All rights reserved.

High concentration biotherapeutic formulation and ultrafiltration: Part 1 pressure limits.

PubMed

Lutz, Herb; Arias, Joshua; Zou, Yu

2017-01-01

High therapeutic dosage requirements and the desire for ease of administration drive the trend to subcutaneous administration using delivery systems such as subcutaneous pumps and prefilled syringes. Because of dosage volume limits, prefilled syringe administration requires higher concentration liquid formulations, limited to about 30 cP or roughly 100-300 g L -1 for mAb's. Ultrafiltration (UF) processes are routinely used to formulate biological therapeutics. This article considers pressure constraints on the UF process that may limit its ability to achieve high final product concentrations. A system hardware analysis shows that the ultrafiltration cassette pressure drop is the major factor limiting UF systems. Additional system design recommendations are also provided. The design and performance of a new cassette with a lower feed channel flow resistance is described along with 3D modeling of feed channel pressure drop. The implications of variations in cassette flow channel resistance for scaling up and setting specifications are considered. A recommendation for a maximum pressure specification is provided. A review of viscosity data and theory shows that molecular engineering, temperature, and the use of viscosity modifying excipients including pH adjustment can be used to achieve higher concentrations. The combined use of a low pressure drop cassette with excipients further increased final concentrations by 35%. Guidance is provided on system operation to control hydraulics during final concentration. These recommendations should allow one to design and operate systems to routinely achieve the 30 cP target final viscosity capable of delivery using a pre-filled syringe. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:113-124, 2017. © 2016 American Institute of Chemical Engineers.

Ultrafast Microfluidic Cellular Imaging by Optical Time-Stretch.

PubMed

Lau, Andy K S; Wong, Terence T W; Shum, Ho Cheung; Wong, Kenneth K Y; Tsia, Kevin K

2016-01-01

There is an unmet need in biomedicine for measuring a multitude of parameters of individual cells (i.e., high content) in a large population efficiently (i.e., high throughput). This is particularly driven by the emerging interest in bringing Big-Data analysis into this arena, encompassing pathology, drug discovery, rare cancer cell detection, emulsion microdroplet assays, to name a few. This momentum is particularly evident in recent advancements in flow cytometry. They include scaling of the number of measurable colors from the labeled cells and incorporation of imaging capability to access the morphological information of the cells. However, an unspoken predicament appears in the current technologies: higher content comes at the expense of lower throughput, and vice versa. For example, accessing additional spatial information of individual cells, imaging flow cytometers only achieve an imaging throughput ~1000 cells/s, orders of magnitude slower than the non-imaging flow cytometers. In this chapter, we introduce an entirely new imaging platform, namely optical time-stretch microscopy, for ultrahigh speed and high contrast label-free single-cell (in a ultrafast microfluidic flow up to 10 m/s) imaging and analysis with an ultra-fast imaging line-scan rate as high as tens of MHz. Based on this technique, not only morphological information of the individual cells can be obtained in an ultrafast manner, quantitative evaluation of cellular information (e.g., cell volume, mass, refractive index, stiffness, membrane tension) at nanometer scale based on the optical phase is also possible. The technology can also be integrated with conventional fluorescence measurements widely adopted in the non-imaging flow cytometers. Therefore, these two combinatorial and complementary measurement capabilities in long run is an attractive platform for addressing the pressing need for expanding the "parameter space" in high-throughput single-cell analysis. This chapter provides the

Femtosecond laser fabrication of fiber based optofluidic platform for flow cytometry applications

NASA Astrophysics Data System (ADS)

Serhatlioglu, Murat; Elbuken, Caglar; Ortac, Bulend; Solmaz, Mehmet E.

2017-02-01

Miniaturized optofluidic platforms play an important role in bio-analysis, detection and diagnostic applications. The advantages of such miniaturized devices are extremely low sample requirement, low cost development and rapid analysis capabilities. Fused silica is advantageous for optofluidic systems due to properties such as being chemically inert, mechanically stable, and optically transparent to a wide spectrum of light. As a three dimensional manufacturing method, femtosecond laser scanning followed by chemical etching shows great potential to fabricate glass based optofluidic chips. In this study, we demonstrate fabrication of all-fiber based, optofluidic flow cytometer in fused silica glass by femtosecond laser machining. 3D particle focusing was achieved through a straightforward planar chip design with two separately fabricated fused silica glass slides thermally bonded together. Bioparticles in a fluid stream encounter with optical interrogation region specifically designed to allocate 405nm single mode fiber laser source and two multi-mode collection fibers for forward scattering (FSC) and side scattering (SSC) signals detection. Detected signal data collected with oscilloscope and post processed with MATLAB script file. We were able to count number of events over 4000events/sec, and achieve size distribution for 5.95μm monodisperse polystyrene beads using FSC and SSC signals. Our platform shows promise for optical and fluidic miniaturization of flow cytometry systems.

Ultrafiltration of thin stillage from conventional and e-mill dry grind processes.

PubMed

Arora, Amit; Dien, Bruce S; Belyea, Ronald L; Wang, Ping; Singh, Vijay; Tumbleson, M E; Rausch, Kent D

2011-05-01

We used ultrafiltration (UF) to evaluate membrane filtration characteristics of thin stillage and determine solids and nutrient compositions of filtered streams. To obtain thin stillage, corn was fermented using laboratory methods. UF experiments were conducted in batch mode under constant temperature and flow rate conditions. Two regenerated cellulose membranes (10 and 100 kDa molecular weight cutoffs) were evaluated with the objective of retaining solids as well as maximizing permeate flux. Optimum pressures for 10 and 100 kDa membranes were 207 and 69 kPa, respectively. Total solids, ash, and neutral detergent fiber contents of input TS streams of dry grind and E-Mill processes were similar; however, fat and protein contents were different (p < 0.05). Retentate obtained from conventional thin stillage fractionation had higher mean total solids contents (27.6% to 27.8%) compared to E-Mill (22.2% to 23.4%). Total solids in retentate streams were found similar to those from commercial evaporators used in industry (25% to 35% total solids). Fat contents of retentate streams ranged from 16.3% to 17.5% for the conventional process. A 2% increment in fat concentration was observed in the E-Mill retentate stream. Thin stillage ash content was reduced 60% in retentate streams.

Research on the experiment of reservoir water treatment applying ultrafiltration membrane technology of different processes.

PubMed

Zhang, Liyong; Zhang, Penghui; Wang, Meng; Yang, Kai; Liu, Junliang

2016-09-01

The processes and effects of coagulation-ultrafiltration (C-UF) and coagulation sedimentation-ultrafiltration (CS-UF) process used in the treatment of Dalangdian Reservoir water were compared. The experiment data indicated that 99% of turbidity removal and basically 100% of microorganism and algae removal were achieved in both C-UF and CS-UF process. The organic removal effect of CS-UF? process was slightly better than C-UF process. However, the organic removal effect under different processes was not obvious due to limitation of ultrafiltration membrane aperture. Polyaluminium chloride was taken as a coagulant in water plant. The aluminum ion removal result revealed that coagulant dosage was effectively saved by using membrane technology during megathermal high algae laden period. Within the range of certain reagent concentration and soaking time, air-water backwashing of every filtration cycle of membrane was conducted to effectively reduce membrane pollution. Besides, maintenance cleaning was conducted every 60 min. whether or not restorative cleaning was conducted depends on the pollution extent. After cleaning, recovery of membrane filtration effect was obvious.

Near-ultraviolet laser diodes for brilliant ultraviolet fluorophore excitation.

PubMed

Telford, William G

2015-12-01

Although multiple lasers are now standard equipment on most modern flow cytometers, ultraviolet (UV) lasers (325-365 nm) remain an uncommon excitation source for cytometry. Nd:YVO4 frequency-tripled diode pumped solid-state lasers emitting at 355 nm are now the primary means of providing UV excitation on multilaser flow cytometers. Although a number of UV excited fluorochromes are available for flow cytometry, the cost of solid-state UV lasers remains prohibitively high, limiting their use to all but the most sophisticated multilaser instruments. The recent introduction of the brilliant ultraviolet (BUV) series of fluorochromes for cell surface marker detection and their importance in increasing the number of simultaneous parameters for high-dimensional analysis has increased the urgency of including UV sources in cytometer designs; however, these lasers remain expensive. Near-UV laser diodes (NUVLDs), a direct diode laser source emitting in the 370-380 nm range, have been previously validated for flow cytometric analysis of most UV-excited probes, including quantum nanocrystals, the Hoechst dyes, and 4',6-diamidino-2-phenylindole. However, they remain a little-used laser source for cytometry, despite their significantly lower cost. In this study, the ability of NUVLDs to excite the BUV dyes was assessed, along with their compatibility with simultaneous brilliant violet (BV) labeling. A NUVLD emitting at 375 nm was found to excite most of the available BUV dyes at least as well as a UV 355 nm source. This slightly longer wavelength did produce some unwanted excitation of BV dyes, but at sufficiently low levels to require minimal additional compensation. NUVLDs are compact, relatively inexpensive lasers that have higher power levels than the newest generation of small 355 nm lasers. They can, therefore, make a useful, cost-effective substitute for traditional UV lasers in multicolor analysis involving the BUV and BV dyes. Published 2015 Wiley Periodicals Inc. on

Design of an Autonomous Underwater Vehicle (AUV) Charging System for Underway, Underwater Recharging

DTIC Science & Technology

2014-05-09

again increase the size of the system. A comparison between switching frequency and efficiency for a nominal DC/DC converter was done in an EE ...Choosing the Optimum Switching Frequency of your DC / DC Converter,” EE Times, pp. 1–7, 2006. [19] ON Semiconductors, “Effects of High Switching Frequency...3.1W OUTPUT FILTER CAPACITOR EEE -FC1H101P 100uF ELECTROLYTIC 50V OUTPUT FILTER CAPACITOR C5750X7S2A106M230KB 10uF CERAMIC 100V

Evaluation of commercial ultrafiltration systems for treating automotive oily wastewater

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kim, B.R.; Kalis, E.M.; Florkey, D.L.

1998-11-01

Currently at Ford Motor Company, oily wastewater is batch treated by chemical demulsification whose performance depends on determining optimum chemical dosages and is occasionally inconsistent because of influent fluctuations. Therefore, a pilot study was conducted at the Ford Romeo Engine Plant, Romeo, Michigan, to study treatment of raw oily wastewater and skim oil (from chemical deemulsification) using commercially available ultrafiltration (UF) systems as an alternative to chemical deemulsification. The study found that most UF membranes performed consistently and reliably, producing average permeate oil and grease (O and G) concentrations of less than 100 mg/L, a typical discharge limit for anmore » automotive plant. In addition, tubular membranes typically outperformed spiral-wound membranes in permeate flux and washing frequency. While all UF systems performed consistently well for removing O and G, the treated effluent still had a chemical oxygen demand (COD) of 100 to 2,000 mg/L, which is comparable to that found in typical chemically treated wastewater. This indicates that many dissolved organics are not removed by either chemical or UF treatment. Metals (such as copper and zinc) were found to be effectively removed by UF when the pH was greater than 8. Most membranes used as a second stage produced retentate with O and G of more than 40%. All attempts at UF skim oil treatment were unsuccessful because of high oil viscosity, which made pumping it through a membrane system almost impossible. Chemical reactions during the chemical deemulsification process might have been responsible for the high viscosity.« less

Intergranular tellurium cracking of nickel-based alloys in molten Li, Be, Th, U/F salt mixture

NASA Astrophysics Data System (ADS)

Ignatiev, Victor; Surenkov, Alexander; Gnidoy, Ivan; Kulakov, Alexander; Uglov, Vadim; Vasiliev, Alexander; Presniakov, Mikhail

2013-09-01

In Russia, R&D on Molten Salt Reactor (MSR) are concentrated now on fast/intermediate spectrum concepts which were recognized as long term alternative to solid fueled fast reactors due to their attractive features: strong negative feedback coefficients, easy in-service inspection, and simplified fuel cycle. For high-temperature MSR corrosion of the metallic container alloy in primary circuit is the primary concern. Key problem receiving current attention include surface fissures in Ni-based alloys probably arising from fission product tellurium attack. This paper summarizes results of corrosion tests conducted recently to study effect of oxidation state in selected fuel salt on tellurium attack and to develop means of controlling tellurium cracking in the special Ni-based alloys recently developed for molten salt actinide recycler and tranforming (MOSART) system. Tellurium corrosion of Ni-based alloys was tested at temperatures up to 750 °C in stressed and unloaded conditions in molten LiF-BeF2 salt mixture fueled by about 20 mol% of ThF4 and 2 mol% of UF4 at different [U(IV)]/[U(III)] ratios: 0.7, 4, 20, 100 and 500. Following Ni-based alloys (in mass%): HN80М-VI (Mo—12, Cr—7.6, Nb—1.5), HN80МТY (Mo—13, Cr—6.8, Al—1.1, Ti—0.9), HN80МТW (Mo—9.4, Cr—7.0, Ti—1.7, W—5.5) and ЕМ-721 (W—25.2, Cr—5.7, Ti—0.17) were used for the study in the corrosion facility. If the redox state the fuel salt is characterized by uranium ratio [U(IV)]/[U(III)] < 1 the alloys' specimens get a more negative stationary electrode potential than equilibrium electrode potentials of some uranium intermetallic compounds and alloys with nickel and molybdenum. This leads to spontaneous behavior of alloy formation processes on the specimens' surface and further diffusion of uranium deep into the metallic phase. As consequence of this films of intermetallic compounds and alloys of nickel, molybdenum, tungsten with uranium are formed on the alloys specimens' surface

Performance of an ultrafiltration membrane bioreactor (UF-MBR) as a processing strategy for the synthesis of galacto-oligosaccharides at high substrate concentrations.

PubMed

Córdova, Andrés; Astudillo, Carolina; Vera, Carlos; Guerrero, Cecilia; Illanes, Andrés

2016-04-10

The performance of an ultrafiltration membrane bioreactor for galacto-oligosaccharides (GOS) synthesis using high lactose concentrations (470 g/L) and β-galactosidase from Aspergillus oryzae was assessed. Tested processing variables were: transmembrane-pressure (PT), crossflow-velocity (CFV) and temperature. Results showed that processing variables had significant effect on the yield, the enzyme productivity and the flux but did not on GOS concentration and reaction conversion obtained. As expected, the use of high turbulences improved mass transfer and reduced the membrane fouling, but the use of very high crossflow-velocities caused operational instability due to vortex formation and lactose precipitation. The use of a desirability function allowed determining optimal conditions which were: PT (4.38 bar), CFV (7.35 m/s) and temperature (53.1 °C), optimizing simultaneously flux and specific enzyme productivity Under these optimal processing conditions, shear-stress and temperature did not affect the enzyme but long-term operation was limited by flux decay. In comparison to a conventional batch system, at 12.5h of processing time, the continuous GOS synthesis in the UF-MBR increased significantly the amount of processed substrate and a 2.44-fold increase in the amount of GOS produced per unit mass of catalyst was obtained with respect to a conventional batch system. Furthermore, these results can be improved by far by tuning the membranearea/reactionvolume ratio, showing that the use of an UF-MBR is an attractive alternative for the GOS synthesis at very high lactose concentrations. Copyright © 2016 Elsevier B.V. All rights reserved.

Biological and Physical Forcing of Picophytoplankton on the New England Shelf: New Insights From Submersible Flow Cytometry at the Martha's Vineyard Coastal Observatory

NASA Astrophysics Data System (ADS)

Sosik, H. M.; Lentz, S. J.; Olson, R. J.

2008-12-01

Phytoplankton communities of temperate continental shelf systems typically exhibit dramatic seasonal variations. We are taking advantage of the Martha's Vineyard Coastal Observatory (MVCO), a cabled facility on the New England inner shelf, to better understand the physical and biological processes that interact to produce this variability. Our approach depends on high resolution (~hourly) multi-year time series of taxonomically resolved phytoplankton acquired with FlowCytobot and Imaging FlowCytobot, custom-built automated submersible flow cytometers optimized for measurement of picophytoplankton and microphytoplankton, respectively. These observational capabilities enable new approaches to understanding classical problems in plankton ecology. Our time series document a seasonal shift from microphytoplankton (especially large, often chain-forming diatoms) in fall and winter to pico- and small nanophytoplankton in spring and summer. The shift is associated not only with a decline in large cell abundance but also with a dramatic increase (100- to 1000- fold) in small cells. This seasonal pattern reflects a regional-scale phenomenon that emerges despite affects of advection and spatial patchiness. To begin quantifying the factors that produce these seasonal patterns, we have focused first on the cyanobacterial genus, Synechococcus, the numerically dominant picoplankter in this coastal system. In contrast to prevailing views in picoplankton ecology, the annual Synechococcus bloom in this system suggests a persistent decoupling from grazer control in spring. Synechococcus population growth rates, determined from diel changes in cell size distributions, show strong seasonal variability associated with physiological consequences of environmental conditions. Low growth rates are linked to temperature limitation in winter/early spring and a combination of temperature and light limitation in fall associated with decreasing solar angles, seasonal cooling, and breakdown in

Quantifying particulate matter deposition in Niwot Ridge, Colorado: Collection of dry deposition using marble inserts and particle imaging using the FlowCAM

NASA Astrophysics Data System (ADS)

Goss, Natasha R.; Mladenov, Natalie; Seibold, Christine M.; Chowanski, Kurt; Seitz, Leslie; Wellemeyer, T. Barret; Williams, Mark W.

2013-12-01

Atmospheric wet and dry deposition are important sources of carbon for remote alpine lakes and soils. The carbon inputs from dry deposition in alpine National Atmospheric Deposition Program (NADP) collectors, including aeolian dust and biological material, are not well constrained due to difficulties in retaining particulate matter in the collectors. Here, we developed and tested a marble insert for dry deposition collection at the Niwot Ridge Long Term Ecological Research Station (NWT LTER) Soddie site (3345 m) between 24 May and 8 November 2011. We conducted laboratory tests of the insert's effect on particulate matter (PM) mass and non-purgeable organic carbon (DOC) and found that the insert did not significantly change either measurement. Thus, the insert may enable dry deposition collection of PM and DOC at NADP sites. We then developed a method for enumerating the collected wet and dry deposition with the Flow Cytometer and Microscope (FlowCAM), a dynamic-image particle analysis tool. The FlowCAM has the potential to establish morphology, which affects particle settling and retention, through particle diameter and aspect ratio. Particle images were used to track the abundance of pollen grains over time. Qualitative image examination revealed that most particles were biological in nature, such as intact algal cells and pollen. Dry deposition loading to the Soddie site as determined by FlowCAM measurements was highly variable, ranging from 100 to >230 g ha-1 d-1 in June-August 2011 and peaking in late June. No significant difference in diameter or aspect ratio was found between wet and dry deposition, suggesting fundamental similarities between those deposition types. Although FlowCAM statistics and identification of particle types proved insightful, our total-particle enumeration method had a high variance and underestimated the total number of particles when compared to imaging of relatively large volumes (60-125 mL) from a single sample. We recommend use of

Pore level visualization of foam flow in a silicon micromodel. SUPRI TR 100

DOE Office of Scientific and Technical Information (OSTI.GOV)

Woody, F.; Blunt, M.; Castanier, L.

