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Sample records for cytometric co-immunoprecipitation technique

  1. Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein.

    Science.gov (United States)

    Verhelst, Judith; De Vlieger, Dorien; Saelens, Xavier

    2015-04-21

    Studying the interaction between proteins is key in understanding their function(s). A very powerful method that is frequently used to study interactions of proteins with other macromolecules in a complex sample is called co-immunoprecipitation. The described co-immunoprecipitation protocol allows to demonstrate and further investigate the interaction between the antiviral myxovirus resistance protein 1 (Mx1) and one of its viral targets, the influenza A virus nucleoprotein (NP). The protocol starts with transfected mammalian cells, but it is also possible to use influenza A virus infected cells as starting material. After cell lysis, the viral NP protein is pulled-down with a specific antibody and the resulting immune-complexes are precipitated with protein G beads. The successful pull-down of NP and the co-immunoprecipitation of the antiviral Mx1 protein are subsequently revealed by western blotting. A prerequisite for successful co-immunoprecipitation of Mx1 with NP is the presence of N-ethylmaleimide (NEM) in the cell lysis buffer. NEM alkylates free thiol groups. Presumably this reaction stabilizes the weak and/or transient NP-Mx1 interaction by preserving a specific conformation of Mx1, its viral target or an unknown third component. An important limitation of co-immunoprecipitation experiments is the inadvertent pull-down of contaminating proteins, caused by nonspecific binding of proteins to the protein G beads or antibodies. Therefore, it is very important to include control settings to exclude false positive results. The described co-immunoprecipitation protocol can be used to study the interaction of Mx proteins from different vertebrate species with viral proteins, any pair of proteins, or of a protein with other macromolecules. The beneficial role of NEM to stabilize weak and/or transient interactions needs to be tested for each interaction pair individually.

  2. Nanodisc-based Co-immunoprecipitation for Mass Spectrometric Identification of Membrane-interacting Proteins

    DEFF Research Database (Denmark)

    Borch-Jensen, Jonas; Roepstorff, Peter; Møller-Jensen, Jakob

    2011-01-01

    enterotoxigenic Escherischia coli, GM1-nanodiscs were employed for co-immunoprecipitation. The B subunit of heat labile enterotoxin was identified as a specific interaction partner by mass spectrometry, thus demonstrating that nanodisc technology is useful for highly specific detection and identification...

  3. A flow cytometric technique for quantification and differentiation of bacteria in bulk tank milk

    DEFF Research Database (Denmark)

    Holm, C.; Mathiasen, T.; Jespersen, Lene

    2004-01-01

    AIMS: The present study describes a flow cytometric technique for quantification and differentiation of bacteria in bulk tank milk according to the main cause of elevated counts. METHODS AND RESULTS: A total of 75 Danish bulk tank milk samples exceeding the grading level of 3.0 x 10(4) CFU ml(-1)...

  4. Oligomerization of the UDP-glucuronosyltransferase 1A proteins: homo- and heterodimerization analysis by fluorescence resonance energy transfer and co-immunoprecipitation.

    Science.gov (United States)

    Operaña, Theresa N; Tukey, Robert H

    2007-02-16

    UDP-glucuronosyltransferases (UGTs) are membrane-bound proteins localized to the endoplasmic reticulum and catalyze the formation of beta-d-glucopyranosiduronic acids (glucuronides) using UDP-glucuronic acid and acceptor substrates such as drugs, steroids, bile acids, xenobiotics, and dietary nutrients. Recent biochemical evidence indicates that the UGT proteins may oligomerize in the membrane, but conclusive evidence is still lacking. In the present study, we have used fluorescence resonance energy transfer (FRET) to study UGT1A oligomerization in live cells. This technique demonstrated that UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, and UGT1A10 self-oligomerize (homodimerize). Heterodimer interactions were also explored, and it was determined that UGT1A1 was capable of binding with UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, and UGT1A10. In addition to the in vivo FRET analysis, UGT1A protein-protein interactions were demonstrated through co-immunoprecipitation experiments. Co-expression of hemagglutinin-tagged and cyan fluorescent protein-tagged UGT1A proteins, followed by immunoprecipitation with anti-hemagglutinin beads, illustrated the potential of each UGT1A protein to homodimerize. Co-immunoprecipitation results also confirmed that UGT1A1 was capable of forming heterodimer complexes with all of the UGT1A proteins, corroborating the FRET results in live cells. These preliminary studies suggest that the UGT1A family of proteins form oligomerized complexes in the membrane, a property that may influence function and substrate selectivity.

  5. Proteomics Analysis of Cellular Proteins Co-Immunoprecipitated with Nucleoprotein of Influenza A Virus (H7N9

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    Ningning Sun

    2015-10-01

    Full Text Available Avian influenza A viruses are serious veterinary pathogens that normally circulate among avian populations, causing substantial economic impacts. Some strains of avian influenza A viruses, such as H5N1, H9N2, and recently reported H7N9, have been occasionally found to adapt to humans from other species. In order to replicate efficiently in the new host, influenza viruses have to interact with a variety of host factors. In the present study, H7N9 nucleoprotein was transfected into human HEK293T cells, followed by immunoprecipitated and analyzed by proteomics approaches. A series of host proteins co-immunoprecipitated were identified with high confidence, some of which were found to be acetylated at their lysine residues. Bioinformatics analysis revealed that spliceosome might be the most relevant pathway involved in host response to nucleoprotein expression, increasing our emerging knowledge of host proteins that might be involved in influenza virus replication activities.

  6. Proteomics Analysis of Cellular Proteins Co-Immunoprecipitated with Nucleoprotein of Influenza A Virus (H7N9)

    Science.gov (United States)

    Sun, Ningning; Sun, Wanchun; Li, Shuiming; Yang, Jingbo; Yang, Longfei; Quan, Guihua; Gao, Xiang; Wang, Zijian; Cheng, Xin; Li, Zehui; Peng, Qisheng; Liu, Ning

    2015-01-01

    Avian influenza A viruses are serious veterinary pathogens that normally circulate among avian populations, causing substantial economic impacts. Some strains of avian influenza A viruses, such as H5N1, H9N2, and recently reported H7N9, have been occasionally found to adapt to humans from other species. In order to replicate efficiently in the new host, influenza viruses have to interact with a variety of host factors. In the present study, H7N9 nucleoprotein was transfected into human HEK293T cells, followed by immunoprecipitated and analyzed by proteomics approaches. A series of host proteins co-immunoprecipitated were identified with high confidence, some of which were found to be acetylated at their lysine residues. Bioinformatics analysis revealed that spliceosome might be the most relevant pathway involved in host response to nucleoprotein expression, increasing our emerging knowledge of host proteins that might be involved in influenza virus replication activities. PMID:26528969

  7. Comparison of multidimensional flow cytometric data by a novel data mining technique

    Science.gov (United States)

    Leary, James F.; Smith, Jacob; Szaniszlo, Peter; Reece, Lisa M.

    2007-02-01

    Most flow/image cytometric data analysis methods look for clusters in the data corresponding to specific cell subpopulations. Comparisons between different cytometry datafiles often use human pattern recognition visualization of all the different combinations of variables ("parameters") two at a time in so-called bivariate scattergrams. Not only is this tedious, but it can miss potential clusters due to projection of higher dimensional dataspaces down onto two dimensional planes making them indiscernible as separate clusters. Novel data mining algorithms, implemented in software allow for the comparison of two or more higher dimensional datafiles without the requirement for reduction of dimensionality for human visualization. Of equal importance is the comparison of higher dimensional clusters which may move around slightly in space, yet still be "similar" according to algorithms which provide measures of similarity. This software, written in C/C++ and currently implemented in software with a Windows graphical user interface, allows for direct reading of FCS2.0 format flow cytometry datafiles of any number of parameters. In a few minutes or less, complex multiparameter data of two or more files can be compared on a personal computer or workstation. The software operates in either supervised or unsupervised mode, depending on whether the user wishes to include prior user knowledge or in a data mining discovery mode. Differences between these files can be exported as sub-datafiles which can be further analyzed using any other software that can read FCS2.0 data format.

  8. Effective identification of Akt interacting proteins by two-step chemical crosslinking, co-immunoprecipitation and mass spectrometry.

    Science.gov (United States)

    Huang, Bill X; Kim, Hee-Yong

    2013-01-01

    Akt is a critical protein for cell survival and known to interact with various proteins. However, Akt binding partners that modulate or regulate Akt activation have not been fully elucidated. Identification of Akt-interacting proteins has been customarily achieved by co-immunoprecipitation combined with western blot and/or MS analysis. An intrinsic problem of the method is loss of interacting proteins during procedures to remove non-specific proteins. Moreover, antibody contamination often interferes with the detection of less abundant proteins. Here, we developed a novel two-step chemical crosslinking strategy to overcome these problems which resulted in a dramatic improvement in identifying Akt interacting partners. Akt antibody was first immobilized on protein A/G beads using disuccinimidyl suberate and allowed to bind to cellular Akt along with its interacting proteins. Subsequently, dithiobis[succinimidylpropionate], a cleavable crosslinker, was introduced to produce stable complexes between Akt and binding partners prior to the SDS-PAGE and nanoLC-MS/MS analysis. This approach enabled identification of ten Akt partners from cell lysates containing as low as 1.5 mg proteins, including two new potential Akt interacting partners. None of these but one protein was detectable without crosslinking procedures. The present method provides a sensitive and effective tool to probe Akt-interacting proteins. This strategy should also prove useful for other protein interactions, particularly those involving less abundant or weakly associating partners.

  9. A novel method for purifying bluetongue virus with high purity by co-immunoprecipitation with agarose protein A

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    Ming Dai

    2010-06-01

    Full Text Available Abstract Background Bluetongue virus (BTV is an icosahedral non-enveloped virus within the genus Orbivirus of Reoviridae and exists as 24 distinct serotypes. BTV can infect all ruminant species and causes severe sickness in sheep. Recently, it was reported that BTV can infect some human cancer cells selectively. Because of the important oncolysis of this virus, we developed a novel purifying method for large-scale production. The purifying logic is simple, which is picking out all the components unwanted and the left is what we want. The process can be summarized in 4 steps: centrifugation, pulling down cell debrises and soluble proteins by co-immunoprecipitation with agarose Protein A, dialysis and filtration sterilization after concentration. Results The result of transmission electron microscope (TEM observation showed that the sample of purified virus has a very clear background and the virions still kept intact. The result of 50% tissue culture infective dose (TCID50 assay showed that the bioactivity of purified virus is relatively high. Conclusions This method can purify BTV-10 with high quality and high biological activity on large-scale production. It also can be used for purifying other BTV serotypes.

  10. [THE CYTOMETRIC TECHNIQUE OF BINDING OF EOSIN-5-MALEIMIDE IN DIAGNOSTIC OF INHERENT SPHEROCYTOSIS].

    Science.gov (United States)

    Kuzminova, J A; Plyasunova, S A; Jogov, V V; Smetanina, N S

    2016-03-01

    The laboratory diagnostic of inherent spherocytosis is based on detection of spherocytes in peripheral blood, decreasing of index of sphericity, decreasing of osmotic resistance of erythrocytes. The new test of diagnostic of hereditary spherocytosis build on molecular defect was developed on the basis of binding extracellular fragments of protein of band 3 with eosin-5-maleimide (EMA-test). The study was carried out to implement comparative analysis of sensitivity and specificity of techniques applied to diagnose inherent spherocytosis. The sampling of 94 patients with various forms of anemias was analyzed All patients were applied complex clinical laboratory examination including analysis of osmotic resistance of erythrocytes, erythrocytometry and EMA-test as specific techniques of diagnostic of inherent spherocytosis. In 51 out of 94 patients (54%) decreasing of values of EMA-test was detected and in 47 patients diagnosis of inherent spherocytosis was confirmed. The standard values of EMA-test were established in 43 patients (46%) and 12 patients out of them with established diagnosis of inherent spherocytosis. Therefore, sensitivity of EMA-test made up to 79% and specificity - 80%. The most sensitive techniques of diagnostic remain osmotic resistance of erythrocytes (91%) and index of sphericity (up to 96%). But the highest specificity in this respect has EMA-test (80%). Nowadays, none of implemented techniques of diagnostic of inherent spherocytosis can be applied as a universal one. The implementation of complex examination is needed for proper diagnostic of disease.

  11. Heterotrimeric Galphaq11 co-immunoprecipitates with surface-anchored GRP78 from plasma membranes of alpha2M*-stimulated macrophages.

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    Misra, Uma Kant; Pizzo, Salvatore Vincent

    2008-05-01

    We have previously shown that a fraction of newly expressed GRP78 is translocated to the cell surface in association with the co-chaperone MTJ-1. Proteinase and methylamine-activated alpha(2)M (alpha(2)M*) bind to cell surface-associated GRP78 activating phosphoinositide-specific phospholipase C coupled to a pertussis toxin-insensitive heterotrimeric G protein, generating IP(3)/calcium signaling. We have now studied the association of pertussis toxin-insensitive Galphaq11, with GRP78/MTJ-1 complexes in the plasma membranes of alpha(2)M*-stimulated macrophages. When GRP78 was immunoprecipitated from plasma membranes of macrophages stimulated with alpha(2)M*, Galphaq11, and MTJ-1 were co-precipitated. Likewise Galphaq11 and GRP78 co-immunoprecipitated with MTJ-1 while GRP78 and MTJ-1 co-immunoprecipitated with Galphaq11. Silencing GRP78 expression with GRP78 dsRNA or MTJ-1 with MTJ-1 dsRNA greatly reduced the levels of Galphaq11 co-precipitated with GRP78 or MTJ-1. In conclusion, we show here that plasma membrane-associated GRP78 is coupled to pertussis toxin-insensitive Galphaq11 and forms a ternary signaling complex with MTJ-1.

  12. Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae

    Institute of Scientific and Technical Information of China (English)

    HUANG Jian; WEN Ruobing; BAO Zhenmin; SUI Zhenghong; SUN Ningbo; KANG Kyoungho

    2012-01-01

    Over the last severaldecades,harmful algal blooms (HABs) havebecome a serious environmental problem in many parts of the world.A rapid and accurate detection process for HAB algae has yet to be developed.Heterosigma akashiwo is one of the most important HABs species in China.The objective of this study was to develop an immunologic technique that can rapidly and sensitively count H.akashiwo cells.Five HABs species (Alexandrium catenella,Thalassiosira sp.,Cryptomonas sp.,Alexandrium tamarense and Symbiodinium sp.,) were used in this study to evaluate the analysis process we developed.A polyclonal antibody with high titers against H.akashiwo was obtained by injecting H.akashiwo cells into rabbits.Immuno-magnetic beads (IMB) were produced via conjugated polyclonal antibodies with magnetic beads and applied to isolate and count H.akashiwo cells from the culture.Results show that 66.7%-91.6% of the cells were captured from unialgal culture by IMBs,and only 5.3%-12.5% of the four other HAB microalgae species were captured,indicating that the constructed IMBs combined specifically with the H.akashiwo cells.At the same time,flow cytometry (FCM) sorting was exploited to screen H.akashiwo cells after labeling with FITC conjugated polyclonal antibodies.Using the FCM technique,91.7% of the targeted cells were sorted out from mixed microalgae samples in just a few minutes.These results indicate that both antibody-involved IMB and antibody-based FCM techniques are highly effective at detecting and quantifying HAB species.These techniques,especially immuno-magnetic separation,have low associated cost,and are fast and simple processes compared with other techniques currently in use.

  13. Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae

    Science.gov (United States)

    Huang, Jian; Wen, Ruobing; Bao, Zhenmin; Sui, Zhenghong; Sun, Ningbo; Kang, Kyoungho

    2012-05-01

    Over the last several decades, harmful algal blooms (HABs) have become a serious environmental problem in many parts of the world. A rapid and accurate detection process for HAB algae has yet to be developed. Heterosigma akashiwo is one of the most important HABs species in China. The objective of this study was to develop an immunologic technique that can rapidly and sensitively count H. akashiwo cells. Five HABs species ( Alexandrium catenella, Thalassiosira sp., Cryptomonas sp., Alexandrium tamarense and Symbiodinium sp.), were used in this study to evaluate the analysis process we developed. A polyclonal antibody with high titers against H. akashiwo was obtained by injecting H. akashiwo cells into rabbits. Immuno-magnetic beads (IMB) were produced via conjugated polyclonal antibodies with magnetic beads and applied to isolate and count H. akashiwo cells from the culture. Results show that 66.7%-91.6% of the cells were captured from unialgal culture by IMBs, and only 5.3%-12.5% of the four other HAB microalgae species were captured, indicating that the constructed IMBs combined specifically with the H. akashiwo cells. At the same time, flow cytometry (FCM) sorting was exploited to screen H. akashiwo cells after labeling with FITC conjugated polyclonal antibodies. Using the FCM technique, 91.7% of the targeted cells were sorted out from mixed microalgae samples in just a few minutes. These results indicate that both antibody-involved IMB and antibody-based FCM techniques are highly effective at detecting and quantifying HAB species. These techniques, especially immuno-magnetic separation, have low associated cost, and are fast and simple processes compared with other techniques currently in use.

  14. Constitutive oligomerization of human D2 dopamine receptors expressed in Spodoptera frugiperda 9 (Sf9) and in HEK293 cells. Analysis using co-immunoprecipitation and time-resolved fluorescence resonance energy transfer.

    Science.gov (United States)

    Gazi, Lucien; López-Giménez, Juan F; Rüdiger, Martin P; Strange, Philip G

    2003-10-01

    Human D2Long (D2L) and D2Short (D2S) dopamine receptor isoforms were modified at their N-terminus by the addition of a human immunodeficiency virus (HIV) or a FLAG epitope tag. The receptors were then expressed in Spodoptera frugiperda 9 (Sf9) cells using the baculovirus system, and their oligomerization was investigated by means of co-immunoprecipitation and time-resolved fluorescence resonance energy transfer (FRET). [3H]Spiperone labelled D2 receptors in membranes prepared from Sf9 cells expressing epitope-tagged D2L or D2S receptors, with a pKd value of approximately 10. Co-immunoprecipitation using antibodies specific for the tags showed constitutive homo-oligomerization of D2L and D2S receptors in Sf9 cells. When the FLAG-tagged D2S and HIV-tagged D2L receptors were co-expressed, co-immunoprecipitation showed that the two isoforms can also form hetero-oligomers in Sf9 cells. Time-resolved FRET with europium and XL665-labelled antibodies was applied to whole Sf9 cells and to membranes from Sf9 cells expressing epitope-tagged D2 receptors. In both cases, constitutive homo-oligomers were revealed for D2L and D2S isoforms. Time-resolved FRET also revealed constitutive homo-oligomers in HEK293 cells expressing FLAG-tagged D2S receptors. The D2 receptor ligands dopamine, R-(-)propylnorapomorphine, and raclopride did not affect oligomerization of D2L and D2S in Sf9 and HEK293 cells. Human D2 dopamine receptors can therefore form constitutive oligomers in Sf9 cells and in HEK293 cells that can be detected by different approaches, and D2 oligomerization in these cells is not regulated by ligands.

  15. Flow Cytometric Immunobead Assay for Detection of BCR-ABL1 Fusion Proteins in Chronic Myleoid Leukemia: Comparison with FISH and PCR Techniques.

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    Anna Grazia Recchia

    Full Text Available Chronic Myeloid Leukemia (CML is characterized by a balanced translocation juxtaposing the Abelson (ABL and breakpoint cluster region (BCR genes. The resulting BCR-ABL1 oncogene leads to increased proliferation and survival of leukemic cells. Successful treatment of CML has been accompanied by steady improvements in our capacity to accurately and sensitively monitor therapy response. Currently, measurement of BCR-ABL1 mRNA transcript levels by real-time quantitative PCR (RQ-PCR defines critical response endpoints. An antibody-based technique for BCR-ABL1 protein recognition could be an attractive alternative to RQ-PCR. To date, there have been no studies evaluating whether flow-cytometry based assays could be of clinical utility in evaluating residual disease in CML patients. Here we describe a flow-cytometry assay that detects the presence of BCR-ABL1 fusion proteins in CML lysates to determine the applicability, reliability, and specificity of this method for both diagnosis and monitoring of CML patients for initial response to therapy. We show that: i CML can be properly diagnosed at onset, (ii follow-up assessments show detectable fusion protein (i.e. relative mean fluorescent intensity, rMFI%>1 when BCR-ABL1IS transcripts are between 1-10%, and (iii rMFI% levels predict CCyR as defined by FISH analysis. Overall, the FCBA assay is a rapid technique, fully translatable to the routine management of CML patients.

  16. Flow Cytometric Immunobead Assay for Detection of BCR-ABL1 Fusion Proteins in Chronic Myleoid Leukemia: Comparison with FISH and PCR Techniques.

    Science.gov (United States)

    Recchia, Anna Grazia; Caruso, Nadia; Bossio, Sabrina; Pellicanò, Mariavaleria; De Stefano, Laura; Franzese, Stefania; Palummo, Angela; Abbadessa, Vincenzo; Lucia, Eugenio; Gentile, Massimo; Vigna, Ernesto; Caracciolo, Clementina; Agostino, Antolino; Galimberti, Sara; Levato, Luciano; Stagno, Fabio; Molica, Stefano; Martino, Bruno; Vigneri, Paolo; Di Raimondo, Francesco; Morabito, Fortunato

    2015-01-01

    Chronic Myeloid Leukemia (CML) is characterized by a balanced translocation juxtaposing the Abelson (ABL) and breakpoint cluster region (BCR) genes. The resulting BCR-ABL1 oncogene leads to increased proliferation and survival of leukemic cells. Successful treatment of CML has been accompanied by steady improvements in our capacity to accurately and sensitively monitor therapy response. Currently, measurement of BCR-ABL1 mRNA transcript levels by real-time quantitative PCR (RQ-PCR) defines critical response endpoints. An antibody-based technique for BCR-ABL1 protein recognition could be an attractive alternative to RQ-PCR. To date, there have been no studies evaluating whether flow-cytometry based assays could be of clinical utility in evaluating residual disease in CML patients. Here we describe a flow-cytometry assay that detects the presence of BCR-ABL1 fusion proteins in CML lysates to determine the applicability, reliability, and specificity of this method for both diagnosis and monitoring of CML patients for initial response to therapy. We show that: i) CML can be properly diagnosed at onset, (ii) follow-up assessments show detectable fusion protein (i.e. relative mean fluorescent intensity, rMFI%>1) when BCR-ABL1IS transcripts are between 1-10%, and (iii) rMFI% levels predict CCyR as defined by FISH analysis. Overall, the FCBA assay is a rapid technique, fully translatable to the routine management of CML patients.

  17. Recent advances in flow cytometric cell sorting.

    Science.gov (United States)

    Osborne, Geoffrey W

    2011-01-01

    The classification and separation of one cell type or particle from others is a fundamental task in many areas of science. Numerous techniques are available to perform this task; however, electrostatic cell sorting has gained eminence over others because, when combined with the analysis capabilities of flow cytometry it provides flexible separations based on multiple parameters. Unlike competing technologies, such as gradient or magnetic separations that offer much larger total throughput, flow cytometric cell sorting permits selections based on various levels of fluorescent reporters, rather the complete presence or absence of the reporter. As such, this technology has found application in a huge range of fields. This chapter aims to describe the utility of single-cell sorting with particular emphasis given to index sorting. This is followed by two recently developed novel techniques of sorting cells or particles. The first of these is positional sorting which is useful in cell-based studies where sorting can proceed and produce meaningful results without being inherently dependant on prior knowledge of where gates should be set. Secondly, reflective plate sorting is introduced which positionally links multiwell sample and collection plates in a convenient assay format so that cells in the collection plate "reflect" those in the sample plate.

  18. Flow-cytometric Analysis of Bacillus anthracis Spores

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    D. V. Kamboj

    2006-11-01

    Full Text Available Flow-cytometric technique has been established as a powerful tool for detection andidentification of microbiological agents. Unambiguous and rapid detection of Bacillus anthracisspores has been reported using immunoglobulin G-fluorescein isothiocyanate conjugate againstlive spores. In addition to the high sensitivity, the present technique could differentiate betweenspores of closely related species, eg, Bacillus cereus and Bacillus subtilis using fluorescenceintensity. The technique can be used for detection of live as well as inactivated spores makingit more congenial for screening of suspected samples of bioterrorism.

  19. Cytometric Catheter for Neurosurgical Applications

    Energy Technology Data Exchange (ETDEWEB)

    Evans III, Boyd Mccutchen [ORNL; Allison, Stephen W [ORNL; Fillmore, Helen [ORNL; Broaddus, William C [ORNL; Dyer, Rachel L [ORNL; Gillies, George [ORNL

    2010-01-01

    Implantation of neural progenitor cells into the central nervous system has attracted strong interest for treatment of a variety of pathologies. For example, the replacement of dopamine-producing (DA) neural cells in the brain appears promising for the treatment of patients affected by Parkinson's disease. Previous studies of cell-replacement strategies have shown that less than 90% of implanted cells survive longer than 24 - 48 hours following the implantation procedure. However, it is unknown if these cells were viable upon delivery, or if they were affected by other factors such as brain pathology or an immune response. An instrumented cell-delivery catheter has been developed to assist in answering these questions by facilitating quantification and monitoring of the viability of the cells delivered. The catheter uses a fiber optic probe to perform flourescence-based cytometric measurments on cells exiting the port at the catheter tip. The current implementation of this design is on a 3.2 mm diameter catheter with 245 micrometer diameter optical fibers. Results of fluorescence testing data are presented and show that the device can characterize the quantity of cell densities ranging from 60,000 cells/ml to 600,000 cells/ml with a coefficient of determination of 0.93.

  20. Cluster Analysis of Flow Cytometric List Mode Data on a Personal Computer

    NARCIS (Netherlands)

    Bakker Schut, Tom C.; Bakker schut, T.C.; de Grooth, B.G.; Greve, Jan

    1993-01-01

    A cluster analysis algorithm, dedicated to analysis of flow cytometric data is described. The algorithm is written in Pascal and implemented on an MS-DOS personal computer. It uses k-means, initialized with a large number of seed points, followed by a modified nearest neighbor technique to reduce

  1. Clonal heterogeneity of small-cell anaplastic carcinoma of the lung demonstrated by flow-cytometric DNA analysis

    DEFF Research Database (Denmark)

    Vindeløv, L L; Hansen, H H; Christensen, I J

    1980-01-01

    Flow-cytometric DNA analysis yields information on ploidy and proliferative characteristics of a cell population. The analysis was implemented on small-cell anaplastic carcinoma of the lung using a rapid detergent technique for the preparation of fine-needle aspirates for DNA determination...

  2. Howard University Flow Cytometric Sorter For Research and Education

    Science.gov (United States)

    2015-08-04

    SECURITY CLASSIFICATION OF: Howard University’s newly acquired Fluorescence Activated Cytometric Sorter (FACS) has been integrated into the new flow...Research Administrative Services Washington, DC 20059 -0001 11-Mar-2015 ABSTRACT Number of Papers published in peer-reviewed journals : Number of Papers...published in non peer-reviewed journals : Final Report: Howard University Flow Cytometric Sorter For Research and Education Report Title Howard

  3. Flow cytometric detection of anti-gliadin antibodies.

    Science.gov (United States)

    Presani, G; Perticarari, S; Mangiarotti, M A

    1989-05-12

    A very sensitive solid-phase fluorescent immunoassay to detect anti-alpha-gliadin IgA class antibodies is described. The solid phase consisted of polystyrene carboxylated microspheres, of 5 microns diameter, coated with alpha-gliadin. Serum-specific antibodies bound to the alpha-gliadin were measured by flow cytometry using fluorescein-conjugated anti-human IgA. 41 samples were tested and the results compared with those obtained by a standard method: an enzyme-linked immunosorbent assay (ELISA). A good correlation was found between the two techniques (r = 0.96). The sera of untreated coeliac children showed significantly higher antibody values than the sera of children on a gluten-free diet or healthy control groups. The flow cytometric method was more sensitive when the Kolgomorov/Smirnov test was used to analyse the histograms. This method provides an alternative screening test for coeliac disease and may also be used to confirm borderline results obtained in the ELISA test.

  4. Comparative flow cytometric analysis of immunofunctionalized nanowire and nanoparticle signatures.

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    Prina-Mello, Adriele; Whelan, Aine M; Atzberger, Ann; McCarthy, Joseph E; Byrne, Fiona; Davies, Gemma-Louise; Coey, J M D; Volkov, Yuri; Gun'ko, Yurii K

    2010-01-01

    Flow cytometry is one of the gold-standard techniques used in clinical medicine for quantitative immunoassaying. The continuous development of its probes, commonly fluorescent nanoparticles, is important. Lately, the introduction of quantitative multiplexed immunoassay has challenged the use of nanoparticles as probes. Functionalized fluorescent silica-based magnetic nanowires are investigated under flow cytometry as a novel probe category. The preparation and full characterization of these multimodal nanowires is reported and compared to those of silica-based magnetic nanoparticles by flow cytometry. Full characterization includes transmission electron microscopy and fluorescence microscopy imaging, flow cytometric assaying, superconducting quantum interference device (SQUID) magnetization, and Mössbauer spectroscopy measurements. This work shows that loaded silica nanowires have intrinsic geometrical advantages when compared to similar spherical particles due to their unique "flow cytometry fingerprint" when utilized as magnetic carriers for immunodetection applications. These advantages account for a 17% yield in detecting the functional binding between THP-1 and ICAM-1, by utilizing a much lower concentration than that required for the nanoparticles.

  5. Flow cytometric enumeration of marine viral populations at low abundances

    NARCIS (Netherlands)

    Mojica, K.D.A.; Evans, C.; Brussaard, C.P.D.

    2014-01-01

    Flow cytometric enumeration has advanced our ability to analyze aquatic virus samples and thereby our understanding of the ecological role viruses play in the oceans. However, low virus abundances are underestimated using the current flow cytometry (FCM) protocol. Our results revealed that low

  6. Flow cytometric immunoassay for sulfonamides in raw milk

    NARCIS (Netherlands)

    Keizer, de W.; Bienenmann-Ploum, M.; Bergwerff, A.A.; Haasnoot, W.

    2008-01-01

    Sulfonamide antibiotics are applied in veterinary medicine for the treatment of microbial infections. For the detection of residues of sulfonamides in milk, a multi-sulfonamide flow cytometric immunoassay (FCI) was developed using the Luminex MultiAnalyte Profiling (xMAP) technology. In this automat

  7. Flow cytometric methods to investigate culture heterogeneities for plant metabolic engineering.

    Science.gov (United States)

    Gaurav, Vishal; Kolewe, Martin E; Roberts, Susan C

    2010-01-01

    Plant cell cultures provide an important method for production and supply of a variety of natural products, where conditions can be easily controlled, manipulated, and optimized. Development and optimization of plant cell culture processes require both bioprocess engineering and metabolic engineering approaches. Cultures are generally highly heterogeneous, with significant variability amongst cells in terms of growth, metabolism, and productivity of key metabolites. Taxus cultures produce the important anti-cancer agent Taxol((R)) (i.e., paclitaxel) and have demonstrated significant variability amongst cell populations in culture with regard to paclitaxel accumulation, cell cycle participation, and protein synthesis. To fully understand the link between cellular metabolism and culture behavior and to enable targeted metabolic engineering approaches, cultures need to be studied at a single cell level. This chapter describes the application of plant cell flow cytometric techniques to investigate culture heterogeneity at the single cell level, in order to optimize culture performance through targeted metabolic engineering. Flow cytometric analytical methods are described to study Taxus single cells, protoplasts, and nuclei suspensions with respect to secondary metabolite accumulation, DNA content, cell size, and complexity. Reproducible methods to isolate these single particle suspensions from aggregated Taxus cultures are discussed. Methods to stain both fixed and live cells for a variety of biological markers are provided to enable characterization of cell phenotypes. Fluorescence-activated cell sorting (FACS) methods are also presented to facilitate isolation of certain plant cell culture populations for both analysis and propagation of superior cell lines for use in bioprocesses.

  8. Strategies for immunophenotyping and purifying classical Hodgkin lymphoma cells from lymph nodes by flow cytometry and flow cytometric cell sorting.

    Science.gov (United States)

    Fromm, Jonathan R; Wood, Brent L

    2012-07-01

    Flow cytometry is an established technique to immunophenotype hematopoietic neoplasms. While the diagnosis of classical Hodgkin lymphoma (CHL) has commonly been made using paraffin sections, we have recently demonstrated that the neoplastic Hodgkin and Reed-Sternberg (HRS) cells of CHL can be identified by flow cytometry. Using 6- and 9-color flow cytometric assays, CHL can be immunophenotyped with 85-90% sensitivity and nearly 100% specificity. Analysis of this data requires using established gating strategies to help in the identification of putative HRS cell populations. Interestingly, HRS cells bind to reactive T cells (HRS-T cell rosetting) and this phenomenon can be identified and utilized diagnostically by flow cytometry. In addition, the reactive T cells of CHL show characteristic immunophenotypic changes by flow cytometry and these changes can suggest a diagnosis of CHL. Finally, these principles can be employed to rapidly purify HRS cells using flow cytometric cell sorting. This manuscript provides experimental protocols for immunophenotyping CHL by flow cytometry as well as purifying the HRS cells via flow cytometric cell sorting.

  9. A flow cytometric in vivo chalone assay using retransplanted old murine JB-1 ascites tumour cells.

    Science.gov (United States)

    Barfod, N M

    1981-07-01

    A flow cytometric in vivo chalone assay is described. Transplantation of old JB-1 ascites tumour cells to new hosts induced an influx of tumour cells, with G1 DNA content, to the S phase. This induction could be reversibly and specifically blocked by injections of an ultrafiltrate of old JB-1 ascites fluid. The method described is superior to a previously published in vivo chalone assay using regenerating ascites tumours. Owing to a reduced variability in time of onset of DNA synthesis, a smaller scatter of observations is achieved and thus the number of mice per group may be reduced using the new method. In contrast to the older technique, the present one does not necessitate killing of mice during the observation period.

  10. Cytometric patterns reveal growth states of Shewanella putrefaciens.

    Science.gov (United States)

    Melzer, Susanne; Winter, Gudrun; Jäger, Kathrin; Hübschmann, Thomas; Hause, Gerd; Syrowatka, Frank; Harms, Hauke; Tárnok, Attila; Müller, Susann

    2015-05-01

    Bacterial growth is often difficult to estimate beyond classical cultivation approaches. Low cell numbers, particles or coloured and dense media may disturb reliable growth assessment. Further difficulties appear when cells are attached to surfaces and detachment is incomplete. Therefore, flow cytometry was tested and used for analysis of bacterial growth on the single-cell level. Shewanella putrefaciens was cultivated as a model organism in planktonic or biofilm culture. Materials of smooth and rough surfaces were used for biofilm cultivation. Both aerobic and anaerobic as well as feast and famine conditions were applied. Visualization of growth was also done using Environmental Scanning and Phase Contrast Microscopy. Bioinformatic tools were applied for data interpretation. Cytometric proliferation patterns based on distributions of DNA contents per cell corresponded distinctly to the various lifestyles, electron acceptors and substrates tested. Therefore, cell cycling profiles of S. putrefaciens were found to mirror growth conditions. The cytometric patterns were consistently detectable with exception of some biofilm types whose resolution remained challenging. Corresponding heat maps proved to be useful for clear visualization of growth behaviour under all tested conditions. Therefore, flow cytometry in combination with bioinformatic tools proved to be powerful means to determine various growth states of S. putrefaciens, even in constrained environments. The approach is universal and will also be applicable for other bacterial species. © 2014 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  11. Uncovering Aberrant Mutant PKA Function with Flow Cytometric FRET

    Directory of Open Access Journals (Sweden)

    Shin-Rong Lee

    2016-03-01

    Full Text Available Biology has been revolutionized by tools that allow the detection and characterization of protein-protein interactions (PPIs. Förster resonance energy transfer (FRET-based methods have become particularly attractive as they allow quantitative studies of PPIs within the convenient and relevant context of living cells. We describe here an approach that allows the rapid construction of live-cell FRET-based binding curves using a commercially available flow cytometer. We illustrate a simple method for absolutely calibrating the cytometer, validating our binding assay against the gold standard isothermal calorimetry (ITC, and using flow cytometric FRET to uncover the structural and functional effects of the Cushing-syndrome-causing mutation (L206R on PKA’s catalytic subunit. We discover that this mutation not only differentially affects PKAcat’s binding to its multiple partners but also impacts its rate of catalysis. These findings improve our mechanistic understanding of this disease-causing mutation, while illustrating the simplicity, general applicability, and power of flow cytometric FRET.

  12. Cytometric analysis of mammalian sperm for induced morphologic and DNA content errors

    Energy Technology Data Exchange (ETDEWEB)

    Pinkel, D.

    1983-06-27

    Some flow-cytometric and image analysis procedures under development for quantitative analysis of sperm morphology are reviewed. The results of flow-cytometric DNA-content measurements on sperm from radiation exposed mice are also summarized, the results related to the available cytological information, and their potential dosimetric sensitivity discussed. (ACR)

  13. An automated flow cytometric micronucleus assay for human lymphocytes.

    Science.gov (United States)

    Schreiber, G A; Beisker, W; Braselmann, H; Bauchinger, M; Bögl, K W; Nüsse, M

    1992-12-01

    A new flow cytometric method is presented for scoring micronuclei (MN) in human lymphocytes after in vitro gamma-irradiation. Fifty to fifty-five hours after PHA-stimulation, the frequency of micronuclei per nucleus and the fraction of cells in the second cell cycle were measured using flow cytometry. All data were automatically analysed using our DAS-software package. Eight individual linear-quadratic dose response curves derived from five donors revealed inter- and intra-individual variabilities of all curve parameters. Since also an age dependence was found for spontaneous MN-frequencies and for the linear curve parameter, a combined linear-quadratic age-dose-effect model was used to fit the data. The 90% prediction intervals show that a reliable individual dose estimation for donors aged between 23 and 54 years cannot be achieved for exposures below 1 Gy.

  14. Oral squamous cell carcinoma. Cytometric parameters of prognostic interest.

    Science.gov (United States)

    Saiz-Bustillo, Ramón; Corchero-Martín, Guadalupe; García-Montesinos-Perea, Belén; Gonzalez-Terán, Tomás; Sánchez-Santolino, Sergio

    2005-01-01

    The present study was made in order to find possible prognostic factors in oral squamous cell carcinoma, given that it is a frequent disease (3-4% of all malignant tumors) and is the cause of a high morbidity and mortality which justifies any attempt to contribute something towards the understanding of this pathology. 81 oral squamous cell carcinomas, treated with the same procedure, and retrieved from the archive of the Hospital Universitario Marqués de Valdecilla (Santander) were studied. Flow cytometry was carried out on 67 of the samples. No statistically significant differences were found between the cellular proliferative index and the mitotic index, ploidy and the S-phase factor. Likewise, none of the cytometric variables studied presented any association with the appearance of local relapse, distant metastases or survival. These variables cannot be used as a prognostic factors in squamous cell carcinomas of the oral cavity.

  15. Automated High-Dimensional Flow Cytometric Data Analysis

    Science.gov (United States)

    Pyne, Saumyadipta; Hu, Xinli; Wang, Kui; Rossin, Elizabeth; Lin, Tsung-I.; Maier, Lisa; Baecher-Allan, Clare; McLachlan, Geoffrey; Tamayo, Pablo; Hafler, David; de Jager, Philip; Mesirov, Jill

    Flow cytometry is widely used for single cell interrogation of surface and intracellular protein expression by measuring fluorescence intensity of fluorophore-conjugated reagents. We focus on the recently developed procedure of Pyne et al. (2009, Proceedings of the National Academy of Sciences USA 106, 8519-8524) for automated high- dimensional flow cytometric analysis called FLAME (FLow analysis with Automated Multivariate Estimation). It introduced novel finite mixture models of heavy-tailed and asymmetric distributions to identify and model cell populations in a flow cytometric sample. This approach robustly addresses the complexities of flow data without the need for transformation or projection to lower dimensions. It also addresses the critical task of matching cell populations across samples that enables downstream analysis. It thus facilitates application of flow cytometry to new biological and clinical problems. To facilitate pipelining with standard bioinformatic applications such as high-dimensional visualization, subject classification or outcome prediction, FLAME has been incorporated with the GenePattern package of the Broad Institute. Thereby analysis of flow data can be approached similarly as other genomic platforms. We also consider some new work that proposes a rigorous and robust solution to the registration problem by a multi-level approach that allows us to model and register cell populations simultaneously across a cohort of high-dimensional flow samples. This new approach is called JCM (Joint Clustering and Matching). It enables direct and rigorous comparisons across different time points or phenotypes in a complex biological study as well as for classification of new patient samples in a more clinical setting.

  16. Flow cytometric detection of immunoglobulin light chain in hematolymphoid immunophenotyping

    Institute of Scientific and Technical Information of China (English)

    Xu Dongsheng

    2011-01-01

    During B cell development and maturation, the antigen receptor,which is encoded by the immunoglobulin heavy-(IgH)and light chain genes,rearrange to associate one of a number of variable, diverse, and joining gene segments. A single mature Bcell expresses an IgH chain and either a kappa or lambda light chain, which is known as allelic or isotypic exclusion. In normal or reactive conditions, lymphoid cells comprise the mixtures of lymphocytes with either kappa or lambda expression. The norreal proportion of kappa to lambda (κ/λ ratio) is within the range of 0. 5 - 3. 0 in peripheral blood or bone marrow and 1.2 2.7 in lymph nodes[1].The most useful feature for diagnosing mature B cell neoplasm is light chain restriction or monotypic staining with κ or λ light chain. Currently, it is a common assumption that demonstration of light chain restriction in a B lymphocyte population is generally considered proof of monoclonality and indicates malignancy although monotypic B cell populations have been infrequently demonstrated in patients with no definitive evidence ofB cell malignancy[2-4]. The most common flow cytometric analysis for determining B cell monotype is the percent κ and λimmunoglobulin light chains.Because of the importance of light chain restriction in the diagnosis of B cell neoplasm, anti immunoglobulin antibodies (e. g. anti-κ and anti-λ) are vital tools in the detection of monotypic B cell populations. Accurate determination of surface light chain expression depends on many factors, such as proper washing procedure, lysing solution, type of antibody used, specimen type or lymphoma type[5]. This article will discuss some common problems encountered in flow cytometric(FCM)deterruination of surface immunoglobulin light chain expression inhematolymphoid immunophenotyping.%@@ During B-cell development and maturation,the antigen receptor,which is encoded by the immunoglobulin heavy-(IgH) and light-chain genes,rearrange to associate one of a number of

  17. Flow cytometric and laser scanning microscopic approaches in epigenetics research.

    Science.gov (United States)

    Szekvolgyi, Lorant; Imre, Laszlo; Minh, Doan Xuan Quang; Hegedus, Eva; Bacso, Zsolt; Szabo, Gabor

    2009-01-01

    Our understanding of epigenetics has been transformed in recent years by the advance of technological possibilities based primarily on a powerful tool, chromatin immunoprecipitation (ChIP). However, in many cases, the detection of epigenetic changes requires methods providing a high-throughput (HTP) platform. Cytometry has opened a novel approach for the quantitative measurement of molecules, including PCR products, anchored to appropriately addressed microbeads (Pataki et al. 2005. Cytometry 68, 45-52). Here we show selected examples for the utility of two different cytometry-based platforms of epigenetic analysis: ChIP-on-beads, a flow-cytometric test of local histone modifications (Szekvolgyi et al. 2006. Cytometry 69, 1086-1091), and the laser scanning cytometry-based measurement of global epigenetic modifications that might help predict clinical behavior in different pathological conditions. We anticipate that such alternative tools may shortly become indispensable in clinical practice, translating the systematic screening of epigenetic tags from basic research into routine diagnostics of HTP demand.

  18. Flow cytometric quantitation of phagocytosis in heparinized complete blood with latex particles and Candida albicans

    Directory of Open Access Journals (Sweden)

    Jesús M. Egido

    1997-12-01

    Full Text Available We report a rapid method for the flow cytometric quantitation of phagocytosis in heparinized complete peripherial blood (HCPB, using commercially available phycoerythrin-conjugated latex particles of 1µm diameter. The method is faster and shows greater reproducibility than Bjerknes' (1984 standard technique using propidium iodide-stained Candida albicans, conventionally applied to the leukocytic layer of peripherial blood but here modified for HCPB. We also report a modification of Bjerknes' Intracellular Killing Test to allow its application to HCPB.Se da cuenta de un método rápido para la cuantización del flujo citométrico de la fagocitosis en sangre periférica completamente heparinizada (HCPB, mediante la utilización de partículas de látex phycoerythrin-conjugadas de 1µm de diámetro disponibles comercialmente. El método es más rápido y presenta mayor reproducibilidad que la técnica estandar de Bjerknes' (1984 utilizando propidium iodide-teñida Candida albicans, aplicada convencionalmente a la capa leucocitica de sangre periférica pero modificada por HCPB. Tambien damos cuenta de una modificación de Bjerknes' Intracellular Killing Test para permitir su aplicación a HCPB.

  19. Comparison of monoclonal antibodies reactive with lymphocyte subsets in routinely fixed paraffin-embedded material: flow cytometric analyses, immunoperoxidase staining and influence of fixatives.

    Directory of Open Access Journals (Sweden)

    Yoshino,Tadashi

    1990-10-01

    Full Text Available We have attempted to clarify the characteristics of monoclonal antibodies (MAbs detecting lymphocyte subsets in fixed materials. We examined by means of flow cytometric technique influences of fixatives and reactivity with malignant lymphomas (MLs. Specific markers for T-cells were UCHL1 and OPD4, which reacted especially with helper/inducer T-cells. MT1 recognized almost all of T-cells from peripheral blood and tonsils, but reacted with a part of B-MLs. As for B-cell markers, L26 was the most reliable marker for B-MLs. L26 and MB1 antigens could not be detected on living cells flow cytometrically. LN1 reacted with a part of T-cells as well as B-cells, but fluorescent intensity of the former was apparently stronger than that of the latter. Although LN2 antigen was located mainly in the cytoplasm close to the nuclear membrane immunohistochemically, it could be detected on living cells flow cytometrically. LN2 positive cells belonged to B-cells in peripheral blood and tonsils. When fixed for relatively short time, B5 and buffered formalin were better for examining MAbs than non-buffered formalin and ethanol.