This paper is concerned with the behavior of foam in porous media at the pore level. Identical, heterogeneous silicon micromodels, two dimensionally etched to replicate flow in Berea Sandstone, were used. The models, already saturated with varying concentrations of surfactant and, at times, oil were invaded with air. Visual observations were made of these air displacement events in an effort to determine foam flow characteristics with varying surfactant concentrations, and differing surfactants in the presence of oil. These displacement events were recorded on video tape. These tapes are available at the Stanford University Petroleum Research Institute, Stanford, California. The observedmore » air flow characteristics can be broadly classified into two: continuous and discontinuous. Continuous air flow was observed in two phase runs when the micromodel contained no aqueous surfactant solution. Air followed a tortuous path to the outlet, splitting and reconnecting around grains, isolating water located in dead-end or circumvented pores, all without breaking and forming bubbles. No foam was created. Discontinuous air flow occurred in runs containing surfactant - with smaller bubble sizes appearing with higher surfactant concentrations. Air moved through the medium by way of modified bubble train flow where bubbles travel through pore throats and tend to reside more statically in larger pore bodies until enough force is applied to move them along. The lamellae were stable, and breaking and reforming events by liquid drainage and corner flow were observed in higher surfactant concentrations. However, the classic snap-off process, as described by Roof (1973) was not seen at all.« less

Influence of arsenic flow on the crystal structure of epitaxial GaAs grown at low temperatures on GaAs (100) and (111)A substrates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Galiev, G. B.; Klimov, E. A.; Vasiliev, A. L.

The influence of arsenic flow in a growth chamber on the crystal structure of GaAs grown by molecular-beam epitaxy at a temperature of 240°C on GaAs (100) and (111)A substrates has been investigated. The flow ratio γ of arsenic As4 and gallium was varied in the range from 16 to 50. GaAs films were either undoped, or homogeneously doped with silicon, or contained three equidistantly spaced silicon δ-layers. The structural quality of the annealed samples has been investigated by transmission electron microscopy. It is established for the first time that silicon δ-layers in “low-temperature” GaAs serve as formation centers ofmore » arsenic precipitates. Their average size, concentration, and spatial distribution are estimated. The dependence of the film structural quality on γ is analyzed. Regions 100–150 nm in size have been revealed in some samples and identified (by X-ray microanalysis) as pores. It is found that, in the entire range of γ under consideration, GaAs films on (111)A substrates have a poorer structural quality and become polycrystalline beginning with a thickness of 150–200 nm.« less

Interference of blood leucocytes in the measurements of immature red cells (reticulocytes) by two different (semi-) automated flow-cytometry technologies.

PubMed

Villamor, N; Kirsch, A; Huhn, D; Vives-Corrons, J L; Serke, S

1996-06-01

Flow cytometrical methods have been introduced recently as an alternative to the enumeration of reticulocytes by microscopy. Two of these methods have gained widespread use in haematological practice; the multiparametric flow cytometer using thiazole orange staining (Retic-Count, FACScan) and the single-application reticulocyte counter using auramine-O staining (R-series, Sysmex). Several studies have emphasized the excellent correlations between microscopy and these techniques. The purpose of our study has been to examine the specificity of these automated devices with regard to cells classified as 'reticulocytes' and the effect that this may have on measures of reticulocyte maturity. Our results indicate that the specificity of reticulocyte measurements by both the Sysmex R-1000/-3000 and the Retic-Count system is relatively low. This is due to the presence of leucocytes amongst cells classified as reticulocytes. These leucocytes display intense staining with either dye, leading to an erroneous estimation of RMI (thiazole orange) and high fluorescence count (R-1000/-3000). This error is directly correlated with the leucocyte count. The basis for reticulocyte identification should be improved before automated estimation of reticulocyte maturation can be used in clinical practice.

Protein crystallization in a 100 nl solution with new stirring equipment

PubMed Central

Maki, S.; Murai, R.; Yoshikawa, H. Y.; Kitatani, T.; Nakata, S.; Kawahara, H.; Hasenaka, H.; Kobayashi, A.; Okada, S.; Sugiyama, S.; Adachi, H.; Matsumura, H.; Takano, K.; Murakami, S.; Inoue, T.; Sasaki, T.; Mori, Y.

2008-01-01

To investigate quantitatively the effects of stirring on protein crystallization, a new stirring system which can agitate a protein solution, ∼100 nl, by providing Hagen–Poiseuille flow has been successfully developed. In addition, this new stirring system provides flow with a well defined pattern and velocity. Using this system, hen egg-white lysozyme was crystallized in 100–200 nl solutions while being stirred. The optimum stirring conditions for lysozyme crystals have been explored by evaluating the Reynolds (Re) number and the crystals obtained. Intermittent flow, as well as a low Re number, was found to contribute significantly to the growth of a smaller number of larger crystals. PMID:18421156

Nerve-sparing technique and urinary control after robot-assisted laparoscopic prostatectomy.

PubMed

Choi, Wesley W; Freire, Marcos P; Soukup, Jane R; Yin, Lei; Lipsitz, Stuart R; Carvas, Fernando; Williams, Stephen B; Hu, Jim C

2011-02-01

To characterize determinants of 4-, 12-, and 24-month urinary control after robot-assisted laparoscopic prostatectomy (RALP). Adjusted comparative study using prospectively collected, patient self-reported urinary control for 602 consecutive RALPs. Urinary control defined as: (1) EPIC urinary function (UF) scored from 0 to 100 and (2) continence (zero pads per day). Both UF (62.8 vs. 42.4, P<0.001) and continence rates (47.2 vs. 26.7%, P=0.043) were better for bilateral nerve-sparing (BNS) vs. non-nerve-sparing (NNS) at 4 months, but only UF scores were significantly better at 12- (80.9 vs. 70.7, P=0.014) and 24-month (89.2 vs. 77.4, P=0.024) post-RALP. No difference in positive margin rates was observed. In multivariate analysis, older age (parameter estimate -0.42, 95% CI -0.80 to -0.04) and increasing gland volume (-0.13, CI -0.26 to -0.01) resulted in lower UF scores at 4 months, while higher pre-operative UF (0.25, CI 0.05-0.46), bladder neck-sparing technique (10.1, CI 3.79-16.35), BNS (19.1, CI 9.37-28.82), and unilateral nerve-sparing (19.00, CI 7.88-30.11) resulted in higher UF scores at 4 months. At 12 months, higher pre-operative UF (0.24, CI 0.083-0.40) and BNS (9.54, CI 1.92-17.16) resulted in higher UF scores. At 24 months, higher pre-operative UF (0.20, CI 0.06-0.33), bladder neck-sparing technique (7.80, CI 3.48-12.10), and BNS (7.86, CI 1.04-14.68) resulted in higher UF scores. BNS, bladder neck-sparing technique, and higher pre-operative UF score result in improved 24-month urinary control after RALP.

Hybrid MF and membrane bioreactor process applied towards water and indigo reuse from denim textile wastewater.

PubMed

Couto, Carolina Fonseca; Marques, Larissa Silva; Balmant, Janine; de Oliveira Maia, Andreza Penido; Moravia, Wagner Guadagnin; Santos Amaral, Miriam Cristina

2018-03-01

This work investigates the application of a microfiltration (MF)-membrane bioreactor (MBR) hybrid process for textile dyeing process wastewater reclamation. The indigo blue dye was efficiently retained by the MF membrane (100%), which allows its recovery from the concentrate stream. MF promotes 100% of colour removal, and reduces the chemical oxygen demand (COD) and conductivity by about 65% and 25%, respectively, and improves the wastewater biodegradability. MF flux decline was mostly attributed to concentration polarization and the chemical cleaning was efficient enough to recover initial hydraulic resistance. The MBR provides to be a stable process maintaining its COD and ammonia removal efficiency (73% and 100%, respectively) mostly constant throughout and producing a permeate that meets the reuse criteria for some industry activities, such as washing-off and equipment washdown. The use of an MF or ultrafiltration (UF) membrane in the MBR does not impact the MBR performance in terms of COD removal. Although the membrane of MBR-UF shows permeability lower than MBR-MF membrane, the UF membrane contributes to a more stable operation in terms of permeability.

Examination of a high resolution laser optical plankton counter and FlowCAM for measuring plankton concentration and size

NASA Astrophysics Data System (ADS)

Kydd, Jocelyn; Rajakaruna, Harshana; Briski, Elizabeta; Bailey, Sarah

2018-03-01

Many commercial ships will soon begin to use treatment systems to manage their ballast water and reduce the global transfer of harmful aquatic organisms and pathogens in accordance with upcoming International Maritime Organization regulations. As a result, rapid and accurate automated methods will be needed to monitoring compliance of ships' ballast water. We examined two automated particle counters for monitoring organisms ≥ 50 μm in minimum dimension: a High Resolution Laser Optical Plankton Counter (HR-LOPC), and a Flow Cytometer with digital imaging Microscope (FlowCAM), in comparison to traditional (manual) microscopy considering plankton concentration, size frequency distributions and particle size measurements. The automated tools tended to underestimate particle concentration compared to standard microscopy, but gave similar results in terms of relative abundance of individual taxa. For most taxa, particle size measurements generated by FlowCAM ABD (Area Based Diameter) were more similar to microscope measurements than were those by FlowCAM ESD (Equivalent Spherical Diameter), though there was a mismatch in size estimates for some organisms between the FlowCAM ABD and microscope due to orientation and complex morphology. When a single problematic taxon is very abundant, the resulting size frequency distribution curves can become skewed, as was observed with Asterionella in this study. In particular, special consideration is needed when utilizing automated tools to analyse samples containing colonial species. Re-analysis of the size frequency distributions with the removal of Asterionella from FlowCAM and microscope data resulted in more similar curves across methods with FlowCAM ABD having the best fit compared to the microscope, although microscope concentration estimates were still significantly higher than estimates from the other methods. The results of our study indicate that both automated tools can generate frequency distributions of particles

The use of fluorescent indoline dyes for side population analysis.

PubMed

Kohara, Hiroshi; Watanabe, Kohei; Shintou, Taichi; Nomoto, Tsuyoshi; Okano, Mie; Shirai, Tomoaki; Miyazaki, Takeshi; Tabata, Yasuhiko

2013-01-01

Dye efflux assay evaluated by flow cytometry is useful for stem cell studies. The side population (SP) cells, characterized by the capacity to efflux Hoechst 33342 dye, have been shown to be enriched for hematopoietic stem cells (HSCs) in bone marrow. In addition, SP cells are isolated from various tissues and cell lines, and are also potential candidates for cancer stem cells. However, ultra violet (UV) light, which is not common for every flow cytometer, is required to excite Hoechst 33342. Here we showed that a fluorescent indoline dye ZMB793 can be excited by 488-nm laser, equipped in almost all the modern flow cytometers, and ZMB793-excluding cells showed SP phenotype. HSCs were exclusively enriched in the ZMB793-excluding cells, while ZMB793 was localized in cytosol of bone marrow lineage cells. The efflux of ZMB793 dye was mediated by ATP binding cassette (ABC) transporter Abcg2. Moreover, staining properties were affected by the side-chain structure of the dyes. These data indicate that the fluorescent dye ZMB793 could be used for the SP cell analysis. Copyright © 2012 Elsevier Ltd. All rights reserved.

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

Dynamics of lava flow - Thickness growth characteristics of steady two-dimensional flow

NASA Technical Reports Server (NTRS)

Park, S.; Iversen, J. D.

1984-01-01

The thickness growth characteristics of flowing lava are investigated using a heat balance model and a two-dimensional model for flow of a Bingham plastic fluid down an inclined plane. It is found that yield strength plays a crucial role in the thickening of a lava flow of given flow rate. To illustrate this point, downstream thickness profiles and yield strength distributions were calculated for flows with mass flow rates of 10,000 and 100,000 kg/m-sec. Higher flow rates led to slow cooling rates which resulted in slow rate of increase of yield strength and thus greater flow lengths.

19 F(α,n) thick target yield from 3.5 to 10.0 MeV

DOE PAGES

Norman, E.B.; Chupp, T.E.; Lesko, K.T.; ...

2015-09-01

Using a target of PbF2, the thick-target yield from the 19F(α,n) reaction was measured from Eα=3.5–10 MeV. From these results, we infer the thick-target neutron yields from targets of F2 and UF6 over this same alpha-particle energy range.

Flow Cytometric Quantification of Peripheral Blood Cell β-Adrenergic Receptor Density and Urinary Endothelial Cell-Derived Microparticles in Pulmonary Arterial Hypertension.

PubMed

Rose, Jonathan A; Wanner, Nicholas; Cheong, Hoi I; Queisser, Kimberly; Barrett, Patrick; Park, Margaret; Hite, Corrine; Naga Prasad, Sathyamangla V; Erzurum, Serpil; Asosingh, Kewal

2016-01-01

Pulmonary arterial hypertension (PAH) is a heterogeneous disease characterized by severe angiogenic remodeling of the pulmonary artery wall and right ventricular hypertrophy. Thus, there is an increasing need for novel biomarkers to dissect disease heterogeneity, and predict treatment response. Although β-adrenergic receptor (βAR) dysfunction is well documented in left heart disease while endothelial cell-derived microparticles (Ec-MPs) are established biomarkers of angiogenic remodeling, methods for easy large clinical cohort analysis of these biomarkers are currently absent. Here we describe flow cytometric methods for quantification of βAR density on circulating white blood cells (WBC) and Ec-MPs in urine samples that can be used as potential biomarkers of right heart failure in PAH. Biotinylated β-blocker alprenolol was synthesized and validated as a βAR specific probe that was combined with immunophenotyping to quantify βAR density in circulating WBC subsets. Ec-MPs obtained from urine samples were stained for annexin-V and CD144, and analyzed by a micro flow cytometer. Flow cytometric detection of alprenolol showed that βAR density was decreased in most WBC subsets in PAH samples compared to healthy controls. Ec-MPs in urine was increased in PAH compared to controls. Furthermore, there was a direct correlation between Ec-MPs and Tricuspid annular plane systolic excursion (TAPSE) in PAH patients. Therefore, flow cytometric quantification of peripheral blood cell βAR density and urinary Ec-MPs may be useful as potential biomarkers of right ventricular function in PAH.

Flow Cytometric Quantification of Peripheral Blood Cell β-Adrenergic Receptor Density and Urinary Endothelial Cell-Derived Microparticles in Pulmonary Arterial Hypertension

PubMed Central

Rose, Jonathan A.; Wanner, Nicholas; Cheong, Hoi I.; Queisser, Kimberly; Barrett, Patrick; Park, Margaret; Hite, Corrine; Naga Prasad, Sathyamangla V.; Erzurum, Serpil; Asosingh, Kewal

2016-01-01

Pulmonary arterial hypertension (PAH) is a heterogeneous disease characterized by severe angiogenic remodeling of the pulmonary artery wall and right ventricular hypertrophy. Thus, there is an increasing need for novel biomarkers to dissect disease heterogeneity, and predict treatment response. Although β-adrenergic receptor (βAR) dysfunction is well documented in left heart disease while endothelial cell-derived microparticles (Ec-MPs) are established biomarkers of angiogenic remodeling, methods for easy large clinical cohort analysis of these biomarkers are currently absent. Here we describe flow cytometric methods for quantification of βAR density on circulating white blood cells (WBC) and Ec-MPs in urine samples that can be used as potential biomarkers of right heart failure in PAH. Biotinylated β-blocker alprenolol was synthesized and validated as a βAR specific probe that was combined with immunophenotyping to quantify βAR density in circulating WBC subsets. Ec-MPs obtained from urine samples were stained for annexin-V and CD144, and analyzed by a micro flow cytometer. Flow cytometric detection of alprenolol showed that βAR density was decreased in most WBC subsets in PAH samples compared to healthy controls. Ec-MPs in urine was increased in PAH compared to controls. Furthermore, there was a direct correlation between Ec-MPs and Tricuspid annular plane systolic excursion (TAPSE) in PAH patients. Therefore, flow cytometric quantification of peripheral blood cell βAR density and urinary Ec-MPs may be useful as potential biomarkers of right ventricular function in PAH. PMID:27270458

Analysis of fluid fuel flow to the neutron kinetics on molten salt reactor FUJI-12

DOE Office of Scientific and Technical Information (OSTI.GOV)

Aji, Indarta Kuncoro, E-mail: indartaaji@s.itb.ac.id; Waris, Abdul, E-mail: awaris@fi.itb.ac.id; Permana, Sidik

Molten Salt Reactor is a reactor are operating with molten salt fuel flowing. This condition interpret that the neutron kinetics of this reactor is affected by the flow rate of the fuel. This research analyze effect by the alteration velocity of the fuel by MSR type Fuji-12, with fuel composition LiF-BeF{sub 2}-ThF{sub 4}-{sup 233}UF{sub 4} respectively 71.78%-16%-11.86%-0.36%. Calculation process in this study is performed numerically by SOR and finite difference method use C programming language. Data of reactivity, neutron flux, and the macroscopic fission cross section for calculation process obtain from SRAC-CITATION (Standard thermal Reactor Analysis Code) and JENDL-4.0 datamore » library. SRAC system designed and developed by JAEA (Japan Atomic Energy Agency). This study aims to observe the effect of the velocity of fuel salt to the power generated from neutron precursors at fourth year of reactor operate (last critical condition) with number of multiplication effective; 1.0155.« less

Properties of radio-frequency heated argon confined uranium plasmas

NASA Technical Reports Server (NTRS)

1976-01-01

Pure uranium hexafluoride (UF6) was injected into an argon confined, steady state, rf-heated plasma within a fused silica peripheral wall test chamber. Exploratory tests conducted using an 80 kW rf facility and different test chamber flow configurations permitted selection of the configuration demonstrating the best confinement characteristics and minimum uranium compound wall coating. The overall test results demonstrated applicable flow schemes and associated diagnostic techniques were developed for the fluid mechanical confinement and characterization of uranium within an rf plasma discharge when pure UF6 is injected for long test times into an argon-confined, high-temperature, high-pressure, rf-heated plasma.

[Formation and Variation of Brominated Disinfection By-products in A Combined Ultrafiltration and Reverse Osmosis Process for Seawater Desalination].

PubMed

Yang, Zhe; Sun, Ying-xue; Shi, Na; Hu, Hong-ying

2015-10-01

The characteristics of dissolved organic matter (DOM) and brominated disinfection by-products ( Br-DBPs ) during a seawater desalination ultrafiltration (UF) combined reverse osmosis (RO) process were studied. The seawater contained high level of bromide ion (45.6-50.9 mg x L(-1)) and aromatic compounds with specific ultraviolet absorbance ( SUVA) of 3.6-6.0 L x (mg x m)(-1). The tryptophan-like aromatic protein, fulvic acid-like and soluble microbial by-product-like were the main fluorescent DOM in the seawater. After pre-chlorination of the seawater, the concentrations of DBPs was significantly increased in the influent of UF, which was dominantly the Br-DBPs. Bromoform (CHBr3) accounted for 70.48% - 91.50% of total trihalomethanes (THMs), dibromoacetic acid (Br2CHCO2H) occupied 81.14% - 100% of total haloacetic acids (HAAs) and dibromoacetonitrile (C2HBr2N) occupied 83.77% - 87.45% of total haloacetonitriles ( HANs). The removal efficiency of THMs, HAAs and HANs by the UF membrane was 36.63% - 40.39%, 73.83% - 95.38% and 100%, respectively. The RO membrane could completely remove the HAAs, while a little of the THMs was penetrated. The antiestrogenic activity in the seawater was 0.35 - 0.44 mg x L(-1), which was increased 32% - 69% after the pre-chlorination. The DBPs and other bio-toxic organics which formed during the UF-RO process were finally concentrated in the UF concentrate and RO concentrate.