  20. Cytometric Approach for Detection of Encephalitozoon intestinalis, an Emergent Agent▿

    Science.gov (United States)

    Barbosa, Joana; Rodrigues, Acácio Gonçalves; Pina-Vaz, Cidália

    2009-01-01

    Encephalitozoon intestinalis is responsible for intestinal disease in patients with AIDS and immunocompetent patients. The infectious form is a small spore that is resistant to water treatment procedures. Its detection is very important, but detection is very cumbersome and time-consuming. Our main objective was to develop and optimize a specific flow cytometric (FC) protocol for the detection of E. intestinalis in hospital tap water and human feces. To determine the optimal specific antibody (Microspor-FA) concentration, a known concentration of E. intestinalis spores (Waterborne, Inc.) was suspended in hospital tap water and stool specimens with different concentrations of Microspor-FA, and the tap water and stool specimens were incubated under different conditions. The sensitivity limit and specificity were also evaluated. To study spore infectivity, double staining with propidium iodide (PI) and Microspor-FA was undertaken. Distinct approaches for filtration and centrifugation of the stool specimens were used. E. intestinalis spores stained with 10 μg/ml of Microspor-FA at 25°C overnight provided the best results. The detection limit was 5 × 104 spores/ml, and good specificity was demonstrated. Simultaneous staining with Microspor-FA and PI ensured that the E. intestinalis spores were dead and therefore noninfectious. With the stool specimens, better spore recovery was observed with a saturated solution of NaCl and centrifugation at 1,500 × g for 15 min. A new approach for the detection of E. intestinalis from tap water or human feces that ensures that the spores are not viable is now available and represents an important step for the prevention of this threat to public health. PMID:19439525

  1. Ultra-Fast and Optimized Method for the Preparation of Rodent Testicular Cells for Flow Cytometric Analysis

    Directory of Open Access Journals (Sweden)

    López-Carro Beatriz

    2009-01-01

    Full Text Available Abstract Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the complex organization of the mammalian testicular tissue, various methods have been used to obtain enriched or purified cell populations, including flow cell sorting. Current protocols are usually time-consuming and may imply loss of short-lived RNAs, which is undesirable for expression profiling. We describe an optimized method to speed up the preparation of suitable testicular cell suspensions for cytometric analysis of different spermatogenic stages from rodents. The procedure takes only 15 min including testis dissection, tissue cutting, and processing through the Medimachine System (Becton Dickinson. This method could be a substitute for the more tedious and time-consuming cell preparation techniques currently in use.

  2. Ultra-Fast and Optimized Method for the Preparation of Rodent Testicular Cells for Flow Cytometric Analysis

    Directory of Open Access Journals (Sweden)

    Rodríguez-Casuriaga Rosana

    2009-03-01

    Full Text Available Abstract Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the complex organization of the mammalian testicular tissue, various methods have been used to obtain enriched or purified cell populations, including flow cell sorting. Current protocols are usually time-consuming and may imply loss of short-lived RNAs, which is undesirable for expression profiling. We describe an optimized method to speed up the preparation of suitable testicular cell suspensions for cytometric analysis of different spermatogenic stages from rodents. The procedure takes only 15 min including testis dissection, tissue cutting, and processing through the Medimachine System (Becton Dickinson. This method could be a substitute for the more tedious and time-consuming cell preparation techniques currently in use.

  3. Flow cytometric immunophenotypic characteristics of plasma cell leukemia

    Directory of Open Access Journals (Sweden)

    Barbara Kruk

    2011-04-01

    Full Text Available The aim of this prospective study was to define the flow cytometric characteristics of simultaneously investigated bone marrow and peripheral blood plasma cells antigens expression in 36 plasma cell leukemia (PCL patients. The immunophenotypic profile of plasma cells was determined with a panel of monoclonal antibodies. The antigen expression intensity was calculated as relative fluorescence intensity (RFI. Bone marrow plasma cells showed expression of particular antigens in the following proportion of cases: CD49d 100%, CD29 94%, CD54 93%, CD44 83%, CD56 60%, CD18 26%, CD11b 29%, CD11a 19%, CD117 27%, CD71 30%, CD126 100% and CD19 0%, while the expression of those antigens on peripheral blood plasma cells was present in the following percentage of patients: CD49d 100%, CD29 96%, CD54 93%, CD44 95%, CD56 56%, CD18 50%, CD11b 53%, CD11a 29%, CD117 26%, CD71 28%, CD126 100% and CD19 0%. The expression of CD54 was significantly higher than that of adhesion molecules belonging to the integrin b2 family: CD11a, CD18 and CD11b, on both bone marrow and peripheral blood cells (p < 0.01. Expression of CD18, CD11a and CD11b was differential between two cell compartments: lower on bone marrow and higher on peripheral blood cells. We found that plasma cells in the bone marrow of patients with plasma cell leukaemia showed significantly greater granularity and size than those in the peripheral blood (p = 0.0001 and p = 0.04, respectively. However, no differences in cell size or granularity were revealed between bone marrow plasma cells from patients with PCL and multiple myeloma. In conclusion, impaired expression of adhesion molecules such as CD11a/CD18 (LFA-1 or CD56 may explain hematogenic dissemination characterizing PCL. The following pattern of adhesion molecule expression according to the proportion of plasma cells expressing a given antigen in peripheral blood and bone marrow and arranged in diminishing order may be established: CD49d > CD44 > CD54

  4. An imaging flow cytometric method for measuring cell division history and molecular symmetry during mitosis.

    Science.gov (United States)

    Filby, Andrew; Perucha, Esperanza; Summers, Huw; Rees, Paul; Chana, Prabhjoat; Heck, Susanne; Lord, Graham M; Davies, Derek

    2011-07-01

    Asymmetric cell division is an important mechanism for generating cellular diversity, however, techniques for measuring the distribution of fate-regulating molecules during mitosis have been hampered by a lack of objectivity, quantitation, and statistical robustness. Here we describe a novel imaging flow cytometric approach that is able to report a cells proliferative history and cell cycle position using dye dilution, pH3, and PI staining to then measure the spatial distribution of fluorescent signals during mitosis using CCD-derived imagery. Using Jurkat cells, resolution of the fluorescently labeled populations was comparable to traditional PMT based cytometers thus eliminating the need to sort cells with specific division histories for microscopy. Subdividing mitotic stages by morphology allowed us to determine the time spent in each cell cycle phase using mathematical modeling approaches. Furthermore high sample throughput allowed us to collect statistically relevant numbers of cells without the need to use blocking agents that artificially enrich for mitotic events. The fluorescent imagery was used to measure PKCζ protein and EEA-1+ endosome distribution during different mitotic phases in Jurkat cells. While telophase cells represented the favorable population for measuring asymmetry, asynchronously dividing cells spent approximately 43 seconds in this stage, explaining why they were present at such low frequencies. This necessitated the acquisition of large cell numbers. Interestingly we found that PKCζ was inherited asymmetrically in 2.5% of all telophasic events whereas endosome inheritance was significantly more symmetrical. Furthermore, molecular polarity at early mitotic phases was a poor indicator of asymmetry during telophase highlighting that, though rare, telophasic events represented the best candidates for asymmetry studies. In summary, this technique combines the spatial information afforded by fluorescence microscopy with the statistical

  5. Cell kinetics in a model of artificial skin. An immunohistochemical and flow cytometric analysis

    Directory of Open Access Journals (Sweden)

    A Casasco

    2009-12-01

    Full Text Available Bioengineered organs raised in vitro are candidate substitutes for natural organs in biological, pharmacological and clinical applications. We have studied cell kinetics in a human skin equivalent (HSE using a combined immunohistochemical and flow cytometric approach. Morphological analysis has shown that, relative to unstimulated natural skin, cell proliferation mainly occurs in the basal layer of the epidermal equivalent. Immunohistochemical and flow cytometric measurements of the growth fraction suggested a cell turnover comparable to that of natural skin. Immunohistochemical labelling indices matched well with flow cytometric data. These observations are consistent with morphological and histochemical data demonstrating normal cell differentiation and tissue architecture in HSE and suggest that such HSE may be a usefull substitute for human skin.

  6. Flow cytometric method for measuring chromatin fragmentation in fixed sperm from yellow perch (Perca flavescens).

    Science.gov (United States)

    Jenkins, J A; Draugelis-Dale, R O; Pinkney, A E; Iwanowicz, L R; Blazer, V S

    2015-03-15

    Declining harvests of yellow perch, Perca flavescens, in urbanized watersheds of Chesapeake Bay have prompted investigations of their reproductive fitness. The purpose of this study was to establish a flow cytometric technique for DNA analysis of fixed samples sent from the field to provide reliable gamete quality measurements. Similar to the sperm chromatin structure assay, measures were made on the susceptibility of nuclear DNA to acid-induced denaturation, but used fixed rather than live or thawed cells. Nuclei were best exposed to the acid treatment for 1 minute at 37 °C followed by the addition of cold (4 °C) propidium iodide staining solution before flow cytometry. The rationale for protocol development is presented graphically through cytograms. Field results collected in 2008 and 2009 revealed DNA fragmentation up to 14.5%. In 2008, DNA fragmentation from the more urbanized watersheds was significantly greater than from reference sites (P = 0.026) and in 2009, higher percentages of haploid testicular cells were noted from the less urbanized watersheds (P = 0.032) indicating better reproductive condition at sites with less urbanization. For both years, total and progressive live sperm motilities by computer-assisted sperm motion analysis ranged from 19.1% to 76.5%, being significantly higher at the less urbanized sites (P fragmented DNA to be denatured under standard conditions, or 1 minute at 37 °C with 10% buffered formalin-fixed cells. The study of fixed sperm makes possible the restrospective investigation of germplasm fragmentation, spermatogenic ploidy patterns, and chromatin compaction levels from samples translocated over distance and time. The protocol provides an approach that can be modified for other species across taxa.

  7. Flow cytometric method for measuring chromatin fragmentation in fixed sperm from yellow perch (Perca flavescens)

    Science.gov (United States)

    Jenkins, Jill A.; Draugelis-Dale, Rassa O.; Pinkney, Alfred E.; Iwanowicz, Luke R.; Blazer, Vicki

    2015-01-01

    Declining harvests of yellow perch, Perca flavescens, in urbanized watersheds of Chesapeake Bay have prompted investigations of their reproductive fitness. The purpose of this study was to establish a flow cytometric technique for DNA analysis of fixed samples sent from the field to provide reliable gamete quality measurements. Similar to the sperm chromatin structure assay, measures were made on the susceptibility of nuclear DNA to acid-induced denaturation, but used fixed rather than live or thawed cells. Nuclei were best exposed to the acid treatment for 1 minute at 37 °C followed by the addition of cold (4 °C) propidium iodide staining solution before flow cytometry. The rationale for protocol development is presented graphically through cytograms. Field results collected in 2008 and 2009 revealed DNA fragmentation up to 14.5%. In 2008, DNA fragmentation from the more urbanized watersheds was significantly greater than from reference sites (P = 0.026) and in 2009, higher percentages of haploid testicular cells were noted from the less urbanized watersheds (P = 0.032) indicating better reproductive condition at sites with less urbanization. For both years, total and progressive live sperm motilities by computer-assisted sperm motion analysis ranged from 19.1% to 76.5%, being significantly higher at the less urbanized sites (P fragmented DNA to be denatured under standard conditions, or 1 minute at 37 °C with 10% buffered formalin–fixed cells. The study of fixed sperm makes possible the restrospective investigation of germplasm fragmentation, spermatogenic ploidy patterns, and chromatin compaction levels from samples translocated over distance and time. The protocol provides an approach that can be modified for other species across taxa.

  8. Long-term storage of samples for flow cytometric DNA analysis

    DEFF Research Database (Denmark)

    Vindeløv, L L; Christensen, I J; Keiding, N

    1983-01-01

    A simple procedure for long-term storage of cells for flow cytometric DNA analysis was developed and tested. The cells were stored as single cells or fine-needle aspirates suspended in a citrate buffer with dimethylsulfoxide (DMSO), or as small blocks of tissue from solid tumors. The cells were...

  9. Flow cytometric sorting of paraffin-embedded tumor tissues considerably improves molecular genetic analysis

    NARCIS (Netherlands)

    Jordanova, ES; Corver, WE; Vonk, MJ; Leers, MPG; Riemersma, SA; Schuuring, E; Kluin, PM

    2003-01-01

    The characterization of genetic aberrations in paraffin-embedded tumor material is impaired by contaminating normal cells. In the present study on the genetic causes of loss of HLA expression in diffuse large B-cell lymphoma (DLBCL), we compared the efficacy of microdissection with flow cytometric s

  10. Flow cytometric determination of circulating immune complexes with the indirect granulocyte phagocytosis test

    NARCIS (Netherlands)

    Terstappen, L.W.M.M.; Grooth, de B.G.; Nolten, G.M.J.; Napel, ten C.H.H.; Berkel, van W.; Greve, J.

    1985-01-01

    A method for the determination of circulating immune complexes (CIC) was adapted for flow cytometric analysis. Human granulocytes were used to phagocytose IgG-bearing CIC of serum from systemic lupus erythematosus (SLE) patients. A method for labeling the phagocytosed CIC with FITC-conjugated anti-h

  11. Discrimination of bromodeoxyuridine labelled and unlabelled mitotic cells in flow cytometric bromodeoxyuridine/DNA analysis

    DEFF Research Database (Denmark)

    Jensen, P O; Larsen, J K; Christensen, I J

    1994-01-01

    Bromodeoxyuridine (BrdUrd) labelled and unlabelled mitotic cells, respectively, can be discriminated from interphase cells using a new method, based on immunocytochemical staining of BrdUrd and flow cytometric four-parameter analysis of DNA content, BrdUrd incorporation, and forward and orthogona...

  12. A flow-cytometric method to evaluate eosinophil-mediated uptake of probiotic Lactobacillus reuteri.

    Science.gov (United States)

    Kraemer, Laura S; Brenner, Todd A; Krumholz, Julia O; Rosenberg, Helene F

    2017-03-27

    Eosinophils are resident leukocytes of gut mucosa. Here we present a combined flow cytometric-antibiotic protection assay to identify mouse eosinophils capable of bacterial uptake, specifically, Gram-positive Lactobacillus reuteri, in studies performed ex vivo. The assay may be adapted for use in vivo.

  13. Cytometric analysis of shape and DNA content in mammalian sperm

    Energy Technology Data Exchange (ETDEWEB)

    Gledhill, B.L.

    1983-10-10

    Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. Sperm were analyzed by flow cytometry and slit-scan flow analysis for injury following the exposure of testes to mutagens. The utility of flow cytometry in genotoxin screening and monitoring of occupational exposure was evaluated. The technique proved valuable in separation of X- and Y-chromosome bearing sperm and the potential applicability of this technique in artificial insemination and a solution, of accurately assessing the DNA content of sperm were evaluated-with reference to determination of X- and Y-chromosome bearing sperm.

  14. Results of external quality control study in flow cytometric acute lymphoblastic leukemia diagnostics

    Directory of Open Access Journals (Sweden)

    A. M. Popov

    2016-01-01

    Full Text Available Comparison of interpretation of acute lymphoblastic leukemia (ALL flow cytometric diagnostics data was the aim of the study. Immunophenotyping data obtained from 10 patients with ALL were analysed separately in 26 laboratories from Russian Federation and Kazahstan. Results comparison showed four main type of discordance: B-lineage ALL diagnostics during heavy bone marrow regeneration, great variability of T-ALL interpretation, complexity of ambiguous lineage acute leukemia and, finally, very different report types, unique for each laboratory. All these problems are the serious obstacles for standardization of flow cytometric ALL diagnostics in multicenter setting. Continuation of similar QC rounds following by consecutive discussions with further development of consensus diagnostic algorithm could be the first step for standardization of ALL immunophenotyping in Russian Federation and CIS countries.

  15. Downregulation of pro-inflammatory cytokines by lupeol measured using cytometric bead array immunoassay.

    Science.gov (United States)

    Ahmad, Sheikh Fayaz; Pandey, Anjali; Kour, Kiranjeet; Bani, Sarang

    2010-01-01

    The objective of the study was to investigate the activity of Lupeol (LUP) on proinflammatory and anti-inflammatory cytokines in the pleural exudate from male swiss albino mice. We applied Cytometric bead array technology for simultaneously measurement of these cytokines in pleurisy induced mice treated with lupeol in graded oral doses. Cytometric bead array uses the sensitivity of amplified fluorescence detection by flowcytometer to measure soluble analytes in a particle based immune assay. This assay can accurately quantitate 5 cytokines in a 50 microlitre sample volume. Oral administration of LUP at doses of 25, 50, 100 and 200 mg/kg p.o. produced dose related inhibition of IL-2, IFN-gamma and TNF-alpha in the pleural exudate with the most significant effect at 100 mg/kg oral dose. LUP had a non significant inhibitory effect on the levels of IL-4 and IL-5.

  16. Simultaneous preparation of RNA and nuclei for Northern blot and flow cytometric analysis

    Energy Technology Data Exchange (ETDEWEB)

    Tesfaigzi, J.; Jaramillo, R. [Inhalation Toxicology Research Inst., Albuquerque, NM (United States)

    1996-11-01

    Several methods have been developed to quantify RNA synthesis during the progression of the cell cycle. The rate of RNA synthesis can be detected during different stages of the cell cycle by staining cells with agents that intercalate with nucleic acids. For example, following staining of mammalian cells with acridin orange, the green and red fluorescence that correlates with DNA and RNA content, respectively, can be analyzed by flow cytometry. Increase in RNA content during the progression of cells through the cell cycle can be measured after staining with acridin orange. RNA synthesis resulting from the stimulation of quiescent cells with various growth factors has also been demonstrated by labeling cells with bromo-uridine and using the anti-bromo-deoxyuridine antibody. These methods allow measurement of the overall RNA content in cells; however, they do not allow the measurement of the levels of specific mRNAs throughout the cell cycle. Current methods to quantify specific mRNAs generally require the preparation of a large number of cells (5--10 {times} 10{sup 6} cells) to carry out flow cytometric analyses and to isolate RNA for Northern blot analysis or solution hybridization. In this report, the authors describe a method of simultaneously preparing RNA and nuclei for Northern blot and flow cytometric analyses, respectively. The minimum number of nuclei required to obtain flow cytometric data and the effect of conserving nuclei in methanol for several days are also presented.

  17. Postlarval muscle growth in fish: a DNA flow cytometric and morphometric analysis.

    Science.gov (United States)

    Alfei, L; Maggi, F; Parvopassu, F; Bertoncello, G; De Vita, R

    1989-01-01

    The mechanism of postlarval fish myotomal growth was investigated in trout (Salmo gairdneri) by means of morphometric and cytofluorometric analysis. The mechanism by which new fibres are added during postlarval growth (hyperplasia) is not fully understood. In histological cross sections these new fibres have a small diameter which give the muscle a "mosaic" appearance. One hypothesis suggested that they could be derived from the proliferative activity of satellite cells. DNA cytofluorometric analysis of nuclei suspensions obtained from trout white myotomal muscle during different developmental stages (eleutherembyronic; alevin; yearling and adult) showed a consistently low S-cytometric phase during all stage in which myofibres of small diameters were present. The percentage of such small fibres, determined by morphometric analysis, suggested that satellite cells are the proliferative population. In fact, their percentages, as determined by morphometric analysis in histological section, bear a linear relationship with the S-cytometric phase percent nuclei (R = 0.927). Only in adults (67 cm in size) there was a significant decrease in the S-cytometric phase. At this stage, in histological sections, the myotomal muscle no longer had a "mosaic" appearance because of the disappearance of the small fibres. It may, therefore, be supposed that in the cm 67 adult specimens, the proliferative population is entering the G0 phase. It is known, in fact, that muscle growth proceeds only by fibre hypertrophy in trout longer than 70 cm in length (Stickland, 1983).

  18. Laser-based flow cytometric analysis of genotoxicity of humans exposed to ionizing radiation during the Chernobyl accident

    Science.gov (United States)

    Jensen, Ronald H.; Bigbee, William L.; Langlois, Richard G.; Grant, Stephen G.; Pleshanov, Pavel G.; Chirkov, Andre A.; Pilinskaya, Maria A.

    1991-05-01

    An analytical technique has been developed that allows laser-based flow cytometric measurement of the frequency of red blood cells that have lost allele-specific expression of a cell surface antigen due to genetic toxicity in bone marrow precursor cells. Previous studies demonstrated a correlation of such effects with the exposure of each individual to mutagenic phenomena, such as ionizing radiation, and the effects can persist for the lifetime of each individual. During the emergency response to the nuclear power plant accidert at Chemobyl, Ukraine, USSR, a number of people were exposed to whole body doses of ioniing radiation. Some of these individuals were tested with this laser-based assay and found to express a dose-dependent increase in the frequency of variant red blood cells that appears to be a persistent biological effect. This effect is similar to that which was previously observed in individuals who were exposed to ionizing radiation at Hiroshima in 1945 because of the A-bomb explosion. All data indicate that this assay might well be used as a biodosimeter to estimate radiation dose and also as an element to be used for estimating the risk of each individual to develop cancer due to radiation exposure.

  19. Cytologic and DNA-Cytometric Early Diagnosis of Oral Cancer

    Directory of Open Access Journals (Sweden)

    Torsten W. Remmerbach

    2001-01-01

    Full Text Available Objective. The aim of this prospective study was to report on the diagnostic accuracy of conventional oral exfoliative cytology taken from white‐spotted, ulcerated or other suspicious oral lesions in our clinic. In addition we checked DNA‐image cytometry as an adjuvant diagnostic tool. Our hypothesis is that DNA‐aneuploidy is a sensitive and specific marker for the early identification of tumor cells in oral brushings. Study design. 251 cytological diagnoses obtained from exfoliative smears of 181 patients from macroscopically suspicious lesions of the oral mucosa and from clinically seemingly benign oral lesions which were exisiced for establishing histological diagnoses were compared with histological and/or clinical follow‐ups of the respective patients. Additionally nuclear DNA‐contents were measured after Feulgen restaining using a TV image analysis system. Results. Sensitivity of our cytological diagnosis on oral smears for the detection of cancer cells was 94.6%, specificity 99.5%, positive predictive value 98.1% and negative predictive value 98.5%. DNA‐aneuploidy was assumed if abnormal DNA‐stemlines or cells with DNA‐content greater 9c were observed. On this basis the prevalence of DNA‐aneuploidy in smears of oral squamous cell carcinomas in situ or invasive carcinomas was 96.4%. Sensitivity of DNA‐aneuploidy in oral smears for the detection of cancer cells was 96.4%, specificity 100%, positive predictive value 100% and negative 99.0%. The combination of both techniques increased the sensivity to 98.2%, specificity to 100%, positive predictive value to 100% and negative to 99.5%. Conclusions. Brush cytology of all visible oral lesions, if they are clinically considered as suspicious for cancer, are an easily practicable, cheap, non‐invasive, painless, safe and accurate screening method for detection of oral precancerous lesions, carcinoma in situ or invasive squamous cell carcinoma in all stages. We conclude that

  20. Karyological and flow cytometric evidence of triploid specimens in Bufo viridis (Amphibia Anura

    Directory of Open Access Journals (Sweden)

    D Cavallo

    2010-01-01

    Full Text Available Karyological and flow cytometric (FCM analyses were performed on a group of 14 green toads of the Bufo viridis species from seven Eurasian populations. Both approaches gave concordant results concerning the DNA ploidy level. All the populations examined were represented exclusively by diploid or tetraploid specimens, except one, where triploids were found. Results evidenced an interpopulation variability in DNA content against the same ploidy level, as well as an unusually high number of triploids in a particular reproductive place. The origin of polyploidy and the presence and persistence of a high number of triploids in a particular population are discussed.

  1. Two and three-color fluorescence flow cytometric analysis of immunoidentified viable bacteria.

    Science.gov (United States)

    Barbesti, S; Citterio, S; Labra, M; Baroni, M D; Neri, M G; Sgorbati, S

    2000-07-01

    Traditional culture methods well established in the past and still in use are not able to detect the environmental microorganisms that exist in a viable but not culturable state. A number of different fluorescence-based assays have been developed over the past decade to detect and identify viable bacteria in the environment. We have developed a simple and rapid method for measuring the number and viability of immunolabeled bacteria by means of a two/three color fluorescence flow cytometric analysis. After washing, cultured bacteria in suspension were labeled with a rabbit polyclonal antibody recognizing the wall lipopolysaccharide complex. A secondary biotinylated anti-rabbit polyclonal antibody was added allowing the cells to be labeled with the streptavidin R-phycoerythrin-Cyanine 5 (RPE-Cy5) fluorochrome. Before flow cytometric analysis, bacterial suspensions were stained with SYBR Green I and propidium iodide which stain all of the cells and the non viable ones, respectively. With the appropriate filter sets of both Bryte-HS (Bio-Rad, Hercules, CA) and FACScan (Becton Dickinson, San Jose, CA) flow cytometers, the measurement of separated green (SYBR Green I), orange-red (propidium iodide), and far red (RPE-Cy5) fluorescence was possible, allowing the enumeration of viable immunodetected bacteria. The entire protocol is completed in less than 3 h, offering numerous possibilities for rapid and precise analyses in sanitary, industrial, and environmental microbiology. Copyright 2000 Wiley-Liss, Inc.

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

    Directory of Open Access Journals (Sweden)

    Katarzyna Czaczyk

    2012-06-01

    Full Text Available Background. Flow cytometry constitutes an alternative for traditional methods of microorganisms identifi cation 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 signifi cant 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 fl ow cytometry seems to constitute an ideal tool for the assessment of microbial contamination on the surface of technological lines. Material and methods. 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 fl ow cytometer. The measured parameters were forward and side scatter of laser light signals allowing the estimation of microbial cell contents in each sample. Results. 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. Conclusions. 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 laborsaving detection of microbial contamination in food industry.

  3. Multiplex competitive microbead-based flow cytometric immunoassay using quantum dot fluorescent labels

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Hye-Weon; Kim, In S. [School of Environmental Science and Engineering, Gwangju Institute of Science and Technology (GIST), 261 Cheomdan-gwagiro, Buk-gu, Gwangju (Korea, Republic of); Niessner, Reinhard [Chair for Analytical Chemistry, Institute of Hydrochemistry, Technische Universitaet Muenchen, Marchioninistrasse 17, 81377 Muenchen (Germany); Knopp, Dietmar, E-mail: dietmar.knopp@ch.tum.de [Chair for Analytical Chemistry, Institute of Hydrochemistry, Technische Universitaet Muenchen, Marchioninistrasse 17, 81377 Muenchen (Germany)

    2012-10-31

    Highlights: Black-Right-Pointing-Pointer First time, duplex competitive bead-based flow cytometric immunoassay was developed using ODs. Black-Right-Pointing-Pointer Antibody-coated QD detection probes and antigen-immobilized microspheres were synthesized. Black-Right-Pointing-Pointer The two model target analytes were low molecular weight compounds of microbial and chemical origin. Black-Right-Pointing-Pointer The determination of different water types was possible after simple filtration of samples. - Abstract: In answer to the ever-increasing need to perform the simultaneous analysis of environmental hazards, microcarrier-based multiplex technologies show great promise. Further integration with biofunctionalized quantum dots (QDs) creates new opportunities to extend the capabilities of multicolor flow cytometry with their unique fluorescence properties. Here, we have developed a competitive microbead-based flow cytometric immunoassay using QDs fluorescent labels for simultaneous detection of two analytes, bringing the benefits of sensitive, rapid and easy-of-manipulation analytical tool for environmental contaminants. As model target compounds, the cyanobacterial toxin microcystin-LR and the polycyclic aromatic hydrocarbon compound benzo[a]pyrene were selected. The assay was carried out in two steps: the competitive immunological reaction of multiple targets using their exclusive sensing elements of QD/antibody detection probes and antigen-coated microsphere, and the subsequent flow cytometric analysis. The fluorescence of the QD-encoded microsphere was thus found to be inversely proportional to target analyte concentration. Under optimized conditions, the proposed assay performed well within 30 min for the identification and quantitative analysis of the two environmental contaminants. For microcystin-LR and benzo[a]pyrene, dose-response curves with IC{sub 50} values of 5 {mu}g L{sup -1} and 1.1 {mu}g L{sup -1} and dynamic ranges of 0.52-30 {mu}g L{sup -1} and 0

  4. Biotinylation of interleukin-2 (IL-2) for flow cytometric analysis of IL-2 receptor expression. Comparison of different methods

    NARCIS (Netherlands)

    M.O. de Jong (Marg); H. Rozemuller (Henk); J.G.J. Bauman (J. G J); J.W.M. Visser (Jan)

    1995-01-01

    textabstractThe main prerequisites for the use of biotinylated ligands to study the expression of growth factor receptors on heterogeneous cell populations, such as peripheral blood or bone marrow, by flow cytometric methods, are that the biotinylated ligand retains its binding ability and that bind

  5. Seasonality in molecular and cytometric diversity of marine bacterioplankton: the reshuffling of bacterial taxa by vertical mixing

    KAUST Repository

    García, Francisca C.

    2015-07-17

    The ’cytometric diversity’ of phytoplankton communities has been studied based on single-cell properties, but the applicability of this method to characterize bacterioplankton has been unexplored. Here, we analysed seasonal changes in cytometric diversity of marine bacterioplankton along a decadal time-series at three coastal stations in the Southern Bay of Biscay. Shannon-Weaver diversity estimates and Bray-Curtis similarities obtained by cytometric and molecular (16S rRNA tag sequencing) methods were significantly correlated in samples from a 3.5-year monthly time-series. Both methods showed a consistent cyclical pattern in the diversity of surface bacterial communities with maximal values in winter. The analysis of the highly resolved flow cytometry time-series across the vertical profile showed that water column mixing was a key factor explaining the seasonal changes in bacterial composition and the winter increase in bacterial diversity in coastal surface waters. Due to its low cost and short processing time as compared to genetic methods, the cytometric diversity approach represents a useful complementary tool in the macroecology of aquatic microbes.

  6. Biotinylation of interleukin-2 (IL-2) for flow cytometric analysis of IL-2 receptor expression. Comparison of different methods

    NARCIS (Netherlands)

    M.O. de Jong (Marg); H. Rozemuller (Henk); J.G.J. Bauman (J. G J); J.W.M. Visser (Jan)

    1995-01-01

    textabstractThe main prerequisites for the use of biotinylated ligands to study the expression of growth factor receptors on heterogeneous cell populations, such as peripheral blood or bone marrow, by flow cytometric methods, are that the biotinylated ligand retains its binding ability and that bind

  7. A short-term in vitro test for tumour sensitivity to adriamycin based on flow cytometric DNA analysis

    DEFF Research Database (Denmark)

    Engelholm, S A; Spang-Thomsen, M; Vindeløv, L L

    1983-01-01

    A new method to test the sensitivity of tumour cells to chemotherapy is presented. Tumour cells were incubated in vitro on agar, and drug-induced cell cycle perturbation was monitored by flow cytometric DNA analysis. In the present study the method was applied to monitor the effect of adriamycin...

  8. Simple flow cytometric detection of haemozoin containing leukocytes and erythrocytes for research on diagnosis, immunology and drug sensitivity testing

    NARCIS (Netherlands)

    Frita, R.; Rebelo, M.; Pamplona, A.; Vigario, A.M.; Mota, M.M.; Grobusch, M.P.; Haenscheid, T.

    2011-01-01

    Background: Malaria pigment (haemozoin, Hz) has been the focus of diverse research efforts. However, identification of Hz-containing leukocytes or parasitized erythrocytes is usually based on microscopy, with inherent limitations. Flow cytometric detection of depolarized Side-Scatter is more accurat

  9. A simple and sensitive flow cytometric assay for determination of the cytotoxic activity of human natural killer cells

    NARCIS (Netherlands)

    Radosevic, Katarina; Radosevic, K.; Garritsen, Henk S.P.; Garritsen, H.S.P.; van Graft, M.; van Graft, Marja; de Grooth, B.G.; Greve, Jan

    1990-01-01

    A new, simple and sensitive flow cytometric assay for the determination of the cytotoxic activity of human natural killer cells is described. The assay is based on the use of two fluorochromes. The target cell population is stained with one fluorochrome (octadecylamine-fluorescein isothiocyanate,

  10. Flow-Cytometric Isolation of Human Antibodies from a Nonimmune Saccharomyces cerevisiae Surface Display Library

    Energy Technology Data Exchange (ETDEWEB)

    Feldhaus, Michael (BATTELLE (PACIFIC NW LAB)); Siegel, Robert W.(BATTELLE (PACIFIC NW LAB)); Opresko, Lee (BATTELLE (PACIFIC NW LAB)); Coleman, James R.(BATTELLE (PACIFIC NW LAB)); Feldhaus, Jane M.(BATTELLE (PACIFIC NW LAB)); Yeung, Yik A.(Massachusetts Institute Of Tec); Cochran, Jennifer R.(Massachusetts Institute Of Tec); Heinzelman, Peter (Massachusetts Institute Of Tec); Colby, David (Massachusetts Institute Of Tec); Swers, Jeffrey (Massachusetts Institute Of Tec); Graff, Christilyn (Massachusetts Institute Of Tec); Wiley, H Steven (BATTELLE (PACIFIC NW LAB)); Wittrup, K D.(Massachusetts Institute Of Tec)

    2003-02-28

    A nonimmune library of 109 human antibody scFv fragments has been cloned and expressed on the surface of yeast, and nanomolar-affinity scFvs routinely obtained by magnetic bead screening and flow cytometric sorting. The yeast library can be amplified 1010-fold without measurable loss of clonal diversity, enabling effectively indefinite expansion of the library. The expression, stability, and antigen binding properties of more than 50 isolated scFv clones were assessed directly on the yeast cell surface by immunofluorescent labeling and flow cytometry, obviating separate subcloning, expression, and purification steps and thereby expediting the isolation of novel affinity reagents. The ability to use multiplex library screening demonstrates the utility of this approach for high throughput antibody isolation for proteomics applications.

  11. A flow cytometric method for characterization of circulating cell-derived microparticles in plasma

    DEFF Research Database (Denmark)

    Nielsen, Morten Hjuler; Beck-Nielsen, Henning; Andersen, Morten Nørgaard;

    2014-01-01

    BACKGROUND AND AIM: Previous studies on circulating microparticles (MPs) indicate that the majority of MPs are of a size below the detection limit of most standard flow cytometers. The objective of the present study was to establish a method to analyze MP subpopulations above the threshold...... of detection of a new generation BD FACSAria™ III digital flow cytometer. METHODS: We analyzed MP subpopulations in plasma from 24 healthy individuals (9 males and 15 females). MPs were identified according to their size (.... The sensitivity of the flow cytometer was tested against that of a previous-generation instrument FC500. Reproducibility of the FACSAria and our set-up was investigated, and the percentage of phosphatidylserine (PS) exposing MPs binding Lactadherin was determined. RESULTS: By using a flow cytometric approach we...

  12. Standardizing flow cytometric assays in long-term population-based studies

    Science.gov (United States)

    Melzer, Susanne; Bocsi, Jozsef; Tárnok, Attila

    2015-03-01

    Quantification of leukocyte subpopulations and characterization of antigen-expression pattern on the cellular surface can play an important role in diagnostics. The state of cellular immunology on the single-cell level was analyzed by polychromatic flow cytometry in a recent comparative study within the average Leipzig population (LIFE - Leipzig Research Centre for Civilization Diseases). Data of 1699 subjects were recorded over a long-time period of three years (in a total of 1126 days). To ensure compatibility of such huge data sets, quality-controls on many levels (stability of instrumentation, low intra-laboratory variance and reader independent data analysis) are essential. The LIFE study aims to analyze various cytometric pattern to reveal the relationship between the life-style, the environmental effects and the individual health. We therefore present here a multi-step quality control procedure for long-term comparative studies.

  13. Procarbazine effects on spermatogenesis in golden hamster: a flow cytometric evaluation.

    Science.gov (United States)

    Weissenberg, R; Golan, R; Shochat, L; Lewin, L M

    2002-01-01

    The response of hamster testis to the administration of 450mg/kg procarbazine (PCB) over a period of 4 weeks was evaluated. Flow cytometry was used to investigate changes in cell populations in testicular single cell suspensions and to correlate these changes with those observed in histological sections. PCB caused significant decrease in testicular and epididymal weight and a drastic reduction in haploid cells and spermatogenic arrest, demonstrating variation among the test animals. The results obtained confirm previous observations concerning detrimental effects of PCB upon spermatogenesis in species such as the rat and mouse, though its effect on hamster testis is milder and does not include the germinal stem cells. The histological evaluation of the testis showed a good correlation with flow cytometric evaluation, emphasizing the usefulness of this method in providing quantitative and rapid results.

  14. Flow cytometric DNA analysis of ducks accumulating 137Cs on a reactor reservoir.

    Science.gov (United States)

    George, L S; Dallas, C E; Brisbin, I L; Evans, D L

    1991-06-01

    The objective of this study was to detect red blood cell (rbc) DNA abnormalities in male, game-farm mallard ducks as they ranged freely and accumulated 137Cs (radiocesium) from an abandoned nuclear reactor cooling reservoir. Prior to release, the ducks were tamed to enable recapture at will. Flow cytometric measurements conducted at intervals during the first year of exposure yielded cell cycle percentages of DNA (G0/G1, S, G2 + M phases) of rbc, as well as coefficients of variation (CV) in the G0/G1 phase. DNA histograms of exposed ducks were compared with two sets of controls which were maintained 30 and 150 miles from the study site. 137Cs live wholebody burdens were also measured in these animals in a parallel kinetics study, and an approximate steady-state equilibrium was attained after about 8 months. DNA histograms from 2 of the 14 contaminated ducks revealed DNA aneuploid-like patterns after 9 months exposure. These two ducks were removed from the experiment at this time, and when sampled again 1 month later, one continued to exhibit DNA aneuploidy. None of the control DNA histograms demonstrated DNA aneuploid-like patterns. There were no significant differences in cell cycle percentages at any time point between control and exposed animals. A significant increase in CV was observed at 9 months exposure, but after removal of the two ducks with DNA aneuploidy, no significant difference was detected in the group monitored after 12 months exposure. An increased variation in the DNA and DNA aneuploidy could, therefore, be detected in duck rbc using flow cytometric analysis, with the onset of these effects being related to the attainment of maximal levels of 137Cs body burdens in the exposed animals.

  15. A Flow Cytometric and Computational Approaches to Carbapenems Affinity to the Different Types of Carbapenemases

    Science.gov (United States)

    Pina-Vaz, Cidália; Silva, Ana P.; Faria-Ramos, Isabel; Teixeira-Santos, Rita; Moura, Daniel; Vieira, Tatiana F.; Sousa, Sérgio F.; Costa-de-Oliveira, Sofia; Cantón, Rafael; Rodrigues, Acácio G.

    2016-01-01

    The synergy of carbapenem combinations regarding Enterobacteriaceae producing different types of carbapenemases was study through different approaches: flow cytometry and computational analysis. Ten well characterized Enterobacteriaceae (KPC, verona integron-encoded metallo-β-lactamases –VIM and OXA-48-like enzymes) were selected for the study. The cells were incubated with a combination of ertapenem with imipenem, meropenem, or doripenem and killing kinetic curves performed with and without reinforcements of the drugs. A cephalosporin was also used in combination with ertapenem. A flow cytometric assay with DiBAC4-(3), a membrane potential dye, was developed in order to evaluate the cellular lesion after 2 h incubation. A chemical computational study was performed to understand the affinity of the different drugs to the different types of enzymes. Flow cytometric analysis and time-kill assays showed a synergic effect against KPC and OXA-48 producing-bacteria with all combinations; only ertapenem with imipenem was synergic against VIM producing-bacteria. A bactericidal effect was observed in OXA-48-like enzymes. Ceftazidime plus ertapenem was synergic against ESBL-negative KPC producing-bacteria. Ertapenem had the highest affinity for those enzymes according to chemical computational study. The synergic effect between ertapenem and others carbapenems against different carbapenemase-producing bacteria, representing a therapeutic choice, was described for the first time. Easier and faster laboratorial methods for carbapenemase characterization are urgently needed. The design of an ertapenem derivative with similar affinity to carbapenemases but exhibiting more stable bonds was demonstrated as highly desirable. PMID:27555844

  16. A flow cytometric and computational approaches to carbapenems affinity to the different types of carbapenemases

    Directory of Open Access Journals (Sweden)

    Cidália Pina-Vaz

    2016-08-01

    Full Text Available The synergy of carbapenem combinations regarding Enterobacteriaceae producing different types of carbapenemases was study through different approaches: flow cytometry and computa-tional analysis. Ten well characterized Enterobacteriaceae (KPC, verona integron-encoded metallo-β-lactamases –VIM and OXA-48-like enzymes were selected for the study. The cells were incubated with a combination of ertapenem with imipenem, meropenem or doripenem and killing kinetic curves performed with and without reinforments of the drugs. A cephalosporin was also used in combination with ertapenem. A flow cytometric assay with DiBAC4-(3, a membrane potential dye, was developed in order to evaluate the cellular lesion after 2 h incuba-tion. A chemical computational study was performed to understand the affinity of the different drugs to the different types of enzymes. Flow cytometric analysis and time-kill assays showed a synergic effect against KPC and OXA-48 producing-bacteria with all combinations; only ertapenem with imipenem was synergic against VIM producing-bacteria. A bactericidal effect was observed in OXA-48-like enzymes. Ceftazidime plus ertapenem was synergic against ESBL-negative KPC producing-bacteria. Ertapenem had the highest affinity for those enzymes according to chemical computational study. The synergic effect between ertapenem and others carbapenems against different carbapenemase-producing bacteria, representing a therapeutic choice, was described for the first time. Easier and faster laboratorial methods for car-bapenemase characterization are urgently needed. The design of an ertapenem derivative with similar affinity to carbapenemases but exhibiting more stable bonds was demonstrated as highly desirable.

  17. Improved flow cytometric assessment reveals distinct microvesicle (cell-derived microparticle signatures in joint diseases.

    Directory of Open Access Journals (Sweden)

    Bence György

    Full Text Available INTRODUCTION: Microvesicles (MVs, earlier referred to as microparticles, represent a major type of extracellular vesicles currently considered as novel biomarkers in various clinical settings such as autoimmune disorders. However, the analysis of MVs in body fluids has not been fully standardized yet, and there are numerous pitfalls that hinder the correct assessment of these structures. METHODS: In this study, we analyzed synovial fluid (SF samples of patients with osteoarthritis (OA, rheumatoid arthritis (RA and juvenile idiopathic arthritis (JIA. To assess factors that may confound MV detection in joint diseases, we used electron microscopy (EM, Nanoparticle Tracking Analysis (NTA and mass spectrometry (MS. For flow cytometry, a method commonly used for phenotyping and enumeration of MVs, we combined recent advances in the field, and used a novel approach of differential detergent lysis for the exclusion of MV-mimicking non-vesicular signals. RESULTS: EM and NTA showed that substantial amounts of particles other than MVs were present in SF samples. Beyond known MV-associated proteins, MS analysis also revealed abundant plasma- and immune complex-related proteins in MV preparations. Applying improved flow cytometric analysis, we demonstrate for the first time that CD3(+ and CD8(+ T-cell derived SF MVs are highly elevated in patients with RA compared to OA patients (p=0.027 and p=0.009, respectively, after Bonferroni corrections. In JIA, we identified reduced numbers of B cell-derived MVs (p=0.009, after Bonferroni correction. CONCLUSIONS: Our results suggest that improved flow cytometric assessment of MVs facilitates the detection of previously unrecognized disease-associated vesicular signatures.

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Serkan Yazıcı

    2015-01-01

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

  20. The influence of fixation delay on mitotic activity and flow cytometric cell cycle variables.

    Science.gov (United States)

    Bergers, E; Jannink, I; van Diest, P I; Cuesta, M A; Meyer, S; van Mourik, J C; Baak, J P

    1997-01-01

    Proliferation variables such as mitotic activity and the percentage of S-phase cells have been shown to be of prognostic value in many tumors, especially in breast cancer. However, some studies reported a decrease in mitotic activity caused by delay in fixation of the tissue. In contrast, other studies showed that the identifiability of mitotic figures decreases after fixation delay, but the total number of mitotic figures and also the percentage of S-phase cells remain unchanged. Most studies have been done on small numbers of experimental tumors, thus introducing the risk of selection bias. The aim of this study was to reinvestigate the influence of fixation delay on mitotic activity and cell cycle variables assessed by flow cytometry in an adequate number of resected human tissues to reach firmer conclusions. Resection specimens of 19 and 21 cases, respectively, for the mitotic activity estimate and the flow cytometric percentage of S-phase calculation were collected directly from the operating theater using lung, breast, and intestinal cancers and normal intestinal mucosa. The tissues were cut in pieces, and from each specimen, pieces were fixed in 4% buffered formaldehyde (for mitosis counting) as well as snap frozen (for flow cytometry) immediately after excision, as well as after a fixation delay of 1, 2, 4, 6, 8, 18, and 24 hours. Moreover, during the fixation delay, one series from each specimen was kept in the refrigerator and the second at room temperature. Thus, a total of 304 (19 X 16) and 336 (21 X 16) specimens were investigated for the mitotic activity estimate and the percentage of S-phase cells calculation, respectively. With regard to the estimation of the mitotic activity, both clear and doubtful mitotic figures were registered separately, obtaining an "uncorrected" and "corrected" (for doubtful mitotic figures) mitotic activity estimate. The percentage of S-phase cells was obtained by cell cycle analysis of flow cytometric DNA-histograms. The

  1. Flow cytometric sorting coupled with exon capture sequencing identifies somatic mutations in archival lymphoma tissues.