Digital Analysis and Sorting of Fluorescence Lifetime by Flow Cytometry

PubMed Central

Houston, Jessica P.; Naivar, Mark A.; Freyer, James P.

2010-01-01

Frequency-domain flow cytometry techniques are combined with modifications to the digital signal processing capabilities of the Open Reconfigurable Cytometric Acquisition System (ORCAS) to analyze fluorescence decay lifetimes and control sorting. Real-time fluorescence lifetime analysis is accomplished by rapidly digitizing correlated, radiofrequency modulated detector signals, implementing Fourier analysis programming with ORCAS’ digital signal processor (DSP) and converting the processed data into standard cytometric list mode data. To systematically test the capabilities of the ORCAS 50 MS/sec analog-to-digital converter (ADC) and our DSP programming, an error analysis was performed using simulated light scatter and fluorescence waveforms (0.5–25 ns simulated lifetime), pulse widths ranging from 2 to 15 µs, and modulation frequencies from 2.5 to 16.667 MHz. The standard deviations of digitally acquired lifetime values ranged from 0.112 to >2 ns, corresponding to errors in actual phase shifts from 0.0142° to 1.6°. The lowest coefficients of variation (<1%) were found for 10-MHz modulated waveforms having pulse widths of 6 µs and simulated lifetimes of 4 ns. Direct comparison of the digital analysis system to a previous analog phase-sensitive flow cytometer demonstrated similar precision and accuracy on measurements of a range of fluorescent microspheres, unstained cells and cells stained with three common fluorophores. Sorting based on fluorescence lifetime was accomplished by adding analog outputs to ORCAS and interfacing with a commercial cell sorter with a radiofrequency modulated solid-state laser. Two populations of fluorescent microspheres with overlapping fluorescence intensities but different lifetimes (2 and 7 ns) were separated to ~98% purity. Overall, the digital signal acquisition and processing methods we introduce present a simple yet robust approach to phase-sensitive measurements in flow cytometry. The ability to simply and inexpensively

Distribution analysis for F100(3) engine

NASA Technical Reports Server (NTRS)

Walter, W. A.; Shaw, M.

1980-01-01

The F100(3) compression system response to inlet circumferential distortion was investigated using an analytical compressor flow model. Compression system response to several types of distortion, including pressure, temperature, and combined pressure/temperature distortions, was investigated. The predicted response trends were used in planning future F100(3) distortion tests. Results show that compression system response to combined temperature and pressure distortions depends upon the relative orientation, as well as the individual amplitudes and circumferential extents of the distortions. Also the usefulness of the analytical predictions in planning engine distortion tests is indicated.

Expanding the potential of standard flow cytometry by extracting fluorescence lifetimes from cytometric pulse shifts

PubMed Central

Cao, Ruofan; Naivar, Mark A; Wilder, Mark; Houston, Jessica P

2014-01-01

Fluorescence lifetime measurements provide information about the fluorescence relaxation, or intensity decay, of organic fluorophores, fluorescent proteins, and other inorganic molecules that fluoresce. The fluorescence lifetime is emerging in flow cytometry and is helpful in a variety of multiparametric, single cell measurements because it is not impacted by nonlinearity that can occur with fluorescence intensity measurements. Yet time-resolved cytometry systems rely on major hardware modifications making the methodology difficult to reproduce. The motivation of this work is, by taking advantage of the dynamic nature of flow cytometry sample detection and applying digital signal processing methods, to measure fluorescence lifetimes using an unmodified flow cytometer. We collect a new lifetime-dependent parameter, referred to herein as the fluorescence-pulse-delay (FPD), and prove it is a valid representation of the average fluorescence lifetime. To verify we generated cytometric pulses in simulation, with light emitting diode (LED) pulsation, and with true fluorescence measurements of cells and microspheres. Each pulse is digitized and used in algorithms to extract an average fluorescence lifetime inherent in the signal. A range of fluorescence lifetimes is measurable with this approach including standard organic fluorophore lifetimes (∼1 to 22 ns) as well as small, simulated shifts (0.1 ns) under standard conditions (reported herein). This contribution demonstrates how digital data acquisition and signal processing can reveal time-dependent information foreshadowing the exploitation of full waveform analysis for quantification of similar photo-physical events within single cells. © 2014 The Authors. Published by Wiley Periodicals, Inc. PMID:25274073

CFD Analysis of Coolant Flow in VVER-440 Fuel Assemblies with the Code ANSYS CFX 10.0

DOE Office of Scientific and Technical Information (OSTI.GOV)

Toth, Sandor; Legradi, Gabor; Aszodi, Attila

2006-07-01

From the aspect of planning the power upgrading of nuclear reactors - including the VVER-440 type reactor - it is essential to get to know the flow field in the fuel assembly. For this purpose we have developed models of the fuel assembly of the VVER-440 reactor using the ANSYS CFX 10.0 CFD code. At first a 240 mm long part of a 60 degrees segment of the fuel pin bundle was modelled. Implementing this model a sensitivity study on the appropriate meshing was performed. Based on the development of the above described model, further models were developed: a 960more » mm long part of a 60-degree-segment and a full length part (2420 mm) of the fuel pin bundle segment. The calculations were run using constant coolant properties and several turbulence models. The impacts of choosing different turbulence models were investigated. The results of the above-mentioned investigations are presented in this paper. (authors)« less

NIST/ISAC standardization study: variability in assignment of intensity values to fluorescence standard beads and in cross calibration of standard beads to hard dyed beads.

PubMed

Hoffman, Robert A; Wang, Lili; Bigos, Martin; Nolan, John P

2012-09-01

Results from a standardization study cosponsored by the International Society for Advancement of Cytometry (ISAC) and the US National Institute of Standards and Technology (NIST) are reported. The study evaluated the variability of assigning intensity values to fluorophore standard beads by bead manufacturers and the variability of cross calibrating the standard beads to stained polymer beads (hard-dyed beads) using different flow cytometers. Hard dyed beads are generally not spectrally matched to the fluorophores used to stain cells, and spectral response varies among flow cytometers. Thus if hard dyed beads are used as fluorescence calibrators, one expects calibration for specific fluorophores (e.g., FITC or PE) to vary among different instruments. Using standard beads surface-stained with specific fluorophores (FITC, PE, APC, and Pacific Blue™), the study compared the measured intensity of fluorophore standard beads to that of hard dyed beads through cross calibration on 133 different flow cytometers. Using robust CV as a measure of variability, the variation of cross calibrated values was typically 20% or more for a particular hard dyed bead in a specific detection channel. The variation across different instrument models was often greater than the variation within a particular instrument model. As a separate part of the study, NIST and four bead manufacturers used a NIST supplied protocol and calibrated fluorophore solution standards to assign intensity values to the fluorophore beads. Values assigned to the reference beads by different groups varied by orders of magnitude in most cases, reflecting differences in instrumentation used to perform the calibration. The study concluded that the use of any spectrally unmatched hard dyed bead as a general fluorescence calibrator must be verified and characterized for every particular instrument model. Close interaction between bead manufacturers and NIST is recommended to have reliable and uniformly assigned

CMOS based image cytometry for detection of phytoplankton in ballast water.

PubMed

Pérez, J M; Jofre, M; Martínez, P; Yáñez, M A; Catalan, V; Parker, A; Veldhuis, M; Pruneri, V

2017-02-01

We introduce an image cytometer (I-CYT) for the analysis of phytoplankton in fresh and marine water environments. A linear quantification of cell numbers was observed covering several orders of magnitude using cultures of Tetraselmis and Nannochloropsis measured by autofluorescence in a laboratory environment. We assessed the functionality of the system outside the laboratory by phytoplankton quantification of samples taken from a marine water environment (Dutch Wadden Sea, The Netherlands) and a fresh water environment (Lake Ijssel, The Netherlands). The I-CYT was also employed to study the effects of two ballast water treatment systems (BWTS), based on chlorine electrolysis and UV sterilization, with the analysis including the vitality of the phytoplankton. For comparative study and benchmarking of the I-CYT, a standard flow cytometer was used. Our results prove a limit of detection (LOD) of 10 cells/ml with an accuracy between 0.7 and 0.5 log, and a correlation of 88.29% in quantification and 96.21% in vitality, with respect to the flow cytometry results.

Rapid quantification of live/dead lactic acid bacteria in probiotic products using high-sensitivity flow cytometry

NASA Astrophysics Data System (ADS)

He, Shengbin; Hong, Xinyi; Huang, Tianxun; Zhang, Wenqiang; Zhou, Yingxing; Wu, Lina; Yan, Xiaomei

2017-06-01

A laboratory-built high-sensitivity flow cytometer (HSFCM) was employed for the rapid and accurate detection of lactic acid bacteria (LAB) and their viability in probiotic products. LAB were stained with both the cell membrane-permeable SYTO 9 green-fluorescent nucleic acid stain and the red-fluorescent nucleic acid stain, propidium iodide, which penetrates only bacteria with compromised membranes. The side scatter and dual-color fluorescence signals of single bacteria were detected simultaneously by the HSFCM. Ultra-high temperature processing milk and skim milk spiked with Lactobacillus casei were used as the model systems for the optimization of sample pretreatment and staining. The viable LAB counts measured by the HSFCM were in good agreement with those of the plate count method, and the measured ratios between the live and dead LAB matched well with the theoretical ratios. The established method was successfully applied to the rapid quantification of live/dead LAB in yogurts and fermented milk beverages of different brands. Moreover, the concentration and viability status of LAB in ambient yogurt, a relatively new yet popular milk product in China, are also reported.

In vivo flow cytometry of circulating clots using negative photothermal and photoacoustic contrasts.

PubMed

Galanzha, Ekaterina I; Sarimollaoglu, Mustafa; Nedosekin, Dmitry A; Keyrouz, Salah G; Mehta, Jawahar L; Zharov, Vladimir P

2011-10-01

Conventional photothermal (PT) and photoacousic (PA) imaging, spectroscopy, and cytometry are preferentially based on positive PT/PA effects, when signals are above background. Here, we introduce PT/PA technique based on detection of negative signals below background. Among various new applications, we propose label-free in vivo flow cytometry of circulating clots. No method has been developed for the early detection of clots of different compositions as a source of thromboembolism including ischemia at strokes and myocardial infarction. When a low-absorbing, platelet-rich clot passes a laser-irradiated vessel volume, a transient decrease in local absorption results in an ultrasharp negative PA hole in blood background. Using this phenomenon alone or in combination with positive contrasts, we demonstrated identification of white, red, and mixed clots on a mouse model of myocardial infarction and human blood. The concentration and size of clots were measured with threshold down to few clots in the entire circulation with size as low as 20 μm. This multiparameter diagnostic platform using portable personal high-speed flow cytometer with negative dynamic contrast mode has potential to real-time defining risk factors for cardiovascular diseases, and for prognosis and prevention of stroke or use clot count as a marker of therapy efficacy. Possibility for label-free detection of platelets, leukocytes, tumor cells or targeting themby negative PA probes (e.g., nonabsorbing beads or bubbles) is also highlighted. Copyright © 2011 International Society for Advancement of Cytometry.

In vivo flow cytometry of circulating clots using negative phototothermal and photoacoustic contrasts

PubMed Central

Galanzha, Ekaterina I.; Sarimollaoglu, Mustafa; Nedosekin, Dmitry A.; Keyrouz, Salah G.; Mehta, Jawahar L.; Zharov, Vladimir P.

2012-01-01

Conventional photothermal (PT) and photoacousic (PA) imaging, spectroscopy, and cytometry are preferentially based on positive PT/PA effects, when signals are above background. Here, we introduce PT/PA technique based on detection of negative signals below background. Among various new applications, we propose label-free in vivo flow cytometry of circulating clots. No method has been developed for the early detection of clots of different compositions as a source of severe thromboembolisms including ischemia at strokes and myocardial dysfunction at heart attack. When a low-absorbing, platelet-rich clot passes a laser-irradiated vessel volume, a transient decrease in local absorption results in an ultrasharp negative PA hole in blood background. Using this phenomenon alone or in combination with positive contrasts, we demonstrated identification of white, red and mixed clots on a mouse model of myocardial infarction and human blood. The concentration and size of clots were measured with threshold down to few clots in the entire circulation with size as low as 20 µm. This multiparameter diagnostic platform using portable personal high-speed flow cytometer with negative dynamic contrast mode has potential to real-time defining risk factors for cardiovascular diseases, and for prognosis and prevention of stroke or use clot count as a marker of therapy efficacy. Possibility for label-free detection of platelets, leukocytes, tumor cells or targeting them by negative PA probes (e.g., nonabsorbing beads or bubbles) is also highlighted. PMID:21976458

CD68 on rat macrophages binds tightly to S100A8 and S100A9 and helps to regulate the cells' immune functions.

PubMed

Okada, Kohki; Arai, Satoshi; Itoh, Hiroshi; Adachi, Souichi; Hayashida, Masahiko; Nakase, Hiroshi; Ikemoto, Masaki

2016-11-01

S100A8 and S100A9 (S100 proteins) are regulators of immune cells of myeloid origin. Whereas S100 proteins are found at high concentrations in such cells, their immunologic roles remain unclear. We focused on cluster of differentiation 68 (CD68). The aim of this study is to investigate whether CD68 binds to extracellular S100A8 and/or S100A9 and subsequently participates in the regulation of the cells' immune functions. ELISA and affinity chromatography showed that both recombinant rat S100A8 (r-S100A8) and r-S100A9 bound to r-CD68, but not to r-CD14. Flow cytometry clearly showed evidences supporting above the 2 results. As analyzed by flow cytometry, a less amount of r-S100A8 or r-S100A9 bound to the macrophages treated with some deglycosylation enzymes. In an in vitro assay, the expression levels of S100A8 and S100A9 were significantly suppressed after the macrophages had been treated with an anti-CD68 antibody (ED1). As stimulated macrophages with r-S100A9, the expression of IL-1β mRNA in macrophages, which were treated with anti-TLR4 or -RAGE antibodies, was significantly suppressed. r-S100A8 up-regulated IL-6 and IL-10 mRNAs, while r-S100A9 did TNF-α and IL-6 mRNAs, although these regulations were not statistically significant. Small interfering CD68 also significantly suppressed activation of macrophages through an autocrine pathway by r-S100A8 or r-S100A9. In macrophages stimulated with LPS, fluorescent immunologic staining showed that most CD68 colocalized with S100A8 or S100A9 and that the levels of all 3 molecules were markedly increased. In conclusion, CD68 on macrophages binds to S100A8 and S100A9 and thereby, plays a role in the regulation of the cells' immune functions. © Society for Leukocyte Biology.

Biomass measurement by flow cytometry during solid-state fermentation of basidiomycetes.

PubMed

Steudler, Susanne; Böhmer, Ulrike; Weber, Jost; Bley, Thomas

2015-02-01

Solid-state fermentation (SSF) is a robust process that is well suited to the on-site cultivation of basidiomycetes that produce enzymes for the treatment of lignocellulosics. Reliable methods for biomass quantification are essential for the analysis of fungal growth kinetics. However, direct biomass determination is not possible during SSF because the fungi grow into the substrate and use it as a nutrient source. This necessitates the use of indirect methods that are either very laborious and time consuming or can only provide biomass measurements during certain growth periods. Here, we describe the development and optimization of a new rapid method for fungal biomass determination during SSF that is based on counting fungal nuclei by flow cytometry. Fungal biomass was grown on an organic substrate and its concentration was measured by isolating the nuclei from the fungal hyphae after cell disruption, staining them with SYTOX(®) Green, and then counting them using a flow cytometer. A calibration curve relating the dry biomass of the samples to their concentrations of nuclei was established. Multiple buffers and disruption methods were tested. The results obtained were compared with values determined using the method of ergosterol determination, a classical technique for fungal biomass measurement during SSF. Our new approach can be used to measure fungal biomass on a range of different scales, from 15 mL cultures to a laboratory reactor with a working volume of 10 L (developed by the Research Center for Medical Technology and Biotechnology (fzmb GmbH)). © 2014 International Society for Advancement of Cytometry. © 2014 International Society for Advancement of Cytometry.

Ultrahigh temperature vapor core reactor-MHD system for space nuclear electric power

NASA Technical Reports Server (NTRS)

Maya, Isaac; Anghaie, Samim; Diaz, Nils J.; Dugan, Edward T.

1991-01-01

The conceptual design of a nuclear space power system based on the ultrahigh temperature vapor core reactor with MHD energy conversion is presented. This UF4 fueled gas core cavity reactor operates at 4000 K maximum core temperature and 40 atm. Materials experiments, conducted with UF4 up to 2200 K, demonstrate acceptable compatibility with tungsten-molybdenum-, and carbon-based materials. The supporting nuclear, heat transfer, fluid flow and MHD analysis, and fissioning plasma physics experiments are also discussed.

A multicolor flow cytometric assay for measurement of platelet-derived microparticles.

PubMed

Mobarrez, Fariborz; Antovic, Jovan; Egberg, Nils; Hansson, Mona; Jörneskog, Gun; Hultenby, Kjell; Wallén, Håkan

2010-03-01

Flow cytometry (FCM) is the most commonly used method for detection of platelet-derived microparticles (PDMPs), but it is poorly standardized and mainly used for "bedside" analyses in fresh samples. If PDMPs could be analyzed in previously frozen samples it would increase the usefulness of the method. However, cell membrane fragments from contaminating cells created during freezing/thawing may cause artifacts and disturb measurements. PDMPs were labeled with monoclonal antibodies directed against CD42a and CD62P, or CD42a and CD142. The PDMP gate was determined using forward scatter (FSC) and CD42a expression. The mean fluorescence intensities (MFIs) of CD62P or CD142 positive particles were translated into MESF -values (Molecules of Equivalent Soluble Fluorochrome) using a standard curve. FITC-labeled phalloidin (which binds to intracellular actin) was used to detect destroyed cells/cell fragments. Phalloidin-positive particles were significantly more common in supernatants of frozen/thawed platelet rich and platelet poor plasma samples compared with supernatants of platelet free plasma. High-speed centrifugation was then used to obtain PDMP samples with low contamination of cell fragments. Electron microscopy showed that these samples contained numerous round stained particles with cellular membranes of a size of 100-700 nm. Reproducibility experiments using plasma samples from healthy individuals showed that the coefficients of variation (CVs) of MESF values of CD62P and CD142 (both intra- and interassay) were <10%, and the variation between two cytometers in two different laboratories was <5%. We also found that PDMP expression of CD142 (i.e. tissue factor [TF]) and CD62P (i.e P-selectin) was around two times higher in samples from type 1-diabetes patients compared with those from healthy controls (p<0.001). The use of MESF values to quantify PDMP expression of P-selectin and TF yields reproducible data and enables comparison of data between laboratories. If

Evaluation of Ginger (Zingiber officinale Roscoe) Bioactive Compounds in Increasing the Ratio of T-cell Surface Molecules of CD3+CD4+:CD3+CD8+ In-Vitro.

PubMed

Tejasari, Dr

2007-09-01

The potential ability of ginger bioactive compounds in increasing the ratio of T-cell surface molecules of CD3+CD4+:CD3+CD8+ was investigated using dual tagging FITC and PE of monoclonal antibody anti-human with its fluorescence measured by flow cytometer. Oleoresin was extracted using sinkhole distillation technique. Its components namely, gingerol in fraction-1, shogaol in fraction 2 and zingeron in fraction-3 were separated by column vacuum chromatography method. The doses of oleoresin, gingerol, shogaol, and zingeron tested were 50, 100,150, 200, and 250 μg/ml. Lymphocytes (2x106 cell/ml) from human peripheral blood were isolated using ficoll density gradient technique, and cultured in the presence of the compounds in RPMI-1640 medium and phytohemaglutinin (PHA) mitogen for 96 h under normal conditions. Percentages of T-cell surface molecules (CD4+ and CD8+) were determined using dual-tagging FITC and PE fluorescents labeled on monoclonal antibody anti human. The fluorescence-labeled bands on the T-cell surface molecules were counted using flow cytometer. The experiment revealed that oleoresin and its three fractions increased the percentage of CD3+CD4+. The compound in fraction 3 of oleoresin at 200 μg/ml increased by the highest percentage of CD3+CD4+ of 9%, but slightly decreased the percentage of CD3+CD8+. These ginger bioactive compounds increased the ratio of CD3+CD4:CD3+CD8+ T-cells with the highest increment of 30% from effects of 200 μg/ml fraction 3 of oleoresin. This in vitro finding revealed that ginger bioactive compounds potentially increased cellular and humoral immune response. Further clinical studies are needed to confirm the benefits of these ginger bioactive compounds as a potential functional food for testing on HIV infected patients.

Cancer Detection Device

NASA Technical Reports Server (NTRS)

1998-01-01

Ratcom, Inc., has joined NASA Johnson Space Center in an active program to develop cytometry capabilities for space station freedom. This agreement results from a cooperative program that NASA entered into with the American Cancer Society to aid in cancer prevention, diagnosis, and treatment. The flow cytometer is used by cancer researchers to make cellular measurements.

Quality assessment program for EuroFlow protocols: summary results of four-year (2010-2013) quality assurance rounds.

PubMed

Kalina, Tomas; Flores-Montero, Juan; Lecrevisse, Quentin; Pedreira, Carlos E; van der Velden, Vincent H J; Novakova, Michaela; Mejstrikova, Ester; Hrusak, Ondrej; Böttcher, Sebastian; Karsch, Dennis; Sędek, Łukasz; Trinquand, Amelie; Boeckx, Nancy; Caetano, Joana; Asnafi, Vahid; Lucio, Paulo; Lima, Margarida; Helena Santos, Ana; Bonaccorso, Paola; van der Sluijs-Gelling, Alita J; Langerak, Anton W; Martin-Ayuso, Marta; Szczepański, Tomasz; van Dongen, Jacques J M; Orfao, Alberto

2015-02-01

Flow cytometric immunophenotyping has become essential for accurate diagnosis, classification, and disease monitoring in hemato-oncology. The EuroFlow Consortium has established a fully standardized "all-in-one" pipeline consisting of standardized instrument settings, reagent panels, and sample preparation protocols and software for data analysis and disease classification. For its reproducible implementation, parallel development of a quality assurance (QA) program was required. Here, we report on the results of four consecutive annual rounds of the novel external QA EuroFlow program. The novel QA scheme aimed at monitoring the whole flow cytometric analysis process (cytometer setting, sample preparation, acquisition and analysis) by reading the median fluorescence intensities (MedFI) of defined lymphocytes' subsets. Each QA participant applied the predefined reagents' panel on blood cells of local healthy donors. A uniform gating strategy was applied to define lymphocyte subsets and to read MedFI values per marker. The MedFI values were compared with reference data and deviations from reference values were quantified using performance score metrics. In four annual QA rounds, we analyzed 123 blood samples from local healthy donors on 14 different instruments in 11 laboratories from nine European countries. The immunophenotype of defined cellular subsets appeared sufficiently standardized to permit unified (software) data analysis. The coefficient of variation of MedFI for 7 of 11 markers performed repeatedly below 30%, average MedFI in each QA round ranged from 86 to 125% from overall median. Calculation of performance scores was instrumental to pinpoint standardization failures and their causes. Overall, the new EuroFlow QA system for the first time allowed to quantify the technical variation that is introduced in the measurement of fluorescence intensities in a multicentric setting over an extended period of time. EuroFlow QA is a proficiency test specific for

An in vitro AChE inhibition assay combined with UF-HPLC-ESI-Q-TOF/MS approach for screening and characterizing of AChE inhibitors from roots of Coptis chinensis Franch.

PubMed

Zhao, Hengqiang; Zhou, Siduo; Zhang, Minmin; Feng, Jinhong; Wang, Shanshan; Wang, Daijie; Geng, Yanling; Wang, Xiao

2016-02-20

In this study, an in vitro acetylcholinesterase (AChE) inhibition assay based on microplate reader combined with ultrafiltration high performance liquid chromatography-electrospray quadrupole time of flight mass (UF-HPLC-ESI-Q-TOF/MS) was developed for the rapid screening and identification of acetylcholinesterase inhibitors (AChEI) from roots of Coptis chinensis Franch. Incubation conditions such as enzyme concentration, incubation time, incubation temperature and co-solvent was optimized so as to get better screening results. Five alkaloids including columbamine, jatrorrhizine, coptisine, palmatine and berberine were found with AChE inhibition activity in the 80% ethanol extract of C. chinensis Franch. The screened compounds were identified by HPLC-DAD-ESI-Q-TOF/MS compared with the reference stands and literatures. The screened results were verified by in vitro AChE inhibition assays, palmatine showed the best AChE inhibitory activities with IC50 values of 36.6μM among the five compounds. Results of the present study indicated that the combinative method using in vitro AChE inhibition assay and UF-HPLC-ESI-Q-TOF/MS could be widely applied for rapid screening and identification of AChEI from complex TCM extract. Copyright © 2015 Elsevier B.V. All rights reserved.

AGT100 turbomachinery. [for automobiles

NASA Technical Reports Server (NTRS)

Tipton, D. L.; Mckain, T. F.

1982-01-01

High-performance turbomachinery components have been designed and tested for the AGT100 automotive engine. The required wide range of operation coupled with the small component size, compact packaging, and low cost of production provide significant aerodynamic challenges. Aerodynamic design and development testing of the centrifugal compressor and two radial turbines are described. The compressor achieved design flow, pressure ratio, and surge margin on the initial build. Variable inlet guide vanes have proven effective in modulating flow capacity and in improving part-speed efficiency. With optimum use of the variable inlet guide vanes, the initial efficiency goals have been demonstrated in the critical idle-to-70% gasifier speed range. The gasifier turbine exceeded initial performance goals and demonstrated good performance over a wide range. The radial power turbine achieved 'developed' efficiency goals on the first build.

Magnetic Separation and Antibiotics Selection Enable Enrichment of Cells with ZFN/TALEN-Induced Mutations

PubMed Central

Lee, Choong-il; Kim, Hyongbum; Kim, Jin-Soo

2013-01-01

The ability to enrich cells with targeted mutations greatly facilitates the process of using engineered nucleases, including zinc-finger nucleases and transcription activator-like effector nucleases, to construct such cells. We previously used surrogate reporters to enrich cells containing nuclease-induced mutations via flow cytometry. This method is, however, limited by the availability of flow cytometers. Furthermore, sorted cells occasionally fail to form colonies after exposure to a strong laser and hydrostatic pressure. Here we describe two different types of novel reporters that enable mutant cell enrichment without the use of flow cytometers. We designed reporters that express H-2Kk, a surface antigen, and the hygromycin resistance protein (HygroR), respectively, when insertions or deletions are generated at the target sequences by the activity of engineered nucleases. After cotransfection of these reporters and the engineered nuclease-encoding plasmids, H-2Kk- and HygroR-expressing cells were isolated using magnetic separation and hygromycin treatment, respectively. We found that mutant cells were drastically enriched in the isolated cells, suggesting that these two reporters enable efficient enrichment of mutants. We propose that these two reporters will greatly facilitate the use of engineered nucleases in a wider range of biomedical research. PMID:23441197

Evaluation of Plasma Platelet Microparticles in Thrombotic Thrombocytopenic Purpura.

PubMed

Tahmasbi, Leila; Karimi, Mehran; Kafiabadi, Sedigheh Amini; Nikougoftar, Mahin; Haghpanah, Sezaneh; Ranjbaran, Reza; Moghadam, Mohamad

2017-01-01

Platelet microparticles (PMPs) have a procoagulant activity about 50-100 times greater than active platelets due to high expression of negatively charged phospholipids on their surfaces. In this study, we evaluated microparticle immunophenotyping and also plasma PMPs level in patients with Thrombotic Thrombocytopenic Purpura (TTP) in Southern Iran. We had two study groups: 15 TTP patients and 15 healthy control group and PMPs from platelet concentrate (PC) at the 5 th day of storage. Microparticles were prepared in two steps, by low and high centrifugation followed by size confirmation via 'Dynamic Light Scattering (DLS)' Zetasizer. Immunophenotyping of PMPs was done via flow cytometry, using a FACS Calibur flow cytometer (BD, USA). PMPs counts were obtained using Partec-cyflow and Polysciences Microbeads (1 micron in diameter). Results were analyzed using FlowJo 7.6 (Treestar, USA) and Partec FlowMax software. Our results showed that the majority of microparticles in TTP patients and normal individuals were PMPs and also demonstrated that the plasma PMPs level in TTP patients was higher than the normal control group ( P -value<0.001). It seems that elevated PMPs level in TTP patients could be related to thrombotic events. Nevertheless, more studies are needed to confirm these results. © 2017 by the Association of Clinical Scientists, Inc.

Reverse-engineering flow-cytometry gating strategies for phenotypic labelling and high-performance cell sorting.

PubMed

Becht, Etienne; Simoni, Yannick; Coustan-Smith, Elaine; Maximilien, Evrard; Cheng, Yang; Ng, Lai Guan; Campana, Dario; Newell, Evan

2018-06-21

Recent flow and mass cytometers generate datasets of dimensions 20 to 40 and a million single cells. From these, many tools facilitate the discovery of new cell populations associated with diseases or physiology. These new cell populations require the identification of new gating strategies, but gating strategies become exponentially more difficult to optimize when dimensionality increases. To facilitate this step, we developed Hypergate, an algorithm which given a cell population of interest identifies a gating strategy optimized for high yield and purity. Hypergate achieves higher yield and purity than human experts, Support Vector Machines and Random-Forests on public datasets. We use it to revisit some established gating strategies for the identification of innate lymphoid cells, which identifies concise and efficient strategies that allow gating these cells with fewer parameters but higher yield and purity than the current standards. For phenotypic description, Hypergate's outputs are consistent with fields' knowledge and sparser than those from a competing method. Hypergate is implemented in R and available on CRAN. The source code is published at http://github.com/ebecht/hypergate under an Open Source Initiative-compliant licence. Supplementary data are available at Bioinformatics online.

A high-throughput flow cytometry-on-a-CMOS platform for single-cell dielectric spectroscopy at microwave frequencies.

PubMed

Chien, Jun-Chau; Ameri, Ali; Yeh, Erh-Chia; Killilea, Alison N; Anwar, Mekhail; Niknejad, Ali M

2018-06-06

This work presents a microfluidics-integrated label-free flow cytometry-on-a-CMOS platform for the characterization of the cytoplasm dielectric properties at microwave frequencies. Compared with MHz impedance cytometers, operating at GHz frequencies offers direct intracellular permittivity probing due to electric fields penetrating through the cellular membrane. To overcome the detection challenges at high frequencies, the spectrometer employs on-chip oscillator-based sensors, which embeds simultaneous frequency generation, electrode excitation, and signal detection capabilities. By employing an injection-locking phase-detection technique, the spectrometer offers state-of-the-art sensitivity, achieving a less than 1 aFrms capacitance detection limit (or 5 ppm in frequency-shift) at a 100 kHz noise filtering bandwidth, enabling high throughput (>1k cells per s), with a measured cellular SNR of more than 28 dB. With CMOS/microfluidics co-design, we distribute four sensing channels at 6.5, 11, 17.5, and 30 GHz in an arrayed format whereas the frequencies are selected to center around the water relaxation frequency at 18 GHz. An issue in the integration of CMOS and microfluidics due to size mismatch is also addressed through introducing a cost-efficient epoxy-molding technique. With 3-D hydrodynamic focusing microfluidics, we perform characterization on four different cell lines including two breast cell lines (MCF-10A and MDA-MB-231) and two leukocyte cell lines (K-562 and THP-1). After normalizing the higher frequency signals to the 6.5 GHz ones, the size-independent dielectric opacity shows a differentiable distribution at 17.5 GHz between normal (0.905 ± 0.160, mean ± std.) and highly metastatic (1.033 ± 0.107) breast cells with p ≪ 0.001.

Comparative Analysis of Various Aspects of Plateletpheresis on the Fenwal Amicus and Fresenius COM.TEC Cell Separator Instruments.

PubMed

Philip, Joseph; Biswas, Amit Kumar; Chatterjee, Tathagata; Mallhi, Rajiv Singh

2014-01-01

To compare the Fenwal Amicus and the Fresenius COM.TEC apheresis instruments regarding donor peripheral blood parameters, operational variables of the instruments, and quality control parameters of the product obtained. We performed 100 platelet collections from 100 voluntary donors using the 2 studied devices. We measured platelet count using an automated analyzer and analyzed the activation statuses using a flow cytometer. The median time needed to perform the procedures was significantly longer with the COM.TEC. However, the product we obtained using the Amicus instrument showed higher degrees of platelet-activation. All products we obtained with both instruments had white blood cell counts of less than 5 × 10(6) per bag. We observed no statistical difference regarding collection efficiency and collection rates between the devices. Both instruments collected platelets efficiently, with minimal donor discomfort. Compared with the COM.TEC instrument, the Amicus reached the platelet target yield more quickly; however, it displayed an increase in platelet activation. Copyright© by the American Society for Clinical Pathology (ASCP).

Inlet flow field investigation. Part 1: Transonic flow field survey

NASA Technical Reports Server (NTRS)

Yetter, J. A.; Salemann, V.; Sussman, M. B.

1984-01-01

A wind tunnel investigation was conducted to determine the local inlet flow field characteristics of an advanced tactical supersonic cruise airplane. A data base for the development and validation of analytical codes directed at the analysis of inlet flow fields for advanced supersonic airplanes was established. Testing was conducted at the NASA-Langley 16-foot Transonic Tunnel at freestream Mach numbers of 0.6 to 1.20 and angles of attack from 0.0 to 10.0 degrees. Inlet flow field surveys were made at locations representative of wing (upper and lower surface) and forebody mounted inlet concepts. Results are presented in the form of local inlet flow field angle of attack, sideflow angle, and Mach number contours. Wing surface pressure distributions supplement the flow field data.

Influence of gas-liquid two-phase flow on angiotensin-I converting enzyme inhibitory peptides separation by ultra-filtration.

PubMed

Charoenphun, Narin; Youravong, Wirote

2017-01-01

Membrane fouling is a major problem in ultra-filtration systems and two-phase flow is a promising technique for permeate flux enhancement. The objective of this research was to study the use of an ultra-filtration (UF) system to enrich angiotensin-I converting enzyme (ACE) inhibitory peptides from tilapia protein hydrolysate. To select the most appropriate membrane and operating condition, the effects of membrane molecular weight cut-off (MWCO), transmembrane pressure (TMP) and cross-flow velocity (CFV) on permeate flux and ACE inhibitory peptide separation were studied. Additionally, the gas-liquid two-phase flow technique was applied to investigate its effect on the process capability. The results showed that the highest ACE inhibitory activity was obtained from permeate of the 1 kDa membrane. In terms of TMP and CFV, the permeate flux tended to increase with TMP and CFV. The use of gas-liquid two-phase flow as indicated by shear stress number could reduce membrane fouling and increase the permeate flux up to 42%, depending on shear stress number. Moreover, the use of a shear stress number of 0.039 led to an augmentation in ACE inhibitory activity of permeates. Operating conditions using a shear stress number of 0.039 were recommended for enrichment of ACE inhibitory peptides. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

The UF Family of hybrid phantoms of the pregnant female for computational radiation dosimetry

NASA Astrophysics Data System (ADS)

Maynard, Matthew R.; Long, Nelia S.; Moawad, Nash S.; Shifrin, Roger Y.; Geyer, Amy M.; Fong, Grant; Bolch, Wesley E.

2014-08-01

Efforts to assess in utero radiation doses and related quantities to the developing fetus should account for the presence of the surrounding maternal tissues. Maternal tissues can provide varying levels of protection to the fetus by shielding externally-emitted radiation or, alternatively, can become sources of internally-emitted radiation following the biokinetic uptake of medically-administered radiopharmaceuticals or radionuclides located in the surrounding environment—as in the case of the European Union’s SOLO project (Epidemiological Studies of Exposed Southern Urals Populations). The University of Florida had previously addressed limitations in available computational phantom representation of the developing fetus by constructing a series of hybrid computational fetal phantoms at eight different ages and three weight percentiles. Using CT image sets of pregnant patients contoured using 3D-DOCTORTM, the eight 50th percentile fetal phantoms from that study were systematically combined in RhinocerosTM with the UF adult non-pregnant female to yield a series of reference pregnant female phantoms at fetal ages 8, 10, 15, 20, 25, 30, 35 and 38 weeks post-conception. Deformable, non-uniform rational B-spline surfaces were utilized to alter contoured maternal anatomy in order to (1) accurately position and orient each fetus and surrounding maternal tissues and (2) match target masses of maternal soft tissue organs to reference data reported in the literature.

Static Flow Characteristics of a Mass Flow Injecting Valve

NASA Technical Reports Server (NTRS)

Mattern, Duane; Paxson, Dan

1995-01-01

A sleeve valve is under development for ground-based forced response testing of air compression systems. This valve will be used to inject air and to impart momentum to the flow inside the first stage of a multi-stage compressor. The valve was designed to deliver a maximum mass flow of 0.22 lbm/s (0.1 kg/s) with a maximum valve throat area of 0.12 sq. in (80 sq. mm), a 100 psid (689 KPA) pressure difference across the valve and a 68 F, (20 C) air supply. It was assumed that the valve mass flow rate would be proportional to the valve orifice area. A static flow calibration revealed a nonlinear valve orifice area to mass flow relationship which limits the maximum flow rate that the valve can deliver. This nonlinearity was found to be caused by multiple choking points in the flow path. A simple model was used to explain this nonlinearity and the model was compared to the static flow calibration data. Only steady flow data is presented here. In this report, the static flow characteristics of a proportionally controlled sleeve valve are modelled and validated against experimental data.