    Science.gov (United States)

    Jiang, Nenggang; Chen, Christopher; Gong, Qiang; Shields, Kristen; Li, Yuping; Chen, YuanYuan; Song, Joo; McKeithan, Timothy W; Chan, Wing C

    2017-08-07

    The enormous number of archived formalin-fixed paraffin-embedded (FFPE) tissues available are a valuable resource of material for research. However, the use of such tissues poses many challenges, among which is the difficulty of isolating different cell populations within the tissue. In this study, we used tissue from two types of non-Hodgkin lymphoma as a model to demonstrate a method we have established and optimized to separate FFPE samples into distinct tumor and nonmalignant populations. Using FFPE reactive tonsil sections, various approaches for antigen retrieval and labeling, and the effectiveness of flow cytometric sorting were tested. We found that, among the 11 cell surface or intracellular antigen markers investigated, CD3ɛ, CD79A, LAT, PD-1, and PAX5 could be successfully labeled after antigen retrieval in Tris-EDTA buffer (pH 8.0) at 65 °C for 60 min, and 1.8-2.7 μg DNA per million cells could be extracted after sorting with DNA quality similar to that of tissue without staining or sorting. To test whether we could perform next-generation sequencing using a custom capture platform on sorted cells, we used three lymphoma cases with FFPE tissues which had been stored for 1 to 4 years. We demonstrated that the DNA from sorted cells was adequate for exon capture sequencing. By comparing the sequencing results between neoplastic and normal populations, somatic mutations could be clearly identified in the tumor population with variant frequencies as low as 11.7%.The corresponding normal fraction clearly helps in the analysis of somatic mutations and the exclusion of artifacts. This study provides an approach using flow cytometric sorting to separate different cellular populations in paraffin-embedded tissues and to unambiguously distinguish somatic mutations from germline variants or artifacts. This approach is also useful in enriching the tumor component in samples with heterogeneous components and low tumor content.Laboratory Investigation advance

  2. A new spreadsheet method for the analysis of bivariate flow cytometric data

    Directory of Open Access Journals (Sweden)

    Isacke Clare M

    2004-03-01

    Full Text Available Abstract Background A useful application of flow cytometry is the investigation of cell receptor-ligand interactions. However such analyses are often compromised due to problems interpreting changes in ligand binding where the receptor expression is not constant. Commonly, problems are encountered due to cell treatments resulting in altered receptor expression levels, or when cell lines expressing a transfected receptor with variable expression are being compared. To overcome this limitation we have developed a Microsoft Excel spreadsheet that aims to automatically and effectively simplify flow cytometric data and perform statistical tests in order to provide a clearer graphical representation of results. Results To demonstrate the use and advantages of this new spreadsheet method we have investigated the binding of the transmembrane adhesion receptor CD44 to its ligand hyaluronan. In the first example, phorbol ester treatment of cells results in both increased CD44 expression and increased hyaluronan binding. By applying the spreadsheet method we effectively demonstrate that this increased ligand binding results from receptor activation. In the second example we have compared AKR1 cells transfected either with wild type CD44 (WT CD44 or a mutant with a truncated cytoplasmic domain (CD44-T. These two populations do not have equivalent receptor expression levels but by using the spreadsheet method hyaluronan binding could be compared without the need to generate single cell clones or FACS sorting the cells for matching CD44 expression. By this method it was demonstrated that hyaluronan binding requires a threshold expression of CD44 and that this threshold is higher for CD44-T. However, at high CD44-T expression, binding was equivalent to WT CD44 indicating that the cytoplasmic domain has a role in presenting the receptor at the cell surface in a form required for efficient hyaluronan binding rather than modulating receptor activity. Conclusion

  3. Flow cytometric assessment of specific leucine incorporation in the open Mediterranean

    Science.gov (United States)

    Talarmin, A.; van Wambeke, F.; Catala, P.; Courties, C.; Lebaron, P.

    2011-02-01

    The surface of the Mediterranean Sea is a low-phosphate-low-chlorophyll marine area where marine heterotrophic prokaryotes significantly contribute to the biogeochemical cycles of all biogenic elements such as carbon, notably through the mineralization of dissolved organic compounds. Cell-specific leucine incorporation rates were determined in early summer in the open stratified Mediterranean Sea. The bulk leucine incorporation rate was on average 5 ± 4 pmol leu l-1 h-1 (n=30). Cell-specific 3H-leucine incorporation rates were assayed using flow cytometry coupled to cell sorting. Heterotrophic prokaryotes (Hprok) were divided into cytometric groups according to their side scatter and green fluorescence properties: high nucleic acid containing cells (HNA) with high scatter (HNA-hs) and low scatter (HNA-ls) and low nucleic acid containing cells (LNA). Cell-specific leucine incorporation rates of these cytometric groups ranged from 2 to 54, 0.9 to 11, and 1 to 12 × 10-21 mol cell-1 h-1, respectively. LNA cells represented 45 to 63% of the Hprok abundance, and significantly contributed to the bulk leucine incorporation rates, from 12 to 43%. HNA/LNA ratios of cell-specific leucine incorporation were on average 2.0 ± 0.7 (n=30). In surface layers (from 0 m down to the deep chlorophyll depth, DCM), cell-specific rates of HNA-hs were elevated (7 and 13 times greater than LNA and HNA-ls, respectively). Nevertheless, on average HNA-hs (26%) and LNA (27%) equally contributed to the bulk leucine incorporation in these layers. Prochlorococcus cells were easily sorted near the DCM and displayed cell-specific leucine incorporation rates ranging from 3 to 55 × 10-21 mol leu cell-1 h-1, i.e. as high as HNA-hs'. These sorted groups could therefore be defined as key-players in the process of leucine incorporation into proteins. The mixotrophic features of certain photosynthetic prokaryotes and the high contribution of LNA cells to leucine incorporation within the microbial

  4. Flow cytometric assessment of specific leucine incorporation in the open Mediterranean

    Directory of Open Access Journals (Sweden)

    A. Talarmin

    2011-02-01

    Full Text Available The surface of the Mediterranean Sea is a low-phosphate-low-chlorophyll marine area where marine heterotrophic prokaryotes significantly contribute to the biogeochemical cycles of all biogenic elements such as carbon, notably through the mineralization of dissolved organic compounds. Cell-specific leucine incorporation rates were determined in early summer in the open stratified Mediterranean Sea. The bulk leucine incorporation rate was on average 5 ± 4 pmol leu l−1 h−1 (n=30. Cell-specific 3H-leucine incorporation rates were assayed using flow cytometry coupled to cell sorting. Heterotrophic prokaryotes (Hprok were divided into cytometric groups according to their side scatter and green fluorescence properties: high nucleic acid containing cells (HNA with high scatter (HNA-hs and low scatter (HNA-ls and low nucleic acid containing cells (LNA. Cell-specific leucine incorporation rates of these cytometric groups ranged from 2 to 54, 0.9 to 11, and 1 to 12 × 10-21 mol cell−1 h−1, respectively. LNA cells represented 45 to 63% of the Hprok abundance, and significantly contributed to the bulk leucine incorporation rates, from 12 to 43%. HNA/LNA ratios of cell-specific leucine incorporation were on average 2.0 ± 0.7 (n=30. In surface layers (from 0 m down to the deep chlorophyll depth, DCM, cell-specific rates of HNA-hs were elevated (7 and 13 times greater than LNA and HNA-ls, respectively. Nevertheless, on average HNA-hs (26% and LNA (27% equally contributed to the bulk leucine incorporation in these layers. Prochlorococcus cells were easily sorted near the DCM and displayed cell-specific leucine incorporation rates ranging from 3 to 55 × 10-21 mol leu cell−1 h−1, i.e. as high as HNA-hs'. These sorted groups could therefore be defined as key-players in the process of leucine incorporation into proteins. The

  5. The effects of orange juice clarification on the physiology of Escherichia coli; growth-based and flow cytometric analysis.

    Science.gov (United States)

    Anvarian, Amir H P; Smith, Madeleine P; Overton, Tim W

    2016-02-16

    Orange juice (OJ) is a food product available in various forms which can be processed to a greater or lesser extent. Minimally-processed OJ has a high consumer perception but presents a potential microbiological risk due to acid-tolerant bacteria. Clarification of OJ (such as removal of cloud) is a common processing step in many OJ products. However, many of the antimicrobial components of OJ such as essential oils are present in the cloud fraction. Here, the effect of clarification by filtration on the viability and physiology of Escherichia coli K-12 was tested using total viable count (TVC) and flow cytometric (FCM) analysis. The latter technique was also used to monitor intracellular pH during incubation in OJ. Removal of the OJ cloud fraction was shown to have dramatic effects on bacterial viability and physiology during storage at a range of incubation temperatures. For instance, at 4 °C, a significantly lower number of healthy cells and a significantly higher number of injured cells were observed in 0.22 μm-filtered OJ at 24h post-inoculation, compared to filtered OJ samples containing particles between 0.22 μm and 11 μm in size. Similarly, there was a significant difference between the number of healthy bacteria in the 0.7 μm-filtered OJ and both 0.22 μm-filtered and 1.2 μm-filtered OJ after 24 hour incubation at 22.5 °C. This indicated that OJ cloud between 0.7 μm and 0.22 μm in size might have an adverse effect on the viability of E. coli K-12. Furthermore, FCM allowed the rapid analysis of bacterial physiology without the requirement for growth on agar plates, and revealed the extent of the viable but non-culturable (VBNC) population. For example, at 4 °C, while the FCM viable count did not substantially decrease until 48 h, decreases in TVC were observed between 0 and 48 hour incubation, due to a subset of injured bacteria entering the VBNC state, hence being unable to grow on agar plates. This study highlights the application of FCM in

  6. A novel flow cytometric assay for measurement of In Vivo pulmonary neutrophil phagocytosis

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    Gentry-Nielsen Martha J

    2006-07-01

    Full Text Available Abstract Background Phagocytosis assays are traditionally performed in vitro using polymorphonuclear leukocytes (PMNs isolated from peripheral blood or the peritoneum and heat-killed, pre-opsonized organisms. These assays may not adequately mimic the environment within the infected lung. Our laboratory therefore has developed a flow cytometric in vivo phagocytosis assay that enables quantification of PMN phagocytosis of viable bacteria within the lungs of rats. In these studies, rats are injected transtracheally with lipopolysaccharide (LPS to recruit PMNs to their lungs. They are then infected with live 5(-and 6 carboxyfluorescein diacetate succinimidyl ester (CFDA/SE labeled type 3 Streptococcus pneumoniae. Bronchoalveolar lavage is performed and resident alveolar macrophages and recruited PMNs are labeled with monoclonal antibodies specific for surface epitopes on each cell type. Three color flow cytometry is utilized to identify the cell types, quantify recruitment, and determine uptake of the labeled bacteria. Results The viability of the alveolar macrophages and PMNs isolated from the lavage fluid was >95%. The values of the percentage of PMNs in the lavage fluid as well as the percentage of PMNs associated with CFSE-labeled S. pneumoniae as measured through flow cytometry showed a high degree of correlation with the results from manual counting of cytospin slides. Conclusion This assay is suitable for measuring bacterial uptake within the infected lung. It can be adapted for use with other organisms and/or animal model systems.

  7. [Flow cytometric analysis of ICRF-193 influence on cell passage through mitosis].

    Science.gov (United States)

    Shatrova, A N; Aksenov, N D; Zenin, V V

    2002-01-01

    Studying the effect of topoisomerase II (topo II) inhibitors on cell passage through mitosis seems to be important for understanding the role of this enzyme during chromosome condensation and segregation. A flow cytometric assay (Zenin et al., 2001) allowed to determine the mitotic index, and to discriminate between not only cells in G2 and M phases (including metaphase and anaphase cells), but also cells in pseudo-G1 with 4c DNA content. It is shown that topo II catalytic inhibitor ICRF-193 blocks G2-M transition in a lymphoblastoid cell line GM-130. Addition of caffeine to cells abrogated a block of their entering mitosis but not the inhibitor action. Cells entered mitosis, which was proven by the presence of chromosomes in the examined specimen, and, bypassing anaphase, appeared in pseudo-G1 with 4c DNA content. We have found that in the presence of ICRF-193 cells, GM-130 and Hep-2 lines, previously blocked by nocodazole when in mitosis and then washed, pass through metaphase, enter anaphase and leave it to pass to pseudo-G1 with the 4c DNA content. Thus, by inhibiting topo II activity ICRF-193 causes abnormal mitotic transition.

  8. Evaluation of Red Cell Membrane Cytoskeletal Disorders Using a Flow Cytometric Method in South Iran

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    Habib Alah Golafshan

    2014-03-01

    Full Text Available OBJECTIVE: The diagnosis of hereditary red blood cell (RBC membrane disorders, and in particular hereditary spherocytosis (HS and Southeast Asian ovalocytosis (SAO, is based on clinical history, RBC morphology, and other conventional tests such as osmotic fragility. However, there are some milder cases of these disorders that are difficult to diagnose. The application of eosin-5’-maleimide (EMA was evaluated for screening of RBC membrane defects along with some other anemias. We used EMA dye, which binds mostly to band 3 protein and to a lesser extent some other membrane proteins, for screening of some membrane defects such as HS. METHODS: Fresh RBCs from hematologically normal controls and patients with HS, SAO, hereditary elliptocytosis, hereditary spherocytosis with pincered cells, severe iron deficiency, thalassemia minor, and autoimmune hemolytic anemia were stained with EMA dye and analyzed for mean fluorescent intensity (MFI using a flow cytometer. RESULTS: RBCs from patients with HS and iron deficiency showed a significant reduction in MFI compared to those from normal controls (p<0.0001 and p<0.001, respectively, while macrocytic RBCs showed a significant increase in MFI (p<0.01. A significant correlation was shown between mean corpuscular volume and MFI, with the exceptions of HS and thalassemia minor. CONCLUSION: Our results showed that the flow cytometric method could be a reliable diagnostic method for screening and confirmation, with higher sensitivity and specificity (95% and 93%, respectively than conventional routine tests for HS patients prior to further specific membrane protein molecular tests.

  9. Establishment of multiplexed, microsphere-based flow cytometric assay for multiple human tumor markers

    Institute of Scientific and Technical Information of China (English)

    Kai SUN; Qian WANG; Xiao-hui HUANG; Mao-chuan ZHEN; Wen LI; Long-juan ZHANG

    2007-01-01

    Aim: The multiplexed, microsphere-based flow cytometric assay (MFCA) for mul- tiple human tumor markers was established for the early screening and detection of suspected cancer patients. Methods: Covalent coupling of capture antibodies directed against their respective tumor markers to fluorescent microspheres was performed by following the protocols recommended by a commercial corporation with some modifications. The coupling efficiency and cross-reactivity were iden- tified by the Luminex 100 system and associated software. The standard curve was constructed by using serial dilution of recombinant tumor marker standards and was validated by comparison with ELISA for quantifying the tumor markers in serum samples. Results: The identifications revealed that the coupling proce- dures were successful without non-specific cross-reactivity and the standard curve was highly efficient. However, it was necessary to ensure the quality con- trol of the coupling process since slight variations in the coupling procedures could profoundly affect the density of capture reagents coupled to the microspheres and consequently adversely affect the assay precision. In addition to its multi-analyte capability, the MFCA system had definite advantages, such as higher reproducibility, greater dynamic range of measurement, and considerably less preparation time and labor over the conventional "gold standard", which was the ELISA. Conclusion: The successful establishment of the MFCA system for the simultaneous detection of multiple tumor markers will provide the foundation for the further study of clinical applications.

  10. Flow cytometric analysis of lymphocyte proliferative responses to food allergens in dogs with food allergy.

    Science.gov (United States)

    Fujimura, Masato; Masuda, Kenichi; Hayashiya, Makio; Okayama, Taro

    2011-10-01

    Two different allergy tests, antigen-specific immunoglobulin E quantification (IgE test) and flow cytometric analysis of antigen-specific proliferation of peripheral lymphocytes (lymphocyte proliferation test), were performed to examine differences in allergic reactions to food allergens in dogs with food allergy (FA). Thirteen dogs were diagnosed as FA based on clinical findings and elimination diet trials. Seven dogs clinically diagnosed with canine atopic dermatitis (CAD) were used as a disease control group, and 5 healthy dogs were used as a negative control group. In the FA group, 19 and 33 allergen reactions were identified using the serum IgE test and the lymphocyte proliferation test, respectively. Likewise, in the CAD group, 12 and 6 allergen reactions and in the healthy dogs 3 and 0 allergen reactions were identified by each test, respectively. A significant difference was found between FA and healthy dogs in terms of positive allergen detection by the lymphocyte proliferation test, suggesting that the test can be useful to differentiate FA from healthy dogs but not from CAD. Both tests were repeated in 6 of the dogs with FA after a 1.5- to 5-month elimination diet trial. The IgE concentrations in 9 of 11 of the positive reactions decreased by 20-80%, whereas all the positive reactions in the lymphocyte proliferation test decreased to nearly zero (Pfood allergens may be involved in the pathogenesis of canine FA.

  11. Flow cytometric quantification of T cell proliferation and division kinetics in woodchuck model of hepatitis B.

    Science.gov (United States)

    Gujar, Shashi A; Michalak, Tomasz I

    2005-01-01

    Woodchucks infected with woodchuck hepatitis virus (WHV) represent the closest natural animal model to study the immunopathogenesis of liver injury caused by essentially noncytopathic, highly human specific hepatitis B virus (HBV). The importance of antiviral T cell response in induction of hepatitis and in control of HBV replication has been demonstrated. However, the understanding of how these responses contribute to the development of different immunomorphological forms of liver disease and their outcomes remain elusive. In this study, we established and standardized a flow cytometry assay using peripheral blood mononuclear cells labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) to assess WHV-specific and mitogen-driven T lymphocyte proliferative responses in woodchucks. The assay is of significantly greater sensitivity than the adenine incorporation assay currently used when applied to measure either WHV-specific T cell responses in acute (P measuring cell division rates. The study shows that woodchuck PBMC labeled with CFSE exhibit light scatter and fluorescence profiles compatible to those of human PBMC, allowing quantitation and deconvolution of the flow cytometric data by applying the existing analytical softwares. The availability of this novel assay should facilitate a more precise and comprehensive evaluation of hepadnavirus-specific and generalized T cell responses in experimental WHV hepatitis.

  12. A Flow Cytometric Clonogenic Assay Reveals the Single-Cell Potency of Doxorubicin

    Science.gov (United States)

    Maass, Katie F.; Kulkarni, Chethana; Quadir, Mohiuddin A.; Hammond, Paula T.; Betts, Alison M.; Wittrup, K. Dane

    2015-01-01

    Standard cell proliferation assays use bulk media drug concentration to ascertain the potency of chemotherapeutic drugs; however, the relevant quantity is clearly the amount of drug actually taken up by the cell. To address this discrepancy, we have developed a flow cytometric clonogenic assay to correlate the amount of drug in a single cell with the cell’s ability to proliferate using a cell tracing dye and doxorubicin, a naturally fluorescent chemotherapeutic drug. By varying doxorubicin concentration in the media, length of treatment time, and treatment with verapamil, an efflux pump inhibitor, we introduced 105 – 1010 doxorubicin molecules per cell; then used a dye-dilution assay to simultaneously assess the number of cell divisions. We find that a cell’s ability to proliferate is a surprisingly conserved function of the number of intracellular doxorubicin molecules, resulting in single-cell IC50 values of 4 – 12 million intracellular doxorubicin molecules. The developed assay is a straightforward method for understanding a drug’s single-cell potency and can be used for any fluorescent or fluorescently-labeled drug, including nanoparticles or antibody-drug conjugates. PMID:26344409

  13. Flow cytometric analysis of oil palm: a preliminary analysis for cultivars and genomic DNA alteration

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    Warawut Chuthammathat

    2005-12-01

    Full Text Available DNA contents of oil palm (Elaeis guineensis Jacq. cultivars were analyzed by flow cytometry using different external reference plant species. Analysis using corn (Zea mays line CE-777 as a reference plant gave the highest DNA content of oil palm (4.72±0.23 pg 2C-1 whereas the DNA content was found to be lower when using soybean (Glycine max cv. Polanka (3.77±0.09 pg 2C-1 or tomato (Lycopersicon esculentum cv. Stupicke (4.25±0.09 pg 2C-1 as a reference. The nuclear DNA contents of Dura (D109, Pisifera (P168 and Tenera (T38 cultivars were 3.46±0.04, 3.24±0.03 and 3.76±0.04 pg 2C-1 nuclei, respectively, using soybean as a reference. One haploid genome of oil palm therefore ranged from 1.56 to 1.81±109 base pairs. DNA contents from one-year-old calli and cell suspension of oil palm were found to be significantly different from those of seedlings. It thus should be noted that genomic DNA alteration occurred in these cultured tissues. We therefore confirm that flow cytometric analysis could verify cultivars, DNA content and genomic DNA alteration of oil palm using soybean as an external reference standard.

  14. A novel flow cytometric hemozoin detection assay for real-time sensitivity testing of Plasmodium falciparum.

    Directory of Open Access Journals (Sweden)

    Maria Rebelo

    Full Text Available Resistance of Plasmodium falciparum to almost all antimalarial drugs, including the first-line treatment with artemisinins, has been described, representing an obvious threat to malaria control. In vitro antimalarial sensitivity testing is crucial to detect and monitor drug resistance. Current assays have been successfully used to detect drug effects on parasites. However, they have some limitations, such as the use of radioactive or expensive reagents or long incubation times. Here we describe a novel assay to detect antimalarial drug effects, based on flow cytometric detection of hemozoin (Hz, which is rapid and does not require any additional reagents. Hz is an optimal parasite maturation indicator since its amount increases as the parasite matures. Due to its physical property of birefringence, Hz depolarizes light, hence it can be detected using optical methods such as flow cytometry. A common flow cytometer was adapted to detect light depolarization caused by Hz. Synchronized in vitro cultures of P. falciparum were incubated for 48 hours with several antimalarial drugs. Analysis of depolarizing events, corresponding to parasitized red blood cells containing Hz, allowed the detection of parasite maturation. Moreover, chloroquine resistance and the inhibitory effect of all antimalarial drugs tested, except for pyrimethamine, could be determined as early as 18 to 24 hours of incubation. At 24 hours incubation, 50% inhibitory concentrations (IC50 were comparable to previously reported values. These results indicate that the reagent-free, real-time Hz detection assay could become a novel assay for the detection of drug effects on Plasmodium falciparum.

  15. Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells

    Directory of Open Access Journals (Sweden)

    Coignoul Freddy

    2007-09-01

    Full Text Available Abstract Background The morphopathological picture of a subset of equine myopathies is compatible with a primary mitochondrial disease, but functional confirmation in vivo is still pending. The cationic dye JC-1 exhibits potential-dependent accumulation in mitochondria that is detectable by a fluorescence shift from green to orange. As a consequence, mitochondrial membrane potential can be optically measured by the orange/green fluorescence intensity ratio. A flow cytometric standardized analytic procedure of the mitochondrial function of equine peripheral blood mononuclear cells is proposed along with a critical appraisal of the crucial questions of technical aspects, reproducibility, effect of time elapsed between blood sampling and laboratory processing and reference values. Results The JC-1-associated fluorescence orange and green values and their ratio were proved to be stable over time, independent of age and sex and hypersensitive to intoxication with a mitochondrial potential dissipator. Unless time elapsed between blood sampling and laboratory processing does not exceed 5 hours, the values retrieved remain stable. Reference values for clinically normal horses are given. Conclusion Whenever a quantitative measurement of mitochondrial function in a horse is desired, blood samples should be taken in sodium citrate tubes and kept at room temperature for a maximum of 5 hours before the laboratory procedure detailed here is started. The hope is that this new test may help in confirming, studying and preventing equine myopathies that are currently imputed to mitochondrial dysfunction.

  16. A new flow cytometric method for quantitative assessment of lymphocyte mitogenic potentials.

    Science.gov (United States)

    Yamamura, Y; Rodriguez, N; Schwartz, A; Eylar, E; Bagwell, B; Yano, N

    1995-01-01

    A new flow cytometric method was developed to quantitatively assess lymphocyte proliferation simultaneously for different subsets. The cells were stained with a fluorescent dye, PKH-26 and were stimulated with mitogens. The fluorescence intensities (FL2) of proliferating cells were measured by flow cytometry; and each subset was identified by the use of a monoclonal antibody (Mab)-fluorescein-isothiocyanate (FITC) (FL1). FL2 histograms were then analyzed by the cell proliferation model based on the ModFit software (Verity). This new method revealed information which could not be obtained by conventional mitogen assays. For example, the CD4+ and the CD4- T-subsets responded to phytohemagglutinin (PHA) quite differently from each other and it was indicated that activation of one population could significantly alter the response of the other. In addition, even within a subset, all activated cells did not proliferate uniformly. Some cells divided only once while others underwent further cellular division during the same time period. The method is, therefore, invaluable for studying the nature and the extent of interactions between different cellular subsets within a culture.

  17. Flow cytometric evaluation of the intracellular bacterium, Wolbachia pipientis, in mosquito cells

    Science.gov (United States)

    Fallon, Ann M

    2014-01-01

    Wolbachia is an obligate intracellular bacterium (Anaplasmataceae, Rickettisales) that occurs in arthropods and filarial worms, and spreads by vertical transmission in the oocyte cytoplasm. In insects, reproductive distortions associated with Wolbachia, such as cytoplasmic incompatibility in mosquitoes, have potential value for controlling pests, including species that transmit human, animal and plant diseases. Wolbachia strains that propagate as a persistent infection in insect cell lines provide an important resource for developing the genetic tools that will facilitate these applications. Here I describe conditions for flow cytometric evaluation of Wolbachia growth in persistently infected mosquito cells. Cytometry parameters were established using uninfected mosquito cells and Escherichia coli as a surrogate for Wolbachia, and quantitation was correlated with cell counts determined with a Coulter electronic cell counter and bacterial counts based on optical density. The protocol was validated by showing depletion of Wolbachia in medium containing tetracycline and rifampicin, and sensitivity of Wolbachia to treatment of host cells with paraquat, an oxidizing agent, and lumiflavin, an inhibitor of riboflavin uptake. The Wolbachia peak on the flow cytometry histogram was shown to contain Wolbachia by DNA analysis using the polymerase chain reaction, and by infection of naive recipient cells. This approach will streamline investigation of Wolbachia growth in insect cell lines and facilitate identification of culture conditions that select for Wolbachia-infected cells. PMID:25300665

  18. Antiphospholipid antibody syndrome: the flow cytometric annexin A5 competition assay as a diagnostic tool.

    Science.gov (United States)

    Tomer, A; Bar-Lev, S; Fleisher, S; Shenkman, B; Friger, M; Abu-Shakra, M

    2007-10-01

    The mechanism underlying hypercoagulability in antiphospholipid antibody syndrome (APS) is uncertain. Here, we present a flow-cytometric assay (FCA) based on the hypothesis that anti-platelet-anionic-phospholipid autoantibodies (aPL) interfere with the activity of the natural anticoagulant protein annexin A5, thereby accelerating platelet procoagulant activity. This study assessed the clinical utility of the feasible FCA, which demonstrates the competition of the patient's aPL with the binding of annexin A5 to the platelet-anionic-phospholipids, in the diagnosis of APS. Sixty-two (94%) of 66 APS patients, 20 (51%) of 39 patients with systemic lupus erythematosus and two (4%) of 49 healthy individuals were positive by FCA. Compared with the anticardiolipin (aCL) assay, the relative sensitivity was 82% and the specificity 73.3%. However, 19 (25%) aCL-negative patients were positive by FCA; 12 were positive for lupus-anticoagulant (LA). Compared with LA assay, the relative sensitivity was 85% and the specificity 72.2%. However, 21 (26%) LA-negative patients were FCA-positive, 12 were positive for aCL. The FCA was particularly sensitive for APS patients with arterial (97.0%) and gestational vascular complications (100%) with overall sensitivity of 95% and specificity of 97%. Our findings suggest that the FCA is practical, sensitive and specific for the detection of clinically relevant aPL in the diagnosis of APS.

  19. Flow cytometric analysis using SYBR Green I for genome size estimation in coffee.

    Science.gov (United States)

    Ronildo Clarindo, Wellington; Roberto Carvalho, Carlos

    2011-02-01

    Plant genome size has been measured by flow cytometry using propidium iodide as a dye for nuclear DNA staining. However, some authors have reported the occurrence of genome size estimation errors, especially in plants rich in secondary metabolites, such as the coffee tree. In this context, we tested an alternative cytometric protocol using the SYBR Green I as a fluorochrome for stoichiometrically staining nuclear double-stranded DNA in Coffea canephora (2x) and Coffea arabica (4x). The results showed that the respective mean genome size measured from nuclei stained with SYBR Green I and propidium iodide was statistically identical. However, the G(0)/G(1) peaks of nuclei stained with SYBR Green I exhibited lower coefficient variations (1.57-2.85%) compared to those stained with propidium iodide (2.75-4.80%). Coefficient variation statistical data suggest that SYBR Green I is adequate for stoichiometric nuclei staining using this methodology. Our results provide evidence that SYBR Green I can be used in flow cytometry measurements of plants, with the advantages of minimizing errors in nuclear DNA content quantification, staining relatively quicker, with high affinity, and being less mutagenic than propidium iodide.

  20. Dissociation of skeletal muscle for flow cytometric characterization of immune cells in macaques.

    Science.gov (United States)

    Liang, Frank; Ploquin, Aurélie; Hernández, José DelaO; Fausther-Bovendo, Hugues; Lindgren, Gustaf; Stanley, Daphne; Martinez, Aiala Salvador; Brenchley, Jason M; Koup, Richard A; Loré, Karin; Sullivan, Nancy J

    2015-10-01

    The majority of vaccines and several treatments are administered by intramuscular injection. The aim is to engage and activate immune cells, although they are rare in normal skeletal muscle. The phenotype and function of resident as well as infiltrating immune cells in the muscle after injection are largely unknown. While methods for obtaining and characterizing murine muscle cell suspensions have been reported, protocols for nonhuman primates (NHPs) have not been well defined. NHPs comprise important in vivo models for studies of immune cell function due to their high degree of resemblance with humans. In this study, we developed and systematically compared methods to collect vaccine-injected muscle tissue to be processed into single cell suspensions for flow cytometric characterization of immune cells. We found that muscle tissue processed by mechanical disruption alone resulted in significantly lower immune cell yields compared to enzymatic digestion using Liberase. Dendritic cell subsets, monocytes, macrophages, neutrophils, B cells, T cells and NK cells were readily detected in the muscle by the classic human markers. The methods for obtaining skeletal muscle cell suspension established here offer opportunities to increase the understanding of immune responses in the muscle, and provide a basis for defining immediate post-injection vaccine responses in primates.

  1. Successful low dose insemination of flow cytometrically sorted ram spermatozoa in sheep.

    Science.gov (United States)

    de Graaf, S P; Evans, G; Maxwell, W M C; Downing, J A; O'Brien, J K

    2007-12-01

    The fertility of ram spermatozoa that had undergone flow cytometric sorting (MoFlo SX) and cryopreservation was assessed after low-dose insemination of synchronized Merino ewes. Oestrus was synchronized with progestagen-impregnated pessaries, PMSG and GnRH treatment. Ewes (n = 360) were inseminated with 1 x 10(6), 5 x 10(6) or 15 x 10(6) motile sorted frozen-thawed (S(1), S(5), or S(15) respectively) or non-sorted frozen-thawed (C(1), C(5) or C(15) respectively) spermatozoa from three rams. An additional group of ewes were inseminated with 50 x 10(6) motile non-sorted frozen-thawed spermatozoa (C(50)) to provide a commercial dose control. The percentage of ewes lambing after insemination was similar for C(50) (24/38, 63.2%), C(15) (37/54, 68.5%), S(15) (38/57, 66.7%), S(5) (37/56, 66.1%) and S(1) (32/52, 61.5%) groups (p > 0.05), but lower for C(5) (19/48, 39.6%) and C(1) (19/55, 34.5%) treatments (p sheep as a reduction in the minimum effective sperm number will allow a corresponding decrease in the associated cost per dose.

  2. Flow cytometric scoring of micronucleated erythrocytes: an efficient platform for assessing in vivo cytogenetic damage.

    Science.gov (United States)

    Dertinger, Stephen D; Torous, Dorothea K; Hayashi, Makoto; MacGregor, James T

    2011-01-01

    The relative simplicity of the micronucleated erythrocyte endpoint has made it amenable to automated scoring approaches. Flow cytometry is one such scoring platform that has been employed successfully. This review describes the evolution and properties of flow cytometry-based scoring of micronucleated erythrocytes. The methodology has become widely applied to rodent blood specimens and the high throughput nature of the technology provides a number of advantages over manual microscopic scoring. For instance, the ability to efficiently survey many dose levels and many more cells per specimen relative to microscopy benefits studies that are designed to identify no observable effect levels or lowest observable effect levels. Furthermore, flow cytometry makes it practical to study species with low spontaneous reticulocyte (RET) counts and micronucleus (MN) frequencies, thereby facilitating integration of blood-based micronucleated reticulocyte (MN-RET) frequency measurements into experiments conducted across species of toxicological interest. This capability enhances genotoxicity assessments that have historically been made in dedicated MN tests performed in one species. Importantly, the feasibility of using MN-RET frequencies in blood from humans as an index of genetic damage in bone marrow opens a critical area of application that had not been practical previously. We conclude with recommendations for additional work that is needed to more fully realise the potential of flow cytometric in vivo MN scoring.

  3. Effects of Hoechst33342 staining on the viability and flow cytometric sex-sorting of frozen-thawed ram sperm.

    Science.gov (United States)

    Quan, Guo Bo; Ma, Yuan; Li, Jian; Wu, Guo Quan; Li, Dong Jiang; Ni, Yi Na; Lv, Chun Rong; Zhu, Lan; Hong, Qiong Hua

    2015-02-01

    Cytometric sorting of frozen-thawed sperm can overcome difficulties caused by the unavailability of sorting facilities on farms where semen is collected from male livestock. In order to optimize the cytometric sex-sorting procedure, effects of Hoechst33342 staining on the viability and cytometric sorting efficiency of frozen-thawed ram sperm were evaluated. The frozen-thawed sperm were stained with Hoechst33342 at various dye concentrations (80 μM, 120 μM, 160 μM, 200 μM, 240 μM, or 320 μM) for 45 min to evaluate effects of dye dose. The frozen-thawed sperm were stained with 160 μM Hoechst33342 for various durations (0 min, 15 min, 30 min, 45 min, 60 min, 75 min, or 90 min) to evaluate effects of staining duration. Sperm motility and moving velocity were analyzed using a computer-assisted sperm analysis system (CASAS). Acrosome status, membrane integrity, and distribution of phosphatidylserine (PS) in Hoechst33342-stained sperm were analyzed using flow cytometry after staining with fluorescein isothiocyanate-labeled lectin from pisum sativum (FITC-PSA), Annexin V, or propidium iodide (PI). The fertility of Hoechst33342-stained sperm was analyzed by in vitro fertilization (IVF). A high-speed cell sorter was used to evaluate effects of Hoechst33342 staining on cytometric sex-sorting of frozen-thawed sperm. The motility, moving velocity, membrane integrity, and PS distribution of Hoechst33342-stained sperm were significantly different from that of immediately thawed sperm (Pram sperm. Results of cytometric sorting indicated that frozen-thawed sperm can be efficiently sorted into two sperm populations with X and Y chromosome when the Hoechst33342 concentration was 160 μM. Moreover, when the staining duration was equal to or longer than 45 min, the frozen-thawed sperm can be successfully sorted in the presence of 160μM Hoechst33342. In conclusion, Hoechst33342 staining can detrimentally influence viability of frozen-thawed ram sperm except acrosome and in vitro

  4. Picoplankton Community Composition by CARD-FISH and Flow Cytometric Techniques: A Preliminary Study in Central Adriatic Sea Water

    Directory of Open Access Journals (Sweden)

    Anita Manti

    2012-01-01

    Full Text Available Data concerning picoplanktonic community composition and abundance in the Central Adriatic Sea are presented in an effort to improve the knowledge of bacterioplankton and autotrophic picoplankton and their seasonal changes. Flow cytometry analyses revealed the presence of two distinct bacteria populations: HNA and LNA cells. HNA cells showed an explicit correlation with viable and actively respiring cells. The study of viability and activity may increase our knowledge of the part that contributes really to the remineralization and bacterial biomass production. Authotrophic picoplankton abundance, especially picocyanobacteria, was strongly influenced by seasonality, indicating that light availability and water temperature are very important regulating factors. In terms of total carbon biomass, the main contribution came from heterotrophic bacteria with a lower contribution from autotrophic picoplankton. CARD-FISH evidenced, within the Eubacteria domain, the dominance of members of the phyla Alphaproteobacteria, with a strong contribution from SAR11clade, followed by Cytophaga-Flavobacterium and Gammaproteobacteria. The bacterial groups detected contributed differently depending when the sample was taken, suggesting possible seasonal patterns. This study documents for the first time picoplankton community composition in the Central Adriatic Sea using two different approaches, FCM and CARD-FISH, and could provide preliminary data for future studies.

  5. Flow cytometric immunobead assay for fast and easy detection of PML-RARA fusion proteins for the diagnosis of acute promyelocytic leukemia.

    Science.gov (United States)

    Dekking, E H A; van der Velden, V H J; Varro, R; Wai, H; Böttcher, S; Kneba, M; Sonneveld, E; Koning, A; Boeckx, N; Van Poecke, N; Lucio, P; Mendonça, A; Sedek, L; Szczepański, T; Kalina, T; Kanderová, V; Hoogeveen, P; Flores-Montero, J; Chillón, M C; Orfao, A; Almeida, J; Evans, P; Cullen, M; Noordijk, A L; Vermeulen, P M; de Man, M T; Dixon, E P; Comans-Bitter, W M; van Dongen, J J M

    2012-09-01

    The PML-RARA fusion protein is found in approximately 97% of patients with acute promyelocytic leukemia (APL). APL can be associated with life-threatening bleeding complications when undiagnosed and not treated expeditiously. The PML-RARA fusion protein arrests maturation of myeloid cells at the promyelocytic stage, leading to the accumulation of neoplastic promyelocytes. Complete remission can be obtained by treatment with all-trans-retinoic acid (ATRA) in combination with chemotherapy. Diagnosis of APL is based on the detection of t(15;17) by karyotyping, fluorescence in situ hybridization or PCR. These techniques are laborious and demand specialized laboratories. We developed a fast (performed within 4-5 h) and sensitive (detection of at least 10% malignant cells in normal background) flow cytometric immunobead assay for the detection of PML-RARA fusion proteins in cell lysates using a bead-bound anti-RARA capture antibody and a phycoerythrin-conjugated anti-PML detection antibody. Testing of 163 newly diagnosed patients (including 46 APL cases) with the PML-RARA immunobead assay showed full concordance with the PML-RARA PCR results. As the applied antibodies recognize outer domains of the fusion protein, the assay appeared to work independently of the PML gene break point region. Importantly, the assay can be used in parallel with routine immunophenotyping for fast and easy diagnosis of APL.

  6. A Simple Flow-Cytometric Method Measuring B Cell Surface Immunoglobulin Avidity Enables Characterization of Affinity Maturation to Influenza A Virus

    Science.gov (United States)

    Frank, Gregory M.; Ince, William L.; Gibbs, James S.; Khurana, Surender; Wheatley, Adam K.; Max, Edward E.; McDermott, Adrian B.; Golding, Hana; Stevens, James; Bennink, Jack R.

    2015-01-01

    ABSTRACT Antibody (Ab) affinity maturation enables an individual to maintain immunity to an increasing number of pathogens within the limits of a total Ig production threshold. A better understanding of this process is critical for designing vaccines that generate optimal Ab responses to pathogens. Our study describes a simple flow-cytometric method that enumerates virus-specific germinal center (GC) B cells as well as their AC50, a measure of Ab avidity, defined as the antigen concentration required to detect 50% of specific B cells. Using a model of mouse Ab responses to the influenza A virus hemagglutinin (IAV HA), we obtained data indicating that AC50 decreases with time postinfection in an affinity maturation-dependent process. As proof of principle of the utility of the method, our data clearly show that relative to intranasal IAV infection, intramuscular immunization against inactivated IAV in adjuvant results in a diminished GC HA B cell response, with increased AC50 correlating with an increased serum Ab off-rate. Enabling simultaneous interrogation of both GC HA B cell quantity and quality, this technique should facilitate study of affinity maturation and rational vaccine design. PMID:26242629

  7. Evaluation of the S phase distribution of flow cytometric DNA histograms by autoradiography and computer algorithms.

    Science.gov (United States)

    Sheck, L E; Muirhead, K A; Horan, P K

    1980-09-01

    Cell sorting and tritiated thymidine autoradiography were used to define the distribution of S phase cells in flow cytometric DNA histograms obtained from exponential mouse lymphoma cells (L5178Y). The numbers of labeled S phase cells, autoradiographically determined from cells sorted at 2-channel intervals in the G1/early S and late S/G2M regions of the histogram, were compared with the numbers of computed S phase cells in comparable 2-channel intervals as predicted by several computer algorithms used to extract cell cycle phase distributions from DNA histograms. Polynomial and multirectangle algorithms gave computed estimates of total %S in close agreement with the tritiated thymidine labeling index for the cell population, while multi-Gaussian algorithms underestimated %S. Interval autoradiographic and algorithm studies confirmed these results in that no significant differences were found between the autoradiographic S phase distribution and S phase distributions calculated by the polynomial and multirectangle models. However, S phase cells were significantly underestimated in G1/early S by a constrained multi-Gaussian model and in both G1/early S and late S/G2 by an unconstrained multi-Gaussian model. For the particular cell line (L5178Y), staining protocol (mithramycin following ethanol fixation) and instrumentation (Coulter TPS-2 cell sorter) used in this study, close agreement between computed %S and tritiated thymidine labeling index was found to be a reliable indicator of an algorithm's success in resolving S phase cells in the G1/S and S/G2 transition regions of the DNA histograms.

  8. Flow cytometric analysis of lectin binding to in vitro-cultured Perkinsus marinus surface carbohydrates

    Science.gov (United States)

    Gauthier, J.D.; Jenkins, J.A.; La Peyre, Jerome F.

    2004-01-01

    Parasite surface glycoconjugates are frequently involved in cellular recognition and colonization of the host. This study reports on the identification of Perkinsus marinus surface carbohydrates by flow cytometric analyses of fluorescein isothiocyanate-conjugated lectin binding. Lectin-binding specificity was confirmed by sugar inhibition and Kolmogorov-Smirnov statistics. Clear, measurable fluorescence peaks were discriminated, and no parasite autofluorescence was observed. Parasites (GTLA-5 and Perkinsus-1 strains) harvested during log and stationary phases of growth in a protein-free medium reacted strongly with concanavalin A and wheat germ agglutinin, which bind to glucose-mannose and N-acetyl-D-glucosamine (GlcNAc) moieties, respectively. Both P. marinus strains bound with lower intensity to Maclura pomifera agglutinin, Bauhinia purpurea agglutinin, soybean agglutinin (N-acetyl-D-galactosamine-specific lectins), peanut agglutinin (PNA) (terminal galactose specific), and Griffonia simplicifolia II (GlcNAc specific). Only background fluorescence levels were detected with Ulex europaeus agglutinin I (L-fucose specific) and Limulus polyphemus agglutinin (sialic acid specific). The lectin-binding profiles were similar for the 2 strains except for a greater relative binding intensity of PNA for Perkinsus-1 and an overall greater lectin-binding capacity of Perkinsus-1 compared with GTLA-5. Growth stage comparisons revealed increased lectin-binding intensities during stationary phase compared with log phase of growth. This is the first report of the identification of surface glycoconjugates on a Perkinsus spp. by flow cytometry and the first to demonstrate that differential surface sugar expression is growth phase and strain dependent. ?? American Society of Parasitologists 2004.

  9. Flow cytometric viability assessment of lactic acid bacteria starter cultures produced by fluidized bed drying.

    Science.gov (United States)

    Bensch, Gerald; Rüger, Marc; Wassermann, Magdalena; Weinholz, Susann; Reichl, Udo; Cordes, Christiana

    2014-06-01

    For starter culture production, fluidized bed drying is an efficient and cost-effective alternative to the most frequently used freeze drying method. However, fluidized bed drying also poses damaging or lethal stress to bacteria. Therefore, investigation of impact of process variables and conditions on viability of starter cultures produced by fluidized bed drying is of major interest. Viability of bacteria is most frequently assessed by plate counting. While reproductive growth of cells can be characterized by the number of colony-forming units, it cannot provide the number of viable-but-nonculturable cells. However, in starter cultures, these cells still contribute to the fermentation during food production. In this study, flow cytometry was applied to assess viability of Lactobacillus plantarum starter cultures by membrane integrity analysis using SYBR®Green I and propidium iodide staining. The enumeration method established allowed for rapid, precise and sensitive determination of viable cell concentration, and was used to investigate effects of fluidized bed drying and storage on viability of L. plantarum. Drying caused substantial membrane damage on cells, most likely due to dehydration and oxidative stress. Nevertheless, high bacterial survival rates were obtained, and granulates contained in the average 2.7 × 10(9) viable cells/g. Furthermore, increased temperatures reduced viability of bacteria during storage. Differences in results of flow cytometry and plate counting suggested an occurrence of viable-but-nonculturable cells during storage. Overall, flow cytometric viability assessment is highly feasible for rapid routine in-process control in production of L. plantarum starter cultures, produced by fluidized bed drying.

  10. Flow Cytometric Assessment of Bacterial Abundance in Soils, Sediments and Sludge.

    Science.gov (United States)

    Frossard, Aline; Hammes, Frederik; Gessner, Mark O

    2016-01-01

    Bacterial abundance is a fundamental measure in microbiology, but its assessment is often tedious, especially for soil, and sediment samples. To overcome this limitation, we adopted a time-efficient flow-cytometric (FCM) counting method involving cell detachment and separation from matrix particles by centrifugation in tubes receiving sample suspensions and Histodenz(®) solution. We used this approach to assess bacterial abundances in diverse soils (natural and agricultural), sediments (streams and lakes) and sludge from sand-filters in a drinking water treatment plant and compared the results to bacterial abundances determined by two established methods, epifluorescence microscopy (EM) and adenosine triphosphate (ATP) quantification. Cell abundances determined by FCM and EM correlated fairly well, although absolute cell abundances were generally lower when determined by FCM. FCM also showed significant relations with cell counts converted from ATP concentrations, although estimates derived from ATP determinations were typically higher, indicating the presence of ATP sources other than bacteria. Soil and sediment organic matter (OM) content influenced the goodness of fit between counts obtained with EM and FCM. In particular, bacterial abundance determined by FCM in samples containing less than 10% OM, such as stream sediment, was particularly well correlated with the cell counts assessed by EM. Overall, these results suggest that FCM following cell detachment and purification is a useful approach to increase sample throughput for determining bacterial abundances in soils, sediments and sludge. However, notable scatter and only partial concordance among the FCM and reference methods suggests that protocols require further improvement for assessments requiring high precision, especially when OM contents in samples are high.