Micro-Electro-Mechanical Systems (MEMS) Fabrication Course Projects Review for FY15

DTIC Science & Technology

2015-09-01

TECHNICAL DOCUMENT 3298 September 2015 Micro-Electro-Mechanical Systems (MEMS) Fabrication Course Projects Review for FY15 Paul D. Swanson...Naval Warfare Systems Center Pacific (SSC Pacific), San Diego, CA. SSC Pacific’s Naval Innovative Science and Engineering (NISE) Program provided...for Miniaturized Flow Cytometer o Howard Dyckman: 71730 Infrared Waveguides o Teresa Emery: 55360 Bistable MEMS systems for Energy

Resuscitation With 100% Oxygen Causes Intestinal Glutathione Oxidation and Reoxygenation Injury in Asphyxiated Newborn Piglets

PubMed Central

Haase, Erika; Bigam, David L.; Nakonechny, Quentin B.; Jewell, Laurence D.; Korbutt, Gregory; Cheung, Po-Yin

2004-01-01

Objective: To compare mesenteric blood flow, oxidative stress, and mucosal injury in piglet small intestine during hypoxemia and reoxygenation with 21%, 50%, or 100% oxygen. Summary Background Data: Necrotizing enterocolitis is a disease whose pathogenesis likely involves hypoxia-reoxygenation and the generation of oxygen-free radicals, which are known to cause intestinal injury. Resuscitation of asphyxiated newborns with 100% oxygen has been shown to increase oxidative stress, as measured by the glutathione redox ratio, and thus may predispose to free radical-mediated tissue injury. Methods: Newborn piglets subjected to severe hypoxemia for 2 hours were resuscitated with 21%, 50%, or 100% oxygen while superior mesenteric artery (SMA) flow and hemodynamic parameters were continuously measured. Small intestinal tissue samples were analyzed for histologic injury and levels of oxidized and reduced glutathione. Results: SMA blood flow decreased to 34% and mesenteric oxygen delivery decreased to 9% in hypoxemic piglets compared with sham-operated controls. With reoxygenation, SMA blood flow increased to 177%, 157%, and 145% of baseline values in piglets resuscitated with 21%, 50%, and 100% oxygen, respectively. Mesenteric oxygen delivery increased to more than 150% of baseline values in piglets resuscitated with 50% or 100% oxygen, and this correlated significantly with the degree of oxidative stress, as measured by the oxidized-to-reduced glutathione ratio. Two of eight piglets resuscitated with 100% oxygen developed gross and microscopic evidence of pneumatosis intestinalis and severe mucosal injury, while all other piglets were grossly normal. Conclusions: Resuscitation of hypoxemic newborn piglets with 100% oxygen is associated with an increase in oxygen delivery and oxidative stress, and may be associated with the development of small intestinal hypoxia-reoxygenation injury. Resuscitation of asphyxiated newborns with lower oxygen concentrations may help to decrease

Treatment of cosmetic effluent in different configurations of ceramic UF membrane based bioreactor: Toxicity evaluation of the untreated and treated wastewater using catfish (Heteropneustes fossilis).

PubMed

Banerjee, Priya; Dey, Tanmoy Kumar; Sarkar, Sandeep; Swarnakar, Snehasikta; Mukhopadhyay, Aniruddha; Ghosh, Sourja

2016-03-01

Extensive usage of pharmaceutical and personal care products (PPCPs) and their discharge through domestic sewage have been recently recognized as a new generation environmental concern which deserves more scientific attention over the classical environmental pollutants. The major issues of this type of effluent addressed in this study were its colour, triclosan and anionic surfactant (SDS) content. Samples of cosmetic effluent were collected from different beauty treatment salons and spas in and around Kolkata, India and treated in bioreactors containing a bacterial consortium isolated from activated sludge samples collected from a common effluent treatment plant. Members of the consortium were isolated and identified as Klebsiella sp., Pseudomonas sp., Salmonella sp. and Comamonas sp. The biotreated effluent was subjected to ultrafiltration (UF) involving indigenously prepared ceramic membranes in both side-stream and submerged mode. Analysis of the MBR treated effluent revealed 99.22%, 98.56% and 99.74% removal of colour, triclosan and surfactant respectively. Investigation of probable acute and chronic cyto-genotoxic potential of the untreated and treated effluents along with their possible participation in triggering oxidative stress was carried out with Heteropneustes fossilis (Bloch). Comet formation recorded in both liver and gill cells and micronucleus count in peripheral erythrocytes of individuals exposed to untreated effluent increased with duration of exposure and was significantly higher than those treated with UF permeates which in turn neared control levels. Results of this study revealed successful application of the isolated bacterial consortium in MBR process for efficient detoxification of cosmetic effluent thereby conferring the same suitable for discharge and/or reuse. Copyright © 2015 Elsevier Ltd. All rights reserved.

Green fiber lasers: An alternative to traditional DPSS green lasers for flow cytometry

PubMed Central

Telford, William G.; Babin, Sergey A.; Khorev, Serge V.; Rowe, Stephen H.

2009-01-01

Green and yellow diode-pumped solid state (DPSS) lasers (532 and 561 nm) have become common fixtures on flow cytometers, due to their efficient excitation of phycoerythrin (PE) and its tandems, and their ability to excite an expanding array of expressible red fluorescent proteins. Nevertheless, they have some disadvantages. DPSS 532 nm lasers emit very close to the fluorescein bandwidth, necessitating optical modifications to permit detection of fluorescein and GFP. DPSS 561 nm lasers likewise emit very close to the PE detection bandwidth, and also cause unwanted excitation of APC and its tandems, requiring high levels of crossbeam compensation to reduce spectral overlap into the PE tandems. In this paper, we report the development of a new generation of green fiber lasers that can be engineered to emit in the range between 532 and 561 nm. A 550 nm green fiber laser was integrated into both a BD LSR II™ cuvette and FACSVantage DiVa™ jet-in-air cell sorter. This laser wavelength avoided both the fluorescein and PE bandwidths, and provided better excitation of PE and the red fluorescent proteins DsRed and dTomato than a power-matched 532 nm source. Excitation at 550 nm also caused less incidental excitation of APC and its tandems, reducing the need for crossbeam compensation. Excitation in the 550 nm range therefore proved to be a good compromise between 532 and 561 nm sources. Fiber laser technology is therefore providing the flexibility necessary for precisely matching laser wavelengths to our flow cytometry applications. PMID:19777600

The impact of dialysis solution biocompatibility on ultrafiltration and on free water transport in rats.

PubMed

Aubertin, Gaëlle; Choquet, Philippe; Dheu, Céline; Constantinesco, André; Ratomponirina, Charline; Zaloszyc, Ariane; Passlick-Deetjen, Jutta; Fischbach, Michel

2012-01-01

This study compares different peritoneal dialysis fluids (PDF) in rats over a short contact time. For greater accuracy, net ultrafiltration (UF) and peritoneal transport indices, mass transfer area coefficient (MTAC) were scaled for the in vivo peritoneal surface area recruited (ivPSA) measured by microcomputerized tomography. Wistar rats underwent nephrectomy (5/6ths), were randomized into two groups and given 1.5% glucose PDF, either conventional acidic lactate (n = 14) or pH neutral bicarbonate (BicaVera) (n = 13); MTAC and UF were measured using a 90-min peritoneal equilibrium test (PET), fill volume (IPV) of 10 ml/100 g; small pore fluid transport was determined from sodium balance and used to calculate free water transport (FWT). Each ivPSA value was significantly correlated with the actual IPV, which varied from one rat to another. At 90 min of contact, there was no difference in recruited ivPSA in relation to PDFs. There was a difference (p < 0.01) in net UF/ivPSA 0.45 vs. 1.41 cm(2)/ml for bicarbonate versus lactate, as there was in the proportion of FWT with bicarbonate (42 ± 5% of net UF) compared to lactate (29 ± 4% of net UF). Net UF for individual values of ivPSA differs between conventional PDF and more biocompatible solutions, such as bicarbonate PDF. This observed change in UF cannot be fully explained by differences in glucose transport. The changes in FWT may be explained by the impact of the PDF biocompatibility on aquaporin function.

Binding of fluoresceinated epidermal growth factor to A431 cell sub-populations studied using a model-independent analysis of flow cytometric fluorescence data.

PubMed Central

Chatelier, R C; Ashcroft, R G; Lloyd, C J; Nice, E C; Whitehead, R H; Sawyer, W H; Burgess, A W

1986-01-01

A method is developed for determining ligand-cell association parameters from a model-free analysis of data obtained with a flow cytometer. The method requires measurement of the average fluorescence per cell as a function of ligand and cell concentration. The analysis is applied to data obtained for the binding of fluoresceinated epidermal growth factor to a human epidermoid carcinoma cell line, A431. The results indicate that the growth factor binds to two classes of sites on A431 cells: 4 X 10(4) sites with a dissociation constant (KD) of less than or equal to 20 pM, and 1.5 X 10(6) sites with a KD of 3.7 nM. A derived plot of the average fluorescence per cell versus the average number of bound ligands per cell is used to construct binding isotherms for four sub-populations of A431 cells fractionated on the basis of low-angle light scatter. The four sub-populations bind the ligand with equal affinity but differ substantially in terms of the number of binding sites per cell. We also use this new analysis to critically evaluate the use of 'Fluorotrol' as a calibration standard in flow cytometry. PMID:3015587

Calcium-Binding Proteins S100A8 and S100A9: Investigation of Their Immune Regulatory Effect in Myeloid Cells.

PubMed

Yang, Jianxin; Anholts, Jacqueline; Kolbe, Ulrike; Stegehuis-Kamp, Janine A; Claas, Frans H J; Eikmans, Michael

2018-06-21

High expression levels of the calcium-binding proteins S100A8 and S100A9 in myeloid cells in kidney transplant rejections are associated with a favorable outcome. Here we investigated the myeloid cell subset expressing these molecules, and their function in inflammatory reactions. Different monocyte subsets were sorted from buffy coats of healthy donors and investigated for S100A8 and S100A9 expression. To characterize S100A9high and S100A9low subsets within the CD14+ classical monocyte subset, intracellular S100A9 staining was combined with flow cytometry (FACS) and qPCR profiling. Furthermore, S100A8 and S100A9 were overexpressed by transfection in primary monocyte-derived macrophages and the THP-1 macrophage cell line to investigate the functional relevance. Expression of S100A8 and S100A9 was primarily found in classical monocytes and to a much lower extent in intermediate and non-classical monocytes. All S100A9+ cells expressed human leukocyte antigen—antigen D related (HLA-DR) on their surface. A small population (<3%) of CD14+ CD11b+ CD33+ HLA-DR− cells, characterized as myeloid derived suppressor cells (MDSCs), also expressed S100A9 to high extent. Overexpression of S100A8 and S00A9 in macrophages led to enhanced extracellular reactive oxygen species (ROS) production, as well as elevated mRNA expression of anti-inflammatory IL-10 . The results suggest that the calcium-binding proteins S100A8 and S100A9 in myeloid cells have an immune regulatory effect.

In vivo flow cytometry for blood cell analysis using differential epi-detection of forward scattered light

NASA Astrophysics Data System (ADS)

Paudel, Hari P.; Jung, Yookyung; Raphael, Anthony; Alt, Clemens; Wu, Juwell; Runnels, Judith; Lin, Charles P.

2018-02-01

The present standard of blood cell analysis is an invasive procedure requiring the extraction of patient's blood, followed by ex-vivo analysis using a flow cytometer or a hemocytometer. We are developing a noninvasive optical technique that alleviates the need for blood extraction. For in-vivo blood analysis we need a high speed, high resolution and high contrast label-free imaging technique. In this proceeding report, we reported a label-free method based on differential epi-detection of forward scattered light, a method inspired by Jerome Mertz's oblique back-illumination microscopy (OBM) (Ford et al, Nat. Meth. 9(12) 2012). The differential epi-detection of forward light gives phase contrast image at diffraction-limited resolution. Unlike reflection confocal microscopy (RCM), which detects only sharp refractive index variation and suffers from speckle noise, this technique is suitable for detection of subtle variation of refractive index in biological tissue and it provides the shape and the size of cells. A custom built high speed electronic detection circuit board produces a real-time differential signal which yields image contrast based on phase gradient in the sample. We recorded blood flow in-vivo at 17.2k lines per second in line scan mode, or 30 frames per second (full frame), or 120 frame per second (quarter frame) in frame scan mode. The image contrast and speed of line scan data recording show the potential of the system for noninvasive blood cell analysis.

Protistan Grazing Analysis by Flow Cytometry Using Prey Labeled by In Vivo Expression of Fluorescent Proteins

PubMed Central

Fu, Yutao; O'Kelly, Charles; Sieracki, Michael; Distel, Daniel L.

2003-01-01

Selective grazing by protists can profoundly influence bacterial community structure, and yet direct, quantitative observation of grazing selectivity has been difficult to achieve. In this investigation, flow cytometry was used to study grazing by the marine heterotrophic flagellate Paraphysomonas imperforata on live bacterial cells genetically modified to express the fluorescent protein markers green fluorescent protein (GFP) and red fluorescent protein (RFP). Broad-host-range plasmids were constructed that express fluorescent proteins in three bacterial prey species, Escherichia coli, Enterobacter aerogenes, and Pseudomonas putida. Micromonas pusilla, an alga with red autofluorescence, was also used as prey. Predator-prey interactions were quantified by using a FACScan flow cytometer and analyzed by using a Perl program described here. Grazing preference of P. imperforata was influenced by prey type, size, and condition. In competitive feeding trials, P. imperforata consumed algal prey at significantly lower rates than FP (fluorescent protein)-labeled bacteria of similar or different size. Within-species size selection was also observed, but only for P. putida, the largest prey species examined; smaller cells of P. putida were grazed preferentially. No significant difference in clearance rate was observed between GFP- and RFP-labeled strains of the same prey species or between wild-type and GFP-labeled strains. In contrast, the common chemical staining method, 5-(4,6-dichloro-triazin-2-yl)-amino fluorescein hydrochloride, depressed clearance rates for bacterial prey compared to unlabeled or RFP-labeled cells. PMID:14602649

Proinflammatory Proteins S100A8/S100A9 Activate NK Cells via Interaction with RAGE.

PubMed

Narumi, Kenta; Miyakawa, Reina; Ueda, Ryosuke; Hashimoto, Hisayoshi; Yamamoto, Yuki; Yoshida, Teruhiko; Aoki, Kazunori

2015-06-01

S100A8/A9, a proinflammatory protein, is upregulated in inflammatory diseases, and also has a tumor-promoting activity by the recruitment of myeloid cells and tumor cell invasion. However, whether the expression of S100A8/A9 in tumors predicts a good or poor prognosis is controversial in the clinical setting. In this study, to clarify the in vivo role of S100A8/A9 in the tumor microenvironment, we s.c. inoculated Pan02 cells stably expressing S100A8 and S100A9 proteins (Pan02-S100A8/A9) in syngeneic C57BL/6 mice. Unexpectedly, after small tumor nodules were once established, they rapidly disappeared. Flow cytometry showed that the number of NK cells in the tumors was increased, and an administration of anti-asialoGM1 Ab for NK cell depletion promoted the growth of Pan02-S100A8/A9 s.c. tumors. Although the S100A8/A9 proteins alone did not change the IFN-γ expression of NK cells in vitro, a coculture with Pan02 cells, which express Rae-1, induced IFN-γ production, and Pan02-S100A8/A9 cells further increased the number of IFN-γ(+) NK cells, suggesting that S100A8/A9 enhanced the NK group 2D ligand-mediated intracellular activation pathway in NK cells. We then examined whether NK cell activation by S100A8/A9 was via their binding to receptor of advanced glycation end product (RAGE) by using the inhibitors. RAGE antagonistic peptide and anti-RAGE Ab inhibited the IFN-γ production of NK cells induced by S100A8/A9 proteins, and an administration of FPS-ZM1, a RAGE inhibitor, significantly enhanced the in vivo growth of Pan02-S100A8/A9 tumors. We thus found a novel activation mechanism of NK cells via S100A8/A9-RAGE signaling, which may open a novel perspective on the in vivo interaction between inflammation and innate immunity. Copyright © 2015 by The American Association of Immunologists, Inc.

Quantum-dot-based suspension microarray for multiplex detection of lung cancer markers: preclinical validation and comparison with the Luminex xMAP® system

NASA Astrophysics Data System (ADS)

Bilan, Regina; Ametzazurra, Amagoia; Brazhnik, Kristina; Escorza, Sergio; Fernández, David; Uríbarri, María; Nabiev, Igor; Sukhanova, Alyona

2017-03-01

A novel suspension multiplex immunoassay for the simultaneous specific detection of lung cancer markers in bronchoalveolar lavage fluid (BALF) clinical samples based on fluorescent microspheres having different size and spectrally encoded with quantum dots (QDEM) was developed. The designed suspension immunoassay was validated for the quantitative detection of three lung cancer markers in BALF samples from 42 lung cancer patients and 10 control subjects. Tumor markers were detected through simultaneous formation of specific immune complexes consisting of a capture molecule, the target antigen, and biotinylated recognition molecule on the surface of the different QDEM in a mixture. The immune complexes were visualized by fluorescently labeled streptavidin and simultaneously analyzed using a flow cytometer. Preclinical validation of the immunoassay was performed and results were compared with those obtained using an alternative 3-plex immunoassay based on Luminex xMAP® technology, developed on classical organic fluorophores. The comparison showed that the QDEM and xMAP® assays yielded almost identical results, with clear discrimination between control and clinical samples. Thus, developed QDEM technology can become a good alternative to xMAP® assays permitting analysis of multiple protein biomarkers using conventional flow cytometers.

Quantum-dot-based suspension microarray for multiplex detection of lung cancer markers: preclinical validation and comparison with the Luminex xMAP® system

PubMed Central

Bilan, Regina; Ametzazurra, Amagoia; Brazhnik, Kristina; Escorza, Sergio; Fernández, David; Uríbarri, María; Nabiev, Igor; Sukhanova, Alyona

2017-01-01

A novel suspension multiplex immunoassay for the simultaneous specific detection of lung cancer markers in bronchoalveolar lavage fluid (BALF) clinical samples based on fluorescent microspheres having different size and spectrally encoded with quantum dots (QDEM) was developed. The designed suspension immunoassay was validated for the quantitative detection of three lung cancer markers in BALF samples from 42 lung cancer patients and 10 control subjects. Tumor markers were detected through simultaneous formation of specific immune complexes consisting of a capture molecule, the target antigen, and biotinylated recognition molecule on the surface of the different QDEM in a mixture. The immune complexes were visualized by fluorescently labeled streptavidin and simultaneously analyzed using a flow cytometer. Preclinical validation of the immunoassay was performed and results were compared with those obtained using an alternative 3-plex immunoassay based on Luminex xMAP® technology, developed on classical organic fluorophores. The comparison showed that the QDEM and xMAP® assays yielded almost identical results, with clear discrimination between control and clinical samples. Thus, developed QDEM technology can become a good alternative to xMAP® assays permitting analysis of multiple protein biomarkers using conventional flow cytometers. PMID:28300171

Electronic sputtering of LiF, CaF2, LaF3 and UF4 with 197 MeV Au ions. Is the stoichiometry of atom emission preserved?

NASA Astrophysics Data System (ADS)

Toulemonde, M.; Assmann, W.; Muller, D.; Trautmann, C.