  11. Intestinal intraepithelial lymphocyte cytometric pattern is more accurate than subepithelial deposits of anti-tissue transglutaminase IgA for the diagnosis of celiac disease in lymphocytic enteritis.

    Directory of Open Access Journals (Sweden)

    Fernando Fernández-Bañares

    Full Text Available BACKGROUND & AIMS: An increase in CD3+TCRγδ+ and a decrease in CD3- intraepithelial lymphocytes (IEL is a characteristic flow cytometric pattern of celiac disease (CD with atrophy. The aim was to evaluate the usefulness of both CD IEL cytometric pattern and anti-TG2 IgA subepithelial deposit analysis (CD IF pattern for diagnosing lymphocytic enteritis due to CD. METHODS: Two-hundred and five patients (144 females who underwent duodenal biopsy for clinical suspicion of CD and positive celiac genetics were prospectively included. Fifty had villous atrophy, 70 lymphocytic enteritis, and 85 normal histology. Eight patients with non-celiac atrophy and 15 with lymphocytic enteritis secondary to Helicobacter pylori acted as control group. Duodenal biopsies were obtained to assess both CD IEL flow cytometric (complete or incomplete and IF patterns. RESULTS: Sensitivity of IF, and complete and incomplete cytometric patterns for CD diagnosis in patients with positive serology (Marsh 1+3 was 92%, 85 and 97% respectively, but only the complete cytometric pattern had 100% specificity. Twelve seropositive and 8 seronegative Marsh 1 patients had a CD diagnosis at inclusion or after gluten free-diet, respectively. CD cytometric pattern showed a better diagnostic performance than both IF pattern and serology for CD diagnosis in lymphocytic enteritis at baseline (95% vs 60% vs 60%, p = 0.039. CONCLUSIONS: Analysis of the IEL flow cytometric pattern is a fast, accurate method for identifying CD in the initial diagnostic biopsy of patients presenting with lymphocytic enteritis, even in seronegative patients, and seems to be better than anti-TG2 intestinal deposits.

  12. Novel nuclei isolation buffer for flow cytometric genome size estimation of Zingiberaceae: a comparison with common isolation buffers.

    Science.gov (United States)

    Sadhu, Abhishek; Bhadra, Sreetama; Bandyopadhyay, Maumita

    2016-11-01

    Cytological parameters such as chromosome numbers and genome sizes of plants are used routinely for studying evolutionary aspects of polyploid plants. Members of Zingiberaceae show a wide range of inter- and intrageneric variation in their reproductive habits and ploidy levels. Conventional cytological study in this group of plants is severely hampered by the presence of diverse secondary metabolites, which also affect their genome size estimation using flow cytometry. None of the several nuclei isolation buffers used in flow cytometry could be used very successfully for members of Zingiberaceae to isolate good quality nuclei from both shoot and root tissues. The competency of eight nuclei isolation buffers was compared with a newly formulated buffer, MB01, in six different genera of Zingiberaceae based on the fluorescence intensity of propidium iodide-stained nuclei using flow cytometric parameters, namely coefficient of variation of the G0/G1 peak, debris factor and nuclei yield factor. Isolated nuclei were studied using fluorescence microscopy and bio-scanning electron microscopy to analyse stain-nuclei interaction and nuclei topology, respectively. Genome contents of 21 species belonging to these six genera were determined using MB01. Flow cytometric parameters showed significant differences among the analysed buffers. MB01 exhibited the best combination of analysed parameters; photomicrographs obtained from fluorescence and electron microscopy supported the superiority of MB01 buffer over other buffers. Among the 21 species studied, nuclear DNA contents of 14 species are reported for the first time. Results of the present study substantiate the enhanced efficacy of MB01, compared to other buffers tested, in the generation of acceptable cytograms from all species of Zingiberaceae studied. Our study facilitates new ways of sample preparation for further flow cytometric analysis of genome size of other members belonging to this highly complex polyploid family.

  13. Clonal evolution demonstrated by flow cytometric DNA analysis of a human colonic carcinoma grown in nude mice

    DEFF Research Database (Denmark)

    Vindeløv, L L; Spang-Thomsen, M; Visfeldt, J

    1982-01-01

    A spontaneous change in DNA content of a human colonic carcinoma grown in nude mice was observed fortuitously. The tumor initially had a G1 cell DNA content of 1.3 times that of normal cells. Flow cytometric DNA analysis showed in transplant generation 56 the appearance of a new subpopulation whi...... evolution of a tumor would be less pronounced if old subpopulations often become extinct as new ones emerge. Heterogeneity of human tumors is of clinical importance because the individual subpopulations may have different sensitivity patterns to antineoplastic drugs....

  14. Rapid detection of BCR-ABL fusion genes using a novel combined LUX primer, in-cell RT-PCR and flow cytometric method.

    Science.gov (United States)

    Shi, Yan; Li, Li-Zhen; Sun, Jian-Zhi; Zhang, Ti; Peng, Jun; Xu, Cong-Gao

    2008-01-01

    Currently, quantitative and semiquantitative assays for minimal residual disease detection include fluorescence in situ hybridisation, multiparameter flow cytometric immunophenotyping and real-time quantitative polymerase chain reaction (RQ-PCR). We have developed a new approach to detect hybrid breakpoint cluster region and Abelson proto-oncogene (BCR-ABL) transcripts inside suspension cells using in situ RT-PCR and light upon extension (LUX) primer, followed by rapid quantitative analysis with flow cytometry. After cellular permeabilization and fixation of single cell suspension, the neoplastic mRNA was reverse transcribed and amplified by PCR with LUX primer. The results demonstrated that a strong positive yellow-green signal was observed in 99-100% cells of K562 cell line, only the red nucleus was detected in NB4 cell line and normal controls. The technique has been utilised to study 12 patients with chronic myeloid leukemia, and the results were compared with those of BCR-ABL fusion mRNA by RT-PCR and BCR-ABL fusion gene of the interphase cells by fluorescence in situ hybridization (FISH). In the five diagnosed patients, 90-98% cells were strongly positive. Four patients, including three patients treated with interferon-alpha and hydroxyurea and one patient treated with imatinib mesylate, had 26-82.5% positive cells. Three patients treated with imatinib mesylate were negative. The in situ RT-PCR results demonstrated complete concordance with the results of I-FISH and RT-PCR. A fluorescence signal was detectable at 1/10(4) cells and became negative below this threshold with flow cytometry. The results of the present study suggest that (1) LUX primers can be used to efficiently detect BCR-ABL fusion mRNA by in-cell RT-PCR; (2) the novel technique is a specific and sensitive way of detecting fusion gene with potential clinical usefulness.

  15. Correlation between histological grading and ploidy status in potentially malignant disorders of the oral mucosa: A flow cytometric analysis

    Directory of Open Access Journals (Sweden)

    T Vijayavel

    2013-01-01

    Full Text Available Background: Histopathological grading of oral dysplastic lesions is the method of choice for evaluating malignant and potentially malignant disorders. Owing to inter- and intra-observer variability, determination of the DNA ploidy status of lesions may serve as an adjunct in the prediction of malignant transformation. Aim: To correlate histopathological grading and ploidy status in potentially malignant and malignant disorders of the oral mucosa. Settings and Design: A pilot study was done with 30 patients (10 patients with oral potentially malignant disorders predominantly leukoplakia, 10 patients with oral malignant lesions and 10 patients with normal mucosa. Materials and Methods: Incisional biopsy was done after isolating the biopsy site with 1% Toluidine blue staining. Two sections of the tissue were removed and sent for histopathological and Flow-cytometric analysis respectively. Histopathological diagnosis was obtained and compared with Flow-cytometric results which were graded as diploid and aneuploid. Further, the S - phase fraction, DNA index were also calculated to evaluate the severity of malignant transformation or malignancy. Statistical Analysis: The results were analyzed using Pearson Chi-Square Test. Results: There exists a significant correlation between histopathology and ploidy status in both potentially malignant and malignant group. (P = 0.002. Conclusion: The data from this study has shown that DNA Ploidy analysis can be used as a valuable tool in assessing the carcinomatous progression of potentially malignant and malignant lesions.

  16. Identification and purification of classical Hodgkin cells from lymph nodes by flow cytometry and flow cytometric cell sorting.

    Science.gov (United States)

    Fromm, Jonathan R; Kussick, Steven J; Wood, Brent L

    2006-11-01

    We demonstrate the feasibility of using flow cytometry (FC) to identify the Hodgkin and Reed-Sternberg (HRS) cells of classical Hodgkin lymphoma (CHL). Initial flow cytometric studies of the HRS cell line L1236 demonstrated potentially useful antigens for identifying HRS cells. L1236 cells spontaneously bound normal T cells, analogous to the T-cell rosetting of HRS cells seen in tissue sections of CHL, but these interactions could be blocked by using a cocktail of unlabeled antibodies to 4 adhesion molecules. Among 27 lymph nodes involved by CHL, FC enabled HRS cells to be identified in 89%, whereas none of 29 non-CHL neoplasms or 23 reactive lymph nodes demonstrated HRS populations. Of the CHL cases, 82% demonstrated interactions between HRS cells and T cells that could be disrupted with blocking antibodies. Flow cytometric cell sorting experiments demonstrated typical HRS cytomorphologic features among the purified cells. FC may offer an alternative to immunohistochemical analysis in confirming the diagnosis of CHL in certain cases, and, through cell sorting, it provides a means of rapidly isolating pure HRS cells.

  17. Effect of intra-cellular trafficking on flow cytometric measurement of neutrophil's oxidative status in iron deficient pregnant females.

    Science.gov (United States)

    Youssef, Soha R; Hendawy, Sherif F; Boshnak, Noha H; Sedhom, Mariana S

    2017-03-27

    Iron deficiency and iron deficiency anemia are prevalent among pregnant women particularly in developing countries. This study aimed to evaluate the iron status among Egyptian pregnant women and its impact on their neutrophil's count and antimicrobial functions. Ninety pregnant females underwent complete blood count, iron profile, flow cytometric studies for neutrophil myeloperoxidase expression & oxidative burst using dihydrorhodamine 123 (DHR) after phorbol-12-myristate-13-acetate (PMA) stimulation as well as neutrophil phagocytic and lytic indices. According to percent saturation 54/90 women (60%) were iron deficient (women were in their third trimester compared to controls. No significant difference was found between the iron deficient & sufficient groups as regards anemia despite a positive correlation between haemoglobin level and percent saturation (P=.02). Both the phagocytic and lytic indices were significantly lower among the cases compared to controls (P=.014 & .002 respectively). Cases and controls were comparable as regards flow cytometric studies of neutrophils' myeloperoxidase and oxidative burst (P>.05). No significant correlation was found between any of the iron profile parameters and the oxidative burst by flow cytometry. Functional microphage assay (phagocytic and lytic indices) may be more relevant and cost effective than flow cytometry assays of myeloperoxidase and oxidative burst in reflecting either iron status or cellular immunity in pregnancy. © 2017 Wiley Periodicals, Inc.

  18. Flow cytometric measurement of calcium influx in murine T cell hybrids using Fluo-3 and an organic-anion transport inhibitor.

    Science.gov (United States)

    Baus, E; Urbain, J; Leo, O; Andris, F

    1994-07-12

    A method is described to facilitate flow cytometric analysis of calcium mobilization upon stimulation of murine T cell hybrids. In these transformed cell lines, the accuracy of cytometric measurement of free cytoplasmic calcium with Fluo-3 is compromised by the rapid loss of the intracellular dye. We have found that the addition of sulfinpyrazone, a known organic-anion transporter inhibitor in epithelial cells and in macrophages, severely impairs the leakage of the Fluo-3 probe from the cytoplasmic matrix. Under appropriate conditions, sulfinpyrazone has little effect on the cell physiology and permits the detection of calcium influx in a variety of murine T cell hybrids.

  19. Quantitation of minimal disease levels in chronic lymphocytic leukemia using a sensitive flow cytometric assay improves the prediction of outcome and can be used to optimize therapy.

    Science.gov (United States)

    Rawstron, A C; Kennedy, B; Evans, P A; Davies, F E; Richards, S J; Haynes, A P; Russell, N H; Hale, G; Morgan, G J; Jack, A S; Hillmen, P

    2001-07-01

    Previous studies have suggested that the level of residual disease at the end of therapy predicts outcome in chronic lymphocytic leukemia (CLL). However, available methods for detecting CLL cells are either insensitive or not routinely applicable. A flow cytometric assay was developed that can differentiate CLL cells from normal B cells on the basis of their CD19/CD5/CD20/CD79b expression. The assay is rapid and can detect one CLL cell in 10(4) to 10(5) leukocytes in all patients. We have compared this assay to conventional assessment in 104 patients treated with CAMPATH-1H and/or autologous transplant. During CAMPATH-1H therapy, circulating CLL cells were rapidly depleted in responding patients, but remained detectable in nonresponders. Patients with more than 0.01 x 10(9)/L circulating CLL cells always had significant (> 5%) marrow disease, and blood monitoring could be used to time marrow assessments. In 25 out of 104 patients achieving complete remission by National Cancer Institute (NCI) criteria, the detection of residual bone marrow disease at more than 0.05% of leukocytes in 6 out of 25 patients predicted significantly poorer event-free (P =.0001) and overall survival (P =.007). CLL cells are detectable at a median of 15.8 months (range, 5.5-41.8) posttreatment in 9 out of 18 evaluable patients with less than 0.05% CLL cells at end of treatment. All patients with detectable disease have progressively increasing disease levels on follow-up. The use of sensitive techniques, such as the flow assay described here, allow accurate quantitation of disease levels and provide an accurate method for guiding therapy and predicting outcome. These results suggest that the eradication of detectable disease may lead to improved survival and should be tested in future studies.

  20. Flow cytometric measurement of RNA synthesis based on bromouridine labelling and combined with measurement of DNA content or cell surface antigen

    DEFF Research Database (Denmark)

    Jensen, P O; Larsen, J; Larsen, J K

    1993-01-01

    that RNA synthesis increased within the first 24 hours of phytohemagglutinin (PHA) stimulation, reaching a maximum at 48 hours, when cells had entered the cell cycle. Using a new method for flow cytometric dual parameter analysis of BrUrd incorporation and a cell surface antigen, spontaneous RNA synthesis...

  1. Effect of 17 beta-oestradiol on growth curves and flow cytometric DNA distribution of two human breast carcinomas grown in nude mice

    DEFF Research Database (Denmark)

    Brünner, N; Spang-Thomsen, M; Vindeløv, L

    1983-01-01

    The effect of 17 beta-oestradiol on a "receptor positive" and on a "receptor negative" human breast carcinoma grown in nude mice was studied. Experimental growth data were used to determine the effect on tumour growth. Flow cytometric DNA analysis (FCM) performed on tumour tissue obtained...

  2. THE EFFECT OF LABELING INTENSITY, ESTIMATED BY REAL-TIME CONFOCAL LASER SCANNING MICROSCOPY, ON FLOW CYTOMETRIC APPEARANCE AND IDENTIFICATION OF IMMUNOCHEMICALLY LABELED MARINE DINOFLAGELLATES

    NARCIS (Netherlands)

    VRIELING, EG; DRAAIJER, A; VANZEIJL, WJM; PEPERZAK, L; GIESKES, WWC; VEENHUIS, M; Zeijl, Wilhelmus J.M. van

    1993-01-01

    Two different fluorescein isothiocyanate (FITC) conjugates were used to analyze the effect of labeling intensity on the flow cytometric appearance of marine dinoflagellates labeled with antibodies that specifically recognized the outer cell wall. Location of the labeling was revealed by epifluoresce

  3. Flow Cytometric Measurement of [Ca2+]i and pHi in Conjugated Natural Killer Cells and K562 Target Cells during the Cytotoxic Process1,2

    NARCIS (Netherlands)

    van Graft, Marja; van Graft, M.; Kraan, Yvonne M.; Segers-Nolten, Gezina M.J.; Radosevic, K.; Radosevic, Katarina; de Grooth, B.G.; Greve, Jan

    1993-01-01

    We describe a flow cytometric assay that enables one to follow conjugate formation between cytotoxic cells and their target cells during the cytotoxic process. In addition, the internal calcium concentration ([Ca2+]i) and internal pH (pHi) of the conjugated cells can be monitored and directly

  4. Flow cytometric analyses of the viability, surface marker expression and function of lymphocytes from children following cryopreservation.

    Science.gov (United States)

    Chen, Xi; Zhang, Hui; Mou, Wenjun; Qi, Zhan; Ren, Xiaoya; Wang, Guoliang; Jiao, Hong; Kong, Xiaohui; Gui, Jingang

    2016-11-01

    Flow cytometric analysis is important for the investigation and clinical preparation of lymphocytes from children. However, the strict requirement of cell freshness and inter‑assay variability limits the application of this methodology for pediatric investigations. Therefore, it is necessary to identify a reliable cryopreservative method capable of maintaining high cell viability and proper cell function in lymphocytes from children. In the present study, eight commonly‑used cell cyropreservative methods were used, and their effects on cell viability, surface marker expression and cell function were examined. In addition, how these methods affect the distribution of T‑cell receptor Vβ subfamilies were also determined. The results of the present study provided valuable experimental evidence, based on which the optimal method for the cryopreservation of lymphocytes from children in pediatric investigations and clinical applications can be selected.

  5. A cluster analysis method for identification of subpopulations of cells in flow cytometric list-mode arrays

    Science.gov (United States)

    Li, Z. K.

    1985-01-01

    A specialized program was developed for flow cytometric list-mode data using an heirarchical tree method for identifying and enumerating individual subpopulations, the method of principal components for a two-dimensional display of 6-parameter data array, and a standard sorting algorithm for characterizing subpopulations. The program was tested against a published data set subjected to cluster analysis and experimental data sets from controlled flow cytometry experiments using a Coulter Electronics EPICS V Cell Sorter. A version of the program in compiled BASIC is usable on a 16-bit microcomputer with the MS-DOS operating system. It is specialized for 6 parameters and up to 20,000 cells. Its two-dimensional display of Euclidean distances reveals clusters clearly, as does its 1-dimensional display. The identified subpopulations can, in suitable experiments, be related to functional subpopulations of cells.

  6. Flow cytometric kinetic assay of calcium mobilization in whole blood platelets using Fluo-3 and CD41.

    Science.gov (United States)

    do Céu Monteiro, M; Sansonetty, F; Gonçalves, M J; O'Connor, J E

    1999-04-01

    Platelet activation plays a major role in the physiology and pathology of hemostasis. Flow cytometry is a promising approach for the structural and functional analysis of platelets. However, the choice of adequate biological parameters and most technical issues are still under discussion. A rise in cytosolic free Ca2+ is a key early event that follows platelet stimulation and precedes several activation responses, including shape change, aggregation, secretion, and expression of procoagulant activity. Our objective was to set up a fast and sensitive flow cytometric method to determine the kinetics of intracellular Ca2+ mobilization in platelets, which could be performed with the least artifactual perturbation of platelet function. Anticoagulated blood was diluted in Tyrode's buffer and incubated with Fluo-3-acetoxymethyl ester prior to staining with phycoerytrin-conjugated antiplatelet GPIIb/IIIa complex monoclonal antibody. Platelets were identified by a gate including only CD41+ events. After the determination of baseline Fluo-3 green fluorescence on a flow cytometer (EPICS XL-MCL, Coulter Electronics, Hialeah, FL), adequate agonists were added and time-dependent changes in Fluo-3 fluorescence were recorded on-line for up to 3 min. In these conditions, a very fast and transient increase of cytosolic-free Ca2+ was observed following the addition of thrombin, a strong platelet agonist. Stimulation with adenosine diphosphate (ADP), a weak agonist, also resulted in evident increase of Ca2+ levels. Our results show that this flow cytometric kinetic method provides a simple and sensitive tool to assess in vitro the time course and intensity of signal transduction responses to different platelet agonists under near physiological conditions. In this way, it may be useful to evaluate the degree of platelet reactivity and thus to monitor antiplatelet therapy.

  7. Flow cytometric evaluation of physico-chemical impact on Gram-positive and Gram-negative bacteria

    Directory of Open Access Journals (Sweden)

    Antje eFröhling

    2015-09-01

    Full Text Available Since heat sensitivity of fruits and vegetables limits the application of thermal inactivation processes, new emerging inactivation technologies have to be established to fulfil the requirements of food safety without affecting the produce quality. The efficiency of inactivation treatments has to be ensured and monitored. Monitoring of inactivation effects is commonly performed using traditional cultivation methods which have the disadvantage of the time span needed to obtain results.The aim of this study was to compare the inactivation effects of peracetic acid (PAA, ozonated water (O3 and cold atmospheric pressure plasma (CAPP on Gram-positive and Gram-negative bacteria using flow cytometric methods. E. coli cells were completely depolarized after treatment (15 s with 0.25 % PAA at 10 °C, and after treatment (10 s with 3.8 mg l-1 O3 at 12°C. The membrane potential of CAPP treated cells remained almost constant at an operating power of 20 W over a time period of 3 min, and subsequently decreased within 30 s of further treatment. Complete membrane permeabilization was observed after 10 s O3 treatment, but treatment with PAA and CAPP did not completely permeabilize the cells within 2 min and 4 min, respectively. Similar results were obtained for esterase activity. O3 inactivates cellular esterase but esterase activity was detected after 4 min CAPP treatment and 2 min PAA treatment. L. innocua cells and P. carotovorum cells were also permeabilized instantaneously by O3 treatment at concentrations of 3.8 ± 1 mg l-1. However, higher membrane permeabilization of L. innocua and P. carotovorum than of E. coli was observed at CAPP treatment of 20 W. The degree of bacterial damage due to the inactivation processes is highly dependent on treatment parameters as well as on treated bacteria. Important information regarding the inactivation mechanisms can be obtained by flow cytometric measurements and this enables the definition of critical process

  8. A Protocol for the Comprehensive Flow Cytometric Analysis of Immune Cells in Normal and Inflamed Murine Non-Lymphoid Tissues.

    Science.gov (United States)

    Yu, Yen-Rei A; O'Koren, Emily G; Hotten, Danielle F; Kan, Matthew J; Kopin, David; Nelson, Erik R; Que, Loretta; Gunn, Michael D

    2016-01-01

    Flow cytometry is used extensively to examine immune cells in non-lymphoid tissues. However, a method of flow cytometric analysis that is both comprehensive and widely applicable has not been described. We developed a protocol for the flow cytometric analysis of non-lymphoid tissues, including methods of tissue preparation, a 10-fluorochrome panel for cell staining, and a standardized gating strategy, that allows the simultaneous identification and quantification of all major immune cell types in a variety of normal and inflamed non-lymphoid tissues. We demonstrate that our basic protocol minimizes cell loss, reliably distinguishes macrophages from dendritic cells (DC), and identifies all major granulocytic and mononuclear phagocytic cell types. This protocol is able to accurately quantify 11 distinct immune cell types, including T cells, B cells, NK cells, neutrophils, eosinophils, inflammatory monocytes, resident monocytes, alveolar macrophages, resident/interstitial macrophages, CD11b- DC, and CD11b+ DC, in normal lung, heart, liver, kidney, intestine, skin, eyes, and mammary gland. We also characterized the expression patterns of several commonly used myeloid and macrophage markers. This basic protocol can be expanded to identify additional cell types such as mast cells, basophils, and plasmacytoid DC, or perform detailed phenotyping of specific cell types. In examining models of primary and metastatic mammary tumors, this protocol allowed the identification of several distinct tumor associated macrophage phenotypes, the appearance of which was highly specific to individual tumor cell lines. This protocol provides a valuable tool to examine immune cell repertoires and follow immune responses in a wide variety of tissues and experimental conditions.

  9. Development of a flow cytometric bead immunoassay and its assessment as a possible aid to potency evaluation of enterotoxaemia vaccines

    Directory of Open Access Journals (Sweden)

    Angela Buys

    2014-02-01

    Full Text Available Enterotoxaemia, an economically important disease of sheep, goats and calves, is caused by systemic effects of the epsilon toxin produced by the anaerobic bacterium Clostridium perfringens type D. The only practical means of controlling the occurrence of enterotoxaemia is to immunise animals by vaccination. The vaccine is prepared by deriving a toxoid from the bacterial culture filtrate and the potency of the vaccine is tested with the in vivo mouse neutralisation test (MNT. Due to ethical, economic and technical reasons, alternative in vitro assays are needed. In this study an indirect cytometric bead immunoassay (I-CBA was developed for use in vaccine potency testing and the results were compared with those obtained using an indirect enzyme-linked immunosorbent assay (I-ELISA and the MNT. Sera were collected from guinea pigs immunised with three different production batches of enterotoxaemia vaccine and the levels of anti-epsilon toxin antibodies were determined. Although the intra- and inter-assay variability was satisfactory, epsilon antitoxin levels determined by both the I-ELISA and indirect cytometric bead immunoassay (I-CBA tests were higher than those of the MNT assay. In contrast to the MNT, all of the serum samples were identified as having antitoxin levels above the required minimum (not less than 5 U/mL. These results indicate that the respective in vitro tests in their current formats are not yet suitable alternatives to the in vivo MNT. The growing demand for a more humane, cost-effective and efficient method for testing the potency of enterotoxaemia vaccines, however, provides a strong impetus for further optimisation and standardisation of the I-CBA assay but further analytical research is required.

  10. Clinical flow cytometric screening of SAP and XIAP expression accurately identifies patients with SH2D1A and XIAP/BIRC4 mutations.

    Science.gov (United States)

    Gifford, Carrie E; Weingartner, Elizabeth; Villanueva, Joyce; Johnson, Judith; Zhang, Kejian; Filipovich, Alexandra H; Bleesing, Jack J; Marsh, Rebecca A

    2014-07-01

    X-linked lymphoproliferative disease is caused by mutations in two genes, SH2D1A and XIAP/BIRC4. Flow cytometric methods have been developed to detect the gene products, SAP and XIAP. However, there is no literature describing the accuracy of flow cytometric screening performed in a clinical lab setting. We reviewed the clinical flow cytometric testing results for 656 SAP and 586 XIAP samples tested during a 3-year period. Genetic testing was clinically performed as directed by the managing physician in 137 SAP (21%) and 115 XIAP (20%) samples. We included these samples for analyses of flow cytometric test accuracy. SH2D1A mutations were detected in 15/137 samples. SAP expression was low in 13/15 (sensitivity 87%, CI 61-97%). Of the 122 samples with normal sequencing, SAP was normal in 109 (specificity 89%, CI 82-94%). The positive predictive values (PPVs) and the negative predictive values (NPVs) were 50% and 98%, respectively. XIAP/BIRC4 mutations were detected in 19/115 samples. XIAP expression was low in 18/19 (sensitivity 95%, CI 73-100%). Of the 96 samples with normal sequencing, 59 had normal XIAP expression (specificity 61%, CI 51-71%). The PPVs and NPVs were 33% and 98%, respectively. Receiver-operating characteristic analysis was able to improve the specificity to 75%. Clinical flow cytometric screening tests for SAP and XIAP deficiencies offer good sensitivity and specificity for detecting genetic mutations, and are characterized by high NPVs. We recommend these tests for patients suspected of having X-linked lymphoproliferative disease type 1 (XLP1) or XLP2. © 2014 Clinical Cytometry Society.

  11. Microbial Eco-Physiology of the human intestinal tract: a flow cytometric approach

    NARCIS (Netherlands)

    Amor, Ben K.

    2004-01-01

    This thesis describes a multifaceted approach to further enhance our view of the complex human intestinal microbial ecosystem. This approach combines me advantages of flow cyrometry (FCM), a single cell and high-throughput technology, and molecular techniques that have proven themselves to be invalu

  12. Microbial Eco-Physiology of the human intestinal tract: a flow cytometric approach

    NARCIS (Netherlands)

    Amor, Ben K.

    2004-01-01

    This thesis describes a multifaceted approach to further enhance our view of the complex human intestinal microbial ecosystem. This approach combines me advantages of flow cyrometry (FCM), a single cell and high-throughput technology, and molecular techniques that have proven themselves to be

  13. Microbial Eco-Physiology of the human intestinal tract: a flow cytometric approach

    NARCIS (Netherlands)

    Amor, Ben K.

    2004-01-01

    This thesis describes a multifaceted approach to further enhance our view of the complex human intestinal microbial ecosystem. This approach combines me advantages of flow cyrometry (FCM), a single cell and high-throughput technology, and molecular techniques that have proven themselves to be invalu

  14. Increased electron donor and electron acceptor characters enhance the adhesion between oil droplets and cells of Yarrowia lipolytica as evaluated by a new cytometric assay.

    Science.gov (United States)

    Aguedo, Mario; Waché, Yves; Mazoyer, Virginie; Sequeira-Le Grand, Anabelle; Belin, Jean-Marc

    2003-05-07

    The adhesion of methyl ricinoleate droplets to cells of the yeast Yarrowia lipolytica was investigated. A new cytometric method, relying on the double staining of fatty globules with Nile Red and of cells with Calcofluor, enabled us to quantify methyl ricinoleate droplet adhesion to cells precultured on a hydrophilic or on a hydrophobic carbon source. In this last case, droplet adsorption was enhanced and a MATS (microbial adhesion to solvents) test revealed that this increase was due to Lewis acid-base interactions and not to an increase in the hydrophobic properties of the cell surface. These preliminary results demonstrate that the developed cytometric method is promising for various applications concerning the study of interactions between microorganisms and an emulsified hydrophobic substrates.

  15. Chemosensitivity of human small cell carcinoma of the lung detected by flow cytometric DNA analysis of drug-induced cell cycle perturbations in vitro

    DEFF Research Database (Denmark)

    Engelholm, S A; Spang-Thomsen, M; Vindeløv, L L

    1986-01-01

    A method based on detection of drug-induced cell cycle perturbation by flow cytometric DNA analysis has previously been described in Ehrlich ascites tumors as a way to estimate chemosensitivity. The method is extended to test human small-cell carcinoma of the lung. Three tumors with different...... sensitivities to melphalan in nude mice were used. Tumors were disaggregated by a combined mechanical and enzymatic method and thereafter have incubated with different doses of melphalan. After incubation the cells were plated in vitro on agar, and drug induced cell cycle changes were monitored by flow...... cytometric DNA analysis. Melphalan produced a dose-related S phase accumulation in the two sensitive tumors, whereas no changes in the cell cycle distribution were found in the resistant tumor. The size of S phase accumulation correlated to the chemosensitivity in vivo. For low concentrations of melphalan...

  16. Overview of very small embryonic-like stem cells (VSELs) and methodology of their identification and isolation by flow cytometric methods.

    Science.gov (United States)

    Zuba-Surma, Ewa K; Ratajczak, Mariusz Z

    2010-01-01

    The protocols presented here describe the procedures employed to identify and isolate very small embryonic-like stem cells (VSELs) using flow cytometric technologies including fluorescence-activated cell sorting (FACS). We describe the recommended steps in detail for their successful identification and isolation from adult tissues. These protocols were initially established to isolate such cells from murine bone marrow (BM) and human cord blood (CB) and may also be employed to isolate these primitive cells from other adult organs and embryonic tissues. Here, we focus on some critical parameters/key points required for the successful identification and purification of these rare cells by employing classical flow cytometry. In the last part of this unit, we also discuss a novel flow cytometric tool, ImageStream, an imaging flow cytometer, which allows better identification and morphological analysis of sorted cells.

  17. Application of advanced cytometric and molecular technologies to minimal residual disease monitoring

    Science.gov (United States)

    Leary, James F.; He, Feng; Reece, Lisa M.

    2000-04-01

    Minimal residual disease monitoring presents a number of theoretical and practical challenges. Recently it has been possible to meet some of these challenges by combining a number of new advanced biotechnologies. To monitor the number of residual tumor cells requires complex cocktails of molecular probes that collectively provide sensitivities of detection on the order of one residual tumor cell per million total cells. Ultra-high-speed, multi parameter flow cytometry is capable of analyzing cells at rates in excess of 100,000 cells/sec. Residual tumor selection marker cocktails can be optimized by use of receiver operating characteristic analysis. New data minimizing techniques when combined with multi variate statistical or neural network classifications of tumor cells can more accurately predict residual tumor cell frequencies. The combination of these techniques can, under at least some circumstances, detect frequencies of tumor cells as low as one cell in a million with an accuracy of over 98 percent correct classification. Detection of mutations in tumor suppressor genes requires insolation of these rare tumor cells and single-cell DNA sequencing. Rare residual tumor cells can be isolated at single cell level by high-resolution single-cell cell sorting. Molecular characterization of tumor suppressor gene mutations can be accomplished using a combination of single- cell polymerase chain reaction amplification of specific gene sequences followed by TA cloning techniques and DNA sequencing. Mutations as small as a single base pair in a tumor suppressor gene of a single sorted tumor cell have been detected using these methods. Using new amplification procedures and DNA micro arrays it should be possible to extend the capabilities shown in this paper to screening of multiple DNA mutations in tumor suppressor and other genes on small numbers of sorted metastatic tumor cells.

  18. A whole blood flow cytometric determination of platelet activation by unfractionated and low molecular weight heparin in vitro.

    Science.gov (United States)

    Klein, Bernd; Faridi, Andreé; von Tempelhoff, G F; Heilmann, Lothar; Mittermayer, Christian; Rath, Werner

    2002-12-15

    The influence of unfractionated (Heparin-Natrium) and low-molecular heparin (Fragmin(R)) on platelet activation in whole blood was investigated by FACS analysis in vitro using antibodies against glycoprotein (gp) IIb/IIIa (CD 41), GMP 140 (CD 62P), gp 53 (CD 63) and fibrinogen. Samples were also labeled with anti-gp Ib (CD 42b). Neither unfractionated heparin (UFH) nor low molecular weight heparin (LMWH) led to significant (i.e., p<0.05) changes in fluorescence intensities of platelets labeled with anti-gp IIb/IIIa or anti-gp 53. Significant platelet activation due to unfractionated heparin could be observed by labeling with anti-GMP 140 (UFH: p=0.009; LMWH: p=0.16). The proportion of platelets with surface-bound fibrinogen was significantly increased (UFH: p=0.00006; LMWH: p=0.008). After incubation with heparins, activation ability of platelets by adenosine diphosphate (ADP) was significantly increased. The potentiating action of unfractionated heparin was larger. Therefore, flow cytometric results of platelet activation in patients receiving heparin should be interpreted carefully.

  19. A Simple Flow Cytometric Method to Measure Glucose Uptake and Glucose Transporter Expression for Monocyte Subpopulations in Whole Blood.

    Science.gov (United States)

    Palmer, Clovis S; Anzinger, Joshua J; Butterfield, Tiffany R; McCune, Joseph M; Crowe, Suzanne M

    2016-08-12

    Monocytes are innate immune cells that can be activated by pathogens and inflammation associated with certain chronic inflammatory diseases. Activation of monocytes induces effector functions and a concomitant shift from oxidative to glycolytic metabolism that is accompanied by increased glucose transporter expression. This increased glycolytic metabolism is also observed for trained immunity of monocytes, a form of innate immunological memory. Although in vitro protocols examining glucose transporter expression and glucose uptake by monocytes have been described, none have been examined by multi-parametric flow cytometry in whole blood. We describe a multi-parametric flow cytometric protocol for the measurement of fluorescent glucose analog 2-NBDG uptake in whole blood by total monocytes and the classical (CD14(++)CD16(-)), intermediate (CD14(++)CD16(+)) and non-classical (CD14(+)CD16(++)) monocyte subpopulations. This method can be used to examine glucose transporter expression and glucose uptake for total monocytes and monocyte subpopulations during homeostasis and inflammatory disease, and can be easily modified to examine glucose uptake for other leukocytes and leukocyte subpopulations within blood.

  20. HLA-targeted flow cytometric sorting of blood cells allows separation of pure and viable microchimeric cell populations.

    Science.gov (United States)

    Drabbels, Jos J M; van de Keur, Carin; Kemps, Berit M; Mulder, Arend; Scherjon, Sicco A; Claas, Frans H J; Eikmans, Michael

    2011-11-10

    Microchimerism is defined by the presence of low levels of nonhost cells in a person. We developed a reliable method for separating viable microchimeric cells from the host environment. For flow cytometric cell sorting, HLA antigens were targeted with human monoclonal HLA antibodies (mAbs). Optimal separation of microchimeric cells (present at a proportion as low as 0.01% in artificial mixtures) was obtained with 2 different HLA mAbs, one targeting the chimeric cells and the other the background cells. To verify purity of separated cell populations, flow-sorted fractions of 1000 cells were processed for DNA analysis by HLA-allele-specific and Y-chromosome-directed real-time quantitative PCR assays. After sorting, PCR signals of chimeric DNA markers in the positive fractions were significantly enhanced compared with those in the presort samples, and they were similar to those in 100% chimeric control samples. Next, we demonstrate applicability of HLA-targeted FACS sorting after pregnancy by separating chimeric maternal cells from child umbilical cord mononuclear cells. Targeting allelic differences with anti-HLA mAbs with FACS sorting allows maximal enrichment of viable microchimeric cells from a background cell population. The current methodology enables reliable microchimeric cell detection and separation in clinical specimens.

  1. Estimation of the Whitefly Bemisia tabaci Genome Size Based on k-mer and Flow Cytometric Analyses

    Directory of Open Access Journals (Sweden)

    Wenbo Chen

    2015-07-01

    Full Text Available Whiteflies of the Bemisia tabaci (Hemiptera: Aleyrodidae cryptic species complex are among the most important agricultural insect pests in the world. These phloem-feeding insects can colonize over 1000 species of plants worldwide and inflict severe economic losses to crops, mainly through the transmission of pathogenic viruses. Surprisingly, there is very little genomic information about whiteflies. As a starting point to genome sequencing, we report a new estimation of the genome size of the B. tabaci B biotype or Middle East-Asia Minor 1 (MEAM1 population. Using an isogenic whitefly colony with over 6500 haploid male individuals for genomic DNA, three paired-end genomic libraries with insert sizes of ~300 bp, 500 bp and 1 Kb were constructed and sequenced on an Illumina HiSeq 2500 system. A total of ~50 billion base pairs of sequences were obtained from each library. K-mer analysis using these sequences revealed that the genome size of the whitefly was ~682.3 Mb. In addition, the flow cytometric analysis estimated the haploid genome size of the whitefly to be ~690 Mb. Considering the congruency between both estimation methods, we predict the haploid genome size of B. tabaci MEAM1 to be ~680–690 Mb. Our data provide a baseline for ongoing efforts to assemble and annotate the B. tabaci genome.

  2. Flow Cytometric Evaluation of Human Neutrophil Apoptosis During Nitric Oxide Generation In Vitro: The Role of Exogenous Antioxidants

    Directory of Open Access Journals (Sweden)

    Zofia Sulowska

    2005-01-01

    in vitro. The effect of exogenous supply of NO donors such as SNP, SIN-1, and GEA-3162 on the course of human neutrophil apoptosis and the role of extracellular antioxidants in this process was investigated. Isolated from peripheral blood, neutrophils were cultured in the presence or absence of NO donor compounds and antioxidants for 8, 12, and 20 hours. Apoptosis of neutrophils was determined in vitro by flow cytometric analysis of cellular DNA content and Annexin V protein binding to the cell surface. Exposure of human neutrophils to GEA-3162 and SIN-1 significantly accelerates and enhances their apoptosis in vitro in a time-dependent fashion. In the presence of SNP, intensification of apoptosis has not been revealed until 12 hours after the culture. The inhibition of GEA-3162- and SIN-1-mediated neutrophil apoptosis by superoxide dismutase (SOD but not by catalase (CAT was observed. Our results show that SOD and CAT can protect neutrophils against NO-donors-induced apoptosis and suggest that the interaction of NO and oxygen metabolites signals may determine the destructive or protective role of NO donor compounds during apoptotic neutrophil death.

  3. Flow cytometric applicability to evaluate UV inactivation of phytoplankton in marine water samples.

    Science.gov (United States)

    Olsen, Ranveig Ottoey; Hess-Erga, Ole-Kristian; Larsen, Aud; Thuestad, Gunnar; Tobiesen, August; Hoell, Ingunn Alne

    2015-07-15

    Disinfection of microbes is of importance to prevent the spread of pathogens and non-indigenous species in the environment. Here we test the applicability of using flow cytometry (FCM) to evaluate inactivation of the phytoplankter Tetraselmis suecica after UV irradiation and labeling with the esterase substrate 5-carboxyfluorescein diacetate acetoxymethyl ester (CFDA-AM). Non-irradiated and UV irradiated samples were analyzed with the plate count technique and FCM for 24 days. The numbers of colony forming units were used as a standard to develop a FCM protocol. Our protocol readily distinguishes live and dead cells, but challenges were encountered when determining whether UV damaged cells are dying or repairable. As damaged cells can represent a risk to aquatic organisms and/or humans, this was taken into account when developing the FCM protocol. In spite of the above mentioned challenges we argue that FCM represents an accurate and rapid method to analyze T. suecica samples.

  4. NucleoCounter—An efficient technique for the determination of cell number and viability in animal cell culture processes

    OpenAIRE

    Shah, Dimpalkumar; Naciri, Mariam; Clee, Paul; Al-Rubeai, Mohamed

    2006-01-01

    The NucleoCounter is a novel, portable cell counting device based on the principle of fluorescence microscopy. The present work establishes its use with animal cells and checks its reliability, consistency and accuracy in comparison with other cytometric techniques. The main advantages of this technique are its ability to handle a large number of samples with a high degree of precision and its simplicity and specificity in detecting viable cells quantitatively in a heterogeneous culture. The ...

  5. NucleoCounter—An efficient technique for the determination of cell number and viability in animal cell culture processes

    OpenAIRE

    Shah, Dimpalkumar; Naciri, Mariam; Clee, Paul; Al-Rubeai, Mohamed

    2006-01-01

    The NucleoCounter is a novel, portable cell counting device based on the principle of fluorescence microscopy. The present work establishes its use with animal cells and checks its reliability, consistency and accuracy in comparison with other cytometric techniques. The main advantages of this technique are its ability to handle a large number of samples with a high degree of precision and its simplicity and specificity in detecting viable cells quantitatively in a heterogeneous culture. The ...

  6. Flow cytometric measurement of the cellular propagation of TDP-43 aggregation.

    Science.gov (United States)

    Zeineddine, Rafaa; Whiten, Daniel R; Farrawell, Natalie E; McAlary, Luke; Hanspal, Maya A; Kumita, Janet R; Wilson, Mark R; Yerbury, Justin J

    2017-05-04

    Amyotrophic lateral sclerosis is a devastating neuromuscular degenerative disease characterized by a focal onset of motor neuron loss, followed by contiguous outward spreading of pathology including TAR DNA-binding protein of 43 kDa (TDP-43) aggregates. Previous work suggests that TDP-43 can move between cells. Here we used a novel flow cytometry technique (FloIT) to analyze TDP-43 inclusions and propagation. When cells were transfected to express either mutant G294A TDP-43 fused to GFP or wild type TDP-43fused to tomato red and then co-cultured, flow cytometry detected intact cells containing both fusion proteins and using FloIT detected an increase in the numbers of inclusions in lysates from cells expressing wild type TDP-43-tomato. Furthermore, in this same model, FloIT analyses detected inclusions containing both fusion proteins. These results imply the transfer of TDP-43 fusion proteins between cells and that this process can increase aggregation of wild-type TDP-43 by a mechanism involving co-aggregation with G294A TDP-43.

  7. Development of a novel flow cytometric approach to evaluate fish sperm chromatin using fixed samples

    Science.gov (United States)

    Jenkins, Jill A.

    2013-01-01

    The integrity of the paternal DNA is essential for the accurate transmission of genetic information, yet fertilization is not inhibited by chromatin breakage. Some methods are available for the sensitive detection of DNA damage and can be applied in studies of environmental toxicology, carcinogenesis, aging, and assisted reproduction techniques in both clinical and experimental settings. Because semen samples obtained from remote locations undergo chromatin damage prior to laboratory assessment, the present study was undertaken to evaluate treatments for effective chromatin staining in the development of a DNA fragmentation assay using fixed milt from yellow perch (Perca flavescens). Similar to the sperm chromatin structure assay (SCSA), susceptibility of nuclear DNA to acid-induced denaturation was measured by flow cytometry (FCM). Use of 10% buffered formalin for milt fixation allowed easier peak discrimination than 4% paraformaldehyde. The effects of time and temperature of incubation in 0.08 N HCl were evaluated in order to determine the ideal conditions for promoting DNA decondensation and making strand breaks more available for staining and detection by FCM. The best results were obtained with incubation at 37°C for 1 minute, followed by cold propidium iodide staining for 30 minutes.

  8. Flow Cytometric Analysis of Leishmania Reactive CD4+/CD8+ Lymphocyte Proliferation in Cutaneous Leishmaniasis

    Directory of Open Access Journals (Sweden)

    H Keshavarz

    2008-12-01

    Full Text Available Background: Determination of the division history of T cells in vitro is helpful in the study of effector mechanisms against infections. Technique described here uses the intracellular fluorescent label carboxyfluorescein diacetate succinimidyl ester (CFSE to monitor the proliferation. Methods: In a cross sectional study, blood samples were collected from 7 volunteers with history of cutaneous leishmania­sis (CL and one healthy control from endemic areas in Isfahan province who referred to the Center for Research and Training in Skin Diseases and Leprosy (CRTSDL, then CD4+/CD8+ lymphocytes and CD14+ monocytes were isolated from peri­pheral blood mononuclear cells (PBMC using mAbs and magnetic nanoparticles. CFSE labeled CD4+ or CD8+ lympho­cytes cultured with autologous monocytes in the presence of PHA, SLA, live Leishmania major or as control with­out sti­mulation. Cells were harvested after 7 days and were analyzed using flow cytometry. Results: Five consecutive divisions were monitored separately. Stimulation of CD4+ or CD8+ lymphocytes from CL sub­jects with SLA showed a significant difference in proliferation comparing with unstimulated cells (P< 0.05. The signifi­cant difference in the percentages of CD4+ cells stimulated with SLA was revealed at different divisions for each subject. In CD8+ lymphocyte, significant stronger stimulation of SLA was evident later in the proliferation process. The mean number of divisions in both CD4+/CD8+ lymphocytes stimulated with SLA was significantly greater than when stimulated with live L. major (P=0.007 / P=0.012, respectively Conclusion: The percentage of divided cells might be calculated separately in each division. The cells remained active following CFSE staining and there is possibility of functional analysis simultaneously.

  9. Flow cytometric assessment of microbial abundance in the near-field area of seawater reverse osmosis concentrate discharge

    KAUST Repository

    Van Der Merwe, Riaan

    2014-06-01

    The discharge of concentrate and other process waters from seawater reverse osmosis (SWRO) plant operations into the marine environment may adversely affect water quality in the near-field area surrounding the outfall. The main concerns are the increase in salt concentration in receiving waters, which results in a density increase and potential water stratification near the outfall, and possible increases in turbidity, e.g., due to the discharge of filter backwash waters. Changes in ambient water quality may affect microbial abundance in the area, for example by hindering the photosynthesis process or disrupting biogenesis. It is widely accepted that marine biodiversity is lower in more extreme conditions, such as high salinity environments. As aquatic microbial communities respond very rapidly to changes in their environment, they can be used as indicators for monitoring ambient water quality. The objective of this study was to assess possible changes in microbial abundance as a result of concentrate discharge into the near-field area (<. 25. m) surrounding the outfall of the King Abdullah University of Science and Technology (KAUST) SWRO plant. Flow cytometric (FCM) analysis was conducted in order to rapidly determine microbial abundance on a single-cell level in 107 samples, taken by diving, from the discharge area, the intake area and two control sites. FCM analysis combined the measurement of distinct scatter of cells and particles, autofluorescence of cyanobacteria and algae, and fluorescence after staining of nucleic acids with SYBR® Green for a total bacterial count. The results indicate that changes in microbial abundance in the near-field area of the KAUST SWRO outfall are minor and appear to be the result of a dilution effect rather than a direct impact of the concentrate discharge. © 2014 Elsevier B.V.

  10. CD33 monoclonal antibody conjugated Au cluster nano-bioprobe for targeted flow-cytometric detection of acute myeloid leukaemia

    Energy Technology Data Exchange (ETDEWEB)

    Retnakumari, Archana; Jayasimhan, Jasusri; Chandran, Parwathy; Menon, Deepthy; Nair, Shantikumar; Mony, Ullas; Koyakutty, Manzoor, E-mail: manzoork@aims.amrita.edu, E-mail: ullasmony@aims.amrita.edu [Amrita Centre for Nanoscience and Molecular Medicine, Amrita Institute of Medical Science, Cochin 682 041 (India)

    2011-07-15

    Protein stabilized gold nanoclusters (Au-NCs) are biocompatible, near-infrared (NIR) emitting nanosystems having a wide range of biomedical applications. Here, we report the development of a Au-NC based targeted fluorescent nano-bioprobe for the flow-cytometric detection of acute myeloid leukaemia (AML) cells. Au-NCs with {approx} 25-28 atoms showing bright red-NIR fluorescence (600-750 nm) and average size of {approx} 0.8 nm were prepared by bovine serum albumin assisted reduction-cum-stabilization in aqueous phase. The protein protected clusters were conjugated with monoclonal antibody against CD33 myeloid antigen, which is overexpressed in {approx} 99.2% of the primitive population of AML cells, as confirmed by immunophenotyping using flow cytometry. Au-NC-CD33 conjugates having average size of {approx} 12 nm retained bright fluorescence over an extended duration of {approx} a year, as the albumin protein protects Au-NCs against degradation. Nanotoxicity studies revealed excellent biocompatibility of Au-NC conjugates, as they showed no adverse effect on the cell viability and inflammatory response. Target specificity of the conjugates for detecting CD33 expressing AML cells (KG1a) in flow cytometry showed specific staining of {approx} 95.4% of leukaemia cells within 1-2 h compared to a non-specific uptake of {approx} 8.2% in human peripheral blood cells (PBMCs) which are CD33{sup low}. The confocal imaging also demonstrated the targeted uptake of CD33 conjugated Au-NCs by leukaemia cells, thus confirming the flow cytometry results. This study demonstrates that novel nano-bioprobes can be developed using protein protected fluorescent nanoclusters of Au for the molecular receptor targeted flow cytometry based detection and imaging of cancer cells.

  11. Histopathologic and Flow-Cytometric Analysis of Neoplastic and Benign “background” Tissue in Breast Carcinoma Resections

    Directory of Open Access Journals (Sweden)

    Daniel W. Visscher

    1998-01-01

    Full Text Available Two-color, multiparametric synthesis phase fraction (SPF analysis of cytokeratin-labeled epithelial cells was flow cytometrically performed on both benign (SPFb and malignant tissue samples (if available, SPFt from 132 mastectomy/lumpectomy specimens. These data were then correlated with clinicopathologic features, including (1 tumor differentiation, (2 the proportion of tumor comprised of duct carcinoma-in situ (DCIS, and (3 the histology of accompanying benign breast tissue, classified by predominant microscopic pattern as intact, normal terminal duct lobular units (NTDLU, 34% of cases, atrophic (AT, 33% of cases, proliferative fibrocystic (PFC, 26% of cases, and non-proliferative fibrocystic (NPFC, 7% of cases. SPFt was inversely correlated with extent of DCIS (DCIS =0 – 20% tumor volume – 12.7% mean SPFt, vs. DCIS >20% tumor volume – 6.4% mean SPFt, p = 0.001. SPFt also correlated with the histology of background benign breast tissue (NTDLU – 14.8% mean SPFt vs. AT – 6.9% mean SPFt vs. PFC – 12.7% mean SPFt, p = 0.05 but it did not correlate with patient age or SPFb (overall mean =0.73%. SPFb was correlated with patient age (>56 yr – 0.59% mean SPFb vs. < yr – 0.84% mean SPFb, p = 0.02, with background histology (NTDLU – 1.1% mean SPFb vs. AT – 0.43% mean SPFb vs. PFC – 0.70% mean SPFb, p < 0.02 and with the grade of the neoplasm (well/moderate – 0.58% mean vs. poorly differentiated – 0.85% mean, p = 0.04. Patients having a background of PFC were significantly older than patients with a background of NTDLU (45.2 yr vs. 60.2 yr, p = 0.01.

  12. Performance of the flow cytometric E-screen assay in screening estrogenicity of pure compounds and environmental samples

    Energy Technology Data Exchange (ETDEWEB)

    Vanparys, Caroline, E-mail: caroline.vanparys@ua.ac.be [Laboratory of Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp (Belgium); Depiereux, Sophie; Nadzialek, Stephanie [Research Unit in Organismal Biology (URBO), University of Namur (FUNDP), Namur (Belgium); Robbens, Johan; Blust, Ronny [Laboratory of Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp (Belgium); Kestemont, Patrick [Research Unit in Organismal Biology (URBO), University of Namur (FUNDP), Namur (Belgium); De Coen, Wim [Laboratory of Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp (Belgium); European Chemicals Agency (ECHA), Helsinki (Finland)

    2010-09-15

    In vitro estrogenicity screens are believed to provide a first prioritization step in hazard characterization of endocrine disrupting chemicals. When applied to complex environmental matrices or mixture samples, they have been indicated valuable in estimating the overall estrogen-mimicking load. In this study, the performance of an adapted format of the classical E-screen or MCF-7 cell proliferation assay was profoundly evaluated to rank pure compounds as well as influents and effluents of sewage treatment plants (STPs) according to estrogenic activity. In this adapted format, flow cytometric cell cycle analysis was used to allow evaluation of the MCF-7 cell proliferative effects after only 24 h of exposure. With an average EC{sub 50} value of 2 pM and CV of 22%, this assay appears as a sensitive and reproducible system for evaluation of estrogenic activity. Moreover, estrogenic responses of 17 pure compounds corresponded well, qualitatively and quantitatively, with other in vitro and in vivo estrogenicity screens, such as the classical E-screen (R{sup 2} = 0.98), the estrogen receptor (ER) binding (R{sup 2} = 0.84) and the ER transcription activation assay (R{sup 2} = 0.87). To evaluate the applicability of this assay for complex samples, influents and effluents of 10 STPs covering different treatment processes, were compared and ranked according to estrogenic removal efficiencies. Activated sludge treatment with phosphorus and nitrogen removal appeared most effective in eliminating estrogenic activity, followed by activated sludge, lagoon and filter bed. This is well in agreement with previous findings based on chemical analysis or biological activity screens. Moreover, ER blocking experiments indicated that cell proliferative responses were mainly ER mediated, illustrating that the complexity of the end point, cell proliferation, compared to other ER screens, does not hamper the interpretation of the results. Therefore, this study, among other E-screen studies

  13. Evaluation of Prognostic Factors Following Flow-Cytometric DNA Analysis after Cytokeratin Labelling: II. Cervical and Endometrial Cancer

    Directory of Open Access Journals (Sweden)

    Pauline Wimberger

    2002-01-01

    Full Text Available In gynecologic oncology valid prognostic factors are necessary to define biologically similar subgroups for analysis of therapeutic efficacy. This study is the first published prospective study concerning prognostic significance of DNA ploidy and S‐phase fraction in cervical and endometrial cancer following enrichment of tumor cells by cytokeratin labelling. Epithelial cells were labeled by FITC‐conjugated cytokeratin antibody (CK 5, 6, 8, and CK 17 prior to flow cytometric cell cycle analysis in 91 specimens of cervical cancer and 73 samples of endometrial cancer. In cervical cancer neither DNA‐ploidy nor S‐phase fraction were relevant prognostic parameters. But CV of the G0G1‐peak showed prognostic relevance in cervical cancer cells, even in multivariate analysis. This interesting observation, however, seems to have no therapeutic consequence due to the small discrimination capacity of CV. In endometrial carcinoma, gross DNA‐aneuploidy (DNA‐index > 1.3 and a high percentage of proliferating cells (>75th percentile were univariate and multivariate highly significant prognostic factors for recurrence‐free survival. Especially DNA‐aneuploidy (DI>1.3 is one of the most important independent molecular biological prognostic factors. While diagnostic curettage we could identify risk patients even preoperatively by determination of the prognostic factors like histologic tumor type, grading, cervical involvement and DNA‐ploidy. Thereby these patients could be treated primarily in an oncologic center. In conclusion, our investigations showed that the determination of DNA‐ploidy should be done in endometrial carcinoma. In cervical cancer no clinical significance for determination of DNA‐parameters was found.

  14. Current International Flow Cytometric Practices for the Detection and Monitoring of Paroxysmal Nocturnal Haemoglobinuria (PNH) clones: A UK NEQAS Survey.

    Science.gov (United States)

    Fletcher, Matthew; Whitby, Liam; Whitby, Alison; Barnett, David

    2016-03-02

    Background Paroxysmal Nocturnal Haemoglobinuria (PNH) is a rare acquired genetic disorder, with an incidence of approximately 1.3 new cases per million population per year. Evidence from the UK National External Quality Assessment Service for Leucocyte Immunophenotyping (UK NEQAS LI) programme suggested major discrepancies on how PNH testing is undertaken. To investigate this we surveyed laboratories in the UK NEQAS LI PNH programme and report here the findings. Method A questionnaire was distributed to all centres registered in UK NEQAS LI flow cytometry programmes (n=1587). Comprising several subsections, it covered the majority of clinical flow cytometric practices. Participants completed a general section and then the subsections relevant to their laboratory repertoire. One subsection contained 34 questions regarding practices in PNH clone detection. Results A total of 105 laboratories returned results for the PNH section; the results demonstrated lack of consensus in all areas of PNH testing. Variation was seen in gating and testing strategies, sensitivity levels and final reporting of test results. Several incorrect practices were highlighted such as inappropriate antibody selection and failure to wash the red blood cells (RBCs) prior to analysis. Conclusion Despite the availability of consensus guidelines there appears to be no agreement in the detection and monitoring of PNH. We found only fourteen centres using methods compatible with the International Clinical Cytometry Society guidelines. Of specific note we found that no two laboratories used the same method. This technical variation could lead to incorrect diagnoses, highlighting the need for better adoption and understanding of consensus practices. This article is protected by copyright. All rights reserved.

  15. Antibody affinity maturation through combining display of two-chain paired antibody and precision flow cytometric sorting.

    Science.gov (United States)

    Sun, Shuang; Yang, Xiao; Wang, Haifeng; Zhao, Yun; Lin, Yan; Ye, Chen; Fang, Xiangdong; Hang, Haiying

    2016-07-01

    Recombination of antibody light and heavy chain libraries greatly increases the size of a two-chain paired antibody library, thus easing the construction of large antibody libraries. Here, light and heavy chain variable domains paired by a coiled coil were applied to a bacterial inner membrane display system. However, the probability of the correct pairing of light and heavy chains through random recombination after each round of flow cytometric sorting and cloning was very low in the presence of mostly unmatched light and heavy chain genes, resulting in inefficient enrichment; a target antibody clone in the ratio of 1:100,000 negative control spheroplasts was unable to be enriched by six rounds of sorting and cloning by a conventional sorting strategy (sorting the top 1 %). By just sorting the top 0.000025 % of spheroplasts, we succeeded in enriching the target antibody clone mixed with negative control spheroplasts in a ratio of 1:10(8) by just one round of sorting and cloning. Furthermore, using this gating strategy, we efficiently enriched for an antibody clone with an affinity slightly better than the parent antibody clone from mixed spheroplasts which were present in the ratio of 1 better affinity clone to 10 parent clones to 10(6) negative control clones after just two rounds of sorting and cloning, suggesting that this gating strategy is highly sensitive in distinguishing between clones with a small difference in affinity and also enriching for clones with a higher affinity. Taken together, the combination of the display of a two-chain paired antibody library and the use of stringent gating has significantly increased the efficiency of the antibody maturation system.

  16. Relevance of Flow Cytometric Auto-Crossmatch to the Post-transplant Course of Kidney Transplant Recipients.

    Science.gov (United States)

    Demir, E; Yeğit, O; Erol, A; Akgül, S U; Çalışkan, B; Bayraktar, A; Çalışkan, Y; Türkmen, A; Savran, F O; Sever, M S

    2017-04-01

    The crossmatch test is essential prior to kidney transplantation (tx) to confirm compatibility between the donor and the recipient. However, its results can be misleading due to "undetectable antibodies" in the recipient's serum. To establish if undetectable autoantibodies are responsible for a positive result, an auto-crossmatch test can be performed. In this study, we aim to determine the long-term prognostic value of auto-flow cytometric auto-crossmatch (FCXM) test on kidney survival in kidney tx recipients. The primary outcome variable was reduced renal function. Secondary endpoints were incidence of biopsy-confirmed chronic antibody-mediated rejection (CAMR) and recurrent glomerulonephritis (GN). There were no differences regarding initial serum creatinine levels between the study and control groups (P = .441). Patients who had positive auto-B FCXM had a significantly reduced renal function compared with the control group (P = .016). Four patients developed biopsy-confirmed CAMR in the study group and 1 patient in the control group (P = .047). Five patients had biopsy-confirmed recurrent GN in the GN study group, and only 1 patient had recurrent GN in the GN control group (P = .026). Kidney transplant recipients with positive auto-FCXM test had significantly reduced renal function and a higher incidence of recurrent GN and CAMR compared with the control group. The findings of this study suggest a potential role of auto-antibody causing positive auto-FCXM test result, meanwhile increasing the risk of CAMR, recurrent GN, and new-onset diabetes after tx. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Flow cytometric gating for spleen monocyte and DC subsets: differences in autoimmune NOD mice and with acute inflammation.

    Science.gov (United States)

    Dong, Matthew B; Rahman, M Jubayer; Tarbell, Kristin V

    2016-05-01

    The role of antigen presenting cells (APCs) in the pathogenesis of autoimmune and other inflammatory diseases is now better understood due to advances in multicolor flow cytometry, gene expression analysis of APC populations, and functional correlation of mouse to human APC populations. A simple but informative nomenclature of conventional and plasmacytoid dendritic cell subsets (cDC1, cDC2, pDC) and monocyte-derived populations incorporates these advances, but accurate subset identification is critical. Ambiguous gating schemes and alterations of cell surface markers in inflammatory condition can make comparing results between studies difficult. Both acute inflammation, such as TLR-ligand stimulation, and chronic inflammation as found in mouse models of autoimmunity can alter DC subset gating. Here, we address these issues using in vivo CpG stimulation as an example of acute inflammation and the non-obese diabetic (NOD) mouse as a model of chronic inflammation.We provide a flow cytometric antibody panel and gating scheme that differentiate 2 monocytic and 3DC subsets in the spleen both at steady state and after CpG stimulation. Using this method, we observed differences in the composition of NOD DCs that have been previously reported, and newly identified increases in the number of NOD monocyte-derived DCs. Finally, we established a protocol for DC phosphoflow to measure the phosphorylation state of intracellular proteins, and use it to confirm functional differences in the identified subsets. Therefore, we present optimized methods for distinguishing monocytic and DC populations with and without inflammation and/or autoimmunity associated with NOD mice.

  18. Flow cytometric assay to assess short-term effects of personal care products on the marine microalga Tetraselmis suecica.

    Science.gov (United States)

    Seoane, Marta; Esperanza, Marta; Rioboo, Carmen; Herrero, Concepción; Cid, Ángeles

    2017-03-01

    Large quantities of personal care products (PCPs) are used daily and many of their chemical ingredients are subsequently released into marine environments. Cultures of the marine microalga Tetraselmis suecica were exposed for 24 h to three emerging compounds included in the main classes of PCPs: the UV filter benzophenone-3 (BP-3), the disinfectant triclosan (TCS) and the fragrance tonalide (AHTN). Concentrations tested, expressed as cellular quota (pg cell(-1)), ranged from 5 to 40 for BP-3, from 2 to 16 for TCS and from 1.2 to 2.4 for AHTN. A small cytometric panel was carried out to evaluate key cytotoxicity biomarkers including inherent cell properties, growth and metabolic activity and cytoplasmic membrane properties. BP-3 caused a significant increase in growth rate, metabolic activity and chlorophyll a fluorescence from 10 pg cell(-1). However, growth and esterase activity decreased in cells exposed to all TCS and AHTN concentrations, except the lowest ones. Also these two compounds provoked a significant swelling of cells, more pronounced in the case of TCS-exposed cells. Although all treated cells remained viable, changes in membrane potential were observed. BP-3 and AHTN caused a significant depolarization of cells from 10 to 1.6 pg cell(-1), respectively; however all TCS concentrations assayed caused a noticeable hyperpolarization of cells. Metabolic activity and cytoplasmic membrane potential were the most sensitive parameters. It can be concluded that the toxicological model used and the toxicological parameters evaluated are suitable to assess the toxicity of these emerging contaminants.

  19. Cytometric analysis of surface molecules of leucocytes and phagocytic activity of granulocytes and monocytes/macrophages in cows with pyometra.

    Science.gov (United States)

    Brodzki, P; Kostro, K; Brodzki, A; Niemczuk, K; Lisiecka, U

    2014-10-01

    Pyometra is a serious problem in dairy cow herds, causing large economic losses due to infertility. The development of pyometra depends mainly on the immunological status of the cow. The aim of the study was a comparative evaluation of selected indicators involving non-specific and specific immunity in cows with pyometra and in cows without inflammation of the uterus. The study was performed in 20 cows, which were divided into two groups: pyometra group and healthy group, each comprising 10 cows, based on the results of cytological and ultrasonographic tests. A flow cytometric analysis was performed for the surface molecules CD4, CD8, CD14, CD21, CD25 and CD4(+) CD25(+) on leucocytes, and the phagocytic activity was determined from granulocytes and monocytes/macrophages in the peripheral blood and uterine washings, respectively. It was demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower in cows with pyometra compared with the healthy group (p < 0.001). Significantly (p ≤ 0.001) lower percentage of CD4(+) , CD14(+) , CD25(+) and CD4(+) CD25(+) phenotype leucocytes was also observed in the peripheral blood of cows from the pyometra group, along with a significantly higher (p < 0.001) percentage of CD8(+) and CD21(+) lymphocytes as compared to the healthy group. The results of work indicate that disfunction of cell immunity coexisting with pyometra may be caused by a bacterial infection and the presence of blocking agents (IL-10), released by the increasing number of CD8(+) lymphocytes what leads to the advanced inflammation of uterus.

  20. Flow Cytometric Quantification of Peripheral Blood Cell β-Adrenergic Receptor Density and Urinary Endothelial Cell-Derived Microparticles in Pulmonary Arterial Hypertension.

    Directory of Open Access Journals (Sweden)

    Jonathan A Rose

    Full Text Available 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.

  1. Simple flow cytometric detection of haemozoin containing leukocytes and erythrocytes for research on diagnosis, immunology and drug sensitivity testing

    Directory of Open Access Journals (Sweden)

    Grobusch Martin P

    2011-03-01

    Full Text Available Abstract Background Malaria pigment (haemozoin, Hz has been the focus of diverse research efforts. However, identification of Hz-containing leukocytes or parasitized erythrocytes is usually based on microscopy, with inherent limitations. Flow cytometric detection of depolarized Side-Scatter is more accurate and its adaptation to common bench top flow cytometers might allow several applications. These can range from the ex-vivo and in-vitro detection and functional analysis of Hz-containing leukocytes to the detection of parasitized Red-Blood-Cells (pRBCs to assess antimalarial activity. Methods A standard benchtop flow cytometer was adapted to detect depolarized Side-Scatter. Synthetic and Plasmodium falciparum Hz were incubated with whole blood and PBMCs to detect Hz-containing leukocytes and CD16 expression on monocytes. C5BL/6 mice were infected with Plasmodium berghei ANKA or P. berghei NK65 and Hz-containing leukocytes were analysed using CD11b and Gr1 expression. Parasitized RBC from infected mice were identified using anti-Ter119 and SYBR green I and were analysed for depolarized Side Scatter. A highly depolarizing RBC population was monitored in an in-vitro culture incubated with chloroquine or quinine. Results A flow cytometer can be easily adapted to detect depolarized Side-Scatter and thus, intracellular Hz. The detection and counting of Hz containing leukocytes in fresh human or mouse blood, as well as in leukocytes from in-vitro experiments was rapid and easy. Analysis of CD14/CD16 and CD11b/Gr1 monocyte expression in human or mouse blood, in a mixed populations of Hz-containing and non-containing monocytes, appears to show distinct patterns in both types of cells. Hz-containing pRBC and different maturation stages could be detected in blood from infected mice. The analysis of a highly depolarizing population that contained mature pRBC allowed to assess the effect of chloroquine and quinine after only 2 and 4 hours, respectively

  2. Innovations in diagnosis and post-therapeutic monitoring of Chagas disease: Simultaneous flow cytometric detection of IgG1 antibodies anti-live amastigote, anti-live trypomastigote, and anti-fixed epimastigote forms of Trypanosoma cruzi.

    Science.gov (United States)

    Alessio, Glaucia Diniz; Côrtes, Denise Fonseca; Machado de Assis, Girley Francisco; Júnior, Policarpo Ademar Sales; Ferro, Eloisa Amália Vieira; Antonelli, Lis Ribeiro do Valle; Teixeira-Carvalho, Andréa; Martins-Filho, Olindo Assis; de Lana, Marta

    2014-11-01

    This study developed a remarkable methodological innovation (FC-ATE) which enables simultaneous detection of antibodies specific to the three evolutive forms of Trypanosoma cruzi: live amastigote (AMA), live trypomastigote (TRYPO), and fixed epimastigote (EPI) using a differential fluorescence staining as low (AMA), intermediate (TRYPO), and high (EPI). An outstanding performance (100%) was observed in the discrimination of the chagasic (CH) and non-chagasic (NCH) patients. In the applicability of FC-ATE in the diagnosis of Chagas disease, 100% of the CH samples presented positivity in the percentage of positive fluorescent parasites (PPFP) for all the three forms of T. cruzi. Moreover, 94% of the samples of NCH presented negative values of PPFP with AMA and TRYPO, and 88% with EPI. Samples from the NCH group with false-positive results were those belonging to the leishmaniasis patients. Considering the applicability of this technique in post-therapeutic monitoring of Chagas disease, 100% of non-treated (NT) and treated non-cured (TNC) samples were positive with the three T. cruzi evolutive forms, while a percentage of 100% from samples of the treated cured (TC) patients were negative with AMA, 93% with TRYPO and 96% with EPI. The comparison between FC-ATE and two other flow cytometric tests using the same samples of patients NT, TNC and TC showed that the three techniques presented different reactivities, although categorical correlation between the methodologies was observed. Taken together, the results obtained with the novel FC-ATE method have shown an outstanding performance in the diagnosis and post-therapeutic monitoring of Chagas disease. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Adaptation of ubiquitin-PNA based sperm quality assay for semen evaluation by a conventional flow cytometer and a dedicated platform for flow cytometric semen analysis.

    Science.gov (United States)

    Odhiambo, J F; Sutovsky, M; DeJarnette, J M; Marshall, C; Sutovsky, P

    2011-10-01

    The purpose of semen quality evaluation is to predict the fertility potential of the sample in an objective, rapid and inexpensive manner. However, utilization of sperm quality biomarkers such as ubiquitin and lectin Arachis hypogaea agglutinin (PNA) for flow cytometric semen evaluation might eliminate the need for visual assessment by microscopy. Herein, we demonstrate a robust ubiquitin and PNA-based semen evaluation conducted on a simple, easy to operate, dedicated sperm flow cytometer, EasyCyte Plus (IMV Technologies, L'Aigle, France). Semen samples were collected periodically from two dairy bulls, which were subjected to temporary scrotal insults to induce variable semen quality. Samples were labeled with fluorescently-conjugated anti-ubiquitin antibodies (bind exclusively to the surface of defective sperm) and lectin PNA (binds to acrosomal surface in prematurely capacitated and acrosome-damaged sperm). Fluorescent properties of the samples were measured with a conventional flow cytometer (Becton Dickinson FACScan; Becton Dickinson Corp., Franklin Lakes, NJ, USA) and by the EasyCyte (IMV Technologies) instrument. Data from the two flow cytometers were positively correlated for the percentage of PNA-positive sperm with a damaged acrosome (r = 0.47; P flow cytometric semen evaluation.

  4. Dose-dependent effect of 17 beta-estradiol determined by growth curves and flow cytometric DNA analysis of a human breast carcinoma (T61) grown in nude mice

    DEFF Research Database (Denmark)

    Brünner, N; Spang-Thomsen, M; Vindeløv, L

    1985-01-01

    of the treatment was evaluated using growth curves and flow cytometric DNA analysis. The treatment induced a dose-dependent growth delay and dose-dependent changes in the cell cycle distribution. The cell cycle changes comprised a decrease in the G1 phase, an accumulation of cells in the S phase, and an increasing...

  5. Flow cytometric method for in situ preparation of standard materials of a small defined number of microbial cells with colony-forming potentiality.

    Science.gov (United States)

    Matsuoka, Hideaki; Nakano, Koichiro; Takatani, Norimasa; Yoshida, Tomonori; Igimi, Shizunobu; Saito, Mikako

    2014-01-01

    Standard materials of a small defined number of cells with colony-forming potentiality are essential for the rational validation of food microbiological methods. An in situ flow cytometric method using viable staining with 6-carboxyfluorescein diacetate (CFDA) and tryptic soy agar (TSA) was previously proposed and its feasibility was demonstrated with five strains. In this study, this method was applied to 16 strains to support its broad applicability. The cell sorting gate was previously determined based on the CFDA stainability alone. Now the structural properties of cells designated by forward and side-scattering intensities have been introduced as the second gating criteria. Under the optimum gate condition, 100 cells have been selected and sorted on TSA. Consequently, a 95% or higher colony-forming rate has been attained for every strain. A successful application to microaerophilic Campylobacter spp. is especially of great importance because it suggests further broader applicability.

  6. Detection of P-glycoprotein with a rapid flow cytometric functional assay using Fluo-3: evaluation of sensitivity, specificity and feasibility in multiparametric analysis.

    Science.gov (United States)

    Van Acker, K L; De Greef, C; Eggermont, J; Zhang, P; Vandenberghe, P; Boogaerts, M A

    1995-08-01

    The specificity and sensitivity of a flow cytometric assay simultaneously measuring expression and transport function of the multidrug resistance associated P-glycoprotein (Pgp) was evaluated. The monoclonal antibody (mAb), MRK16 was used to detect phenotypic Pgp expression while Fluo-3-AM was used as a fluorescent substrate in a Pgp functional transport assay. The specificity of the functional assay was examined in two vinblastine selected human leukemic cell lines (K562/VLB2.5 and CCRF-CEM/VLB50) with acquired Pgp overexpression. Downmodulation of Pgp function in these cell lines could be demonstrated with different substances (verapamil, vinblastine, trifluoperazine, cyclosporin A, progesterone and quinidine) and was proven to be consistently higher in the vinblastine selected cells than in their non-selected drug sensitive counterparts. Unexpectedly, modulator activity was also observed in drug sensitive K562 and CCRF-CEM cell lines despite the inability to detect Pgp in those cells by MRK16 flow cytometrically. Low level expression of the MDR1 gene encoding Pgp in sensitive K562 cells was however demonstrated with a sensitive RT-PCR procedure. The small effect of Pgp modulators in non-drug selected cells could therefore be attributed to low level basal expression of Pgp and illustrates the sensitivity of the functional assay. Also, the effect of various Pgp modulators on Pgp function was more pronounced in a subpopulation of Pgp expressing lymphocytes than in lymphocytes which did not express Pgp. Finally, a correlation was found between discrete variations in Pgp expression and Pgp function of CD4+ lymphocytes, underscoring the feasibility of the functional assay in a triple parametric procedure. The triple parametric assay holds promise to detect Pgp expression and function in clinical samples containing mixtures of malignant and non-malignant cells.

  7. Flow cytometric 96-well microplate-based in vitro micronucleus assay with human TK6 cells: protocol optimization and transferability assessment.

    Science.gov (United States)

    Bryce, Steven M; Avlasevich, Svetlana L; Bemis, Jeffrey C; Tate, Matthew; Walmsley, Richard M; Saad, Frédéric; Van Dijck, Kris; De Boeck, Marlies; Van Goethem, Freddy; Lukamowicz-Rajska, Magdalena; Elhajouji, Azeddine; Dertinger, Stephen D

    2013-04-01

    An automated approach for scoring in vitro micronuclei (MN) has been described in which flow cytometric analysis is combined with compound exposure, processing, and sampling in a single 96-well plate (Bryce SM et al. [2010]: Mutat Res 703:191-199). The current report describes protocol optimization and an interlaboratory assessment of the assay's transferability and reproducibility. In a training phase, the methodology was refined and collaborating laboratories were qualified by repeatedly testing three compounds. Second, a set of 32 chemicals comprised of reference genotoxicants and presumed non-genotoxicants was tested at each of four sites. TK6 cells were exposed to 10 closely spaced compound concentrations for 1.5- to 2-cell population doublings, and were then stained and lysed for flow cytometric analysis. MN frequencies were determined by evaluating ≥ 5,000 cells per replicate well, and several indices of cytotoxicity were acquired. The prevalence of positive results varied according to the MN-fold increase used to signify a genotoxic result, as well as the endpoint used to define a cytotoxicity limit. By varying these parameters, assay sensitivity and specificity values ranged from 82 to 98%, and 86 to 97%, respectively. In a third phase, one laboratory tested a further six genotoxicants and five non-genotoxic apoptosis inducers. In these experiments assay specificity was markedly improved when top concentration selection was based on two cytotoxicity endpoints-relative survival and quantification of ethidium monoazide-positive events. Collectively, the results indicate that the miniaturized assay is transferable across laboratories. The 96-well format consumes considerably less compound than conventional in vitro MN test methods, and the high information content provided by flow cytometry helps guard against irrelevant positive results arising from overt toxicity.

  8. Flow cytometric analysis with a fluorescently labeled formyl peptide receptor ligand as a new method to study the pharmacological profile of the histamine H2 receptor.

    Science.gov (United States)

    Werner, Kristin; Kälble, Solveig; Wolter, Sabine; Schneider, Erich H; Buschauer, Armin; Neumann, Detlef; Seifert, Roland

    2015-10-01

    The histamine H2 receptor (H2R) is a Gs protein-coupled receptor. Its activation leads to increases in the second messenger adenosine-3',5'-cyclic monophosphate (cAMP). Presently, several systems are established to characterize the pharmacological profile of the H2R, mostly requiring radioactive material, animal models, or human blood cells. This prompted us to establish a flow cytometric analysis with a fluorescently labeled formyl peptide receptor (FPR) ligand in order to investigate the H2R functionally and pharmacologically. First, we stimulated U937 promonocytes, which mature in a cAMP-dependent fashion upon H2R activation, with histamine (HA) or selective H2R agonists and measured increases in cAMP concentrations by mass spectrometry. Next, indicative for the maturation of U937 promonocytes, we assessed the FPR expression upon incubation with HA or H2R agonists. FPR expression was measured either indirectly by formyl peptide-induced changes in intracellular calcium concentrations ([Ca(2+)]i) or directly with the fluorescein-labeled FPR ligand fNleLFNleYK-Fl. HA and H2R agonists concentration-dependently induced FPR expression, and potencies and efficacies of fMLP-induced increases in [Ca(2+)]i and FPR density correlated linearly. Accordingly, flow cytometric analysis of FPR expression constitutes a simple, inexpensive, sensitive, and reliable method to characterize the H2R pharmacologically. Furthermore, we evaluated FPR expression at the mRNA level. Generally, quantitative real-time polymerase chain reaction confirmed functional data. Additionally, our study supports the concept of functional selectivity of the H2R, since we observed dissociations in the efficacies of HA and H2R agonists in cAMP accumulation and FPR expression.

  9. Detection of chromosome aneuploidy in breast lesions with fluorescence in situ hybridization: Comparison of whole nuclei to thin tissue sections and correlation with flow cytometric DNA analysis

    Energy Technology Data Exchange (ETDEWEB)

    Visscher, D.W.; Wallis, T.; Ritchie, C.A. [Wayne State Univ., Detroit, MI (United States)

    1995-09-01

    We compared flow-cytometric DNA histogram pattern to counts of 4 fluorescent-labelled centromeric probes (chromosomes 1, 7, 8, and 17) in whole nuclei (WN) and in nuclei from formalin-fixed deparaffinized thin tissue section (TS) in 25 breast lesions. In benign lesions, signal gains (i.e., trisomic nuclei) were never observed in greater than 10% of nuclei from either WN or TS preparations. Loss of signal in benign breast lesions, however, varied considerably (0-43%) between individual case and between chromosome probes. The mean incidence of signal loss in WN of benign lesions ranged from 8.9% (chromosome 7) to 14.4 % (chromosome 1) of nuclei. These signal loss frequencies exceeded those of benign lymphoid control cells. In three benign lesions, signal loss in WN (with one probe) was observed in at least 25% of nuclei. Signal losses in benign TS, on average, were 50-150% greater than in matched WN preparations (chromosome 1: 21.7%, chromosome 7: 21.5%). Malignant lesions generally, but not always, displayed fewer monosomic nuclei and more trisomic nuclei in compared to TS, compatible with a slicing (i.e., nuclear truncation) artifact. Signal counts in carcinomas correlated well with flow cytometric DNA index; however, they were also characterized by evidence of genetic instability, manifest as signal gains in a subset of nuclei (10-25%) with individual probes in diploid range cases, as well as intratumoral heterogeneity, reflected as discrepancies in probe counts between WN and TS samples. We conclude that signal losses with centromeric probes are largely, but not entirely, explained by nuclear slicing. The minimum signal loss threshold for establishment of monosomy using interphase cytogenetics is thus unclear, even in WN. Signal gains indicative of trisomy, in contrast, are reliably associated with malignancy and may reflect gross DNA aneuploidy as well as genetic instability. 10 refs., 1 fig., 3 tabs.

  10. Flow Cytometric DNA Analysis Using Cytokeratin Labeling for Identification of Tumor Cells in Carcinomas of the Breast and the Female Genital Tract

    Directory of Open Access Journals (Sweden)

    Rainer Kimmig

    2001-01-01

    Full Text Available Flow cytometric assessment of DNA‐ploidy and S‐phase fraction in malignant tumors is compromised by the heterogeneity of cell subpopulations derived from the malignant and surrounding connective tissue, e.g., tumor, stromal and inflammatory cells. To evaluate the effect on quality of DNA cell cycle analysis and determination of DNA ploidy, cytokeratin labeling of epithelial cells was used for tumor cell enrichment in breast, ovarian, cervical and endometrial cancer prior to DNA analysis. In a prospective study, tumor cell subpopulations of 620 malignant tumors were labeled by a FITC‐conjugated cytokeratin antibody (CK 5, 6, CK18 and CK 5, 6, 8 and CK 17, respectively prior to flow cytometric cell cycle analysis. Compared to total cell analysis, detection rate of DNA‐aneuploid tumors following cytokeratin labeling was increased from 62% to 76.5% in breast cancer, from 68% to 77% in ovarian cancer, from 60% to 80% in cervical cancer and from 30% to 53% in endometrial cancer. Predominantly in DNA‐diploid tumors, a significantly improved detection of S‐phase fraction of the tumor cells was shown due to the elimination of contaminating nonproliferating “normal cells”. S‐phase fraction following tumor cell enrichment was increased by 10% (mean following cytokeratin staining in ovarian and endometrial cancer, by 30% in breast cancer and even by 70% in cervical cancer compared to total cell analysis. Thus, diagnostic accuracy of DNA‐analysis was enhanced by cytokeratin labeling of tumor cells for all tumor entities investigated.

  11. Experimental techniques; Techniques experimentales

    Energy Technology Data Exchange (ETDEWEB)

    Roussel-Chomaz, P. [GANIL CNRS/IN2P3, CEA/DSM, 14 - Caen (France)

    2007-07-01

    This lecture presents the experimental techniques, developed in the last 10 or 15 years, in order to perform a new class of experiments with exotic nuclei, where the reactions induced by these nuclei allow to get information on their structure. A brief review of the secondary beams production methods will be given, with some examples of facilities in operation or under project. The important developments performed recently on cryogenic targets will be presented. The different detection systems will be reviewed, both the beam detectors before the targets, and the many kind of detectors necessary to detect all outgoing particles after the reaction: magnetic spectrometer for the heavy fragment, detection systems for the target recoil nucleus, {gamma} detectors. Finally, several typical examples of experiments will be detailed, in order to illustrate the use of each detector either alone, or in coincidence with others. (author)

  12. Establishment of flow cytometric in micronucleus assay in vitro%流式细胞术检测体外微核方法的建立

    Institute of Scientific and Technical Information of China (English)

    欧红梅; 周长慧; 涂宏刚; 黄鹏程; 常艳

    2015-01-01

    OBJECTIVE:Establish the flow cytometric 96-well microplate-basedin vitro micronucleus assay in CHO-K1 cells,and explore the possibility of this method for early genetic toxicity screening during drug discovery. MEHTODS:The test included treatment with and without metabolic activation. For the treatment with metabolic activation,CHO-K1 cells were treated with three different concentrations of cyclophosphamide in the S9 mixmedium for 4 h,then incubated with S9-free fresh medium for 20 h. For the treatment without metabolic activation,cells were incubated with three different concentrations of mitomycin C continuously for 24 h. In all cases,after a total of 24 h since initiation of the treatment,cells were processed for microscopic scoring or flow cytometric MN analysis. A flow cytometric method for scoring MN used EMA and SYTOX Green to label the cells in 96-well microplate,and then compared with cytokinesis-block micronucleus assay in cell culture disks based on microscopy.RESULTS:Mitomycin C and cyclophosphamide at different concerntrations caused statistically significant and dose-dependent increasess in micronucleus assay . Non-parametric Spearman's coefficients (rs) is 1.000.CONCLUSION:Similar to literature published,mitomycin C and cyclophosphamide induced positive results in flow cytometric based in vitro micronucleus assay. So the method of flow cytometric 96-well microplate-based in vitro micronucleus assay in CHO-K1 cells was established. The concordance between microscopic scoring and flow cytometricwas good,therefore this method is promising for screening and evaluating genetic toxicity of chemicals.%目的:建立96孔板流式细胞术体外微核自动化检测的方法,并探讨其用于药物早期遗传毒性筛选和遗传毒性评价的可能性。方法:试验分为+S9短时处理组(4 h)和-S9持续处理组(24 h),分别选择3个不同浓度的环磷酰胺和丝裂霉素C处理CHO-K1细胞,24 h后收获细胞。采用EMA和SYTOX Green

  13. Development of a flow cytometric method to analyze subpopulations of bacteria in probiotic products and dairy starters

    NARCIS (Netherlands)

    Bunthof, C.J.; Abee, T.

    2002-01-01

    Flow cytometry (FCM) is a rapid and sensitive technique that can determine cell numbers and measure various physiological characteristics of individual cells by using appropriate fluorescent probes. Previously, we developed an FCM assay with the viability probes carboxyfluorescein diacetate (cFDA) a

  14. Expression of Cyclins A, E and Topoisomerase II α correlates with centrosome amplification and genomic instability and influences the reliability of cytometric S-phase determination

    Directory of Open Access Journals (Sweden)

    Laytragoon-Lewin Nongnit

    2003-07-01

    Full Text Available Abstract Background The progression of normal cells through the cell cycle is meticulously regulated by checkpoints guaranteeing the exact replication of the genome during S-phase and its equal division at mitosis. A prerequisite for this achievement is synchronized DNA-replication and centrosome duplication. In this context the expression of cyclins A and E has been shown to play a principal role. Results Our results demonstrated a correlation between centrosome amplification, cell cycle fidelity and the level of mRNA and protein expression of cyclins A and E during the part of the cell cycle defined as G1-phase by means of DNA content based histogram analysis. It is shown that the normal diploid breast cell line HTB-125, the genomically relatively stable aneuploid breast cancer cell line MCF-7, and the genomically unstable aneuploid breast cancer cell line MDA-231 differ remarkably concerning both mRNA and protein expression of the two cyclins during G1-phase. In MDA-231 cells the expression of e.g. cyclin A mRNA was found to be ten times higher than in MCF-7 cells and about 500 times higher than in HTB-125 cells. Topoisomerase II α showed high mRNA expression in MDA compared to MCF-7 cells, but the difference in protein expression was small. Furthermore, we measured centrosome aberrations in 8.4% of the MDA-231 cells, and in only 1.3% of the more stable aneuploid cell line MCF-7. MDA cells showed 27% more incorporation of BrdU than reflected by S-phase determination with flow cytometric DNA content analysis, whereas these values were found to be of the same size in both HTB-125 and MCF-7 cells. Conclusions Our data indicate that the breast cancer cell lines MCF-7 and MDA-231, although both DNA-aneuploid, differ significantly regarding the degree of cell cycle disturbance and centrosome aberrations, which partly could explain the different genomic stability of the two cell lines. The results also question the reliability of cytometric DNA

  15. A flow cytometric comparison of Indo-1 to fluo-3 and Fura Red excited with low power lasers for detecting Ca(2+) flux.

    Science.gov (United States)

    Bailey, Sheree; Macardle, Peter J

    2006-04-20

    Indo-1 and high-power water-cooled lasers have been the standard for flow cytometric based Ca(2+) flux measurements. With advances in technology and the availability of low-power air-cooled lasers, there is interest in alternative protocols. Here, we have compared Indo-1 with the combination of fluo-3 and Fura Red calcium indicator dyes using low-power air-cooled lasers as the excitation source. The reagents were examined in parallel to detect Ca(2+) flux in peripheral blood T lymphocytes and in a T lymphoblastoid cell line. Ca(2+) flux was detected with a FACSVantage SE equipped with an Omnichrome Series 74 Helium-Cadmium, or a Spectra Physics 177-G1202 Argon ion air-cooled laser. Following determination of optimal loading conditions, Ca(2+) flux was examined in response to membrane receptor stimulation or intracellular Ca(2+) mobilization. Dose dependent Ca(2+) flux to anti-CD3 and thapsigargin was detected with either Indo-1 or with fluo-3 and Fura Red. The profile of the Ca(2+) flux detected by Indo-1 or with fluo-3 and Fura Red appeared similar, with the combination of fluo-3 and Fura Red more sensitive under the particular test conditions. The results clearly demonstrated that Indo-1 could be usefully excited with a low-power air-cooled laser. The alternative use of fluo-3 and Fura Red does not require the availability of a UV capable laser and produced equivalent data.

  16. Flow cytometric analysis of p21 protein expression on irradiated human lymphocytes; Analise por citometria de fluxo da expressao da proteina p21 em linfocitos humanos irradiados

    Energy Technology Data Exchange (ETDEWEB)

    Santos, N.F.G.; Amaral, A., E-mail: neyliane@gmail.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Energia Nuclear. Laboratorio de Modelagem e Biodosimetria Aplicada; Freitas-Silva, R. [Universidade Federal de Pernambuco (UFPE), Garanhuns, PE (Brazil). Departamento de Ciencias Naturais e Exatas; Pereira, V.R.A. [Fundacao Oswaldo Cruz (FIOCRUZ), Recife, PE (Brazil). Centro de Pesquisas Aggeu Magalhaes. Departamento de Imunologia. Lab. de Imunoparasitologia; Tasat, D.R. [Universidad Nacional de General San Martin, Buenos Aires (Argentina). Escuela de Ciencia y Tecnologia. Laboratorio de Biologia Celular del Pulmon

    2013-08-15

    Cell cycle blockage in G1 is a mechanism p21 protein-regulated and coupled to DNA damage response to permit genetic content analysis, damage repair and cell death. Analysis of proteins that participates of this response has progressed with new analytic tools, and data contributes to comprehension of radioinduced molecular events as well as to new approaches on practices that employ ionizing radiation. On this perspective, the aim of this research was to evaluate, by flow cytometry, p21 expression on irradiated human lymphocytes, maintained under different experimental conditions. Peripheral blood samples from 10 healthy subjects were irradiated with doses of 0 (non-irradiated), 1, 2 and 4 Gy. Lymphocytes were processed to analysis on ex vivo (no cultured) condition and after 24; 48 and 72 hours culture, with and without phytohemagglutinin stimulation. p21 protein expression levels were measured by flow cytometry, as percentage values. Results indicate that flow cytometric assay allows detection of changes on p21 expression, since it was detected significant increase on phytohemagglutinin-stimulated samples, for all times, against basal expression (ex vivo). However, it was not observed significant alterations on p21 protein radioinduced levels, for all doses, times and culture conditions analyzed. These results not indicate so p21 protein as bioindicator of ionizing radiation exposure. Nevertheless, data confirmation may to require analysis of a more numerous population. (author)

  17. Fluorescence in situ hybridization and sequential catalysed reporter deposition (2C-FISH for the flow cytometric sorting of freshwater ultramicrobacteria

    Directory of Open Access Journals (Sweden)

    Stefan M Neuenschwander

    2015-03-01

    Full Text Available Flow cytometric sorting is a powerful tool to physically separate cells within mixed microbial communities. If combined with phylogenetic staining (fluorescence in situ hybridization, FISH it allows to specifically sort defined genotypic microbial populations from complex natural samples. However, the targeted enrichment of freshwater ultramicrobacteria, such as members of the LD12 clade of Alphaproteobacteria (SAR11-IIIb, is still challenging. Current FISH protocols, even in combination with signal amplification by catalysed reporter deposition (CARD, are not sufficiently sensitive for the distinction of these bacteria from background noise by flow cytometry, presumably due to their low ribosome content and small cell sizes. We, therefore, modified a CARD based flow sorting protocol with the aim of increasing its sensitivity to a level sufficient for ultramicrobacteria. This was achieved by a second signal amplification step mediated by horseradish peroxidase labelled antibodies targeted to the fluorophores that were previously deposited by CARD-FISH staining. The protocol was tested on samples from an oligo-mesotrophic lake. Ultramicrobacteria affiliated with LD12 Alphaproteobacteria could be successfully sorted to high purity by flow cytometry. The ratios of median fluorescence signal to background ranged around 20, and hybridization rates determined by flow cytometry were comparable to those obtained by fluorescence microscopy. Potential downstream applications of our modified cell staining approach range from the analysis of microdiversity within 16S rRNA-defined populations to that of functional properties, such as the taxon-specific incorporation rates of organic substrates.