2017-09-01

Sputtering experiments with swift heavy ions in the electronic energy loss regime were performed by using the catcher technique in combination with elastic recoil detection analysis. Four different fluoride targets, LiF, CaF2, LaF3 and UF4 were irradiated in the electronic energy loss regime using 197 MeV Au ions. The angular distribution of particles sputtered from the surface of freshly cleaved LiF and CaF2 single crystals is composed of a broad cosine distribution superimposed by a jet-like peak that appears perpendicular to the surface independent of the angle of beam incidence. For LiF, the particle emission in the entire angular distribution (jet plus broad cosine component) is stoichiometric, whereas for CaF2 the ratio of the sputtered F to Ca particles is at large angles by a factor of two smaller than the stoichiometry of the crystal. For single crystalline LaF3 no jet component is observed and the angular distribution is non-stoichiometric with the number of sputtered F particles being slightly larger than the number of sputtered La particles. In the case of UF4, the target was polycrystalline and had a much rougher surface compared to cleaved crystals. This destroys the appearance of a possible jet component leading to a broad angular distribution. The ratio of sputtered U atoms compared to F atoms is in the order of 1-2, i.e. the number of collected particles on the catcher is also non-stoichiometric. Such unlike behavior of particles sputtered from different fluoride crystals creates new questions.

An open-source solution for advanced imaging flow cytometry data analysis using machine learning.

PubMed

Hennig, Holger; Rees, Paul; Blasi, Thomas; Kamentsky, Lee; Hung, Jane; Dao, David; Carpenter, Anne E; Filby, Andrew

2017-01-01

Imaging flow cytometry (IFC) enables the high throughput collection of morphological and spatial information from hundreds of thousands of single cells. This high content, information rich image data can in theory resolve important biological differences among complex, often heterogeneous biological samples. However, data analysis is often performed in a highly manual and subjective manner using very limited image analysis techniques in combination with conventional flow cytometry gating strategies. This approach is not scalable to the hundreds of available image-based features per cell and thus makes use of only a fraction of the spatial and morphometric information. As a result, the quality, reproducibility and rigour of results are limited by the skill, experience and ingenuity of the data analyst. Here, we describe a pipeline using open-source software that leverages the rich information in digital imagery using machine learning algorithms. Compensated and corrected raw image files (.rif) data files from an imaging flow cytometer (the proprietary .cif file format) are imported into the open-source software CellProfiler, where an image processing pipeline identifies cells and subcellular compartments allowing hundreds of morphological features to be measured. This high-dimensional data can then be analysed using cutting-edge machine learning and clustering approaches using "user-friendly" platforms such as CellProfiler Analyst. Researchers can train an automated cell classifier to recognize different cell types, cell cycle phases, drug treatment/control conditions, etc., using supervised machine learning. This workflow should enable the scientific community to leverage the full analytical power of IFC-derived data sets. It will help to reveal otherwise unappreciated populations of cells based on features that may be hidden to the human eye that include subtle measured differences in label free detection channels such as bright-field and dark-field imagery

Acoustic Microfluidics for Bioanalytical Application

NASA Astrophysics Data System (ADS)

Lopez, Gabriel

2013-03-01

This talk will present new methods the use of ultrasonic standing waves in microfluidic systems to manipulate microparticles for the purpose of bioassays and bioseparations. We have recently developed multi-node acoustic focusing flow cells that can position particles into many parallel flow streams and have demonstrated the potential of such flow cells in the development of high throughput, parallel flow cytometers. These experiments show the potential for the creation of high throughput flow cytometers in applications requiring high flow rates and rapid detection of rare cells. This talk will also present the development of elastomeric capture microparticles and their use in acoustophoretic separations. We have developed simple methods to form elastomeric particles that are surface functionalized with biomolecular recognition reagents. These compressible particles exhibit negative acoustic contrast in ultrasound when suspended in aqueous media, blood serum or diluted blood. These particles can be continuously separated from cells by flowing them through a microfluidic device that uses an ultrasonic standing wave to align the blood cells, which exhibit positive acoustic contrast, at a node in the acoustic pressure distribution while aligning the negative acoustic contrast elastomeric particles at the antinodes. Laminar flow of the separated particles to downstream collection ports allows for collection of the separated negative contrast particles and cells. Separated elastomeric particles were analyzed via flow cytometry to demonstrate nanomolar detection for prostate specific antigen in aqueous buffer and picomolar detection for IgG in plasma and diluted blood samples. This approach has potential applications in the development of rapid assays that detect the presence of low concentrations of biomarkers (including biomolecules and cells) in a number of biological sample types. We acknowledge support through the NSF Research Triangle MRSEC.

Effect of perforation on flow past a conic cylinder at \\varvec{Re} = 100 : wavy vortex and sign laws

NASA Astrophysics Data System (ADS)

Lin, L. M.; Zhong, X. F.; Wu, Y. X.

2018-04-01

In order to find the intrinsic physical mechanism of the original Kármán vortex wavily distorted across the span due to the introduction of three-dimensional (3-D) geometric disturbances, a flow past a peak-perforated conic shroud is numerically simulated at a Reynolds number of 100. Based on previous work by Meiburg and Lasheras (1988), the streamwise and vertical interactions with spanwise vortices are introduced and analyzed. Then vortex-shedding patterns in the near wake for different flow regimes are reinspected and illustrated from the view of these two interactions. Generally, in regime I, spanwise vortices are a little distorted due to the weak interaction. Then in regime II, spanwise vortices, even though curved obviously, are still shed synchronously with moderate streamwise and vertical interactions. But in regime III, violently wavy spanwise vortices in some vortex-shedding patterns, typically an Ω -type vortex, are mainly attributed to the strong vertical interactions, while other cases, such as multiple vortex-shedding patterns in sub-regime III-D, are resulted from complex streamwise and vertical interactions. A special phenomenon, spacial distribution of streamwise and vertical components of vorticity with specific signs in the near wake, is analyzed based on two models of streamwise and vertical vortices in explaining physical reasons of top and bottom shear layers wavily varied across the span. Then these two models and above two interactions are unified. Finally two sign laws are summarized: the first sign law for streamwise and vertical components of vorticity is positive in the upper shear layer, but negative in the lower shear layer, while the second sign law for three vorticity components is always negative in the wake.

Uranium Enrichment Safeguards

DOE Office of Scientific and Technical Information (OSTI.GOV)

Demuth, Scott F.; Trahan, Alexis Chanel

2017-06-26

DIV of facility layout, material flows, and other information provided in the DIQ. Material accountancy through an annual PIV and a number of interim inventory verifications, including UF6 cylinder identification and counting, NDA of cylinders, and DA on a sample collection of UF6. Application of C/S technologies utilizing seals and tamper-indicating devices (TIDs) on cylinders, containers, storage rooms, and IAEA instrumentation to provide continuity of knowledge between inspection. Verification of the absence of undeclared material and operations, especially HEU production, through SNRIs, LFUA of cascade halls, and environmental swipe sampling

{100}<100> or 45.degree.-rotated {100}<100>, semiconductor-based, large-area, flexible, electronic devices

DOEpatents

Goyal, Amit [Knoxville, TN

2012-05-15

Novel articles and methods to fabricate the same resulting in flexible, {100}<100> or 45.degree.-rotated {100}<100> oriented, semiconductor-based, electronic devices are disclosed. Potential applications of resulting articles are in areas of photovoltaic devices, flat-panel displays, thermophotovoltaic devices, ferroelectric devices, light emitting diode devices, computer hard disc drive devices, magnetoresistance based devices, photoluminescence based devices, non-volatile memory devices, dielectric devices, thermoelectric devices and quantum dot laser devices.

Micron2 Lab: Microfluidic Microbiology Lab Project

NASA Technical Reports Server (NTRS)

Burton, Aaron; Botkin, Douglas; Castro, Sarah; Crucian, Brian

2015-01-01

Microbial monitoring during spaceflight is crucial to maintain crew health and ensure water purifications systems are functioning properly. Current protocols for in-flight enumeration of bacteria in potable water systems require culture based methods. In this project, we aim to develop a flight- and microgravity-compatible flow cytometer capable of counting total microbial counts in the water supply and differentiating live from dead bacteria.

Recent measurements concerning uranium hexafluoride-electron collision processes

NASA Technical Reports Server (NTRS)

Trajmar, S.; Chutjian, A.; Srivastava, S.; Williams, W.; Cartwright, D. C.

1976-01-01

Scattering of electrons by UF6 molecules was studied at impact energies ranging from 5 to 100 eV and momentum transfer, elastic and inelastic scattering cross sections were determined. The measurements also yielded spectroscopic information which made possible to extend the optical absorption cross sections from 2000 angstroms to 435 angstroms. It was found that UF6 is a very strong absorber in the vacuum UV region. No transitions were found to lie below the onset of the optically detected 3.0 eV feature.

Super-resolved calibration-free flow cytometric characterization of platelets and cell-derived microparticles in platelet-rich plasma.

PubMed

Konokhova, Anastasiya I; Chernova, Darya N; Moskalensky, Alexander E; Strokotov, Dmitry I; Yurkin, Maxim A; Chernyshev, Andrei V; Maltsev, Valeri P

2016-02-01

Importance of microparticles (MPs), also regarded as extracellular vesicles, in many physiological processes and clinical conditions motivates one to use the most informative and precise methods for their characterization. Methods based on individual particle analysis provide statistically reliable distributions of MP population over characteristics. Although flow cytometry is one of the most powerful technologies of this type, the standard forward-versus-side-scattering plots of MPs and platelets (PLTs) overlap considerably because of similarity of their morphological characteristics. Moreover, ordinary flow cytometry is not capable of measurement of size and refractive index (RI) of MPs. In this study, we 1) employed the potential of the scanning flow cytometer (SFC) for identification and characterization of MPs from light scattering; 2) suggested the reference method to characterize MP morphology (size and RI) with high precision; and 3) determined the lowest size of a MP that can be characterized from light scattering with the SFC. We equipped the SFC with 405 and 488 nm lasers to measure the light-scattering profiles and side scattering from MPs, respectively. The developed two-stage method allowed accurate separation of PLTs and MPs in platelet-rich plasma. We used two optical models for MPs, a sphere and a bisphere, in the solution of the inverse light-scattering problem. This solution provides unprecedented precision in determination of size and RI of individual spherical MPs-median uncertainties (standard deviations) were 6 nm and 0.003, respectively. The developed method provides instrument-independent quantitative information on MPs, which can be used in studies of various factors affecting MP population. © 2015 International Society for Advancement of Cytometry.

Heterogeneous distribution of antigens on human platelets demonstrated by fluorescence flow cytometry.

PubMed

Dunstan, R A; Simpson, M B

1985-12-01

We have used fluorescence flow cytometry to analyse cell-to-cell variability in the density of platelet ABH, Ii, Lewis, P, P1A1, Bak,a and HLA class I antigens. Human IgG and IgM antibodies were used in a two-stage assay with goat FITC-conjugated antihuman IgG (H&L) antibody as the label, followed by single cell analysis of 10 000 platelets per sample using a 256-channel fluorescence flow cytometer (Becton-Dickinson FACS Analyser). Computer analysis of fluorescence intensity histograms for mean and peak channel and coefficient of variation shows that the degree of heterogeneity in platelet antigen density varies with each particular blood group. The broad fluorescence distribution curves with oligosaccharide antigens (CVs: A = 53, B = 40, I = 44, Lea = 40, P = 40) indicate that these antigens possess a greater variability in the number of sites per cell compared to the more homogeneous distribution of P1,A1 BaK,a and HLA (CVs: P1A1 = 24, HLA = 30). These findings may partly account for the mechanism by which transfusion of ABO-incompatible platelets results in a biphasic survival curve, with a period of early rapid removal of those platelets with a high density of antigen sites, followed by a relatively normal survival curve for those platelets that possess only a few or no antigen sites. In contrast, P1A1 and HLA sites are less variable in number from one platelet to another in a given donor, and immune-mediated removal would be more likely to approximate a single exponential curve.

Scattering of phytoplankton cells from cytometry during a microcosm experiment

NASA Astrophysics Data System (ADS)

Moutier, W.; Duforêt-Gaurier, L.; Loisel, H.; Thyssen, M.; Mériaux, X.; Desailly, D.; Courcot, L.; Dugenne, M.

2016-02-01

This study presents an application of the CytoSense flow cytometer (CytoBuoy b.v., NL) as a powerful tool to analyze optical properties of phytoplankton cells. Recently, Duforêt et al., (2015) developed a methodology to derive the forward, sideward and backward cross section (σFWS, σSWS and σbb, respectively) of individual particles from the CytoSense. For the first time, this methodology was applied to phytoplankton cultures. A 20 day microcosm experiment was conducted on two phytoplankton species (Chlamydomonas concordia and Thalassiosira pseudonana). We realized daily sampling for biogeochemical and flow cytometer analysis and carried out optical measurements. Scanning electron migrographs (SEM) were performed at different life stages to investigate the cells morphology.First, CytoSense estimates were tested against radiative transfer computations. The comparison exercise, is based on radiative transfer simulations because for phytoplankton cultures, in situ measurements of σFWS and σSWS, particle by particle, are not available in literature. For that purpose, we build a database of 590,000 simulations, considering homogeneous and multi-layered spheres, to represent the optical properties of a large diversity of phytoplankton cells. Comparison showed that the CytoSense estimates for the cultures are consistent with values predicted by the theory. Second, the flow cytometer was used to analyze the temporal course of the forward and the sideward efficiency during the entire life-cycle. Results showed differences between the two species. From an ACP analysis, the variation of the optical properties were associated with the chlorophyll-a concentration by living cell, the thickness of the frustule and the aggregate formation. To finish, the bulk backscattering coefficient was rebuilt from σbb of individual cells and compare with the bb measured by a WET Labs ECO-BB9. Relative errors (RE) were between 0.3 and 0.47 and the mean RE was of 0.36. A such work shows

Immunophenotyping by slide-based cytometry and by flow cytometry are comparable

NASA Astrophysics Data System (ADS)

Gerstner, Andreas O.; Laffers, Wiebke; Mittag, Anja; Daehnert, Ingo; Lenz, Domnik; Bootz, Friedrich; Bocsi, Jozsef; Tarnok, Attila

2005-03-01

Immunophenotyping of peripheral blood leukocytes (PBLs) is performed by flow cytometry (FCM) as the golden standard. Slide based cytometry systems for example laser scanning cytometer (LSC) can give additional information (repeated staining and scanning, morphology). In order to adequately judge on the clinical usefulness of immunophenotyping by LSC it is obligatory to compare it with the long established FCM assays. We performed this study to systematically compare the two methods, FCM and LSC for immunophenotyping and to test the correlation of the results. Leucocytes were stained with directly labeled monoclonal antibodies with whole blood staining method. Aliquots of the same paraformaldehyde fixed specimens were analyzed in a FACScan (BD-Biosciences) using standard protocols and parallel with LSC (CompuCyte) after placing to glass slide, drying and fixation by aceton and 7-AAD staining. Calculating the percentage distribution of PBLs obtained by LSC and by FCM shows very good correlation with regression coefficients close to 1.0 for the major populations (neutrophils, lymphocytes, and monocytes), as well as for the lymphocyte sub-populations (T-helper-, T-cytotoxic-, B-, NK-cells). LSC can be recommended for immunophenotyping of PBLs especially in cases where only very limited sample volumes are available or where additional analysis of the cells" morphology is important. There are limitations in the detection of rare leucocytes or weak antigens where appropriate amplification steps for immunofluorescence should be engaged.

Recovery of process water from spent emulsions generated in copper cable factory.

PubMed

Karakulski, K; Morawski, A W

2011-02-28

Treatment of waste emulsions generated in the cable factory from copper wire drawing was investigated using the integrated membrane processes: ultrafiltration (UF) and nanofiltration (NF). The application of UF tubular membranes (MWCO 100 kDa) resulted in 98% retention of oil and lubricants, whereas the degree of passage of copper ions (the major component of effluents from cable factory) was 99%. The average permeate flux amounted to 45 l/m(2) h for the transmembrane pressure of 3.5 bar during the UF pretreatment of waste emulsions. The Silt Density Index (SDI) values of UF permeates were appropriate for the application of spiral wound membranes in the NF process. The complete removal of oil and lubricants was achieved in NF process and the content of TOC was reduced by more than 90%. The rejection of copper ions in the NF process was 90% and 98% for NF270 and NF90 membranes (FILMTEC), respectively. The quality of NF permeates allows a direct reuse of treated water for the preparation of fresh emulsion. Copyright © 2010 Elsevier B.V. All rights reserved.

Solution of a large hydrodynamic problem using the STAR-100 computer

NASA Technical Reports Server (NTRS)

Weilmuenster, K. J.; Howser, L. M.

1976-01-01

A representative hydrodynamics problem, the shock initiated flow over a flat plate, was used for exploring data organizations and program structures needed to exploit the STAR-100 vector processing computer. A brief description of the problem is followed by a discussion of how each portion of the computational process was vectorized. Finally, timings of different portions of the program are compared with equivalent operations on serial machines. The speed up of the STAR-100 over the CDC 6600 program is shown to increase as the problem size increases. All computations were carried out on a CDC 6600 and a CDC STAR 100, with code written in FORTRAN for the 6600 and in STAR FORTRAN for the STAR 100.

The Sperm Chromatin Structure Assay (SCSA(®)) and other sperm DNA fragmentation tests for evaluation of sperm nuclear DNA integrity as related to fertility.

PubMed

Evenson, Donald P

2016-06-01

Thirty-five years ago the pioneering paper in Science (240:1131) on the relationship between sperm DNA integrity and pregnancy outcome was featured as the cover issue showing a fluorescence photomicrograph of red and green stained sperm. The flow cytometry data showed a very significant difference in sperm DNA integrity between fertile and subfertile bulls and men. This study utilized heat (100°C, 5min) to denature DNA at sites of DNA strand breaks followed by staining with acridine orange (AO) and measurements of 5000 individual sperm of green double strand (ds) DNA and red single strand (ss) DNA fluorescence. Later, the heat protocol was changed to a low pH protocol to denature the DNA at sites of strand breaks; the heat and acid procedures produced the same results. SCSA data are very advantageously dual parameter with 1024 channels (degrees) of both red and green fluorescence. Hundreds of publications on the use of the SCSA test in animals and humans have validated the SCSA as a highly useful test for determining male breeding soundness. The SCSA test is a rapid, non-biased flow cytometer machine measurement providing robust statistical data with exceptional precision and repeatability. Many genotoxic experiments showed excellent dose response data with very low coefficient of variation that further validated the SCSA as being a highly powerful assay for sperm DNA integrity. Twelve years following the introduction of the SCSA test, the terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labelling (TUNEL) test (1993) for sperm was introduced as the only other flow cytometric assay for sperm DNA fragmentation. However, the TUNEL test can also be done by light microscopy with much less statistical robustness. The COMET (1998) and Sperm Chromatin Dispersion (SCD; HALO) (2003) tests were introduced as light microscope tests that don't require a flow cytometer. Since these tests measure only 50-200 sperm per sample, they suffer from the lack of

21 CFR 100.100 - Misleading containers.

Code of Federal Regulations, 2011 CFR

2011-04-01

... presentation of the food and has value which is both significant in proportion to the value of the product and... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Misleading containers. 100.100 Section 100.100 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD...

Design flow duration curves for environmental flows estimation in Damodar River Basin, India

NASA Astrophysics Data System (ADS)

Verma, Ravindra Kumar; Murthy, Shankar; Verma, Sangeeta; Mishra, Surendra Kumar

2017-06-01

In this study, environmental flows (EFs) are estimated for six watersheds of Damodar River Basin (DRB) using flow duration curve (FDC) derived using two approaches: (a) period of record and (b) stochastic approaches for daily, 7-, 30-, 60-day moving averages, and 7-daily mean annual flows observed at Tenughat dam, Konar dam, Maithon dam, Panchet dam, Damodar bridge, Burnpur during 1981-2010 and at Phusro during 1988-2010. For stochastic FDCs, 7-day FDCs for 10, 20-, 50- and 100-year return periods were derived for extraction of discharge values at every 5% probability of exceedance. FDCs derived using the first approach show high probability of exceedance (5-75%) for the same discharge values. Furthermore, discharge values of 60-day mean are higher than those derived using daily, 7-, and 30-day mean values. The discharge values of 95% probability of exceedance (Q95) derived from 7Q10 (ranges from 2.04 to 5.56 cumec) and 7Q100 (ranges from 3.4 to 31.48 cumec) FDCs using the second approach are found more appropriate as EFs during drought/low flow and normal precipitation years.

Effects of Nitroglycerin on Regional Myocardial Blood Flow in Coronary Artery Disease

PubMed Central

Horwitz, Lawrence D.; Gorlin, Richard; Taylor, Warren J.; Kemp, Harvey G.

1971-01-01

Regional myocardial blood flow before and after sublingual nitroglycerin was measured in 10 patients with coronary artery disease. During thoracotomy, 133Xe was injected directly into the subepicardium in diseased regions of the anterior left ventricular wall, and washout rates were recorded with a scintillation counter. All disappearance curves were closely approximated by two exponential decays analyzed as two parallel flow systems by the compartmental method. The appearance of a double exponential decay pattern in diseased regions suggests that the slow phase was associated with collateral blood flow, although nonhomogeneous myocardium-to-blood partition coefficients for xenon cannot be excluded. Nitroglycerin increased the rapid phase flow in 9 of 10 patients and the slow flow in 7 of 10 patients. Average flow increased in 9 of the 10 patients (P < 0.01). Mean rapid phase flow in the control state was 110 ml/100 g per min and after nitroglycerin increased to 132 ml/100 g per min (P < 0.01); slow phase flow increased from 12 ml/100 g per min to 15 ml/100 g per min (P < 0.05). It is concluded that, under these conditions, nitroglycerin improves perfusion in regions of diseased myocardium in patients with coronary artery disease. PMID:4999635

The UF/NCI family of hybrid computational phantoms representing the current US population of male and female children, adolescents, and adults—application to CT dosimetry

NASA Astrophysics Data System (ADS)

Geyer, Amy M.; O'Reilly, Shannon; Lee, Choonsik; Long, Daniel J.; Bolch, Wesley E.

2014-09-01

Substantial increases in pediatric and adult obesity in the US have prompted a major revision to the current UF/NCI (University of Florida/National Cancer Institute) family of hybrid computational phantoms to more accurately reflect current trends in larger body morphometry. A decision was made to construct the new library in a gridded fashion by height/weight without further reference to age-dependent weight/height percentiles as these become quickly outdated. At each height/weight combination, circumferential parameters were defined and used for phantom construction. All morphometric data for the new library were taken from the CDC NHANES survey data over the time period 1999-2006, the most recent reported survey period. A subset of the phantom library was then used in a CT organ dose sensitivity study to examine the degree to which body morphometry influences the magnitude of organ doses for patients that are underweight to morbidly obese in body size. Using primary and secondary morphometric parameters, grids containing 100 adult male height/weight bins, 93 adult female height/weight bins, 85 pediatric male height/weight bins and 73 pediatric female height/weight bins were constructed. These grids served as the blueprints for construction of a comprehensive library of patient-dependent phantoms containing 351 computational phantoms. At a given phantom standing height, normalized CT organ doses were shown to linearly decrease with increasing phantom BMI for pediatric males, while curvilinear decreases in organ dose were shown with increasing phantom BMI for adult females. These results suggest that one very useful application of the phantom library would be the construction of a pre-computed dose library for CT imaging as needed for patient dose-tracking.

Intravenous fluid flow meter concept for zero gravity environment

NASA Technical Reports Server (NTRS)

Miller, C. G.

1972-01-01

Measuring chamber, included in infusion-set tubing, and peristaltic flow meter concept can be incorporated into flow meter that measures fluid flow rates between 100 and 600 cu cm per hour and at the same time maintains sterilization.

Work Function of Oxide Ultrathin Films on the Ag(100) Surface.

PubMed

Sementa, Luca; Barcaro, Giovanni; Negreiros, Fabio R; Thomas, Iorwerth O; Netzer, Falko P; Ferrari, Anna Maria; Fortunelli, Alessandro

2012-02-14

Theoretical calculations of the work function of monolayer (ML) and bilayer (BL) oxide films on the Ag(100) surface are reported and analyzed as a function of the nature of the oxide for first-row transition metals. The contributions due to charge compression, charge transfer and rumpling are singled out. It is found that the presence of empty d-orbitals in the oxide metal can entail a charge flow from the Ag(100) surface to the oxide film which counteracts the decrease in the work function due to charge compression. This flow can also depend on the thickness of the film and be reduced in passing from ML to BL systems. A regular trend is observed along first-row transition metals, exhibiting a maximum for CuO, in which the charge flow to the oxide is so strong as to reverse the direction of rumpling. A simple protocol to estimate separately the contribution due to charge compression is discussed, and the difference between the work function of the bare metal surface and a Pauling-like electronegativity of the free oxide slabs is used as a descriptor quantity to predict the direction of charge transfer.

Earth observation image of Von Karman Vortices taken during STS-100

NASA Image and Video Library

2001-04-30

STS100-710-182 (19 April-1 May 2001) --- Easily recognizable in this STS-100 70mm frame are phenomena known as the von Karman vortices, which were forming in the stratocumulus field downwind from the volcanic island of Rishiri-to in the northern Sea of Japan off the northwest coast of Hokkaido, Japan. According to NASA scientists studying the STS-100 photo collection, these features form when a stable atmosphere with low clouds flows past a relatively small obstacle with a height greater than the depth of the cloud layer. Because the atmosphere is stable, a series of eddies form in the moving cloud field.

Multiparameter flow cytometry of a pH sensitive ligand bound to receptors and inside cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fay, S.P.; Habbersett, R.; Posner, R.G.

1993-01-01

Because fluoresceinated ligands of the neutrophil formyl peptide receptor can be protonated either upon binding to the receptor on the cell surface or in acidified intracellular compartments, the authors synthesized a ligand conjugated to the pH sensitive fluorescent probe SNAFL (CHO-Met-Leu-Phe-Phe-Lys-SNAFL). In the three laser flow cytometer at LANL, protonated dye is excited at 488 nm and emits at 530 nm; unprotonated dye is excited at 568 nm and emits at 650 nm. Detection at the isobestic and isoemissive points at 528 and 600 nm is used to keep track of variations in ligand concentration from sample to sample. Themore » SNAFL-ligand bound to HL-60 cells (which overexpress the formyl peptide receptor) was compared to the free ligand in solution over a pH range from 6.5 to 9.0. The results suggest that the ligand bound to cell surface receptors was protonated in the binding pocket, possibly by virtue of its proximity to His 90, based on sequence data. When the cells were raised from 4[degrees] to 37[degrees], they also observed a time-dependent acidification of the ligand, indicative of ligand-receptor processing beginning 3-4 minutes after internalization.« less

Comparison of traditional and molecular analytical methods for detecting biological agents in raw and drinking water following ultrafiltration

USGS Publications Warehouse

Francy, D.S.; Bushon, R.N.; Brady, A.M.G.; Bertke, E.E.; Kephart, C.M.; Likirdopulos, C.A.; Mailot, B.E.; Schaefer, F. W.; Lindquist, H.D. Alan

2009-01-01

Aims: To compare the performance of traditional methods to quantitative polymerase chain reaction (qPCR) for detecting five biological agents in large-volume drinking-water samples concentrated by ultrafiltration (UF). Methods and Results: Drinking-water samples (100 l) were seeded with Bacillus anthracis, Cryptospordium parvum, Francisella tularensis, Salmonella Typhi, and Vibrio cholerae and concentrated by UF. Recoveries by traditional methods were variable between samples and between some replicates; recoveries were not determined by qPCR. Francisella tularensis and V. cholerae were detected in all 14 samples after UF, B. anthracis was detected in 13, and C. parvum was detected in 9 out of 14 samples. Numbers found by qPCR after UF were significantly or nearly related to those found by traditional methods for all organisms except for C. parvum. A qPCR assay for S. Typhi was not available. Conclusions: qPCR can be used to rapidly detect biological agents after UF as well as traditional methods, but additional work is needed to improve qPCR assays for several biological agents, determine recoveries by qPCR, and expand the study to other areas. Significance and Impact of the Study: To our knowledge, this is the first study to compare the use of traditional and qPCR methods to detect biological agents in large-volume drinking-water samples. ?? 2009 The Society for Applied Microbiology.

Berberine displays antitumor activity in esophageal cancer cells in vitro.

PubMed

Jiang, Shu-Xian; Qi, Bo; Yao, Wen-Jian; Gu, Cheng-Wei; Wei, Xiu-Feng; Zhao, Yi; Liu, Yu-Zhen; Zhao, Bao-Sheng

2017-04-14

To investigate the effects of berberine on esophageal cancer (EC) cells and its molecular mechanisms. Human esophageal squamous cell carcinoma cell line KYSE-70 and esophageal adenocarcinoma cell line SKGT4 were used. The effects of berberine on cell proliferation were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. For cell cycle progression, KYSE-70 cells were stained with propidium iodide (PI) staining buffer (10 mg/mL PI and 100 mg/mL RNase A) for 30 min and cell cycle was analyzed using a BD FACSCalibur flow cytometer. For apoptosis assay, cells were stained with an Annexin V-FITC/PI apoptosis detection kit. The rate of apoptotic cells was analyzed using a dual laser flow cytometer and estimated using BD ModFit software. Levels of proteins related to cell cycle and apoptosis were examined by western blotting. Berberine treatment resulted in growth inhibition of KYSE-70 and SKGT4 cells in a dose-dependent and time-dependent manner. KYSE-70 cells were more susceptible to the inhibitory activities of berberine than SKGT4 cells were. In KYSE-70 cells treated with 50 μmol/L berberine for 48 h, the number of cells in G 2 /M phase (25.94% ± 5.01%) was significantly higher than that in the control group (9.77% ± 1.28%, P < 0.01), and berberine treatment resulted in p21 up-regulation in KYSE-70 cells. Flow cytometric analyses showed that berberine significantly augmented the KYSE-70 apoptotic population at 12 and 24 h post-treatment, when compared with control cells (0.83% vs 43.78% at 12 h, P < 0.05; 0.15% vs 81.86% at 24 h, P < 0.01), and berberine-induced apoptotic effect was stronger at 24 h compared with 12 h. Western blotting showed that berberine inhibited the phosphorylation of Akt, mammalian target of rapamycin and p70S6K, and enhanced AMP-activated protein kinase phosphorylation in a sustained manner. Berberine is an inhibitor of human EC cell growth and could be considered as a potential drug for the treatment

Berberine displays antitumor activity in esophageal cancer cells in vitro

PubMed Central

Jiang, Shu-Xian; Qi, Bo; Yao, Wen-Jian; Gu, Cheng-Wei; Wei, Xiu-Feng; Zhao, Yi; Liu, Yu-Zhen; Zhao, Bao-Sheng

2017-01-01

AIM To investigate the effects of berberine on esophageal cancer (EC) cells and its molecular mechanisms. METHODS Human esophageal squamous cell carcinoma cell line KYSE-70 and esophageal adenocarcinoma cell line SKGT4 were used. The effects of berberine on cell proliferation were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. For cell cycle progression, KYSE-70 cells were stained with propidium iodide (PI) staining buffer (10 mg/mL PI and 100 mg/mL RNase A) for 30 min and cell cycle was analyzed using a BD FACSCalibur flow cytometer. For apoptosis assay, cells were stained with an Annexin V-FITC/PI apoptosis detection kit. The rate of apoptotic cells was analyzed using a dual laser flow cytometer and estimated using BD ModFit software. Levels of proteins related to cell cycle and apoptosis were examined by western blotting. RESULTS Berberine treatment resulted in growth inhibition of KYSE-70 and SKGT4 cells in a dose-dependent and time-dependent manner. KYSE-70 cells were more susceptible to the inhibitory activities of berberine than SKGT4 cells were. In KYSE-70 cells treated with 50 μmol/L berberine for 48 h, the number of cells in G2/M phase (25.94% ± 5.01%) was significantly higher than that in the control group (9.77% ± 1.28%, P < 0.01), and berberine treatment resulted in p21 up-regulation in KYSE-70 cells. Flow cytometric analyses showed that berberine significantly augmented the KYSE-70 apoptotic population at 12 and 24 h post-treatment, when compared with control cells (0.83% vs 43.78% at 12 h, P < 0.05; 0.15% vs 81.86% at 24 h, P < 0.01), and berberine-induced apoptotic effect was stronger at 24 h compared with 12 h. Western blotting showed that berberine inhibited the phosphorylation of Akt, mammalian target of rapamycin and p70S6K, and enhanced AMP-activated protein kinase phosphorylation in a sustained manner. CONCLUSION Berberine is an inhibitor of human EC cell growth and could be considered as a

The aerodynamic performance of several flow control devices for internal flow systems

NASA Technical Reports Server (NTRS)

Eckert, W. T.; Wettlaufer, B. M.; Mort, K. W.

1982-01-01

An experimental reseach and development program was undertaken to develop and document new flow-control devices for use in the major modifications to the 40 by 80 Foot wind tunnel at Ames Research Center. These devices, which are applicable to other facilities as well, included grid-type and quasi-two-dimensional flow straighteners, louver panels for valving, and turning-vane cascades with net turning angles from 0 deg to 90 deg. The tests were conducted at model scale over a Reynolds number range from 2 x 100,000 to 17 x 100,000, based on chord. The results showed quantitatively the performance benefits of faired, low-blockage, smooth-surface straightener systems, and the advantages of curved turning-vanes with hinge-line gaps sealed and a preferred chord-to-gap ratio between 2.5 and 3.0 for 45 deg or 90 deg turns.

Ultrafiltration membrane for effective removal of chromium ions from potable water

NASA Astrophysics Data System (ADS)

Muthumareeswaran, M. R.; Alhoshan, Mansour; Agarwal, Gopal Prasad

2017-01-01

The objective of the present work was to investigate the efficacy of indigenously developed polyacrylonitrile (PAN) based ultrafiltration (UF) membrane for chromium ions removal from potable water. The hydrolyzed PAN membranes effectively rejected chromium anions in the feed ranging from 250 ppb to 400 ppm and a rejection of ≥90% was achieved for pH ≥ 7 at low chromate concentration (≤25 ppm) in feed. The rejection mechanism of chromium ions was strongly dependent on Donnan exclusion principle, while size exclusion principle for UF did not play a major role on ions rejection. Feed pH played a vital role in changing porosity of membrane, which influenced the retention behavior of chromate ions. Cross-flow velocity, pressure did not play significant role for ions rejection at low feed concentration. However, at higher feed concentration (≥400 ppm), concentration polarization became important and it reduced the chromate rejection to 32% at low cross flow and high pressure. Donnan steric-partitioning pore and dielectric exclusion model (DSPM-DE) was applied to evaluate the chromate ions transport through PAN UF membrane as a function of flux by using optimized model parameters and the simulated data matched well with experimental results.

Single cell analysis using surface enhanced Raman scattering (SERS) tags

PubMed Central

Nolan, John P.; Duggan, Erika; Liu, Er; Condello, Danilo; Dave, Isha; Stoner, Samuel A.

2013-01-01

Fluorescence is a mainstay of bioanalytical methods, offering sensitive and quantitative reporting, often in multiplexed or multiparameter assays. Perhaps the best example of the latter is flow cytometry, where instruments equipped with multiple lasers and detectors allow measurement of 15 or more different fluorophores simultaneously, but increases beyond this number are limited by the relatively broad emission spectra. Surface enhanced Raman scattering (SERS) from metal nanoparticles can produce signal intensities that rival fluorescence, but with narrower spectral features that allow a greater degree of multiplexing. We are developing nanoparticle SERS tags as well as Raman flow cytometers for multiparameter single cell analysis of suspension or adherent cells. SERS tags are based on plasmonically active nanoparticles (gold nanorods) whose plasmon resonance can be tuned to give optimal SERS signals at a desired excitation wavelength. Raman resonant compounds are adsorbed on the nanoparticles to confer a unique spectral fingerprint on each SERS tag, which are then encapsulated in a polymer coating for conjugation to antibodies or other targeting molecules. Raman flow cytometry employs a high resolution spectral flow cytometer capable of measuring the complete SERS spectra, as well as conventional flow cytometry measurements, from thousands of individual cells per minute. Automated spectral unmixing algorithms extract the contributions of each SERS tag from each cell to generate high content, multiparameter single cell population data. SERS-based cytometry is a powerful complement to conventional fluorescence-based cytometry. The narrow spectral features of the SERS signal enables more distinct probes to be measured in a smaller region of the optical spectrum with a single laser and detector, allowing for higher levels of multiplexing and multiparameter analysis. PMID:22498143

Preliminary study of the use of the STAR-100 computer for transonic flow calculations

NASA Technical Reports Server (NTRS)

Keller, J. D.; Jameson, A.

1977-01-01

An explicit method for solving the transonic small-disturbance potential equation is presented. This algorithm, which is suitable for the new vector-processor computers such as the CDC STAR-100, is compared to successive line over-relaxation (SLOR) on a simple test problem. The convergence rate of the explicit scheme is slower than that of SLOR, however, the efficiency of the explicit scheme on the STAR-100 computer is sufficient to overcome the slower convergence rate and allow an overall speedup compared to SLOR on the CYBER 175 computer.

Detection of Apoptosis and Necrosis in Normal Human Lung Cells Using 1H NMR Spectroscopy

NASA Astrophysics Data System (ADS)

Shih, Chwen-Ming; Ko, Wun-Chang; Yang, Liang-Yo; Lin, Chien-Ju; Wu, Jui-Sheng; Lo, Tsui-Yun; Wang, Shwu-Huey; Chen, Chien-Tsu

2005-05-01

This study aimed to detect apoptosis and necrosis in MRC-5, a normal human lung cell line, by using noninvasive proton nuclear magnetic resonance (1H NMR). Live MRC-5 cells were processed first for 1H NMR spectroscopy; subsequently their types and the percentage of cell death were assessed on a flow cytometer. Cadmium (Cd) and mercury (Hg) induced apoptosis and necrosis in MRC-5 cells, respectively, as revealed by phosphatidylserine externalization on a flow cytometer. The spectral intensity ratio of methylene (CH2) resonance (at 1.3 ppm) to methyl (CH3) resonance (at 0.9 ppm) was directly proportional to the percentage of apoptosis and strongly and positively correlated with PI staining after Cd treatment (r2 = 0.9868, P < 0.01). In contrast, this ratio only increased slightly within 2-h Hg treatment, and longer Hg exposure failed to produce further increase. Following 2-h Hg exposure, the spectral intensity of choline resonance (at 3.2 ppm) was abolished, but this phenomenon was absent in Cd-induced apoptosis. These findings together demonstrate that 1H NMR is a novel tool with a quantitative potential to distinguish apoptosis from necrosis as early as the onset of cell death in normal human lung cells.