  18. Identification of New Rat Bone Marrow-Derived Population of Very Small Stem Cell with Oct-4A and Nanog Expression by Flow Cytometric Platforms

    Directory of Open Access Journals (Sweden)

    Anna Labedz-Maslowska

    2016-01-01

    Full Text Available Very small embryonic-like stem cells (VSELs represent a unique rare population of adult stem cells (SCs sharing several structural, genetic, biochemical, and functional properties with embryonic SCs and have been identified in several adult murine and human tissues. However, rat bone marrow- (BM- derived SCs closely resembling murine or human VSELs have not been described. Thus, we employed multi-instrumental flow cytometric approach including classical and imaging cytometry and we established that newly identified population of nonhematopoietic cells expressing CD106 (VCAM-I antigen contains SCs with very small size, expressing markers of pluripotency (Oct-4A and Nanog on both mRNA and protein levels that indicate VSEL population. Based on our experience in both murine and human VSEL isolation procedures by fluorescence-activated cell sorting (FACS, we also optimized sorting protocol for separation of CD45−/Lin−/CD106+ rat BM-derived VSELs from wild type and eGFP-expressing rats, which are often used as donor animals for cell transplantations in regenerative studies in vivo. Thus, this is a first study identifying multiantigenic phenotype and providing sorting protocols for isolation VSELs from rat BM tissue for further examining of their functional properties in vitro as well as regenerative capacity in distinct in vivo rat models of tissue injury.

  19. Response of Syngonium podophyllum L. ‘White Butterfly’ shoot cultures to alternative media additives and gelling agents, and flow cytometric analysis of regenerants

    Directory of Open Access Journals (Sweden)

    JAIME A. TEIXEIRA DA SILVA

    2015-05-01

    Full Text Available Abstract. Teixeira da Silva JA. 2015. Response of Syngonium podophyllum L. ‘White Butterfly’ shoot cultures to alternative media additives and gelling agents, and flow cytometric analysis of regenerants. Nusantara Bioscience 7: 26-32. Syngonium podophyllum L. (arrowhead vine is a popular leafy indoor pot plant whose tissue culture has been established, primarily through in vitro shoot culture, but several interesting aspects have not yet been explored. In this study, cv. ‘White Butterfly’ was used to investigate the response of shoot formation to alternative gelling agents and media additives. Gellan gum (Gelrite® at 2 g/L resulted in greater leaf production, plantlet fresh weight and higher chlorophyll content (SPAD value than all other gelling agents tested, including agar, Bacto agar, phytagel, oatmeal agar, potato dextrose agar, barley starch and corn starch, when on a basal Hyponex® (NPK = 6.5: 6: 19; 3 g/L medium. Several alternative liquid medium additives tested (low and full fat milk, Coca-Cola®, coffee, Japanese green, Oolong and Darjeeling teas negatively impacted plant growth, stunted roots and decreased chlorophyll content (SPAD value of leaves. Plant growth on medium with refined sucrose or table sugar responded similarly. Poor growth was observed when crude extract from a high rebaudioside-containing stevia (Stevia rebaudiana Bertoni line - an artificial sweetener - was used. Leaf tissue from the control did not show any endopolyploidy but low levels of endopolyploidy (8C were detected in some treatments.

  20. The level of heparin-induced antibodies in correlation with the result of the flow cytometric functional assay in the patients with suspected HIT.

    Science.gov (United States)

    Maličev, Elvira; Maček Kvanka, Marjeta; Klemenc, Polona; Rožman, Primož

    2017-09-13

    Heparin can induce the formation of antibodies against a heparin complex with a platelet factor 4 (PF4), leading to platelet activation and the development of heparin-induced thrombocytopaenia (HIT). Because screening ELISA does not discriminate between platelet activating and non-activating anti-heparin/PF4 antibodies, each positive result is confirmed by an additional functional assay. We analysed 1004 sera of patients with suspected HIT. Optical density (OD) values of ELISA-positive results were correlated with the risk for a positive result with our functional flow cytometric assay. Only 10.7% were ELISA positive and 59.8% of those were positive with the functional assay. The positive functional assay was found in 23.4% of patients with OD2.0. Although our results showed that higher ELISA OD values increasethe possibility of the presence of platelet-activating anti-heparin/PF4 antibodies - , there is no need for improving ELISA cut-off value for positive result. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  1. Identification of microbes from the surfaces of food-processing lines based on the flow cytometric evaluation of cellular metabolic activity combined with cell sorting.

    Science.gov (United States)

    Juzwa, W; Duber, A; Myszka, K; Białas, W; Czaczyk, K

    2016-09-01

    In this study the design of a flow cytometry-based procedure to facilitate the detection of adherent bacteria from food-processing surfaces was evaluated. The measurement of the cellular redox potential (CRP) of microbial cells was combined with cell sorting for the identification of microorganisms. The procedure enhanced live/dead cell discrimination owing to the measurement of the cell physiology. The microbial contamination of the surface of a stainless steel conveyor used to process button mushrooms was evaluated in three independent experiments. The flow cytometry procedure provided a step towards monitoring of contamination and enabled the assessment of microbial food safety hazards by the discrimination of active, mid-active and non-active bacterial sub-populations based on determination of their cellular vitality and subsequently single cell sorting to isolate microbial strains from discriminated sub-populations. There was a significant correlation (r = 0.97; p flow cytometry, despite there being differences in the absolute number of cells detected. The combined approach of flow cytometric CRP measurement and cell sorting allowed an in situ analysis of microbial cell vitality and the identification of species from defined sub-populations, although the identified microbes were limited to culturable cells.

  2. Flow cytometric assessment of circulating platelet and erythrocytes microparticles in young thalassemia major patients: relation to pulmonary hypertension and aortic wall stiffness.

    Science.gov (United States)

    Tantawy, Azza A G; Adly, Amira A M; Ismail, Eman A R; Habeeb, Nevin M

    2013-06-01

    Heart disease is the leading cause of mortality and morbidity in β-thalassemia major (β-TM). Aggregability of abnormal red cells and membrane-derived microparticles (MPs) stemming from activated platelets and erythrocytes are responsible for thrombotic risk. We measured platelet and erythrocyte MPs (PMPs and ErMPs) in 60 young β-TM patients compared with 40 age- and sex-matched healthy controls and assessed their relation to clinicopathological characteristics and aortic elastic properties. Patients were studied stressing on transfusion history, splenectomy, thrombotic events, chelation therapy, hematological and coagulation profiles, flow cytometric measurement of PMPs (CD41b(+) ) and ErMPs (glycophorin A(+) ) as well as echocardiographic assessment of aortic elastic properties. Aortic stiffness index and pulmonary artery pressure were significantly higher, whereas aortic strain and distensibility were lower in TM patients than controls (P 2500 μg/L (P < 0.001). Compliant patients on chelation therapy had lower MPs levels than non-compliant patients (P < 0.001). PMPs and ErMPs were positively correlated to markers of hemolysis, serum ferritin, D-dimer, vWF Ag, and aortic stiffness, whereas negatively correlated to hemoglobin level and aortic distensibility (P < 0.05). We suggest that increased MPs may be implicated in vascular dysfunction, pulmonary hypertension risk, and aortic wall stiffness observed in thalassemia patients. Their quantification could provide utility for early detection of cardiovascular abnormalities and monitoring the biological efficacy of chelation therapy.

  3. Development of flow cytometric protocol for nuclear DNA content estimation and determination of chromosome number in Pongamia pinnata L., a valuable biodiesel plant.

    Science.gov (United States)

    Ramesh, Aadi Moolam; Basak, Supriyo; Choudhury, Rimjhim Roy; Rangan, Latha

    2014-01-01

    The potentiality of Pongamia pinnata L. as a sustainable source of feedstock for the biodiesel industry is dependent on an extensive knowledge of the genome structure of the plant. Flow cytometry, with propidium iodide (PI) as the DNA stain, was used to estimate the nuclear DNA content of P. pinnata, with respect to Zea mays 'CE-777' as standard. The internal and pseudo-internal standardization was followed on account of the inhibitory effect of secondary compounds on PI intercalation. The antioxidants (PVP-40 and β-mercaptoethanol) were added to the nuclear isolation buffer for the reduction of inhibitory effect of P. pinnata cytosol. Nuclear DNA content estimation was done for P. pinnata leaves from different altitudes (37-117 m height from sea level) of Assam. Flow cytometry analysis indicated that the nuclear DNA content of P. pinnata is 2.66 pg with predicted 1C value of 1,300 Mb using Z. mays as standard. Coefficient of variation in flow cytometric analysis was within the limit of 5 % indicating that the results were reliable. Somatic chromosome numbers were counted from root-tip cells and was found to be 2n = 22 corresponding to the diploid level (x = 11). A decreasing trend in the nuclear DNA content was observed for the species of different altitudes.

  4. Flow cytometric analysis of kappa and lambda light chain expression in endoscopic biopsy specimens before the diagnosis of B-cell lymphoma.

    Science.gov (United States)

    Oka, Satoko; Muroi, Kazuo; Sato, Kazuya; Fujiwara, Shin-ichiro; Oh, Iekuni; Matsuyama, Tomohiro; Ohmine, Ken; Suzuki, Takahiro; Ozaki, Katsutoshi; Mori, Masaki; Nagai, Tadashi; Fukushima, Noriyoshi; Fukushima, Noriyoshi; Tanaka, Akira; Ozawa, Keiya

    2012-01-01

    Forty-eight patients with gastrointestinal (GI) tract B-cell lymphoma (BCL) were analyzed retrospectively. The diagnosis was based on the histological examination of specimens obtained by endoscopic biopsy. Before the diagnosis was made, single-color flow cytometry was performed to analyze the expression of light chains and B-cell antigens including CD10 in the specimens. Restricted light chain (RLC) expression, a marker of B-cell clonality, was defined as κ and λ ratios of either more than 3.0 or less than 0.5. The specimens from 30 patients (62.5%) showed RLC expression. No RLC expression or RLC expression not examined was divided into two groups : those showing CD10 positivity in more than 20% of cells (4 patients, 8.3%) and those showing no positivity (14 patients, 29.2%). The cell number analyzed in the latter group was significantly smaller than that in the other two groups. Abnormal karyotypes were found in the specimens from 8 patients (16.7%). These results indicate that the flow cytometric analysis of endoscopic biopsy specimens is useful when BCL is suspected if an adequate number of cells are obtained.

  5. Flow cytometric evaluation of antibiotic effects on viability and mitochondrial function of refrigerated spermatozoa of Nile tilapia

    Science.gov (United States)

    Segovia, M.; Jenkins, J.A.; Paniagua-Chavez, C.; Tiersch, T.R.

    2000-01-01

    Improved techniques for storage and evaluation of fish sperm would enhance breeding programs around the world. The goal of this study was to test the effect of antibiotics on refrigerated sperm from Nile tilapia (Oreochromis niloticus) by use of flow cytometry with 2 dual-staining protocols for objective assessment of sperm quality. Concentrations of 1 x 109 sperm/mL were suspended in Ringer's buffer at 318 mOsmol/kg (pH 8.0). The fluorescent stains Sybr 14 (10 ??M), propidium iodide (2.4 mM), and rhodamine 123 (0.13 ??M) were used to assess cell viability and mitochondrial function. Three concentrations of ampicillin, gentamicin, and an antibiotic/antimycotic solution were added to fresh spermatozoa. Motility estimates and flow cytometry measurements were made daily during 7 d of refrigerated storage (4 ??C). The highest concentrations of gentamicin and antibiotic/antimycotic and all 3 concentrations of ampicillin significantly reduced sperm viability. The highest of each of the 3 antibiotic concentrations significantly reduced mitochondrial function. This study demonstrates that objective sperm quality assessments can be made using flow cytometry and that addition of antibiotics at appropriate concentrations can lengthen refrigerated storage time for tilapia spermatozoa. With minor modifications, these protocols can be adapted for use with sperm from other species and with other tissue types.

  6. Cytometric characterization of circulating tumor cells captured by microfiltration and their correlation to the CellSearch(®) CTC test.

    Science.gov (United States)

    Adams, Daniel L; Stefansson, Steingrimur; Haudenschild, Christian; Martin, Stuart S; Charpentier, Monica; Chumsri, Saranya; Cristofanilli, Massimo; Tang, Cha-Mei; Alpaugh, R Katherine

    2015-02-01

    Recent studies reporting hundreds, to thousands, of circulating tumor cells (CTCs) in the blood of cancer patients have raised questions regarding the prevalence of CTCs, as enumerated by the CellSearch(®) CTC Test. Although CellSearch has been shown to consistently detect clinically relevant CTCs; the ability to only capture EpCAM positive cells has led to speculation that it captures limited subsets of CTCs. In contrast, alternative approaches to CTC isolation are often cited as capturing large numbers of CTCs from patient blood. Not surprisingly the number of cells isolated by alternative approaches show poor correlations when compared to CellSearch, even when accounting for EpCAM presence or absence. In an effort to address this discrepancy, we ran an exploratory method comparison study to characterize and compare the CTC subgroups captured from duplicate blood samples from 30 breast and prostate cancer patients using a microfiltration system (CellSieve™) and CellSearch. We then categorized the CellSieve Cytokeratin(CK)+/CD45-/DAPI+ cells into five morphologically distinct subpopulations for correlative analysis. Like other filtration techniques, CellSieve isolated greater numbers of CK+/CD45- cells than CellSearch. Furthermore, analysis showed low correlation between the total CK+/CD45- cells captured by these two assays, regardless of EpCAM presence. However, subgrouping of CK+/CD45-/DAPI+ cells based on distinct cytokeratin staining patterns and nuclear morphologies elucidated a subpopulation correlative to CellSearch. Using method comparison analyses, we identified a specific CTC morphology which is highly correlative between two distinct capture methods. These data suggests that although various morphologic CTCs with similar phenotypic expressions are present in the blood of cancer patients, the clinically relevant cells isolated by CellSearch can potentially be identified using non-EpCAM dependent isolation. © 2014 The Authors. Published by Wiley

  7. Immune toxicity of TiO₂ under hypoxia in the green-lipped mussel Perna viridis based on flow cytometric analysis of hemocyte parameters.

    Science.gov (United States)

    Wang, Youji; Hu, Menghong; Li, Qiongzhen; Li, Jiale; Lin, Daohui; Lu, Weiqun

    2014-02-01

    The combined effects of DO and TiO2 (mixed rutile/anatase phase, 7/3) on immune responses in Perna viridis were examined. Mussels were exposed to six combinations of oxygen levels (hypoxia: 1.5 mg O2l(-1), normoxia: 6.0 mg O2 l(-1)) and TiO2 concentrations (0, 2.5 mg l(-1) and 10 mg l(-1)) for 216 h. Mussels were sampled after 24h, 48h, 120 h and 216 h, and immune parameters of hemocytes, including mortality, phagocytosis, non-specific esterase, ROS production, lysosomal content and total hemocyte count were investigated using flow cytometric assay. Hemocyte mortality was higher under hypoxia than normoxia, and increased with TiO2 concentrations, but no interaction was found between DO and TiO2. Phagocytosis was reduced under hypoxia and decreased with TiO2 exposure, and the interactive effect between time and TiO2 was observed. The percentage of hemocytes showing non-specific esterase activity was lower under hypoxia, and decreased as TiO2 concentration increased with the significant interactive effect of DO and TiO2. ROS production and lysosomal content were lower under hypoxia and reduced as concentration of TiO2 increased, and interactive effect of DO and TiO2 on ROS was evident. THC was significantly affected by the interactive effect between TiO2 and DO, with higher values under normoxia in the presence of TiO2. The present study demonstrated that immune functions of P. viridis were influenced by both nano-TiO2 and hypoxia with some synergistic effects between the two stressors. This implies that DO has to be considered in the evaluation of the toxicity of nano-materials to bivalves. © 2013.

  8. Equal overall rejection rate in pre-transplant flow-cytometric cross-match negative and positive adult recipients in liver transplantation.

    Science.gov (United States)

    Matinlauri, Irma H; Höckerstedt, Krister A; Isoniemi, Helena M

    2005-10-01

    T cell IgG flow-cytometric cross-matches (FCXM) using 48 stored pre-transplant patient serum samples and 40 stored serum samples collected 3 wk after liver transplantation and frozen spleen cells of cadaveric donors in 48 consecutive liver transplantations were performed retrospectively. T cell IgG FCXM using pre-transplant serum samples was compared with 46 complement-dependent lymphocytotoxic cross-matches (CDCXM) performed at the time of transplantation. Clinical relevance of these tests was evaluated in relation to acute rejection, 1-, 3- and 5-yr graft and patient survival. The incidence of positive FCXM was 33% (16 of 48) and 13% (six of 46) by CDCXM. The median time of acute rejection was 29 d after transplantation in FCXM positive group (range 13-101 d) and 22 d in FCXM negative group (range 7-157 d, NS). Rejection rate was similar in 16 pre-transplant FCXM positive patients (eight of 16, 50%) compared with six pre-transplant CDCXM positive patients (three of six, 50%; NS). Recipients having graft rejection tended to be more often pre-transplant FCXM positive (eight of 21, 38%) than CDCXM positive (three of 21, 14%), but the difference was not significant (p > 0.1). No difference was found in the positive predictive value in relation to acute rejection between positive FCXM and CDCXM (69% vs. 50%; NS). Furthermore there was no correlation between post-transplant positive FCXM and acute rejection. No difference was found between pre-transplant T cell IgG FCXM positive and negative recipients in relation to graft or patient survival. Our findings are supportive for little risk associated with preformed donor-specific antibodies in liver transplantation.

  9. Acquired Techniques

    DEFF Research Database (Denmark)

    Lunde Nielsen, Espen; Halse, Karianne

    2013-01-01

    Acquired Techniques - a Leap into the Archive, at Aarhus School of Architecture. In collaboration with Karianne Halse, James Martin and Mika K. Friis. Following the footsteps of past travelers this is a journey into tools and techniques of the architectural process. The workshop will focus upon...... architectural production as a conglomerate of various analogue and digital methods, and provide the basics, the tips/tricks - and how the tool themselves becomes operational for spatial/thematic investigations. Eventually, this will become a city, exhibition and phamplet inhabited by the (by...

  10. Quality control of flow cytometric immunophenotyping%流式细胞术表型分析的质量控制

    Institute of Scientific and Technical Information of China (English)

    吴丽娟; 许东升

    2011-01-01

    FCM表型分析已经从外周血淋巴细胞的双参数定量测定发展到当今对血液细胞病理学包括骨髓、淋巴结分析的5个或更多参数的定性测定,其中白血病和淋巴瘤的免疫表型分析在血液淋巴系统恶性疾病诊断、分类及病情监测上都是对形态学检验的一种重要补充.FCM 5个或更多参数分析的复杂性以及数据的解释依赖于仪器、试剂、程序的标准化及校准.此外,在美国的流式细胞学实验室还必须保留有关水平测试、样品制备、方法学的准确性、特异性、灵敏度和精密度的相关资料.CLSI和UCCC建议每个流式细胞学实验室需要对定性和定量检测程序进行验证.本文对美国临床流式细胞学实验室FCM检验的相关验证、室内与室间质量控制措施进行介绍.%Flow cytometric immunophenotyping has evolved from two-parameter quantitative measurement of peripheral blood lymphocytes to five-or more parameter qualitative evaluation of bone marrow and lymph node in hematopathology.Leukemia/lymphoma immunophenotyping represents an important addition to histomorphology in the diagnosis,classification and monitoring of hematolymphoid neoplasms.The complexity of five-or more parameter analyses and the interpretation of the data rely on standardization and validation of the instrument,the reagent and the procedure.In addition,clinical flow cytometry laboratories in U.S.A are required to document proficiency testing,sample preparation as well as accuracy,specificity,sensitivity and precision of methodology.CLSI and UCCC recommend that each laboratory should validate its own qualitative and quantitative procedures.This paper introduces the procedures for validation and quality control in a clinical flow cytometry laboratory in the United States.

  11. Flow cytometric evaluation of the contribution of ionic silver to genotoxic potential of nanosilver in human liver HepG2 and colon Caco2 cells.

    Science.gov (United States)

    Sahu, Saura C; Njoroge, Joyce; Bryce, Steven M; Zheng, Jiwen; Ihrie, John

    2016-04-01

    Exposure to nanosilver found in food- and cosmetics-related consumer products is of public concern because of the lack of information about its safety. In this study, two widely used in vitro cell culture models, human liver HepG2 and colon Caco2 cells, and the flow cytometric micronucleus (FCMN) assay were evaluated as tools for rapid predictive screening of the potential genotoxicity of nanosilver. Recently, we reported the genotoxicity of 20 nm nanosilver using these systems. In the current study presented here, we tested the hypothesis that the nanoparticle size and cell types were critical determinants of its genotoxicity. To test this hypothesis, we used the FCMN assay to evaluate the genotoxic potential of 50 nm nanosilver of the same shape, composition, surface charge and obtained from the same commercial source using the same experimental conditions and in vitro models (HepG2 and Caco2) as previously tested for the 20 nm silver. Results of our study show that up to the concentrations tested in these cultured cell test systems, the smaller (20 nm) nanoparticle is genotoxic to both the cell types by inducing micronucleus (MN). However, the larger (50 nm) nanosilver induces MN only in HepG2 cells, but not in Caco2 cells. Also in this study, we evaluated the contribution of ionic silver to the genotoxic potential of nanosilver using silver acetate as the representative ionic silver. The MN frequencies in HepG2 and Caco2 cells exposed to the ionic silver in the concentration range tested are not statistically significant from the control values except at the top concentrations for both the cell types. Therefore, our results indicate that the ionic silver may not contribute to the MN-forming ability of nanosilver in HepG2 and Caco2 cells. Also our results suggest that the HepG2 and Caco2 cell cultures and the FCMN assay are useful tools for rapid predictive screening of a genotoxic potential of food- and cosmetics-related chemicals including nanosilver.

  12. Flow cytometric analysis of the inhibition of human basophil activation by histamine high dilutions – a replication study

    Directory of Open Access Journals (Sweden)

    Chantal Wälchli

    2012-09-01

    perform in order to reduce the possibility of artifacts but was omitted in the former study. Conclusions: Laboratory independent replication of homeopathic basic research experiments is still a challenge. Assuming that the results formerly obtained with this model were not due to systematic errors, the quest identifying the crucial factors for successful reproducibility is open for future research. Keywords: Human basophils; histamine; high dilutions; flow cytometry Reference: [1] Sainte-Laudy J, Belon P. Improvement of flow cytometric analysis of basophil activation inhibition by high histamine dilutions. A novel basophil specific marker: CD 203c. Homeopathy. 2006;95:3-8.

  13. 免疫共沉淀筛选牛卵泡CART相互作用蛋白的研究%Screening Proteins Interacting with Cocaine-and Amphetamine-Regulated Transcript in Bovine Follicles by Co-immunoprecipitation

    Institute of Scientific and Technical Information of China (English)

    李鹏飞; 孟金柱; 刘岩; 黄洋; 陈建伟; 姜晓龙; 曹霞; 姚晓磊; 赵妙妙

    2015-01-01

    旨在研究牛卵泡发育过程中可卡因苯丙胺调节转录肽(Cocaine-and amphetamine-regulated transcript,CART)相互作用蛋白(Protein protein interaction,PPI).选择4头10月龄同期发情的健康母牛,B超检测卵泡达8~12 mm,屠宰并采集、分离卵泡颗粒细胞(Granulosa cells,GCs)后,进行总蛋白提取,兔抗CART一抗沉淀CART及其相互作用蛋白,经Protein G-Agarose磁珠吸附分离,多次洗涤去除盐分和杂蛋白,洗脱蛋白复合体并酶解,LC-ESI-Q-TOF质谱分析(Mass spectrometry,MS)及数据库比对.结果显示:共鉴定蛋白质组分111个;经Cytoscape V3.1.0分析,有81个蛋白符合功能模块,并构建蛋白相互作用网络图;同时对相互作用蛋白进行功能富集性分析,共分为3大类34组:其中分子功能占11.73%,生物学过程占46.93%,细胞组分占41.34%.α2巨球蛋白(A2M)和波形蛋白(VIM)与CART相互作用的相关系数最大;CART与NALP1、SERPINH1和PDIA6具有负调控应答的功能,提示可能参与牛卵泡发育过程的调控.

  14. Translation Techniques

    Directory of Open Access Journals (Sweden)

    Marcia Pinheiro

    2015-05-01

    Full Text Available In this paper, we discuss three translation techniques: literal, cultural, and artistic. Literal translation is a well-known technique, which means that it is quite easy to find sources on the topic. Cultural and artistic translation may be new terms. Whilst cultural translation focuses on matching contexts, artistic translation focuses on matching reactions. Because literal translation matches only words, it is not hard to find situations in which we should not use this technique.  Because artistic translation focuses on reactions, judging the quality of an artistic translation work is one of the most difficult things one can do. We end up having a score of complexity and humanity for each one of the mentioned techniques: Literal translation would be the closest thing we have to the machines world and artistic translation would be the closest thing we have to the purely human world. By creating these classifications and studying the subtleties of each one of them, we are adding degrees of quality to our courses and to translation as a professional field. The main contribution of this paper is then the formalization of such a piece of knowledge. We, however, also lay the foundations for studies of this type.

  15. Experimental Techniques

    CERN Document Server

    Engelfried, J

    1999-01-01

    In this course we will give examples for experimental techniques used in particle physics experiments. After a short introduction, we will discuss applications in silicon microstrip detectors, wire chambers, and single photon detection in Ring Imaging Cherenkov (RICH) counters. A short discussion of the relevant physics processes, mainly different forms of energy loss in matter, is enclosed.

  16. Intensity techniques

    DEFF Research Database (Denmark)

    Jacobsen, Finn

    2008-01-01

    The Handbook of Signal Processing in Acoustics will compile the techniques and applications of signal processing as they are used in the many varied areas of Acoustics. The Handbook will emphasize the interdisciplinary nature of signal processing in acoustics. Each Section of the Handbook...... will present topics on signal processing which are important in a specific area of acoustics. These will be of interest to specialists in these areas because they will be presented from their technical perspective, rather than a generic engineering approach to signal processing. Non-specialists, or specialists...... from different areas, will find the self-contained chapters accessible and will be interested in the similarities and differences between the approaches and techniques used in different areas of acoustics....

  17. Electrochemical Techniques

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Gang; Lin, Yuehe

    2008-07-20

    Sensitive and selective detection techniques are of crucial importance for capillary electrophoresis (CE), microfluidic chips, and other microfluidic systems. Electrochemical detectors have attracted considerable interest for microfluidic systems with features that include high sensitivity, inherent miniaturization of both the detection and control instrumentation, low cost and power demands, and high compatibility with microfabrication technology. The commonly used electrochemical detectors can be classified into three general modes: conductimetry, potentiometry, and amperometry.

  18. Combinatorial techniques

    CERN Document Server

    Sane, Sharad S

    2013-01-01

    This is a basic text on combinatorics that deals with all the three aspects of the discipline: tricks, techniques and theory, and attempts to blend them. The book has several distinctive features. Probability and random variables with their interconnections to permutations are discussed. The theme of parity has been specially included and it covers applications ranging from solving the Nim game to the quadratic reciprocity law. Chapters related to geometry include triangulations and Sperner's theorem, classification of regular polytopes, tilings and an introduction to the Eulcidean Ramsey theory. Material on group actions covers Sylow theory, automorphism groups and a classification of finite subgroups of orthogonal groups. All chapters have a large number of exercises with varying degrees of difficulty, ranging from material suitable for Mathematical Olympiads to research.

  19. Experimental Techniques

    DEFF Research Database (Denmark)

    Wyer, Jean

    2013-01-01

    Gas-phase ion spectroscopy requires specialised apparatus, both when it comes to measuring photon absorption and light emission (fluorescence). The reason is much lower ion densities compared to solution-phase spectroscopy. In this chapter different setups are described, all based on mass spectro...... in data interpretation, and the advantages and disadvantages of the different techniques are clarified. New instrumental developments involving cryo-cooled storage rings, which show great promise for the future, are briefly touched upon.......Gas-phase ion spectroscopy requires specialised apparatus, both when it comes to measuring photon absorption and light emission (fluorescence). The reason is much lower ion densities compared to solution-phase spectroscopy. In this chapter different setups are described, all based on mass...... to circumvent this is discussed based on a chemical approach, namely tagging of ammonium groups by crown ether. Prompt dissociation can sometimes be identified from the total beam depletion differing from that due to statistical dissociation. Special emphasis in this chapter is on the limitations and pitfalls...

  20. Epstein-Barr Virus, Human Papillomavirus, and Flow Cytometric Cell Cycle Kinetics in Nasopharyngeal Carcinoma and Inverted Papilloma among Egyptian Patients

    Directory of Open Access Journals (Sweden)

    S. K. Kassim

    1998-01-01

    Full Text Available It is widely accepted that the Epstein-Barr virus is etiologically associated with the development of nasopharyngeal carcinoma. The human papillomavirus is also associated with inverted papilloma. We used the polymerase chain reaction technique to detect both viruses in both types of tumors. Flow cytometry was also used to study the DNA pattern and proliferative behavior of the tumors in relation to the viruses. EBV was detected in 13/20 (65% of NPC specimens, and in none of IP (n = 10 or control specimens (n = 10. This indicates the contribution of EBV as an etiologic factor in NPC. Five cases of NPC (25% were positive for HPV 16, two of them were EBV positive. Four HPV 16 positive cases were found among cases with inverted papilloma, but none among the control cases. Flow cytometry revealed that all NPC, IP, and control samples were diploid except one aneuploid NPC sample. Proliferative capacity (PC of primary tumors was predictive of tumor recurrence in NPC. Using 13.6% as a cut-off point for PC, we were able to discriminate between high risk and low risk groups with 100% sensitivity and 86% specificity. PC can be used as a baseline prognostic parameter in NPC, making it possible to modify courses of treatment in an attempt to inhibit tumor recurrence.

  1. Lymphocyte profiles in multiple myeloma and monoclonal gammopathy of undetermined significance: flow-cytometric characterization and analysis in a two-dimensional correlation biplot.

    Science.gov (United States)

    Van den Hove, L E; Meeus, P; Derom, A; Demuynck, H; Verhoef, G E; Vandenberghe, P; Boogaerts, M A

    1998-06-01

    The distribution of 27 T-, B-, and natural killer-cell subsets in the peripheral blood of 40 patients with multiple myeloma (MM), ten patients with monoclonal gammopathy of undetermined significance (MGUS), and 40 healthy donors was investigated by means of classical univariate statistics and advanced multivariate data-analytical techniques. The latter approach was used to describe, represent, and analyze lymphocyte subset distribution in a two-dimensional correlation biplot, allowing comparison of complex lymphocyte profiles (i.e., compound lymphocyte subset distributions) of individual subjects rather than isolated subset values of selected patient and/or donor groups. The correlation biplot revealed that, in accordance with the univariate statistics, the MM patients were characterized by marked shifts towards CD8+, CD57+, CD62L-, CD(16+56)+, and HLA-DR+ T cells, suggesting in vivo immune activation. The activation profile was most markedly observed in treated MM patients in the advanced disease stage category. The lymphocyte profiles of MGUS patients were heterogeneous, with approximately half of them located in the swarm of MM patients and the other half in the swarm of healthy donors. Although the univariate statistics revealed significant differences between MGUS patients and healthy donors only within the B-cell compartment, the correlation biplot revealed that two MGUS patients clearly had a typical T-cell activation profile similar to that of the MM patients. One MGUS patient with a T-cell activation profile progressed 13 months later to a stage IA MM and required chemotherapy. A marked lymphocyte profile shift in one MM patient was associated with terminal and aggressive disease transformation. Our study illustrates further the practical use of correlation biplots for the detection of aberrant lymphocyte profiles and/or profile shifts in individual patients.

  2. Stress Management Techniques

    Directory of Open Access Journals (Sweden)

    Czesław Czabała

    2016-05-01

    Of all the stress management techniques people used mostly those included in the categories of “problem solving”, “vicarious gratification” and “distancing”. This points to the use of techniques that require no effort but are not fully effective. It could be so that they have not skills to use other techniques, or are aware that such techniques exist.

  3. Radiologic imaging technique

    Energy Technology Data Exchange (ETDEWEB)

    Bushong, S.C. (Dept. of Radiology, Baylor College of Medicine, Houston, TX (US)); Eastman, T.R. (Agfagavert Inc., Irving, TX (US))

    1990-01-01

    The authors focus on the subject of clinical radiographic technique. Emphasizing correct radiographic technique, it's heavily illustrated with radiographs that demonstrate proper exposure and show what happens when exposure variables are changed. A key feature is a discussion and evaluation of radiographic technique charts. Basic technique charts are provided for every body part examined.

  4. Normal adult ramified microglia separated from other central nervous system macrophages by flow cytometric sorting: Phenotypic differences defined and direct ex vivo antigen presentation to myelin basic protein-reactive CD4{sup +} T cells compared

    Energy Technology Data Exchange (ETDEWEB)

    Ford, A.L.; Goodsall, A.L.; Sedgwick, J.D. [Centenary Institute of Cancer Medicine and Cell Biology, Sydney (Australia)] [and others

    1995-05-01

    Ramified microglia in the adult central nervous system (CNS) are the principal glial element up-regulating MHC class I and II expression in response to inflammatory events or neuronal damage. A proportion of these cells also express MHC class II constitutively in the normal CNS. The role of microglia as APCs for CD4{sup +} cells extravasating into the CNS remains undefined. In this study, using irradiation bone marrow chimeras in CD45-congenic rats, the phenotype CD45{sup low}CD11b/c{sup +} is shown to identify microglial cells specifically within the CNS. Highly purified populations of microglia and nonmicroglial but CNS-associated macrophages (CD45{sup high}CD11b/c{sup +}) have been obtained directly from the adult CNS, by using flow cytometric sorting. Morphologically, freshly isolated microglia vs other CNS macrophages are quite distinct. Of the two populations recovered from the normal CNS, it is the minority CD45{sup high}CD11 b/c{sup +} transitional macrophage population, and not microglia, that is the effective APC for experimental autoimmune encephalomyelitis-inducing CD4{sup +} myelin basic protein (MBP)-reactive T cells. CD45{sup high}CD11b/c{sup +} CNS macrophages also stimulate MBP-reactive T cells without addition of MBP to culture suggesting presentation of endogenous Ag. This is the first study in which microglia vs other CNS macrophages have been analyzed for APC ability directly from the CNS, with substantial cross-contamination between the two populations eliminated. The heterogeneity of these populations in terms of APC function is clearly demonstrated. Evidence is still lacking that adult CNS microglia have the capacity to interact with and stimulate CD4{sup +} T cells to proliferate or secrete IL-2. 60 refs., 6 figs., 1 tab.

  5. Tools & techniques--statistics: propensity score techniques.

    Science.gov (United States)

    da Costa, Bruno R; Gahl, Brigitta; Jüni, Peter

    2014-10-01

    Propensity score (PS) techniques are useful if the number of potential confounding pretreatment variables is large and the number of analysed outcome events is rather small so that conventional multivariable adjustment is hardly feasible. Only pretreatment characteristics should be chosen to derive PS, and only when they are probably associated with outcome. A careful visual inspection of PS will help to identify areas of no or minimal overlap, which suggests residual confounding, and trimming of the data according to the distribution of PS will help to minimise residual confounding. Standardised differences in pretreatment characteristics provide a useful check of the success of the PS technique employed. As with conventional multivariable adjustment, PS techniques cannot account for confounding variables that are not or are only imperfectly measured, and no PS technique is a substitute for an adequately designed randomised trial.

  6. Surface science techniques

    CERN Document Server

    Bracco, Gianangelo

    2013-01-01

    The book describes the experimental techniques employed to study surfaces and interfaces. The emphasis is on the experimental method. Therefore all chapters start with an introduction of the scientific problem, the theory necessary to understand how the technique works and how to understand the results. Descriptions of real experimental setups, experimental results at different systems are given to show both the strength and the limits of the technique. In a final part the new developments and possible extensions of the techniques are presented. The included techniques provide microscopic as well as macroscopic information. They cover most of the techniques used in surface science.

  7. Introduction to perturbation techniques

    CERN Document Server

    Nayfeh, Ali H

    2011-01-01

    Similarities, differences, advantages and limitations of perturbation techniques are pointed out concisely. The techniques are described by means of examples that consist mainly of algebraic and ordinary differential equations. Each chapter contains a number of exercises.

  8. Relaxation Techniques for Health

    Science.gov (United States)

    ... R S T U V W X Y Z Relaxation Techniques for Health Share: On This Page What’s the ... Bottom Line? How much do we know about relaxation techniques? A substantial amount of research has been done ...

  9. Semiconductor Research Experimental Techniques

    CERN Document Server

    Balkan, Naci

    2012-01-01

    The book describes the fundamentals, latest developments and use of key experimental techniques for semiconductor research. It explains the application potential of various analytical methods and discusses the opportunities to apply particular analytical techniques to study novel semiconductor compounds, such as dilute nitride alloys. The emphasis is on the technique rather than on the particular system studied.

  10. Low current beam techniques

    Energy Technology Data Exchange (ETDEWEB)

    Saint, A.; Laird, J.S.; Bardos, R.A.; Legge, G.J.F. [Melbourne Univ., Parkville, VIC (Australia). School of Physics; Nishijima, T.; Sekiguchi, H. [Electrotechnical Laboratory, Tsukuba (Japan).

    1993-12-31

    Since the development of Scanning Transmission Microscopy (STIM) imaging in 1983 many low current beam techniques have been developed for the scanning (ion) microprobe. These include STIM tomography, Ion Beam Induced Current, Ion Beam Micromachining and Microlithography and Ionoluminense. Most of these techniques utilise beam currents of 10{sup -15} A down to single ions controlled by beam switching techniques This paper will discuss some of the low beam current techniques mentioned above, and indicate, some of their recent applications at MARC. A new STIM technique will be introduced that can be used to obtain Z-contrast with STIM resolution. 4 refs., 3 figs.

  11. Identification of Streptococcus pneumoniae serotype 11E, serovariant 11Av and mixed populations by high-resolution magic angle spinning nuclear magnetic resonance (HR-MAS NMR spectroscopy and flow cytometric serotyping assay (FCSA.

    Directory of Open Access Journals (Sweden)

    Romina Camilli

    Full Text Available BACKGROUND: Recent studies have identified Streptococcus pneumoniae serotype 11E and serovariant 11Av among isolates previously typed as 11A by classical serotyping methods. Serotype 11E and serovariant 11Av differ from serotype 11A by having totally or partially inactive wcjE, a gene in cps locus coding for an O-acetyl transferase. Serotype 11E is rare among carriage isolates but common among invasive isolates suggesting that it survives better during invasion. Aim of this work was to investigate the epidemiology of serotype 11A in a pneumococcal collection using a new serotyping approach based on High-Resolution Magic Angle Spinning Nuclear Magnetic Resonance (HR-MAS NMR spectroscopy to distinguish serotypes 11A and 11E. METHODS: A collection of 48 (34 invasive and 14 carriage S. pneumoniae isolates from Italy, previously identified as serotype 11A by the Quellung reaction, were investigated by wcjE sequencing, HR-MAS NMR spectroscopy and the reference flow cytometric serotyping assay (FCSA based on monoclonal antibodies. RESULTS: HR-MAS NMR spectra from serotypes 11A and 11E showed different NMR peaks indicating that HR-MAS NMR could be used to distinguish these serotypes, although HR-MAS NMR could not distinguish serotype 11Av from serotype 11E unambiguously. Thirty-eight isolates were confirmed to be serotype 11A, 8 isolates with a mutated wcjE were serotype 11E, 1 isolate belonged to serovariant 11Av, and 1 isolate was a mixed population 11A/11Av. All 11E isolates were identified among invasive isolates. CONCLUSIONS: We proved that HR-MAS NMR can be of potential use for pneumococcal serotyping. The detection of serotype 11E among invasive isolates in our collection, supports previous epidemiological studies suggesting that mutations in wcjE can represent a mechanism promoting pneumococcal survival during invasion. The discovery of a spectrum of immunochemical diversity within established serotypes should stimulate efforts to develop new

  12. Flow cytometric studies of human osteosarcoma.

    Science.gov (United States)

    Mankin, H J; Gebhardt, M C; Springfield, D S; Litwak, G J; Kusazaki, K; Rosenberg, A E

    1991-09-01

    A number of recent studies have emphasized the potential value of flow cytometry as a "marker" to assess the malignity and therefore to help predict the biologic behavior of neoplasms, including bone tumors. Using propidium iodide and a home-built flow cytometer, the authors have studied the DNA distribution in 95 patients with osteosarcoma and determined the percentage of cells in diploidy, S-phase, tetraploidy, and aneuploidy. Using these values and a derived one, mean DNA concentration, it was possible to demonstrate the extent of the abnormalities observed in this group of neoplasms and show their severity as compared with the normal pattern. When the data are compared against disease-free survival and total survival, correlations were noted that, although weak, suggested that some patterns were predictive of increased risk of metastasis and death. The effect of treatment could also be assessed by evaluating the pattern before and after chemotherapy and correlating these with survival. It seems likely that with some improvement in technology, flow cytometry will be of value in the future in assessing the prognosis for osteosarcoma and predicting whether treatment has been effective.

  13. Flow cytometric detection of aberrant chromosomes

    Energy Technology Data Exchange (ETDEWEB)

    Gray, J.W.; Lucas, J.; Yu, L.C.; Langlois, R.

    1983-05-11

    This report describes the quantification of chromosomal aberrations by flow cytometry. Both homogeneously and heterogeneously occurring chromosome aberrations were studied. Homogeneously occurring aberrations were noted in chromosomes isolated from human colon carcinoma (LoVo) cells, stained with Hoechst 33258 and chromomycin A3 and analyzed using dual beam flow cytometry. The resulting bivariate flow karyotype showed a homogeneously occurring marker chromosome of intermediate size. Heterogeneously occurring aberrations were quantified by slit-scan flow cytometry in chromosomes isolated from control and irradiated Chinese hamster cells and stained with propidium iodide. Heterogeneously occurring dicentric chromosomes were detected by their shapes (two centrometers). The frequencies of such chromosomes estimated by slit-scan flow cytometry correlated well with the frequencies determined by visual microscopy.

  14. Technical discussions II - Flow cytometric analysis

    NARCIS (Netherlands)

    Cunningham, A; Cid, A; Buma, AGJ

    1996-01-01

    In this paper the potencial of flow cytometry as applied to the aquatic life sciences is discussed. The use of flow cytometry for studying the ecotoxicology of phytoplankton was introduced. On the other hand, the new flow cytometer EUROPA was presented. This is a multilaser machine which has been sp

  15. Flow cytometric detection method for DNA samples

    Energy Technology Data Exchange (ETDEWEB)

    Nasarabadi,Shanavaz (Livermore, CA); Langlois, Richard G. (Livermore, CA); Venkateswaran, Kodumudi S. (Round Rock, TX)

    2011-07-05

    Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM.TM. on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA.TM., on the 5' end.

  16. Technical discussions II - Flow cytometric analysis

    NARCIS (Netherlands)

    Cunningham, A; Cid, A; Buma, AGJ

    1996-01-01

    In this paper the potencial of flow cytometry as applied to the aquatic life sciences is discussed. The use of flow cytometry for studying the ecotoxicology of phytoplankton was introduced. On the other hand, the new flow cytometer EUROPA was presented. This is a multilaser machine which has been sp

  17. Clasroom Observation Techniques

    OpenAIRE

    Akbayrak, Burcu

    1999-01-01

    In this article observation techniques as a data collecting tool used in social and educational research are examined and discussed. First the concepts and purposes of the observation technique are explained and later systematic and non-systematic observation techniques are described. Two research projects conducted in the classroom are then summarized. These are ‘ORACLE’ (Observational Research and Classroom Learning Evaluation Project), and ‘PRINDEP’ (Primary Needs Independent Evaluation Pr...

  18. Persian fencing techniques

    Directory of Open Access Journals (Sweden)

    Manouchehr Moshtagh Khorasani

    2012-07-01

    Full Text Available There are numerous manuscripts, poems and stories that describe, specifically and in detail, the different techniques used in Persian swordsmanship. The present article explains the origins and the techniques of Persian swordsmanship. The article also describes the traditional code of conduct for Persian warriors. Additionally, it describes an array of techniques that were deployed in actual combat in Iran’s history. Some of these techniques are represented via the miniatures that are reproduced herein. This is the first article on Persian swordsmanship published in any periodical.

  19. Principles of fluorescence techniques

    CERN Document Server

    2016-01-01

    Fluorescence techniques are being used and applied increasingly in academics and industry. The Principles of Fluorescence Techniques course will outline the basic concepts of fluorescence techniques and the successful utilization of the currently available commercial instrumentation. The course is designed for students who utilize fluorescence techniques and instrumentation and for researchers and industrial scientists who wish to deepen their knowledge of fluorescence applications. Key scientists in the field will deliver theoretical lectures. The lectures will be complemented by the direct utilization of steady-state and lifetime fluorescence instrumentation and confocal microscopy for FLIM and FRET applications provided by leading companies.

  20. UIAGM Ropehandling Techniques.

    Science.gov (United States)

    Cloutier, K. Ross

    The Union Internationale des Associations des Guides de Montagne's (UIAGM) rope handling techniques are intended to form the standard for guiding ropework worldwide. These techniques have become the legal standard for instructional institutions and commercial guiding organizations in UIAGM member countries: Austria, Canada, France, Germany, Great…

  1. Emerging optical nanoscopy techniques

    Directory of Open Access Journals (Sweden)

    Montgomery PC

    2015-09-01

    Full Text Available Paul C Montgomery, Audrey Leong-Hoi Laboratoire des Sciences de l'Ingénieur, de l'Informatique et de l'Imagerie (ICube, Unistra-CNRS, Strasbourg, France Abstract: To face the challenges of modern health care, new imaging techniques with subcellular resolution or detection over wide fields are required. Far field optical nanoscopy presents many new solutions, providing high resolution or detection at high speed. We present a new classification scheme to help appreciate the growing number of optical nanoscopy techniques. We underline an important distinction between superresolution techniques that provide improved resolving power and nanodetection techniques for characterizing unresolved nanostructures. Some of the emerging techniques within these two categories are highlighted with applications in biophysics and medicine. Recent techniques employing wider angle imaging by digital holography and scattering lens microscopy allow superresolution to be achieved for subcellular and even in vivo, imaging without labeling. Nanodetection techniques are divided into four subcategories using contrast, phase, deconvolution, and nanomarkers. Contrast enhancement is illustrated by means of a polarized light-based technique and with strobed phase-contrast microscopy to reveal nanostructures. Very high sensitivity phase measurement using interference microscopy is shown to provide nanometric surface roughness measurement or to reveal internal nanometric structures. Finally, the use of nanomarkers is illustrated with stochastic fluorescence microscopy for mapping intracellular structures. We also present some of the future perspectives of optical nanoscopy. Keywords: microscopy, imaging, superresolution, nanodetection, biophysics, medical imaging

  2. Emerging optical nanoscopy techniques

    Science.gov (United States)

    Montgomery, Paul C; Leong-Hoi, Audrey

    2015-01-01

    To face the challenges of modern health care, new imaging techniques with subcellular resolution or detection over wide fields are required. Far field optical nanoscopy presents many new solutions, providing high resolution or detection at high speed. We present a new classification scheme to help appreciate the growing number of optical nanoscopy techniques. We underline an important distinction between superresolution techniques that provide improved resolving power and nanodetection techniques for characterizing unresolved nanostructures. Some of the emerging techniques within these two categories are highlighted with applications in biophysics and medicine. Recent techniques employing wider angle imaging by digital holography and scattering lens microscopy allow superresolution to be achieved for subcellular and even in vivo, imaging without labeling. Nanodetection techniques are divided into four subcategories using contrast, phase, deconvolution, and nanomarkers. Contrast enhancement is illustrated by means of a polarized light-based technique and with strobed phase-contrast microscopy to reveal nanostructures. Very high sensitivity phase measurement using interference microscopy is shown to provide nanometric surface roughness measurement or to reveal internal nanometric structures. Finally, the use of nanomarkers is illustrated with stochastic fluorescence microscopy for mapping intracellular structures. We also present some of the future perspectives of optical nanoscopy. PMID:26491270

  3. Scientific techniques in Egyptology

    Energy Technology Data Exchange (ETDEWEB)

    Newton, G.W.A.

    1987-04-01

    The paper concerns the contribution made by scientific method to Egyptology, in the areas of dating, provenance and medicine. The dating techniques, the analysis techniques and the use of radiology to determine the lifestyle of ancient Egyptians are all described. (U.K.).

  4. Techniques for Teachers Section

    Science.gov (United States)

    Tait, A., Ed.

    1973-01-01

    Includes a simple technique to demonstrate Millikan's oil drop experiment, an environmental studies experiment to measure dissolved oxygen in water samples, and a technique to demonstrate action-reaction. Science materials described are the Pol-A-Star Tomiscope, Nuffield chemistry film loops, air pucks and pH meters. (JR)

  5. Reliability prediction techniques

    Energy Technology Data Exchange (ETDEWEB)

    Whittaker, B.; Worthington, B.; Lord, J.F.; Pinkard, D.

    1986-01-01

    The paper demonstrates the feasibility of applying reliability assessment techniques to mining equipment. A number of techniques are identified and described and examples of their use in assessing mining equipment are given. These techniques include: reliability prediction; failure analysis; design audit; maintainability; availability and the life cycle costing. Specific conclusions regarding the usefulness of each technique are outlined. The choice of techniques depends upon both the type of equipment being assessed and its stage of development, with numerical prediction best suited for electronic equipment and fault analysis and design audit suited to mechanical equipment. Reliability assessments involve much detailed and time consuming work but it has been demonstrated that the resulting reliability improvements lead to savings in service costs which more than offset the cost of the evaluation.

  6. [New microbiological techniques].

    Science.gov (United States)

    Schubert, S; Wieser, A; Bonkat, G

    2017-06-01

    Microbiological diagnostic procedures have changed rapidly in recent years. This is especially true in the field of molecular diagnostics. Classical culture-based techniques are still the gold standard in many areas; however, they are already complemented by automated and also molecular techniques to guarantee faster and better quality results. The most commonly used techniques include real-time polymerase chain reaction (RT-PCR) based systems and nucleic acid hybridization. These procedures are used most powerfully from direct patient samples or in assays to detect the presence of nonculturable or fastidious organisms. Further techniques such as DNA sequencing are not yet used routinely for urological samples and can be considered experimental. However, in conjunction with dropping prices and further technical developments, these techniques promise to be used much more in the near future. Regarding bacterial identification from culture, mass spectrometry (MALDI-TOF MS) has become the technique of choice in recent years especially in Europe. It has tremendously shortened the time to result. This is now going to be extended to antibiotic susceptibility testing. This is of paramount importance in view of ever rising antimicrobial resistance rates. Techniques described in this review offer a faster and better microbiological diagnosis. Such continuous improvements are critical especially in times of cost pressure and rising antimicrobial resistance rates. It is in our interest to provide the best possible care for patients and in this regard a good and effective communication between the laboratory and the clinician is of vital importance.

  7. GROUP PROFILE Computer Technique

    Directory of Open Access Journals (Sweden)

    Andrey V. Sidorenkov

    2015-01-01

    Full Text Available This article contains a description of the structure, the software and functional capabilities, and the scope and purposes of application of the Group Profile (GP computer technique. This technique rests on a conceptual basis (the microgroup theory, includes 16 new and modified questionnaires, and a unique algorithm, tied to the questionnaires, for identification of informal groups. The GP yields a wide range of data about the group as a whole (47 indices, each informal group (43 indices, and each group member (16 indices. The GP technique can be used to study different types of groups: production (work groups, design teams, military units, etc., academic (school classes, student groups, and sports.

  8. Spectrometric techniques 2

    CERN Document Server

    Vanasse, George A

    2013-01-01

    Spectrometric Techniques, Volume II provides information pertinent to vacuum ultraviolet techniques to complete the demonstration of the diversity of methods available to the spectroscopist interested in the ultraviolet visible and infrared spectral regions. This book discusses the specific aspects of the technique of Fourier transform spectroscopy.Organized into five chapters, this volume begins with an overview of the large number of systematic effects in the recording of an interferogram. This text then examines the design approach for a Fourier transform spectrometer with focus on optics.

  9. Computer techniques for electromagnetics

    CERN Document Server

    Mittra, R

    1973-01-01

    Computer Techniques for Electromagnetics discusses the ways in which computer techniques solve practical problems in electromagnetics. It discusses the impact of the emergence of high-speed computers in the study of electromagnetics. This text provides a brief background on the approaches used by mathematical analysts in solving integral equations. It also demonstrates how to use computer techniques in computing current distribution, radar scattering, and waveguide discontinuities, and inverse scattering. This book will be useful for students looking for a comprehensive text on computer techni

  10. Separation techniques: Chromatography

    Science.gov (United States)

    Coskun, Ozlem

    2016-01-01

    Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis. Proteins can be purified based on characteristics such as size and shape, total charge, hydrophobic groups present on the surface, and binding capacity with the stationary phase. Four separation techniques based on molecular characteristics and interaction type use mechanisms of ion exchange, surface adsorption, partition, and size exclusion. Other chromatography techniques are based on the stationary bed, including column, thin layer, and paper chromatography. Column chromatography is one of the most common methods of protein purification. PMID:28058406

  11. Internal septorhinoplasty technique.

    Science.gov (United States)

    Waite, Peter D

    2012-02-01

    The internal septorhinoplasty was the standard operating procedure until the popularity of the external or open rhinoplasty technique. The internal technique of the rhinoplasty should be in the armamentarium of every cosmetic surgeon. There will always be indications for both internal and external rhinoplasty. The advantages of the internal technique are several. Rhinoplasty should be treatment planned based on the individual diagnosis. With experience, most surgeons find the internal rhinoplasty to be successful and extremely rewarding. A simple internal rhinoplasty will produce a good result and satisfied patients.

  12. Spectrometric techniques 4

    CERN Document Server

    Vanasse, George A

    2013-01-01

    Spectrometric Techniques, Volume IV discusses three widely diversified areas of spectrometric techniques. The book focuses on three spectrometric methods. Chapter 1 discusses the phenomenology and applications of Coherent Anti-Stokes Raman Spectroscopy (CARS), the most commonly used optical technique that exploit the Raman effect. The second chapter is concerned with diffraction gratings and mountings for the Vacuum Ultraviolet Spectral Region. Chapter 3 accounts the uses of mass spectrometry, detectors, types of spectrometers, and ion sources. Physicists and chemists will find the book a go

  13. Approximation techniques for engineers

    CERN Document Server

    Komzsik, Louis

    2006-01-01

    Presenting numerous examples, algorithms, and industrial applications, Approximation Techniques for Engineers is your complete guide to the major techniques used in modern engineering practice. Whether you need approximations for discrete data of continuous functions, or you''re looking for approximate solutions to engineering problems, everything you need is nestled between the covers of this book. Now you can benefit from Louis Komzsik''s years of industrial experience to gain a working knowledge of a vast array of approximation techniques through this complete and self-contained resource.

  14. Spectrometric techniques 3

    CERN Document Server

    Vanasse, George A

    2013-01-01

    Spectrometric Techniques, Volume III presents the applications of spectrometric techniques to atmospheric and space studies. This book reviews the spectral data processing and analysis techniques that are of broad applicability.Organized into five chapters, this volume begins with an overview of the instrumentation used for obtaining field data. This text then reviews the contribution that space-borne spectroscopy in the thermal IR has made to the understanding of the planets. Other chapters consider the instruments that have recorded the planetary emission spectra. This book discusses as well

  15. Flow cytometric analysis of peripheral blood leukemic cells in relapse of acute leukemia%急性白血病复发外周血白血病细胞的流式细胞术检验分析

    Institute of Scientific and Technical Information of China (English)

    杨莉; 何浩明

    2015-01-01

    Objective To analyse status of peripheral blood leukemic cells detected by flow cytometry in patients with acute leu‐kemia(AL) ,and to provide references for evaluating clinical efficacy and prognosis of AL .Methods The peripheral blood specimens of 87 cases of patients with AL ,including 53 cases of patients with acute myelocytic leukemia and 34 cases of patients with acute lymphoblastic leukemia ,were detected by using flow cytometry ,morphological changes in bone marrow cells were detected ,as well . Results The sensitivity ,specificity and positive predictive value in determination of acute myelocytic leukemia was 95 .6% ,34 .5%and 81 .3% respectively ,and those in acute lymphoblastic leukemia was 87 .3% ,45 .6% and 68 .9% respectively ,statistically signif‐icant differences were found in sensitivity ,specificity and positive predictive value (P<0 .05) .A total of 19 cases with negative mini‐mal residual disease had recurrence(26 .31% ) after 24 months ,and 68 cases with positive minimal residual disease had recurrence (86 .76% ) after 24 months ,and the recurrence rate between the two groups was statistically significant (P<0 .05) .Among all pa‐tients with positive minimal residual disease ,the recurrence rate in patients with high expression level of minimal residual disease (88 .23% ) was higher than that in patients with low expression level of minimal residual disease (47 .09% ) ,and the difference was statistically significant (P<0 .05) .Conclusion Flow cytometric analysis of peripheral blood leukemic cells may has significance for diagnosing relapse of AL and guiding clinical medication .%目的:分析急性白血病(AL)患者外周血白血病细胞流式细胞术检测情况,为分析AL的临床疗效和预后提供参考。方法采用流式细胞术检测87例A L患者(急性髓细胞白血病53例、急性淋巴细胞白血病34例)外周血标本,同时检测骨髓细胞形态学变化。结果急性髓细胞白

  16. Plant tissue culture techniques

    Directory of Open Access Journals (Sweden)

    Rolf Dieter Illg

    1991-01-01

    Full Text Available Plant cell and tissue culture in a simple fashion refers to techniques which utilize either single plant cells, groups of unorganized cells (callus or organized tissues or organs put in culture, under controlled sterile conditions.

  17. Symbolic rewriting techniques

    CERN Document Server

    Weispfenning, Volker; Grabmeier, Johannes

    1998-01-01

    Symbolic rewriting techniques are methods for deriving consequences from systems of equations, and are of great use when investigating the structure of the solutions. Such techniques appear in many important areas of research within computer algebra: • the Knuth-Bendix completion for groups, monoids and general term-rewriting systems, • the Buchberger algorithm for Gröbner bases, • the Ritt-Wu characteristic set method for ordinary differential equations, and • the Riquier-Janet method for partial differential equations. This volume contains invited and contributed papers to the Symbolic Rewriting Techniques workshop, which was held at the Centro Stefano Franscini in Ascona, Switzerland, from April 30 to May 4, 1995. That workshop brought together 40 researchers from various areas of rewriting techniques, the main goal being the investigation of common threads and methods. Following the workshops, each contribution was formally refereed and 14 papers were selected for publication.

  18. Plant tissue culture techniques

    OpenAIRE

    Rolf Dieter Illg

    1991-01-01

    Plant cell and tissue culture in a simple fashion refers to techniques which utilize either single plant cells, groups of unorganized cells (callus) or organized tissues or organs put in culture, under controlled sterile conditions.

  19. Reversible Data Hiding Techniques

    Directory of Open Access Journals (Sweden)

    Dhananjay Yadav

    2012-03-01

    Full Text Available Reversible data hiding is a technique that is used to hide data inside an image. The data is hidden in such a way that the exact or original data is not visible. The hidden data can be retrieved as and when required. There are several methods that are used in reversible data hiding techniques like Watermarking, Lossless embedding and encryption. In this paper we present a review of reversible watermarking techniques and show different methods that are used to get reversible data hiding technique with higher embedding capacity and invisible objects. Watermark need not be hidden. Watermarking can be applied to 1. Images, 2. Text, 3. Audio/video, 4. Software.

  20. RFCM Techniques Chamber Facility

    Data.gov (United States)

    Federal Laboratory Consortium — FUNCTION: Provides the capability to develop radio-frequency countermeasure (RFCM) techniques in a controlled environment from 2.0 to 40.0 GHz. The configuration of...

  1. INTERNAL ENVIRONMENT ANALYSIS TECHNIQUES

    National Research Council Canada - National Science Library

    Caescu Stefan Claudiu; Popescu Andrei; Ploesteanu Mara Gabriela

    2011-01-01

    .... Objectives of the Research The main purpose of the study of the analysis techniques of the internal environment is to provide insight on those aspects that are of strategic importance to the organization...

  2. Microwave Breast Imaging Techniques

    DEFF Research Database (Denmark)

    Zhurbenko, Vitaliy; Rubæk, Tonny

    2010-01-01

    This paper outlines the applicability of microwave radiation for breast cancer detection. Microwave imaging systems are categorized based on their hardware architecture. The advantages and disadvantages of various imaging techniques are discussed. The fundamental tradeoffs are indicated between v...

  3. Renal Tumor Biopsy Technique

    Institute of Scientific and Technical Information of China (English)

    Lei Zhang; Xue-Song Li; Li-Qun Zhou

    2016-01-01

    Objective:To review hot issues and future direction of renal tumor biopsy (RTB) technique.Data Sources:The literature concerning or including RTB technique in English was collected from PubMed published from 1990 to 2015.Study Selection:We included all the relevant articles on RTB technique in English,with no limitation of study design.Results:Computed tomography and ultrasound were usually used for guiding RTB with respective advantages.Core biopsy is more preferred over fine needle aspiration because of superior accuracy.A minimum of two good-quality cores for a single renal tumor is generally accepted.The use of coaxial guide is recommended.For biopsy location,sampling different regions including central and peripheral biopsies are recommended.Conclusion:In spite of some limitations,RTB technique is relatively mature to help optimize the treatment of renal tumors.

  4. Microwave Breast Imaging Techniques

    DEFF Research Database (Denmark)

    Zhurbenko, Vitaliy; Rubæk, Tonny

    2010-01-01

    This paper outlines the applicability of microwave radiation for breast cancer detection. Microwave imaging systems are categorized based on their hardware architecture. The advantages and disadvantages of various imaging techniques are discussed. The fundamental tradeoffs are indicated between...

  5. Relaxation techniques for stress

    Science.gov (United States)

    ... problems such as high blood pressure, stomachaches, headaches, anxiety, and depression. Using relaxation techniques can help you feel calm. These exercises can also help you manage stress and ease the effects of stress on your body.

  6. Optimization techniques in statistics

    CERN Document Server

    Rustagi, Jagdish S

    1994-01-01

    Statistics help guide us to optimal decisions under uncertainty. A large variety of statistical problems are essentially solutions to optimization problems. The mathematical techniques of optimization are fundamentalto statistical theory and practice. In this book, Jagdish Rustagi provides full-spectrum coverage of these methods, ranging from classical optimization and Lagrange multipliers, to numerical techniques using gradients or direct search, to linear, nonlinear, and dynamic programming using the Kuhn-Tucker conditions or the Pontryagin maximal principle. Variational methods and optimiza

  7. Apollo Onboard Navigation Techniques

    Science.gov (United States)

    Interbartolo, Michael

    2009-01-01

    This viewgraph presentation reviews basic navigation concepts, describes coordinate systems and identifies attitude determination techniques including Primary Guidance, Navigation and Control System (PGNCS) IMU management and Command and Service Module Stabilization and Control System/Lunar Module (LM) Abort Guidance System (AGS) attitude management. The presentation also identifies state vector determination techniques, including PGNCS coasting flight navigation, PGNCS powered flight navigation and LM AGS navigation.

  8. Gulliver, medium, technique

    OpenAIRE

    Bullard, Paddy

    2016-01-01

    In the four Parts of Gulliver’s Travels the narrator attends closely to the manual skills, crafts and techniques of the different countries visited and to the materials and instruments by which they are mediated. The patterned, motif-like presentation of these observations and their rich contextual background, historical and literary, indicate their special significance. These references to technique play an important, previously underappreciated roll in Gulliver. They form a thematic connect...

  9. Separation techniques: Chromatography

    OpenAIRE

    Coskun, Ozlem

    2016-01-01

    Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis. Proteins can be purified based on characteristics such as size and shape, total charge, hydrophobic groups present on the surface, and binding capacity with the stationary phase. Four separation techniques based on molecular characteristics and interaction type use mechanisms of ion exchange, surface adsorp...

  10. Techniques for colorectal anastomosis

    Institute of Scientific and Technical Information of China (English)

    Yik-Hong; Ho; Mohamed; Ahmed; Tawfi; k; Ashour

    2010-01-01

    Colorectal anastomotic leak remains one of the most feared post-operative complications, particularly after anterior resection of the rectum with, the shift from abdomino-peritoneal resections to total mesorectal excision and primary anastomosis. The literature fails to demonstrate superiority of stapled over hand-sewn techniques in colorectal anastomosis, regardless of the level of anastomosis, although a high stricture rate was noted in the former technique. Thus, improvements in safety aspects of anastom...

  11. Computer assisted audit techniques

    Directory of Open Access Journals (Sweden)

    Dražen Danić

    2008-12-01

    Full Text Available The purpose of this work is to point to the possibilities of more efficient auditing. In the encirclement of more and more intensive use of computer techniques that help to CAAT all the aims and the volume of auditing do not change when the audit is done in the computer-informatics environment. The computer assisted audit technique (CAATs can improve the efficiency and productivity of audit procedures. In the computerized information system, the CAATs are the ways in which an auditor can use computer to gather or as help in gathering auditing evidence. There are more reasons why the auditors apply computer techniques that help in auditing. Most often, they do it to achieve improvement of auditing efficiency when the data volume is large. It depends on several factors whether the auditors will apply the computer techniques that help auditing and to what degree respectively. If they do it, the most important are the computer knowledge, professional skill, experience of auditors, and availability of computer technique, and adequacy of computer supports, infeasibility of hand tests, efficiency and time limit. Through several examples from practice, we showed the possibilities of ACL as one of the CAAT tools.

  12. Radiation techniques for acromegaly

    Directory of Open Access Journals (Sweden)

    Minniti Giuseppe

    2011-12-01

    Full Text Available Abstract Radiotherapy (RT remains an effective treatment in patients with acromegaly refractory to medical and/or surgical interventions, with durable tumor control and biochemical remission; however, there are still concerns about delayed biochemical effect and potential late toxicity of radiation treatment, especially high rates of hypopituitarism. Stereotactic radiotherapy has been developed as a more accurate technique of irradiation with more precise tumour localization and consequently a reduction in the volume of normal tissue, particularly the brain, irradiated to high radiation doses. Radiation can be delivered in a single fraction by stereotactic radiosurgery (SRS or as fractionated stereotactic radiotherapy (FSRT in which smaller doses are delivered over 5-6 weeks in 25-30 treatments. A review of the recent literature suggests that pituitary irradiation is an effective treatment for acromegaly. Stereotactic techniques for GH-secreting pituitary tumors are discussed with the aim to define the efficacy and potential adverse effects of each of these techniques.

  13. Telescopes and Techniques

    CERN Document Server

    Kitchin, C R

    2013-01-01

    Telescopes and Techniques has proved itself in its first two editions, having become probably one of the most widely used astronomy texts, both for amateur astronomers and astronomy and astrophysics undergraduates. Both earlier editions of the book were widely used for introductory practical astronomy courses in many universities. In this Third Edition the author guides the reader through the mathematics, physics and practical techniques needed to use today's telescopes (from the smaller models to the larger instruments installed in many colleges) and how to find objects in the sky. Most of the physics and engineering involved is described fully and requires little prior knowledge or experience. Both visual and electronic imaging techniques are covered, together with an introduction to how data (measurements) should be processed and analyzed. A simple introduction to radio telescopes is also included. Brief coverage of the more advanced topics of photometry and spectroscopy are included, but mainly to enable ...

  14. Vibrating wire alignment technique

    CERN Document Server

    Xiao-Long, Wang; lei, Wu; Chun-Hua, Li

    2013-01-01

    Vibrating wire alignment technique is a kind of method which through measuring the spatial distribution of magnetic field to do the alignment and it can achieve very high alignment accuracy. Vibrating wire alignment technique can be applied for magnet fiducialization and accelerator straight section components alignment, it is a necessary supplement for conventional alignment method. This article will systematically expound the international research achievements of vibrating wire alignment technique, including vibrating wire model analysis, system frequency calculation, wire sag calculation and the relation between wire amplitude and magnetic induction intensity. On the basis of model analysis this article will introduce the alignment method which based on magnetic field measurement and the alignment method which based on amplitude and phase measurement. Finally, some basic questions will be discussed and the solutions will be given.

  15. Neuronavigation. Principles. Surgical technique.

    Science.gov (United States)

    Ivanov, Marcel; Ciurea, Alexandru Vlad

    2009-01-01

    Neuronavigation and stereotaxy are techniques designed to help neurosurgeons precisely localize different intracerebral pathological processes by using a set of preoperative images (CT, MRI, fMRI, PET, SPECT etc.). The development of computer assisted surgery was possible only after a significant technological progress, especially in the area of informatics and imagistics. The main indications of neuronavigation are represented by the targeting of small and deep intracerebral lesions and choosing the best way to treat them, in order to preserve the neurological function. Stereotaxis also allows lesioning or stimulation of basal ganglia for the treatment of movement disorders. These techniques can bring an important amount of confort both to the patient and to the neurosurgeon. Neuronavigation was introduced in Romania around 2003, in four neurosurgical centers. We present our five-years experience in neuronavigation and describe the main principles and surgical techniques.

  16. Communication Analysis modelling techniques

    CERN Document Server

    España, Sergio; Pastor, Óscar; Ruiz, Marcela

    2012-01-01

    This report describes and illustrates several modelling techniques proposed by Communication Analysis; namely Communicative Event Diagram, Message Structures and Event Specification Templates. The Communicative Event Diagram is a business process modelling technique that adopts a communicational perspective by focusing on communicative interactions when describing the organizational work practice, instead of focusing on physical activities1; at this abstraction level, we refer to business activities as communicative events. Message Structures is a technique based on structured text that allows specifying the messages associated to communicative events. Event Specification Templates are a means to organise the requirements concerning a communicative event. This report can be useful to analysts and business process modellers in general, since, according to our industrial experience, it is possible to apply many Communication Analysis concepts, guidelines and criteria to other business process modelling notation...

  17. Novel Foraminal Expansion Technique

    Science.gov (United States)

    Senturk, Salim; Ciplak, Mert; Oktenoglu, Tunc; Sasani, Mehdi; Egemen, Emrah; Yaman, Onur; Suzer, Tuncer

    2016-01-01

    The technique we describe was developed for cervical foraminal stenosis for cases in which a keyhole foraminotomy would not be effective. Many cervical stenosis cases are so severe that keyhole foraminotomy is not successful. However, the technique outlined in this study provides adequate enlargement of an entire cervical foraminal diameter. This study reports on a novel foraminal expansion technique. Linear drilling was performed in the middle of the facet joint. A small bone graft was placed between the divided lateral masses after distraction. A lateral mass stabilization was performed with screws and rods following the expansion procedure. A cervical foramen was linearly drilled medially to laterally, then expanded with small bone grafts, and a lateral mass instrumentation was added with surgery. The patient was well after the surgery. The novel foraminal expansion is an effective surgical method for severe foraminal stenosis. PMID:27559460

  18. Novel food processing techniques

    Directory of Open Access Journals (Sweden)

    Vesna Lelas

    2006-12-01

    Full Text Available Recently, a lot of investigations have been focused on development of the novel mild food processing techniques with the aim to obtain the high quality food products. It is presumed also that they could substitute some of the traditional processes in the food industry. The investigations are primarily directed to usage of high hydrostatic pressure, ultrasound, tribomechanical micronization, microwaves, pulsed electrical fields. The results of the scientific researches refer to the fact that application of some of these processes in particular food industry can result in lots of benefits. A significant energy savings, shortening of process duration, mild thermal conditions, food products with better sensory characteristics and with higher nutritional values can be achieved. As some of these techniques act also on the molecular level changing the conformation, structure and electrical potential of organic as well as inorganic materials, the improvement of some functional properties of these components may occur. Common characteristics of all of these techniques are treatment at ambient or insignificant higher temperatures and short time of processing (1 to 10 minutes. High hydrostatic pressure applied to various foodstuffs can destroy some microorganisms, successfully modify molecule conformation and consequently improve functional properties of foods. At the same time it acts positively on the food products intend for freezing. Tribomechanical treatment causes micronization of various solid materials that results in nanoparticles and changes in structure and electrical potential of molecules. Therefore, the significant improvement of some rheological and functional properties of materials occurred. Ultrasound treatment proved to be potentially very successful technique of food processing. It can be used as a pretreatment to drying (decreases drying time and improves functional properties of food, as extraction process of various components

  19. Clustering Techniques in Bioinformatics

    Directory of Open Access Journals (Sweden)

    Muhammad Ali Masood

    2015-01-01

    Full Text Available Dealing with data means to group information into a set of categories either in order to learn new artifacts or understand new domains. For this purpose researchers have always looked for the hidden patterns in data that can be defined and compared with other known notions based on the similarity or dissimilarity of their attributes according to well-defined rules. Data mining, having the tools of data classification and data clustering, is one of the most powerful techniques to deal with data in such a manner that it can help researchers identify the required information. As a step forward to address this challenge, experts have utilized clustering techniques as a mean of exploring hidden structure and patterns in underlying data. Improved stability, robustness and accuracy of unsupervised data classification in many fields including pattern recognition, machine learning, information retrieval, image analysis and bioinformatics, clustering has proven itself as a reliable tool. To identify the clusters in datasets algorithm are utilized to partition data set into several groups based on the similarity within a group. There is no specific clustering algorithm, but various algorithms are utilized based on domain of data that constitutes a cluster and the level of efficiency required. Clustering techniques are categorized based upon different approaches. This paper is a survey of few clustering techniques out of many in data mining. For the purpose five of the most common clustering techniques out of many have been discussed. The clustering techniques which have been surveyed are: K-medoids, K-means, Fuzzy C-means, Density-Based Spatial Clustering of Applications with Noise (DBSCAN and Self-Organizing Map (SOM clustering.

  20. Advanced Techniques in Biophysics

    CERN Document Server

    Arrondo, José Luis R

    2006-01-01

    Technical advancements are basic elements in our life. In biophysical studies, new applications and improvements in well-established techniques are being implemented every day. This book deals with advancements produced not only from a technical point of view, but also from new approaches that are being taken in the study of biophysical samples, such as nanotechniques or single-cell measurements. This book constitutes a privileged observatory for reviewing novel applications of biophysical techniques that can help the reader enter an area where the technology is progressing quickly and where a comprehensive explanation is not always to be found.

  1. Simple Driving Techniques

    DEFF Research Database (Denmark)

    Rosendahl, Mads

    2002-01-01

    -like language. Our aim is to extract a simple notion of driving and show that even in this tamed form it has much of the power of more general notions of driving. Our driving technique may be used to simplify functional programs which use function composition and will often be able to remove intermediate data......Driving was introduced as a program transformation technique by Valentin Turchin in some papers around 1980. It was intended for the programming language REFAL and used in metasystem transitions based on super compilation. In this paper we present one version of driving for a more conventional lisp...

  2. Radiographic constant exposure technique

    DEFF Research Database (Denmark)

    Domanus, Joseph Czeslaw

    1985-01-01

    The constant exposure technique has been applied to assess various industrial radiographic systems. Different X-ray films and radiographic papers of two producers were compared. Special attention was given to fast film and paper used with fluorometallic screens. Radiographic image quality...... was tested by the use of ISO wire IQI's and ASTM penetrameters used on Al and Fe test plates. Relative speed and reduction of kilovoltage obtained with the constant exposure technique were calculated. The advantages of fast radiographic systems are pointed out...

  3. Surface science techniques

    CERN Document Server

    Walls, JM

    2013-01-01

    This volume provides a comprehensive and up to the minute review of the techniques used to determine the nature and composition of surfaces. Originally published as a special issue of the Pergamon journal Vacuum, it comprises a carefully edited collection of chapters written by specialists in each of the techniques and includes coverage of the electron and ion spectroscopies, as well as the atom-imaging methods such as the atom probe field ion microscope and the scanning tunnelling microscope. Surface science is an important area of study since the outermost surface layers play a crucial role

  4. Brachytherapy applications and techniques

    CERN Document Server

    Devlin, Phillip M

    2015-01-01

    Written by the foremost experts in the field, this volume is a comprehensive text and practical reference on contemporary brachytherapy. The book provides detailed, site-specific information on applications and techniques of brachytherapy in the head and neck, central nervous system, breast, thorax, gastrointestinal tract, and genitourinary tract, as well as on gynecologic brachytherapy, low dose rate and high dose rate sarcoma brachytherapy, vascular brachytherapy, and pediatric applications. The book thoroughly describes and compares the four major techniques used in brachytherapy-intraca

  5. Modern recording techniques

    CERN Document Server

    Huber, David Miles

    2013-01-01

    As the most popular and authoritative guide to recording Modern Recording Techniques provides everything you need to master the tools and day to day practice of music recording and production. From room acoustics and running a session to mic placement and designing a studio Modern Recording Techniques will give you a really good grounding in the theory and industry practice. Expanded to include the latest digital audio technology the 7th edition now includes sections on podcasting, new surround sound formats and HD and audio.If you are just starting out or looking for a step up

  6. Wireless communications algorithmic techniques

    CERN Document Server

    Vitetta, Giorgio; Colavolpe, Giulio; Pancaldi, Fabrizio; Martin, Philippa A

    2013-01-01

    This book introduces the theoretical elements at the basis of various classes of algorithms commonly employed in the physical layer (and, in part, in MAC layer) of wireless communications systems. It focuses on single user systems, so ignoring multiple access techniques. Moreover, emphasis is put on single-input single-output (SISO) systems, although some relevant topics about multiple-input multiple-output (MIMO) systems are also illustrated.Comprehensive wireless specific guide to algorithmic techniquesProvides a detailed analysis of channel equalization and channel coding for wi

  7. Craniospinal irradiation techniques

    Energy Technology Data Exchange (ETDEWEB)

    Scarlatescu, Ioana, E-mail: scarlatescuioana@gmail.com; Avram, Calin N. [Faculty of Physics, West University of Timisoara, Bd. V. Parvan 4, 300223 Timisoara (Romania); Virag, Vasile [County Hospital “Gavril Curteanu” - Oradea (Romania)

    2015-12-07

    In this paper we present one treatment plan for irradiation cases which involve a complex technique with multiple beams, using the 3D conformational technique. As the main purpose of radiotherapy is to administrate a precise dose into the tumor volume and protect as much as possible all the healthy tissues around it, for a case diagnosed with a primitive neuro ectoderm tumor, we have developed a new treatment plan, by controlling one of the two adjacent fields used at spinal field, in a way that avoids the fields superposition. Therefore, the risk of overdose is reduced by eliminating the field divergence.

  8. Process measuring techniques; Prozessmesstechnik

    Energy Technology Data Exchange (ETDEWEB)

    Freudenberger, A.

    2000-07-01

    This introduction into measurement techniques for chemical and process-technical plant in science and industry describes in detail the methods used to measure basic quantities. Most prominent are modern measuring techniques by means of ultrasound, microwaves and the Coriolis effect. Alongside physical and measuring technique fundamentals, the practical applications of measuring devices are described. Calculation examples are given to illustrate the subject matter. The book addresses students of physical engineering, process engineering and environmental engineering at technical schools as well as engineers of other disciplines wishing to familiarize themselves with the subject of process measurement techniques. (orig.) [German] Diese Einfuehrung in die Messtechnik fuer chemische und verfahrens-technische Forschungs- und Produktionsanlagen beschreibt ausfuehrlich die Methoden zur Messung der Basisgroessen. Moderne Messverfahren mit Ultraschall, Mikrowellen und Coriolis-Effekt stehen dabei im Vordergrund. Beruecksichtigung finden sowohl die physikalischen und messtechnischen Grundlagen als auch die praktischen Anwendungen der Geraete. Berechnungsbeispiele dienen der Erlaeuterung und Vertiefung des Stoffes. Angesprochen sind Studenten der Ingenieurstufengaenge Physikalische Technik und Verfahrens- und Umwelttechnik an Fachhochschulen als auch Ingenieure anderer Fachrichtungen, die sich in das Gebiet der Prozessmesstechnik einarbeiten wollen. (orig.)

  9. Problem Solving Techniques Seminar.

    Science.gov (United States)

    Massachusetts Career Development Inst., Springfield.

    This booklet is one of six texts from a workplace literacy curriculum designed to assist learners in facing the increased demands of the workplace. Six problem-solving techniques are developed in the booklet to assist individuals and groups in making better decisions: problem identification, data gathering, data analysis, solution analysis,…

  10. Imaging techniques in microbiology.

    Science.gov (United States)

    Fung, D C; Theriot, J A

    1998-06-01

    Recent advances in optical imaging have dramatically expanded the capabilities of the light microscope and its usefulness in microbiology research. Some of these advances include improved fluorescent probes, better cameras, new techniques such as confocal and deconvolution microscopy, and the use of computers in imaging and image analysis. These new technologies have now been applied to microbiological problems with resounding success.

  11. Assessing Classroom Assessment Techniques

    Science.gov (United States)

    Simpson-Beck, Victoria

    2011-01-01

    Classroom assessment techniques (CATs) are teaching strategies that provide formative assessments of student learning. It has been argued that the use of CATs enhances and improves student learning. Although the various types of CATs have been extensively documented and qualitatively studied, there appears to be little quantitative research…

  12. Electronic waste recycling techniques

    CERN Document Server

    Bernardes, Andréa

    2015-01-01

    This book presents an overview of the characterization of electronic waste. In addition, processing techniques for the recovery of metals, polymers and ceramics are described. This book serves as a source of information and as an educational technical reference for practicing scientists and engineers, as well as for students.

  13. Techniques for Vocal Health.

    Science.gov (United States)

    Wiest, Lori

    1997-01-01

    Outlines a series of simple yet effective practices, techniques, and tips for improving the singing voice and minimizing stress on the vocal chords. Describes the four components for producing vocal sound: respiration, phonation, resonation, and articulation. Provides exercises for each and lists symptoms of sickness and vocal strain. (MJP)

  14. Clustering Techniques in Bioinformatics

    National Research Council Canada - National Science Library

    Muhammad Ali Masood; M. N. A. Khan

    2015-01-01

    ... according to well-defined rules. Data mining, having the tools of data classification and data clustering, is one of the most powerful techniques to deal with data in such a manner that it can help researchers identify the required information...

  15. Merchandising Techniques and Libraries.

    Science.gov (United States)

    Green, Sylvie A.

    1981-01-01

    Proposes that libraries employ modern booksellers' merchandising techniques to improve circulation of library materials. Using displays in various ways, the methods and reasons for weeding out books, replacing worn book jackets, and selecting new books are discussed. Suggestions for learning how to market and 11 references are provided. (RBF)

  16. Art Appreciation and Technique.

    Science.gov (United States)

    Dean, Diane R.; Milam, Debora

    1985-01-01

    Presents examples of independent study units for gifted high school students in a resource room setting. Both art appreciation and technique are covered in activities concerned with media (basics of pencil, India ink, pastels, crayons, oil, acrylics, and watercolors), subject matter (landscapes, animals, the human figure), design and illustration…

  17. Managerial Techniques in Educational Administration.

    Science.gov (United States)

    Lane, John J.

    1983-01-01

    Management techniques developed during the past 20 years assume the rational bureaucratic model. School administration requires contingent techniques. Quality Circle, Theory Z, and the McKenzie 7-Framework are discussed as techniques to increase school productivity. (MD)

  18. Techniques to Bring Up Mucus

    Science.gov (United States)

    ... COPD: Lifestyle Management Techniques to Bring Up Mucus Techniques to Bring Up Mucus Make an Appointment Refer ... breathing may become difficult, and infection may occur. Techniques to remove mucus are often done after using ...

  19. Exoplanet Detection Techniques

    CERN Document Server

    Fischer, Debra A; Laughlin, Greg P; Macintosh, Bruce; Mahadevan, Suvrath; Sahlmann, Johannes; Yee, Jennifer C

    2015-01-01

    We are still in the early days of exoplanet discovery. Astronomers are beginning to model the atmospheres and interiors of exoplanets and have developed a deeper understanding of processes of planet formation and evolution. However, we have yet to map out the full complexity of multi-planet architectures or to detect Earth analogues around nearby stars. Reaching these ambitious goals will require further improvements in instrumentation and new analysis tools. In this chapter, we provide an overview of five observational techniques that are currently employed in the detection of exoplanets: optical and IR Doppler measurements, transit photometry, direct imaging, microlensing, and astrometry. We provide a basic description of how each of these techniques works and discuss forefront developments that will result in new discoveries. We also highlight the observational limitations and synergies of each method and their connections to future space missions.

  20. Whole cell entrapment techniques.

    Science.gov (United States)

    Trelles, Jorge A; Rivero, Cintia W

    2013-01-01

    Microbial whole cells are efficient, ecological, and low-cost catalysts that have been successfully applied in the pharmaceutical, environmental, and alimentary industries, among others. Microorganism immobilization is a good way to carry out the bioprocess under preparative conditions. The main advantages of this methodology lie in their high operational stability, easy upstream separation and bioprocess scale-up feasibility. Cell entrapment is the most widely used technique for whole cell immobilization. This technique-in which the cells are included within a rigid network-is porous enough to allow the diffusion of substrates and products, protects the selected microorganism from the reaction medium, and has high immobilization efficiency (100 % in most cases).

  1. [Seldinger modified technique].

    Science.gov (United States)

    Ramos Cuenca, Francisco; Linares Escudero, Joaquín; Romo García, Raquel; Cubo Amaya, Manuel; Climent Villanueva, Magdalena; Santos Sarria, Remedios

    2008-12-01

    The authors describe the procedure to insert central blood vessel catheters through peripheral blood vessel catheters using the Seldinger modified technique since critically ill patients service at the Carlos Haya Regional University Hospital in Malaga cares for a high number of patients who need to have canalized a central blood vessel catheter to maximize the treatment they receive. In many cases it is not possible to insert a DRUM type central blood vessel catheter due to an insufficient blood vessel caliber and then the nursing team at this hospital opts to carry out a procedure to canalize a central blood vessel using the Seldinger modified technique. This report was presented at the III National Social-Sanitary Nursing Congress.

  2. Oberst beam test technique

    Science.gov (United States)

    Fasana, Alessandro; Garibaldi, Luigi; Giorcelli, Ermanno; Ruzzene, Massimo

    1998-06-01

    The definition of the mechanical properties of viscoelastic materials, i.e. the elastic modulus and the loss factor, is carried out, according to many national and international standards, with many different techniques, both of the resonant and non-resonant type. In this paper we focus our attention on the pros and cons of the resonant technique based on the classical Oberst beam method. When the damping material to be tested is not self-supporting, its properties are determined taking start from the measured modal frequencies and loss factors of a laminated beam, constituted by one or two metallic strips, ideally undamped, and one or two viscoelastic layers. The formulae specified on the standards hold valid under the assumptions of the theory developed by Kerwin, Ungar and Ross and we try in this paper to quantify witch deviation of the results should be expected when moving away from their ideal hypotheses.

  3. [Molecular techniques in mycology].

    Science.gov (United States)

    Rodríguez-Tudela, Juan Luis; Cuesta, Isabel; Gómez-López, Alicia; Alastruey-Izquierdo, Ana; Bernal-Martínez, Leticia; Cuenca-Estrella, Manuel

    2008-11-01

    An increasing number of molecular techniques for the diagnosis of fungal infections have been developed in the last few years, due to the growing prevalence of mycoses and the length of time required for diagnosis when classical microbiological methods are used. These methods are designed to resolve the following aspects of mycological diagnosis: a) Identification of fungi to species level by means of sequencing relevant taxonomic targets; b) early clinical diagnosis of invasive fungal infections; c) detection of molecular mechanisms of resistance to antifungal agents; and d) molecular typing of fungi. Currently, these methods are restricted to highly developed laboratories. However, some of these techniques will probably be available in daily clinical practice in the near future.

  4. Nozzle fabrication technique

    Science.gov (United States)

    Wells, Dennis L. (Inventor)

    1988-01-01

    This invention relates to techniques for fabricating hour glass throat or convergent divergent nozzle shapes, and more particularly to new and improved techniques for forming rocket nozzles from electrically conductive material and forming cooling channels in the wall thereof. The concept of positioning a block of electrically conductive material so that its axis is set at a predetermined skew angle with relation to a travelling electron discharge machine electrode and thereafter revolving the body about its own axis to generate a hyperbolic surface of revolution, either internal or external is novel. The method will generate a rocket nozzle which may be provided with cooling channels using the same control and positioning system. The configuration of the cooling channels so produced are unique and novel. Also the method is adaptable to nonmetallic material using analogous cutting tools, such as, water jet, laser, abrasive wire and hot wire.

  5. SUSY using boosted techniques

    CERN Document Server

    Stark, Giordon; The ATLAS collaboration

    2016-01-01

    In this talk, I present a discussion of techniques used in supersymmetry searches in papers published by the ATLAS Collaboration from late Run 1 to early Run 2. The goal is to highlight concepts the analyses have in common, why/how they work, and possible SUSY searches that could benefit from boosted studies. Theoretical background will be provided for reference to encourage participants to explore in depth on their own time.

  6. NEW EXPLOSIVE WELDING TECHNIQUES

    OpenAIRE

    Lotous, V.; Dragobetskii, V.

    2015-01-01

    Purpose - analysis of the variety of factors of the physical phenomena accompanying the process of the power explosive effect for development of new processes of metal treatment: explosive film coating of hardening and updating of a superficial layer of an item. Industrial approbation of cladding techniques by explosion of item surfaces of complex configuration and determination of parameters of the process of the explosive welding of high-strength pig-iron (graphite of the spherical form) wi...

  7. Remote Attitude Measurement Techniques.

    Science.gov (United States)

    1982-12-01

    are other factors which will influence the selection of the communications means. One major factor is the selection of the technology to be used. Aside...electrooptical devices with partitioned focal planes, and the char- acteristics of these techniques influence how the focal planes are partitioned. G. FOCAL...a televison camera). The incident illumination produces a non-uniformity on the scanned side of the sensitive material which can be modeled as an

  8. Miniaturization Techniques for Accelerators

    Energy Technology Data Exchange (ETDEWEB)

    Spencer, James E.

    2003-05-27

    The possibility of laser driven accelerators [1] suggests the need for new structures based on micromachining and integrated circuit technology because of the comparable scales. Thus, we are exploring fully integrated structures including sources, optics (for both light and particle) and acceleration in a common format--an accelerator-on-chip (AOC). Tests suggest a number of preferred materials and techniques but no technical or fundamental roadblocks at scales of order 1 {micro}m or larger.