Qualitative spectroscopic study of magnetic nozzle flow

NASA Technical Reports Server (NTRS)

Umeki, T.; Turchi, P. J.

1992-01-01

The physics of the magnetic nozzle flow for a 100-kW-level quasi-steady MPD thruster was studied by photographic spectroscopy focusing on the plasma model in the flow and the acceleration mechanism. Spectroscopic visualization for the flow-species analysis indicates that the plasma-exhaust flow dominated by NII species were confined by the magnetic nozzle effect to collimate the flow for the better thruster performance. Inside the nozzle, the plasma flow was found to be in nonhomogeneous collisional-radiative condition. There appears to be a substantial flow acceleration from the magnetic nozzle inlet to the outlet with slight expansion. This suggests that the flow resembles that of constant area supersonic duct flow with cooling.

14 CFR 29.955 - Fuel flow.

Code of Federal Regulations, 2014 CFR

2014-01-01

... STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Powerplant Fuel System § 29.955 Fuel flow. (a) General. The fuel system for each engine must provide the engine with at least 100 percent of the fuel required under all... flow transmitter, if installed, and the critical fuel pump (for pump-fed systems) must be installed to...

14 CFR 29.955 - Fuel flow.

Code of Federal Regulations, 2013 CFR

2013-01-01

... STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Powerplant Fuel System § 29.955 Fuel flow. (a) General. The fuel system for each engine must provide the engine with at least 100 percent of the fuel required under all... flow transmitter, if installed, and the critical fuel pump (for pump-fed systems) must be installed to...

14 CFR 29.955 - Fuel flow.

Code of Federal Regulations, 2012 CFR

2012-01-01

... STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Powerplant Fuel System § 29.955 Fuel flow. (a) General. The fuel system for each engine must provide the engine with at least 100 percent of the fuel required under all... flow transmitter, if installed, and the critical fuel pump (for pump-fed systems) must be installed to...

14 CFR 29.955 - Fuel flow.

Code of Federal Regulations, 2010 CFR

2010-01-01

... STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Powerplant Fuel System § 29.955 Fuel flow. (a) General. The fuel system for each engine must provide the engine with at least 100 percent of the fuel required under all... flow transmitter, if installed, and the critical fuel pump (for pump-fed systems) must be installed to...

14 CFR 29.955 - Fuel flow.

Code of Federal Regulations, 2011 CFR

2011-01-01

... STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Powerplant Fuel System § 29.955 Fuel flow. (a) General. The fuel system for each engine must provide the engine with at least 100 percent of the fuel required under all... flow transmitter, if installed, and the critical fuel pump (for pump-fed systems) must be installed to...

[Evaluation of the performances of the UF-1000i(®) automated urine analyzer].

PubMed

Fabbro, Christelle; Darolles, Jacques; Rault, Jean-Philippe

2011-01-01

The purpose of this study was the evaluation of the UF 1000i(®) automated urine analyzer (bioMérieux(©)).The coefficients of variation (CV) for the repeatability of the red blood cell (RBC), white blood cell (WBC) and bacteria counts were overall concordant with those announced by the supplier. However, for low concentrations, the CV for the repeatability for concentrations of 10(3) RBC/mL, 10(3) WBC/mL and 5×10(3) UA/mL were respectively of 26, 18 and 36% and thus higher than the CV (10%) reported for each of the three parameters by the supplier. Reproducibility results agreed with those given by the supplier (10%). The linearity range was different from that reported by the supplier and was shifted by a factor 10 for WBC and bacteria high concentrations. Cross contamination between samples was prevented by using the washing program recommended by the supplier which however led to a lower analysis frequency (80 samples per hour). Detection limits were of 5,7×10(1) RBC/mL, 5,7×10(1) WBC/mL and 1,6×10(4) UA/mL respectively for the RBC, WBC and bacteria. Quantification limits found in this study were of 1,3×10(3) RBC/mL, 1,7×10(3) WBC/mL, 5,1×10(3) UA/mL. An overestimation of the RBC count was observed within the range of 5×10(3) to 2×10(4) RBC/mL. Beyond this concentration range, the concordance is good. The correlation is poor within this range and good for higher concentrations. For WBC, concordance and correlation were satisfactory over the whole range tested.

Measuring PM2.5, Ultrafine Particles, Nicotine Air and Wipe Samples Following the Use of Electronic Cigarettes.

PubMed

Melstrom, Paul; Koszowski, Bartosz; Thanner, Meridith Hill; Hoh, Eunha; King, Brian; Bunnell, Rebecca; McAfee, Tim

2017-09-01

Few studies have examined the extent of inhalation or dermal contact among bystanders following short-term, secondhand e-cigarette exposure. Measure PM2.5 (particles < 2.5 microns), UF (ultrafine particles < 100 nm), and nicotine in air and deposited on surfaces and clothing pre-/during/post- a short-term (2-hour) e-cigarette exposure. E-cigarettes were used ad libitum by three experienced users for 2 hours during two separate sessions (disposable e-cigarettes, then tank-style e-cigarettes, or "tanks") in a 1858 ft3 room. We recorded: uncorrected PM2.5 (using SidePak); UF (using P-Trak); air nicotine concentrations (using air samplers; SKC XAD-4 canisters); ambient air exchange rate (using an air capture hood). Wipe samples were taken by wiping 100 cm2 room surfaces pre- and post- both sessions, and clean cloth wipes were worn during the exposure and collected at the end. Uncorrected PM2.5 and UF were higher (p < .0001) during sessions than before or after. Median PM2.5 during exposure was higher using tanks (0.515 mg/m3) than disposables (0.035 mg/m3) (p < .0001). Median UF during exposure was higher using disposables (31 200 particles/cm3) than tanks (25 200 particles/cm3)(p < .0001). Median air nicotine levels were higher (p < .05) during both sessions (disposables = 0.697 ng/L, tanks = 1.833 ng/L) than before (disposables = 0.004 ng/L, tanks = 0.010 ng/L) or after (disposables = 0.115 ng/L, tanks = 0.147 ng/L). Median accumulation rates of nicotine on surface samples were 2.1 ng/100 cm2/h using disposables and 4.0 ng/100 cm2/h using tanks; for cloth samples, it was 44.4 ng/100 cm2/h using disposables and 69.6 ng/100 cm2/h using tanks (p < .01). Mean room ventilation rate was ~5 air changes per hour during both sessions. Short-term e-cigarette use can produce: elevated PM2.5; elevated UF; nicotine in the air; and accumulation of nicotine on surfaces and clothing. Short-term indoor e-cigarette use produced accumulation of nicotine on surfaces and clothing, which

Quantification of chemotaxis during pediatric cardiac surgery by flow and laser scanning cytometry

NASA Astrophysics Data System (ADS)

Tarnok, Attila; Schmid, Joerg W.; Osmancik, Pavel; Lenz, Dominik; Pipek, Michal; Hambsch, Joerg; Gerstner, Andreas O.; Schneider, Peter

2002-05-01

Cardiac surgery with cardiopulmonary bypass (CPB) alters the leukocyte composition of the peripheral blood (PB). This response contributes to the sometimes adverse outcome with capillary leakage. Migration of activated cells to sites of inflammation, driven by chemokines is part of this response. In order to determine the chemotactic activity of patients serum during and after surgery we established an assay for PB leukocytes (PBL). PBL from healthy donors were isolated and 250,000 cells were placed into a migration chamber separated by a filter from a second lower chamber filled with patient serum. After incubation cells from top and bottom chamber were removed and stained with a cocktail of monoclonal antibodies for leukocyte subsets and analyzed on a flow cytometer (FCM). Cells at the bottom of the filter belong to the migrating compartment and were quantified by LSC after staining of nucleated cells. Increased chemotactic activity started at onset of anaesthesia followed by a phase of low activity immediately after surgery and a second phase of a high post-operative activity. The in vitro results correlated with results obtained by immunopenotyping of circulating PBL. Manipulation of the chemokine pattern might prove beneficial to prevent extravasation of cells leading to tissue damage. In chemotaxis assays with low amount of available serum the combined use of FCM and Laser Scanning LSC proved as an appropriate analytical tool.

Gaseous slip flow analysis of a micromachined flow sensor for ultra small flow applications

NASA Astrophysics Data System (ADS)

Jang, Jaesung; Wereley, Steven T.

2007-02-01

The velocity slip of a fluid at a wall is one of the most typical phenomena in microscale gas flows. This paper presents a flow analysis considering the velocity slip in a capacitive micro gas flow sensor based on pressure difference measurements along a microchannel. The tangential momentum accommodation coefficient (TMAC) measurements of a particular channel wall in planar microchannels will be presented while the previous micro gas flow studies have been based on the same TMACs on both walls. The sensors consist of a pair of capacitive pressure sensors, inlet/outlet and a microchannel. The main microchannel is 128.0 µm wide, 4.64 µm deep and 5680 µm long, and operated under nearly atmospheric conditions where the outlet Knudsen number is 0.0137. The sensor was fabricated using silicon wet etching, ultrasonic drilling, deep reactive ion etching (DRIE) and anodic bonding. The capacitance change of the sensor and the mass flow rate of nitrogen were measured as the inlet-to-outlet pressure ratio was varied from 1.00 to 1.24. The measured maximum mass flow rate was 3.86 × 10-10 kg s-1 (0.019 sccm) at the highest pressure ratio tested. As the pressure difference increased, both the capacitance of the differential pressure sensor and the flow rate through the main microchannel increased. The laminar friction constant f sdot Re, an important consideration in sensor design, varied from the incompressible no-slip case and the mass sensitivity and resolution of this sensor were discussed. Using the current slip flow formulae, a microchannel with much smaller mass flow rates can be designed at the same pressure ratios.

Ultrafine carbon particles promote rotenone-induced dopamine neuronal loss through activating microglial NADPH oxidase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Yinxi; Liu, Dan; Zhang, Huifeng

Background: Atmospheric ultrafine particles (UFPs) and pesticide rotenone were considered as potential environmental risk factors for Parkinson's disease (PD). However, whether and how UFPs alone and in combination with rotenone affect the pathogenesis of PD remains largely unknown. Methods: Ultrafine carbon black (ufCB, a surrogate of UFPs) and rotenone were used individually or in combination to determine their roles in chronic dopaminergic (DA) loss in neuron-glia, and neuron-enriched, mix-glia cultures. Immunochemistry using antibody against tyrosine hydroxylase was performed to detect DA neuronal loss. Measurement of extracellular superoxide and intracellular reactive oxygen species (ROS) were performed to examine activation of NADPHmore » oxidase. Genetic deletion and pharmacological inhibition of NADPH oxidase and MAC-1 receptor in microglia were employed to examine their role in DA neuronal loss triggered by ufCB and rotenone. Results: In rodent midbrain neuron-glia cultures, ufCB and rotenone alone caused neuronal death in a dose-dependent manner. In particularly, ufCB at doses of 50 and 100 μg/cm{sup 2} induced significant loss of DA neurons. More importantly, nontoxic doses of ufCB (10 μg/cm{sup 2}) and rotenone (2 nM) induced synergistic toxicity to DA neurons. Microglial activation was essential in this process. Furthermore, superoxide production from microglial NADPH oxidase was critical in ufCB/rotenone-induced neurotoxicity. Studies in mix-glia cultures showed that ufCB treatment activated microglial NADPH oxidase to induce superoxide production. Firstly, ufCB enhanced the expression of NADPH oxidase subunits (gp91{sup phox}, p47{sup phox} and p40{sup phox}); secondly, ufCB was recognized by microglial surface MAC-1 receptor and consequently promoted rotenone-induced p47{sup phox} and p67{sup phox} translocation assembling active NADPH oxidase. Conclusion: ufCB and rotenone worked in synergy to activate NADPH oxidase in microglia, leading to

Correlation Between Contrast Time-Density Time on Digital Subtraction Angiography and Flow: An in Vitro Study.

PubMed

Brunozzi, Denise; Shakur, Sophia F; Ismail, Rahim; Linninger, Andreas; Hsu, Chih-Yang; Charbel, Fady T; Alaraj, Ali

2018-02-01

Digital subtraction angiography (DSA) provides an excellent anatomic characterization of cerebral vasculature, but hemodynamic assessment is often qualitative and subjective. Various clinical algorithms have been produced to semiquantify flow from the data obtained from DSA, but few have tested them against reliable flow values. An arched flow model was created and injected with contrast material. Seventeen injections were acquired in anterior-posterior and lateral DSA projections, and 4 injections were acquired in oblique projection. Image intensity change over the angiogram cycle of each DSA run was analyzed through a custom MATLAB code. Time-density plots obtained were divided into 3 components (time-density times, TDTs): TDT 10%-100% (time needed for contrast material to change image intensity from 10% to 100%), TDT 100%-10% (time needed for contrast material to change image intensity from 100% to 10%), and TDT 25%-25% (time needed for contrast material to change from 25% image intensity to 25%). Time-density index (TDI) was defined as model cross-sectional area to TDT ratio, and it was measured against different flow rates. TDI 10%-100% , TDI 100%-10% , and TDI 25%-25% all correlated significantly with flow (P < 0.001). TDI 10%-100% , TDI 100%-10% , and TDI 25%-25% showed, respectively, a correlation coefficient of 0.91, 0.91, and 0.97 in the anterior-posterior DSA projections (P < 0.001). In the lateral DSA projection, TDI 100%-10% showed a weaker correlation (r = 0.57; P = 0.03). Also in the oblique DSA projection, TDIs correlated significantly with flow. TDI on DSA correlates significantly with flow. Although in vitro studies might overlook conditions that occur in patients, this method appears to correlate with the flow and could offer a semiquantitative method to evaluate the cerebral blood flow. Copyright © 2017 Elsevier Inc. All rights reserved.

Dosimetry Modeling for Focal Low-Dose-Rate Prostate Brachytherapy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Al-Qaisieh, Bashar; Mason, Josh, E-mail: joshua.mason@nhs.net; Bownes, Peter

2015-07-15

Purpose: Focal brachytherapy targeted to an individual lesion(s) within the prostate may reduce side effects experienced with whole-gland brachytherapy. The outcomes of a consensus meeting on focal prostate brachytherapy were used to investigate optimal dosimetry of focal low-dose-rate (LDR) prostate brachytherapy targeted using multiparametric magnetic resonance imaging (mp-MRI) and transperineal template prostate mapping (TPM) biopsy, including the effects of random and systematic seed displacements and interseed attenuation (ISA). Methods and Materials: Nine patients were selected according to clinical characteristics and concordance of TPM and mp-MRI. Retrospectively, 3 treatment plans were analyzed for each case: whole-gland (WG), hemi-gland (hemi), and ultra-focalmore » (UF) plans, with 145-Gy prescription dose and identical dose constraints for each plan. Plan robustness to seed displacement and ISA were assessed using Monte Carlo simulations. Results: WG plans used a mean 28 needles and 81 seeds, hemi plans used 17 needles and 56 seeds, and UF plans used 12 needles and 25 seeds. Mean D90 (minimum dose received by 90% of the target) and V100 (percentage of the target that receives 100% dose) values were 181.3 Gy and 99.8% for the prostate in WG plans, 195.7 Gy and 97.8% for the hemi-prostate in hemi plans, and 218.3 Gy and 99.8% for the focal target in UF plans. Mean urethra D10 was 205.9 Gy, 191.4 Gy, and 92.4 Gy in WG, hemi, and UF plans, respectively. Mean rectum D2 cm{sup 3} was 107.5 Gy, 77.0 Gy, and 42.7 Gy in WG, hemi, and UF plans, respectively. Focal plans were more sensitive to seed displacement errors: random shifts with a standard deviation of 4 mm reduced mean target D90 by 14.0%, 20.5%, and 32.0% for WG, hemi, and UF plans, respectively. ISA has a similar impact on dose-volume histogram parameters for all plan types. Conclusions: Treatment planning for focal LDR brachytherapy is feasible. Dose constraints are easily met with a

Rapid differentiation of cocci/mixed bacteria from rods in voided urine culture of women with uncomplicated urinary tract infections.

PubMed

Yang, Chun-Chun; Yang, Stephen Shei-Dei; Hung, Hui-Ching; Chiang, I-Ni; Peng, Chiung-Hui; Chang, Shang-Jen

2017-09-01

To evaluate the ability of laser flow cytometry to predict cocci/mixed growth in the pre-analytical phase of urine specimens. We retrospectively reviewed urine samples from women with uncomplicated urinary tract infections from urologic clinics for study. Urine analyses were performed with laser flow cytometry (UF1000i, Sysmex, Kobe, Japan) and then diagrams were generated (forward scatter vs. fluorescent light scatter). Each specimen (bacteria count >357 BACT/μL) was classified as either cocci bacteria or rods/mixed growth according to the diagrams. Standard urine cultures were performed, and the agreement between cultures and the UF1000i interpretations was analyzed with kappa statistics. Finally, 491 specimens met the criteria for analysis. Among the 376 specimens with single bacteria growth, there were 26 gram-positive cocci (13 Streptococci spp., 7 Staphylococci spp., 6 Enterococci spp.), 1 gram-positive rods (Corynebacterium spp.), and 349 gram-negative rods (273 Escherichia coli, 33 Klebsiella spp., 29 Proteus spp., 6 Citrobacter spp., 4 Enterobacter spp., 3 Pseudomonas spp., and 1 Providencia spp.). There were 115 specimens with two bacteria species or more that were regarded as mixed growth. Agreement of rods or cocci/mixed growth between the laser flow cytometry and urine cultures yielded a kappa value of 0.58. The positive and negative predictive rate of the UF1000i for cocci/mixed growth in voided urine culture was 81.8% and 84.7%, respectively. Through laser flow cytometry, we can predict growth of cocci/mixed growth in the pre-analytical phase of urine culture, thus avoiding unnecessary urine culture and waiting time. © 2016 Wiley Periodicals, Inc.

Obstructing Androgen Receptor Activation in Prostate Cancer Cells Through Post-translational Modification by NEDD8

DTIC Science & Technology

2012-11-01

FACS flow cytometer analysis . In addition, we will measure the steady state protein level of p53, p21, p27, and pRb. In the Jab1 silencing cell...affected by DHT treatment, and the endogenous AR level was not affected by Jab1 silencing. Interestingly, Western blot analysis of immunoprecipitated AR...Avantaggiati, and R. G. Pestell . 2003. Acetylation of androgen receptor enhances coactivator binding and promotes prostate cancer cell growth. Mol

Induction of Type 1 Immune Responses to SIV by IFN-Gamma

DTIC Science & Technology

1997-06-01

of rhesus macaques vaccinated with recombinant vaccinia viruses ( rVVs). Five animals were vaccinated with 107 pfu of vHuy/SIVgen, a Wyeth rVV...predominance of SIVAnef viruses . There is strong evidence to support the critical role of the cell-mediated arm of the immune system in the control of HIV...manufacturers, and imnmunofluorescence was measured with a dual- laser flow cytometer (FACSCAN, Becton Dickinson). -9- Induction of Type I Inmune Responses