  9. Colorimetric protein assay techniques.

    Science.gov (United States)

    Sapan, C V; Lundblad, R L; Price, N C

    1999-04-01

    There has been an increase in the number of colorimetric assay techniques for the determination of protein concentration over the past 20 years. This has resulted in a perceived increase in sensitivity and accuracy with the advent of new techniques. The present review considers these advances with emphasis on the potential use of such technologies in the assay of biopharmaceuticals. The techniques reviewed include Coomassie Blue G-250 dye binding (the Bradford assay), the Lowry assay, the bicinchoninic acid assay and the biuret assay. It is shown that each assay has advantages and disadvantages relative to sensitivity, ease of performance, acceptance in the literature, accuracy and reproducibility/coefficient of variation/laboratory-to-laboratory variation. A comparison of the use of several assays with the same sample population is presented. It is suggested that the most critical issue in the use of a chromogenic protein assay for the characterization of a biopharmaceutical is the selection of a standard for the calibration of the assay; it is crucial that the standard be representative of the sample. If it is not possible to match the standard with the sample from the perspective of protein composition, then it is preferable to use an assay that is not sensitive to the composition of the protein such as a micro-Kjeldahl technique, quantitative amino acid analysis or the biuret assay. In a complex mixture it might be inappropriate to focus on a general method of protein determination and much more informative to use specific methods relating to the protein(s) of particular interest, using either specific assays or antibody-based methods. The key point is that whatever method is adopted as the 'gold standard' for a given protein, this method needs to be used routinely for calibration.

  10. TRAC Innovative Visualization Techniques

    Science.gov (United States)

    2016-11-14

    Design thinking”. In: Harvard business review 86.6 (2008), p. 84. [6] U.S. Department of the Army. ATP 5-0.1 Army Design Methodology. Washington D.C... business review 86.6 (2008), p. 84. 3 4. What other visualization techniques can assist TRAC analysts with addressing these issues? 5. What are the best...is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and

  11. Empirical techniques in finance

    CERN Document Server

    Bhar, Ramaprasad

    2005-01-01

    This book offers the opportunity to study and experience advanced empi- cal techniques in finance and in general financial economics. It is not only suitable for students with an interest in the field, it is also highly rec- mended for academic researchers as well as the researchers in the industry. The book focuses on the contemporary empirical techniques used in the analysis of financial markets and how these are implemented using actual market data. With an emphasis on Implementation, this book helps foc- ing on strategies for rigorously combing finance theory and modeling technology to extend extant considerations in the literature. The main aim of this book is to equip the readers with an array of tools and techniques that will allow them to explore financial market problems with a fresh perspective. In this sense it is not another volume in eco- metrics. Of course, the traditional econometric methods are still valid and important; the contents of this book will bring in other related modeling topics tha...

  12. Covariance mapping techniques

    Science.gov (United States)

    Frasinski, Leszek J.

    2016-08-01

    Recent technological advances in the generation of intense femtosecond pulses have made covariance mapping an attractive analytical technique. The laser pulses available are so intense that often thousands of ionisation and Coulomb explosion events will occur within each pulse. To understand the physics of these processes the photoelectrons and photoions need to be correlated, and covariance mapping is well suited for operating at the high counting rates of these laser sources. Partial covariance is particularly useful in experiments with x-ray free electron lasers, because it is capable of suppressing pulse fluctuation effects. A variety of covariance mapping methods is described: simple, partial (single- and multi-parameter), sliced, contingent and multi-dimensional. The relationship to coincidence techniques is discussed. Covariance mapping has been used in many areas of science and technology: inner-shell excitation and Auger decay, multiphoton and multielectron ionisation, time-of-flight and angle-resolved spectrometry, infrared spectroscopy, nuclear magnetic resonance imaging, stimulated Raman scattering, directional gamma ray sensing, welding diagnostics and brain connectivity studies (connectomics). This review gives practical advice for implementing the technique and interpreting the results, including its limitations and instrumental constraints. It also summarises recent theoretical studies, highlights unsolved problems and outlines a personal view on the most promising research directions.

  13. Computer Assisted Audit Techniques

    Directory of Open Access Journals (Sweden)

    Eugenia Iancu

    2007-01-01

    Full Text Available From the modern point of view, audit takes intoaccount especially the information systems representingmainly the examination performed by a professional asregards the manner for developing an activity by means ofcomparing it to the quality criteria specific to this activity.Having as reference point this very general definition ofauditing, it must be emphasized that the best known segmentof auditing is the financial audit that had a parallel evolutionto the accountancy one.The present day phase of developing the financial audithas as main trait the internationalization of the accountantprofessional. World wide there are multinational companiesthat offer services in the financial auditing, taxing andconsultancy domain. The auditors, natural persons and auditcompanies, take part at the works of the national andinternational authorities for setting out norms in theaccountancy and auditing domain.The computer assisted audit techniques can be classified inseveral manners according to the approaches used by theauditor. The most well-known techniques are comprised inthe following categories: testing data techniques, integratedtest, parallel simulation, revising the program logics,programs developed upon request, generalized auditsoftware, utility programs and expert systems.

  14. Safe monsplasty technique.

    Science.gov (United States)

    Patoué, Audrey; De Runz, Antoine; Carloni, Raphael; Aillet, Sylvie; Watier, Eric; Bertheuil, Nicolas

    2017-07-14

    To improve their health and quality of life, obese patients undergo consultation after weight loss. In these patients, the sub-umbilical abdominal and pubic regions are often characterized by redundant skin, creating aesthetic and functional discomfort. Monsplasty is an important step in abdominoplasty or bodylift procedures. We report on an original technique used to correct deformity in the pubic region following weight loss. All interventions were performed by the same surgeon between April and December 2015. On stretched skin, we drew two lines 5 cm lateral to the median line on both sides and connected them with the arc of a circle placed 7 cm from the vulvar fork. Then, monsplasty marks extend to abdominal fold. During lower abdominal contouring, we performed monsplasty with three separate stitches between the camper fascia and aponeurosis of the abdominal muscle. The aim was to bring tension to the pubic region without additional surgical procedure. We report on 21 consecutive cases of monsplasty following lower trunk contouring. No reoperation was performed due to complication or aesthetic demand (no under- or over-correction occurred). No complication (e.g. edema, seroma, disturbance of sensibility) was observed in the pubic area. The results were stable 1 year after surgery. We report on a simple, rapid, and reproducible monsplasty technique for all stages of Pittsburgh classifications, which achieved favorable results with no complication. We recommend performance of this effective technique simultaneously with abdominoplasty or bodylift procedures. IV.

  15. Scalp imaging techniques

    Science.gov (United States)

    Otberg, Nina; Shapiro, Jerry; Lui, Harvey; Wu, Wen-Yu; Alzolibani, Abdullateef; Kang, Hoon; Richter, Heike; Lademann, Jürgen

    2017-05-01

    Scalp imaging techniques are necessary tools for the trichological practice and for visualization of permeation, penetration and absorption processes into and through the scalp and for the research on drug delivery and toxicology. The present letter reviews different scalp imaging techniques and discusses their utility. Moreover, two different studies on scalp imaging techniques are presented in this letter: (1) scalp imaging with phototrichograms in combination with laser scanning microscopy, and (2) follicular measurements with cyanoacrylate surface replicas and light microscopy in combination with laser scanning microscopy. The experiments compare different methods for the determination of hair density on the scalp and different follicular measures. An average terminal hair density of 132 hairs cm-2 was found in 6 Caucasian volunteers and 135 hairs cm-2 in 6 Asian volunteers. The area of the follicular orifices accounts to 16.3% of the skin surface on average measured with laser scanning microscopy images. The potential volume of the follicular infundibulum was calculated based on the laser scanning measurements and is found to be 4.63 mm3 per cm2 skin on average. The experiments show that hair follicles are quantitatively relevant pathways and potential reservoirs for topically applied drugs and cosmetics.

  16. Techniques for Wireless Applications

    KAUST Repository

    Gaaloul, Fakhreddine

    2012-05-01

    Switching techniques have been first proposed as a spacial diversity techniques. These techniques have been shown to reduce considerably the processing load while letting multi-antenna systems achieve a specific target performance. In this thesis, we take a different look at the switching schemes by implementing them for different other wireless applications. More specifically, this thesis consists of three main parts, where the first part considers a multiuser environment and an adaptive scheduling algorithm based on the switching with post-selection scheme for statistically independent but non-identically distributed channel conditions. The performance of this switched based scheduler is investigated and a multitude of performance metrics are presented. In a second part, we propose and analyze the performance of three switched-based algorithms for interference reduction in the downlink of over-loaded femtocells. For instance, performance metrics are derived in closed-form and these metrics are used to compare these three proposed schemes. Finally in a third part, a switch based opportunistic channel access scheme is proposed for a cognitive radio system and its performance is analyzed in terms of two new proposed metrics namely the average cognitive radio access and the waiting time duration.

  17. Applied ALARA techniques

    Energy Technology Data Exchange (ETDEWEB)

    Waggoner, L.O.

    1998-02-05

    The presentation focuses on some of the time-proven and new technologies being used to accomplish radiological work. These techniques can be applied at nuclear facilities to reduce radiation doses and protect the environment. The last reactor plants and processing facilities were shutdown and Hanford was given a new mission to put the facilities in a safe condition, decontaminate, and prepare them for decommissioning. The skills that were necessary to operate these facilities were different than the skills needed today to clean up Hanford. Workers were not familiar with many of the tools, equipment, and materials needed to accomplish:the new mission, which includes clean up of contaminated areas in and around all the facilities, recovery of reactor fuel from spent fuel pools, and the removal of millions of gallons of highly radioactive waste from 177 underground tanks. In addition, this work has to be done with a reduced number of workers and a smaller budget. At Hanford, facilities contain a myriad of radioactive isotopes that are 2048 located inside plant systems, underground tanks, and the soil. As cleanup work at Hanford began, it became obvious early that in order to get workers to apply ALARA and use hew tools and equipment to accomplish the radiological work it was necessary to plan the work in advance and get radiological control and/or ALARA committee personnel involved early in the planning process. Emphasis was placed on applying,ALARA techniques to reduce dose, limit contamination spread and minimize the amount of radioactive waste generated. Progress on the cleanup has,b6en steady and Hanford workers have learned to use different types of engineered controls and ALARA techniques to perform radiological work. The purpose of this presentation is to share the lessons learned on how Hanford is accomplishing radiological work.

  18. Endoscopic thyroidectomy: Our technique

    Directory of Open Access Journals (Sweden)

    Puntambekar Shailesh

    2007-01-01

    Full Text Available Minimally invasive surgery is widely employed for the treatment of thyroid diseases. Several minimal access approaches to the thyroid gland have been described. The commonly performed surgeries have been endoscopic lobectomies. We have performed endoscopic total thyroidectomy by the anterior chest wall approach. In this study, we have described our technique and evaluated the feasibility and efficacy of this procedure. Materials and Methods: From June 2005 to August 2006, 15 cases of endoscopic thyroidectomy were done at our institute. Five patients were male and 10 were female. Mean age was 45 years. (Range 23 to 71 years. Four patients had multinodular goiter and underwent near-total thyroidectomy; four patients had follicular adenoma and underwent hemithyroidectomy. Out of the seven patients of papillary carcinoma, four were low-risk and so a hemithyroidectomy was performed while three patients in the high risk group underwent total thyroidectomy. A detailed description of the surgical technique is provided. Results : The mean nodule size was 48 mm (range 20-80 mm and the mean operating time was 85 min (range 60-120 min. In all cases, the recurrent laryngeal nerve was identified and preserved intact, the superior and inferior parathyroids were also identified in all patients. No patients required conversion to an open cervicotomy. All patients were discharged the day after surgery. All thyroidectomies were completed successfully. No recurrent laryngeal nerve palsies or postoperative tetany occurred. The postoperative course was significantly less painful and all patients were satisfied with the cosmetic results. Conclusions : It is possible to remove large nodules and perform as well as total thyroidectomies using our endoscopic approach. It is a safe and effective technique in the hands of an appropriately trained surgeon. The patients get a cosmetic benefit without any morbidity.

  19. Submersible Packaging Techniques

    Science.gov (United States)

    1989-07-01

    Butyraldehyde E,K Aniline Hydrochloride E Butyric Acid V,E Fluid codes are defined in paragraph 40.2.1 and are listed in order of recommended use . 20...Furfuryl Alcohol E Ethyl Chloride E,B,N Gallic Acid V,FE Ethyl Chiorocarbonate V,F Gasoline BV,F Ethyl Chioroformate V,F Glycerine B,E Ethylene Chloride...This document describes packaging techniques that would allow sophisticated electronics to be used and transported reliably underwater. 20. DISTRIBUTION

  20. Western Blot Techniques.

    Science.gov (United States)

    Kim, Brianna

    2017-01-01

    The Western blot is an important laboratory technique that allows for specific identification and characterization of proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-separated proteins are electophoretically transferred to a polyvinylidene fluoride (PVDF) membrane which is then incubated with specific antibodies, then developed to show the protein of interest. Here, we describe the transfer and detection of Outer surface protein A (OspA), a protein only found on the surface of Borrelia burgdorferi, the bacteria responsible for Lyme disease.

  1. Pileup Mitigation Techniques

    CERN Document Server

    Klein, Matthew Henry; The ATLAS collaboration

    2016-01-01

    We report on recent progress in the ATLAS experiment in developing tools to mitigate the effects of pile-up. Forward pile-up jet tagging techniques, as well as constituent-level pile-up suppression algorithms are discussed in details. The impacts of these approaches on both jet energy and angular resolution, as well as jet substructure and boosted object tagging performance are discussed. Improvements to various physics channels of interest are discussed and the potential future of such algorithms — both online and offline, and both at the current LHC and a future high-luminosity LHC and beyond — is considered in detail

  2. Diamond Anvil Cell Techniques

    Science.gov (United States)

    Piermarini, Gasper J.

    It has often been said that scientific advances are made either in a dramatic and revolutionary way, or, as in the case of the diamond anvil cell (DAC), in a slow and evolutionary manner over a period of several years. For more than 2 decades, commencing in 1958, the DAC developed stepwise from a rather crude qualitative instrument to the sophisticated quantitative research tool it is today, capable of routinely producing sustained static pressures in the multi-megabar range and readily adaptable to numerous scientific measurement techniques because of its optical accessibility, miniature size, and portability.

  3. A fuzzy disaggregation technique

    Directory of Open Access Journals (Sweden)

    Alessandro Polli

    2013-05-01

    Full Text Available The aim of this paper is to analyze a problem of time series disaggregation in presence of broad information lack. In this framework it is not possible to follow standard methodologies, like those stemming from the Chow and Lin algorithm and based on probabilistic assumptions. In general terms, when information sets are limited, instead of referring to probabilistic measures it could be more appropriate to adopt an uncertainty measure satisfying only some general properties, like the fuzzy one. After a synthetic survey about fuzzy aggregation operators, we introduce a fuzzy disaggregation technique, based on Choquet capacity theory and characterized by De Finetti coherence.

  4. Formulation techniques for nanofluids.

    Science.gov (United States)

    Rivera-Solorio, Carlos I; Payán-Rodríguez, Luis A; García-Cuéllar, Alejandro J; Ramón-Raygoza, E D; L Cadena-de-la-Peña, Natalia; Medina-Carreón, David

    2013-11-01

    Fluids with suspended nanoparticles, commonly known as nanofluids, may be formulated to improve the thermal performance of industrial heat transfer systems and applications. Nanofluids may show enhanced thermal and electrical properties such as thermal conductivity, viscosity, heat transfer coefficient, dielectric strength, etc. However, stability problems may arise as nanoparticles usually have the tendency to agglomerate and sediment producing deterioration in the increment of these properties. In this review, we discuss patents that report advances in the formulation of nanofluids including: production methods, selection of components (nanoparticles, base fluid and surfactants), their chemical compositions and morphologies, and characterization techniques. Finally, current and future directions in the development of nanofluid formulation are discussed.

  5. Automatic Threshold Detector Techniques

    Science.gov (United States)

    1976-07-15

    TECHNIQUES Contract No. DAAH01-76-C-0363 ER76-4208 15 July 1976 Prepared for: HEADQUA RTERS U.S. Army Missile Command Redstone Arsenal, Alabama 35809 j...rain cross section each FFT filter) a MUT, MUCL , KCL, MUN, MUCLF (2 ), MUl, MUWI= MUW2 where MUT = Target cross section (M2), MUCL = total ground...variable is currently not used by the program. Since there is yet no point clutter model, MUCL represents the fluctuating component. Until a point plus

  6. APPLICATION OF NUCLEAR TECHNIQUE

    Institute of Scientific and Technical Information of China (English)

    1996-01-01

    1 Application of Fission Track Technique in Stratigraphy Guo Shilun Hao Xiuhong Chen Baoliu Huang Weiwen (Institute of Vertebrate Paleontology and Paleoanthropology, Chinese Academy of Sciences, Beijing, China) Laterite (red soil) is distributed in the south of China from Qinling mountains to Hainan province and extends to Southeast Asia. The age of the formation and deposition of the laterite is an important subject in geological research. Due to lack of fossils of ancient animals and plants, as well as other objects which could be used for dating , the age of the laterite and its related stratum is unknown in the south of China.

  7. Differential doppler heterodyning technique

    DEFF Research Database (Denmark)

    Lading, Lars

    1971-01-01

    Measuring velocity without disturbing the moving object is possible by use of the laser doppler heterodyning technique. Theoretical considerations on the doppler shift show that the antenna property of the photodetector can solve an apparent conflict between two different ways of calculating...... the detected doppler frequency. It is found that the doppler frequency for this particular setup is independent of the direction of detection. Investigations of the signal-to-noise ratio (SNR) show that the maximum SNR-considering the optical setup-is obtained by measuring the frequency difference between two...

  8. Optical techniques in optogenetics

    Science.gov (United States)

    Mohanty, Samarendra K.; Lakshminarayananan, Vasudevan

    2015-07-01

    Optogenetics is an innovative technique for optical control of cells. This field has exploded over the past decade or so and has given rise to great advances in neuroscience. A variety of applications both from the basic and applied research have emerged, turning the early ideas into a powerful paradigm for cell biology, neuroscience, and medical research. This review aims at highlighting the basic concepts that are essential for a comprehensive understanding of optogenetics and some important biological/biomedical applications. Further, emphasis is placed on advancement in optogenetics-associated light-based methods for controlling gene expression, spatially controlled optogenetic stimulation and detection of cellular activities.

  9. DNA Microarray Technique

    Directory of Open Access Journals (Sweden)

    Thakare SP

    2012-11-01

    Full Text Available DNA Microarray is the emerging technique in Biotechnology. The many varieties of DNA microarray or DNA chip devices and systems are described along with their methods for fabrication and their use. It also includes screening and diagnostic applications. The DNA microarray hybridization applications include the important areas of gene expression analysis and genotyping for point mutations, single nucleotide polymorphisms (SNPs, and short tandem repeats (STRs. In addition to the many molecular biological and genomic research uses, this review covers applications of microarray devices and systems for pharmacogenomic research and drug discovery, infectious and genetic disease and cancer diagnostics, and forensic and genetic identification purposes.

  10. High voltage test techniques

    CERN Document Server

    Kind, Dieter

    2001-01-01

    The second edition of High Voltage Test Techniques has been completely revised. The present revision takes into account the latest international developments in High Voltage and Measurement technology, making it an essential reference for engineers in the testing field.High Voltage Technology belongs to the traditional area of Electrical Engineering. However, this is not to say that the area has stood still. New insulating materials, computing methods and voltage levels repeatedly pose new problems or open up methods of solution; electromagnetic compatibility (EMC) or components and systems al

  11. Techniques de hacking

    CERN Document Server

    Erickson, Jon

    2008-01-01

    Dans cet ouvrage, Jon Erickson présente les bases de la programmation en C du point de vue du hacker et dissèque plusieurs techniques de hacking, passées et actuelles, afin de comprendre comment et pourquoi elles fonctionnent. Même si vous ne savez pas programmer, ce livre vous donnera une vue complète de la programmation, de l'architecture des machines, des communications réseau et des techniques de hacking existantes. Associez ces connaissances à l'environnement Linux fourni et laissez libre cours à votre imagination. Avec ce livre vous apprendrez à : • programmer les ordinateurs en C, en assembleur et avec des scripts shell ; • inspecter les registres du processeur et la mémoire système avec un débogueur afin de comprendre précisément ce qui se passe ; Vous découvrirez comment les hackers parviennent à : • corrompre la mémoire d'un système, en utilisant les débordements de tampons et les chaînes de format, pour exécuter un code quelconque ; • surpasser les mesures de sécurit...

  12. A Technique: Exposure Therapy

    Directory of Open Access Journals (Sweden)

    Serkan AKKOYUNLU

    2013-07-01

    Full Text Available Introduction: Exposure with response prevention is an effective treatment for all anxiety disorders. According to the behavioral learning theories, fears which are conditioned via classical conditioning are reinforced by respondent conditioning. Avoidance and safety seeking behaviors prevent disconfirmation of anxious beliefs. In exposure client faces stimulates or cues that elicit fear or distress, by this avoidance is inhibited. Clients are also encouraged to resists performing safety seeking behaviors or rituals that they utilize to reduce fear or distress. Accomplishing these habituation or extinction is achieved. In addition to this clients learn that feared consequences does not realize or not harmful as they believed by experiencing. Emotional processing is believed to be the mechanism of change in exposure.Objective: The aim of this review is to provide a definition of exposure and its effectiveness briefly, and describe how to implement exposure, its steps and remarkable aspects using. Exposure therapies and treatments that involve exposure are proved to be effective in all anxiety disorders. Exposure therapy can be divided in three parts: Assessment and providing a treatment rationale, creating an exposure hierarchy and response prevention plan, implementing exposure sessions. Clients must also continue to perform exposure between sessions. Therapy transcripts are also provided to exemplify these parts. Conclusion: Exposure with response prevention is a basic and effective technique. Every cognitive behavior therapist must be able to implement this technique and be cognizant of pearls of this procedure.

  13. Decision Analysis Technique

    Directory of Open Access Journals (Sweden)

    Hammad Dabo Baba

    2014-01-01

    Full Text Available One of the most significant step in building structure maintenance decision is the physical inspection of the facility to be maintained. The physical inspection involved cursory assessment of the structure and ratings of the identified defects based on expert evaluation. The objective of this paper is to describe present a novel approach to prioritizing the criticality of physical defects in a residential building system using multi criteria decision analysis approach. A residential building constructed in 1985 was considered in this study. Four criteria which includes; Physical Condition of the building system (PC, Effect on Asset (EA, effect on Occupants (EO and Maintenance Cost (MC are considered in the inspection. The building was divided in to nine systems regarded as alternatives. Expert's choice software was used in comparing the importance of the criteria against the main objective, whereas structured Proforma was used in quantifying the defects observed on all building systems against each criteria. The defects severity score of each building system was identified and later multiplied by the weight of the criteria and final hierarchy was derived. The final ranking indicates that, electrical system was considered the most critical system with a risk value of 0.134 while ceiling system scored the lowest risk value of 0.066. The technique is often used in prioritizing mechanical equipment for maintenance planning. However, result of this study indicates that the technique could be used in prioritizing building systems for maintenance planning

  14. Other imaging techniques.

    Science.gov (United States)

    Isard, H J

    1984-02-01

    Images of the breast can now be produced by five modalities: x-ray, heat, sound, light, and magnetism. X-ray mammography is generally accepted as the most accurate of these in the detection of breast cancer, and the standard by which the others are judged. Despite the obvious attraction of nonionizing techniques, the economic factor attendant on multiple studies requires consideration. Nuclear magnetic resonance (NMR) is currently being investigated in several clinics, but as yet there is no large series of documented cases. This report addresses itself to thermography, ultrasonography and diaphanography (transillumination). The unique characteristics of each and their respective roles in evaluation of the breast, particularly in the detection of breast cancer, will be discussed. When used in conjunction with mammography, potential advantages include: enhanced diagnostic accuracy, reduction of unnecessary surgery, and, in proven cases of breast cancer, prognostic capability. Thus far it has not been demonstrated that any of the nonionizing techniques can serve as a sole screening modality for breast cancer detection in asymptomatic women.

  15. Psoriatic arthritis: imaging techniques

    Directory of Open Access Journals (Sweden)

    E. Lubrano

    2012-06-01

    Full Text Available Imaging techniques to assess psoriatic arthritis (PsA include radiography, ultrasonography (US, magnetic resonance imaging (MRI, computed tomography (CT and bone scintigraphy. The radiographic hallmark of PsA is the combination of destructive changes (joint erosions, tuft resorption, osteolysis with bone proliferation (including periarticular and shaft periostitis, ankylosis, spur formation and non-marginal syndesmophytes. US has an increasing important role in the evaluation of PsA. In fact, power Doppler US is useful mainly for its ability to assess musculoskeletal (joints, tendons, entheses and cutaneous (skin and nails involvement, to monitor efficacy of therapy and to guide steroid injections at the level of inflamed joints, tendon sheaths and entheses. MRI allows direct visualization of inflammation in peripheral and axial joints, and peripheral and axial entheses, and has dramatically improved the possibilities for early diagnosis and objective monitoring of the disease process in PsA. MRI has allowed explaining the relationships among enthesitis, synovitis and osteitis in PsA, supporting a SpA pattern of inflammation where enthesitis is the primary target of inflammation. CT has little role in assessment of peripheral joints, but it may be useful in assessing elements of spine disease. CT accuracy is similar to MRI in assessment of erosions in sacroiliac joint involvement, but CT is not as effective in detecting synovial inflammation. Bone scintigraphy lacks specificity and is now supplanted with US and MRI techniques.

  16. Advanced Coating Removal Techniques

    Science.gov (United States)

    Seibert, Jon

    2006-01-01

    An important step in the repair and protection against corrosion damage is the safe removal of the oxidation and protective coatings without further damaging the integrity of the substrate. Two such methods that are proving to be safe and effective in this task are liquid nitrogen and laser removal operations. Laser technology used for the removal of protective coatings is currently being researched and implemented in various areas of the aerospace industry. Delivering thousands of focused energy pulses, the laser ablates the coating surface by heating and dissolving the material applied to the substrate. The metal substrate will reflect the laser and redirect the energy to any remaining protective coating, thus preventing any collateral damage the substrate may suffer throughout the process. Liquid nitrogen jets are comparable to blasting with an ultra high-pressure water jet but without the residual liquid that requires collection and removal .As the liquid nitrogen reaches the surface it is transformed into gaseous nitrogen and reenters the atmosphere without any contamination to surrounding hardware. These innovative technologies simplify corrosion repair by eliminating hazardous chemicals and repetitive manual labor from the coating removal process. One very significant advantage is the reduction of particulate contamination exposure to personnel. With the removal of coatings adjacent to sensitive flight hardware, a benefit of each technique for the space program is that no contamination such as beads, water, or sanding residue is left behind when the job is finished. One primary concern is the safe removal of coatings from thin aluminum honeycomb face sheet. NASA recently conducted thermal testing on liquid nitrogen systems and found that no damage occurred on 1/6", aluminum substrates. Wright Patterson Air Force Base in conjunction with Boeing and NASA is currently testing the laser remOval technique for process qualification. Other applications of liquid

  17. Kinetic Actviation Relaxation Technique

    CERN Document Server

    Béland, Laurent Karim; El-Mellouhi, Fedwa; Joly, Jean-François; Mousseau, Normand

    2011-01-01

    We present a detailed description of the kinetic Activation-Relaxation Technique (k-ART), an off-lattice, self-learning kinetic Monte Carlo algorithm with on-the-fly event search. Combining a topological classification for local environments and event generation with ART nouveau, an efficient unbiased sampling method for finding transition states, k-ART can be applied to complex materials with atoms in off-lattice positions or with elastic deformations that cannot be handled with standard KMC approaches. In addition to presenting the various elements of the algorithm, we demonstrate the general character of k-ART by applying the algorithm to three challenging systems: self-defect annihilation in c-Si, self-interstitial diffusion in Fe and structural relaxation in amorphous silicon.

  18. Data flow modeling techniques

    Science.gov (United States)

    Kavi, K. M.

    1984-01-01

    There have been a number of simulation packages developed for the purpose of designing, testing and validating computer systems, digital systems and software systems. Complex analytical tools based on Markov and semi-Markov processes have been designed to estimate the reliability and performance of simulated systems. Petri nets have received wide acceptance for modeling complex and highly parallel computers. In this research data flow models for computer systems are investigated. Data flow models can be used to simulate both software and hardware in a uniform manner. Data flow simulation techniques provide the computer systems designer with a CAD environment which enables highly parallel complex systems to be defined, evaluated at all levels and finally implemented in either hardware or software. Inherent in data flow concept is the hierarchical handling of complex systems. In this paper we will describe how data flow can be used to model computer system.

  19. Phaco rolling technique.

    Science.gov (United States)

    Güell, Jose L; Vázquez, Mercedes; Lucena, Jose; Velasco, Fortino; Manero, Felicidad

    2004-10-01

    We describe a phacoemulsification technique for soft and medium-hard cataracts to decrease phaco time and enhance the safety of the procedure. After conventional hydrodissection and hydrodelamination are performed, a 15- or 30-degree phaco tip is positioned on the peripheral lens beside the capsulorhexis edge and in contact with the nucleus-epinucleus interface. The lens is then aspirated onto the phaco tip. Phacoemulsification is started with the ultrasound energy level limited to 15% to 25% depending on the nuclear hardness and with linear aspiration power up to 250 mmHg. The phaco tip is slightly displaced to the vertex of the pupil to keep it occluded, and the lens is rotated. The phaco tip is placed in the same area and a modified manipulator used to keep the lens in a horizontal position during rotational movement. The small, hard, central nucleus is usually emulsified at the end.

  20. Technique murale picturale

    Directory of Open Access Journals (Sweden)

    Claude Coupry

    2009-09-01

    Full Text Available Après des études remarquables publiées, entre autres, dans Peindre à Auxerre au Moyen Âge, IXe-XIVe siècles, l’étude technique et analytique des peintures murales n’avait pas donné lieu à de nouvelles recherches méthodologiques en dépit des travaux de terrain (Souvigny, Stavelot… où l’archéologie du bâti avait permis des rencontres entre archéologues, historiens de l’art et restaurateurs particulièrement impliqués pour la période qui nous intéresse, de la basse Antiquité au haut Moyen Âge.Ce...

  1. [Therapeutic education didactic techniques].

    Science.gov (United States)

    Valverde, Maite; Vidal, Mercè; Jansa, Margarida

    2012-10-01

    This article includes an introduction to the role of Therapeutic Education for Diabetes treatment according to the recommendations of the American Diabetes Association (ADA), the Diabetes Education Study Group (DESG) of the "European Association for Study of Diabetes (EASD) and the clinical Practice Guidelines (CPG) of the Spanish Ministry of Health. We analyze theoretical models and the differences between teaching vs. learning as well as current trends (including Internet), that can facilitate meaningful learning of people with diabetes and their families and relatives. We analyze the differences, similarities, advantages and disadvantages of individual and group education. Finally, we describe different educational techniques (metaplan, case method, brainstorming, role playing, games, seminars, autobiography, forums, chats,..) applicable to individual, group or virtual education and its application depending on the learning objective.

  2. Extended Ewald summation technique

    Science.gov (United States)

    Kylänpää, Ilkka; Räsänen, Esa

    2016-09-01

    We present a technique to improve the accuracy and to reduce the computational labor in the calculation of long-range interactions in systems with periodic boundary conditions. We extend the well-known Ewald method by using a linear combination of screening Gaussian charge distributions instead of only one. This enables us to find faster converging real-space and reciprocal space summations. The combined simplicity and efficiency of our method is demonstrated, and the scheme is readily applicable to large-scale periodic simulations, classical as well as quantum. Moreover, apart from the required a priori optimization the method is straightforward to include in most routines based on the Ewald method within, e.g., density-functional, molecular dynamics, and quantum Monte Carlo calculations.

  3. New measurements techniques

    DEFF Research Database (Denmark)

    Torras Rosell, Antoni

    of the pressure encountered by the ‘sensing’ light when traveling through the acoustic field. Far from being a limitation, this integral principle is exploited for sound field visualization using tomography. The most innovative contribution of this PhD project is the applicability of the acousto-optic measuring......Acoustic measurements are traditionally based on transducers, and in particular, the most advanced measurement techniques are nowadays based on transducer arrays. This poses a fundamental problem, namely the influence of the transducer itself on the actual properties of sound when the transducer...... is immersed into the sound field. Typically, this influence is assumed to be negligible when the size of the transducer is small compared to the wavelength of the sound wave, or is rendered negligible by using a transducerbased correction that depends on the frequency. Either solution introduces additional...

  4. Alternative Analysis Techniques for Needs and Needs Documentation Techniques,

    Science.gov (United States)

    1980-06-20

    Have you previously participated in a brainwriting session! a. Yes b. No 9. Have you previously participated in the Nominal Group Technique process...brainstorming technique for future sessions. Strongly I Strongly disagree !I agree 8. It was easy to present my views using the brainwriting technique...Strongly!i Strongly disagree I , agree * 9. I was satisfied with the brainwriting technique. Strongly i Strongly disagree __ agree . 10. I recommend using

  5. Changes in urological surgical techniques

    Directory of Open Access Journals (Sweden)

    Oktay Üçer

    2010-06-01

    Full Text Available Recently, laparoscopic and afterwards robotic techniques have constituted most of urologic surgery procedures. Open surgery may give place to robotic surgery due to possible widespread use of robots in the future. Studies, that compare these two techniques are usually designed about radical prostatectomy, since it is the most common operation performed by using these techniques. In literature,robotic surgery seems more advantageous than other techniques but the most important disadvantage of this technique is cost-effective problems. In present review,history of open, laparoscopic and robotic surgery, and comparison of advantages, disadvantages and cost of these techniques have been discussed with literature.

  6. INTERNAL ENVIRONMENT ANALYSIS TECHNIQUES

    Directory of Open Access Journals (Sweden)

    Caescu Stefan Claudiu

    2011-12-01

    Full Text Available Theme The situation analysis, as a separate component of the strategic planning, involves collecting and analysing relevant types of information on the components of the marketing environment and their evolution on the one hand and also on the organization’s resources and capabilities on the other. Objectives of the Research The main purpose of the study of the analysis techniques of the internal environment is to provide insight on those aspects that are of strategic importance to the organization. Literature Review The marketing environment consists of two distinct components, the internal environment that is made from specific variables within the organization and the external environment that is made from variables external to the organization. Although analysing the external environment is essential for corporate success, it is not enough unless it is backed by a detailed analysis of the internal environment of the organization. The internal environment includes all elements that are endogenous to the organization, which are influenced to a great extent and totally controlled by it. The study of the internal environment must answer all resource related questions, solve all resource management issues and represents the first step in drawing up the marketing strategy. Research Methodology The present paper accomplished a documentary study of the main techniques used for the analysis of the internal environment. Results The special literature emphasizes that the differences in performance from one organization to another is primarily dependant not on the differences between the fields of activity, but especially on the differences between the resources and capabilities and the ways these are capitalized on. The main methods of analysing the internal environment addressed in this paper are: the analysis of the organizational resources, the performance analysis, the value chain analysis and the functional analysis. Implications Basically such

  7. Programing techniques for CDC equipment

    Science.gov (United States)

    Newsom, J. R.; Tiffany, S. H.

    1979-01-01

    Five techniques reduce core requirements for fast batch turnaround time and interactive-terminal capability. Same techniques increase program versatility, decrease problem-configuration dependence, and facilitate interprogram communication.

  8. Improved Search Techniques

    Science.gov (United States)

    Albornoz, Caleb Ronald

    2012-01-01

    Thousands of millions of documents are stored and updated daily in the World Wide Web. Most of the information is not efficiently organized to build knowledge from the stored data. Nowadays, search engines are mainly used by users who rely on their skills to look for the information needed. This paper presents different techniques search engine users can apply in Google Search to improve the relevancy of search results. According to the Pew Research Center, the average person spends eight hours a month searching for the right information. For instance, a company that employs 1000 employees wastes $2.5 million dollars on looking for nonexistent and/or not found information. The cost is very high because decisions are made based on the information that is readily available to use. Whenever the information necessary to formulate an argument is not available or found, poor decisions may be made and mistakes will be more likely to occur. Also, the survey indicates that only 56% of Google users feel confident with their current search skills. Moreover, just 76% of the information that is available on the Internet is accurate.

  9. Dose Reduction Techniques

    CERN Document Server

    Waggoner, L O

    2000-01-01

    As radiation safety specialists, one of the things we are required to do is evaluate tools, equipment, materials and work practices and decide whether the use of these products or work practices will reduce radiation dose or risk to the environment. There is a tendency for many workers that work with radioactive material to accomplish radiological work the same way they have always done it rather than look for new technology or change their work practices. New technology is being developed all the time that can make radiological work easier and result in less radiation dose to the worker or reduce the possibility that contamination will be spread to the environment. As we discuss the various tools and techniques that reduce radiation dose, keep in mind that the radiological controls should be reasonable. We can not always get the dose to zero, so we must try to accomplish the work efficiently and cost-effectively. There are times we may have to accept there is only so much you can do. The goal is to do the sm...

  10. INTRACRANIAL PRESSURE MONITORING TECHNIQUE

    Directory of Open Access Journals (Sweden)

    Ida Bagus Adi Kayana

    2013-03-01

    Full Text Available Head injury is the most significant cause of increased morbidity and mortality. An estimated 1.4 million head injuries occur each year, with and more than 1.1 million come to the Emergency Unit. On each patient head injury, an increase in intracranial pressure (ICP related to poor outcomes and aggressive therapy to increased ICP can improve the outcomes. ICP monitoring is the most widely used because of the prevention and control of ICP as well as maintain the pressure increase perfusion of cerebral (Cerebral Perfusion Pressure/CPP is the basic purpose of handling head injury. There are two methods of monitoring ICP that is an invasive methods (directly and non-invasive techniques (indirectly. The method commonly used, namely intraventricular and intraparenkimal (microtransducer sensor because it is more accurate but keep attention to the existence of the risk of bleeding and infection resulting from installation. Monitoring of ICT can determine the actions that avoid further brain injury, which can be lethal and irreversibel.

  11. Fenestration obscuration techniques

    Science.gov (United States)

    Smalley, Michael

    2007-10-01

    There are situations where it is advantageous to visually obscure through glass, to an external observer, the movement of people within a well lit room. It may be that the building use has changed or existing measures which had provided obscuration such as 'Bomb-blast' curtains have been discontinued. Recognising that implemented solutions must create the minimum disruption to outward visibility and involve the least procedural effort (be simple to use), the Centre for Protection of National Infrastructure, CPNI, commissioned this study, defining key requirements including: (a) Automatic or simple manual operation (b) Obscuration of movement within the building from outside (c) Varying levels of obscuration depending on the difference in internal and external light levels. (d) Minimum disruption to outward visibility (e) Acceptable for use on heritage and iconic sites (f) Easy to retrofit (g) Low cost This report reviews earlier work carried out into the protection of Guardrooms by the use of lighting techniques coupled with the use of reflective and screen printed films. Other innovative solutions including Electrochromatic controllable glazing which may prove more appropriate to office and commercial buildings are also considered. It is seen that some measures, (window films or blinds), are cost effective and unsophisticated while more complex automatic systems using reactive glazing can offer critical design advantages. It must be noted however that some of the key requirements are mutually exclusive and any solution chosen will always be a compromise based on client needs and circumstances.

  12. Techniques of Electrode Fabrication

    Science.gov (United States)

    Guo, Liang; Li, Xinyong; Chen, Guohua

    Electrochemical applications using many kinds of electrode materials as an advanced oxidation/reduction technique have been a focus of research by a number of groups during the last two decades. The electrochemical approach has been adopted successfully to develop various environmental applications, mainly including water and wastewater treatment, aqueous system monitoring, and solid surface analysis. In this chapter, a number of methods for the fabrication of film-structured electrode materials were selectively reviewed. Firstly, the thermal decomposition method is briefly described, followed by introducing chemical vapor deposition (CVD) strategy. Especially, much attention was focused on introducing the methods to produce diamond novel film electrode owing to its unique physical and chemical properties. The principle and influence factors of hot filament CVD and plasma enhanced CVD preparation were interpreted by refereeing recent reports. Finally, recent developments that address electro-oxidation/reduction issues and novel electrodes such as nano-electrode and boron-doped diamond electrode (BDD) are presented in the overview.

  13. COHERENT-LIGHT RECORDING TECHNIQUES.

    Science.gov (United States)

    The purpose of this report is to summarize, define and demonstrate techniques necessary for the application of coherent light to the problems of...Investigations into such areas as the coherent light source itself, modulation, deflection or scanning techniques, readout techniques and the evaluation of recording media are reported.

  14. Dose Reduction Techniques

    Energy Technology Data Exchange (ETDEWEB)

    WAGGONER, L.O.

    2000-05-16

    As radiation safety specialists, one of the things we are required to do is evaluate tools, equipment, materials and work practices and decide whether the use of these products or work practices will reduce radiation dose or risk to the environment. There is a tendency for many workers that work with radioactive material to accomplish radiological work the same way they have always done it rather than look for new technology or change their work practices. New technology is being developed all the time that can make radiological work easier and result in less radiation dose to the worker or reduce the possibility that contamination will be spread to the environment. As we discuss the various tools and techniques that reduce radiation dose, keep in mind that the radiological controls should be reasonable. We can not always get the dose to zero, so we must try to accomplish the work efficiently and cost-effectively. There are times we may have to accept there is only so much you can do. The goal is to do the smart things that protect the worker but do not hinder him while the task is being accomplished. In addition, we should not demand that large amounts of money be spent for equipment that has marginal value in order to save a few millirem. We have broken the handout into sections that should simplify the presentation. Time, distance, shielding, and source reduction are methods used to reduce dose and are covered in Part I on work execution. We then look at operational considerations, radiological design parameters, and discuss the characteristics of personnel who deal with ALARA. This handout should give you an overview of what it takes to have an effective dose reduction program.

  15. Techniques in Broadband Interferometry

    Energy Technology Data Exchange (ETDEWEB)

    Erskine, D J

    2004-01-04

    This is a compilation of my patents issued from 1997 to 2002, generally describing interferometer techniques that modify the coherence properties of broad-bandwidth light and other waves, with applications to Doppler velocimetry, range finding, imaging and spectroscopy. Patents are tedious to read in their original form. In an effort to improve their readability I have embedded the Figures throughout the manuscript, put the Figure captions underneath the Figures, and added section headings. Otherwise I have resisted the temptation to modify the words, though I found many places which could use healthy editing. There may be minor differences with the official versions issued by the US Patent and Trademark Office, particularly in the claims sections. In my shock physics work I measured the velocities of targets impacted by flyer plates by illuminating them with laser light and analyzing the reflected light with an interferometer. Small wavelength changes caused by the target motion (Doppler effect) were converted into fringe shifts by the interferometer. Lasers having long coherence lengths were required for the illumination. While lasers are certainly bright sources, and their collimated beams are convenient to work with, they are expensive. Particularly if one needs to illuminate a wide surface area, then large amounts of power are needed. Orders of magnitude more power per dollar can be obtained from a simple flashlamp, or for that matter, a 50 cent light bulb. Yet these inexpensive sources cannot practically be used for Doppler velocimetry because their coherence length is extremely short, i.e. their bandwidth is much too wide. Hence the motivation for patents 1 & 2 is a method (White Light Velocimetry) for allowing use of these powerful but incoherent lamps for interferometry. The coherence of the illumination is modified by passing it through a preparatory interferometer.

  16. ADOPTABLE TECHNIQUE(S FOR MANAGING GHANAIAN SALINE SOILS

    Directory of Open Access Journals (Sweden)

    Akwasi Asamoah

    2013-03-01

    Full Text Available : Salinization of Ghanaian soils is on the rise. Organic matter application has not proved an effective and feasible technique for curbing this rise. Hence this paper seeks to review techniques that Ghana is using to manage its saline soils and further recommend a feasible, cost effective and beneficial technique for exhaustive research and possible adoption in the future. Halophytes appear to be the most feasible, cost effective and beneficial technique which could be adopted for the effective management of Ghanaian saline soils. But where halophytes are exotic, care must be taken to avoid competition with native species and allow preservation of agrobiodiversity

  17. Theorists and Techniques: Connecting Education Theories to Lamaze Teaching Techniques.

    Science.gov (United States)

    Podgurski, Mary Jo

    2016-01-01

    Should childbirth educators connect education theory to technique? Is there more to learning about theorists than memorizing facts for an assessment? Are childbirth educators uniquely poised to glean wisdom from theorists and enhance their classes with interactive techniques inspiring participant knowledge and empowerment? Yes, yes, and yes. This article will explore how an awareness of education theory can enhance retention of material through interactive learning techniques. Lamaze International childbirth classes already prepare participants for the childbearing year by using positive group dynamics; theory will empower childbirth educators to address education through well-studied avenues. Childbirth educators can provide evidence-based learning techniques in their classes and create true behavioral change.

  18. Classroom Assessment Techniques

    Science.gov (United States)

    Narayanan, M.

    2003-12-01

    and the learner should be carefully observed and monitored. Forrest says that Student Portfolios, which document learning in more detail, seldom reveal how teaching contributes to students' progress. Cerbin further indicates that a course portfolio is essentially, a like a manuscript of scholarly work in progress. In this example, it is a work that explains what, how, and why students learn or do not learn in a course. In this paper, the author reports on a dozen techniques that could perhaps be used to document assessment of student learning. References : Cerbin, W. (1993). Fostering a culture of teaching as scholarship. The Teaching Professor, 7(3), 1-2. Edgerton, R., Hutchings, P., & Quinlan, P. (1991). The teaching portfolio: Capturing the scholarship in teaching. Washington, DC: American Association for Higher Education. Forrest, A. (1990). Time will tell: Portfolio-assisted assessment of general education. Washington, DC: American Association for Higher Education. Linn, R., Baker, E., & Dunbar, S. (1991). Complex, Performance-based Assessment: Expectations and Validation Criteria. Educational Researcher, 20 (8), 15-21. Narayanan, M. (2003). Assessment in Higher Education: Partnerships in Learning. Paper presented at the 23rd Annual Lilly Conference on College Teaching, Miami University, Oxford, OH. Seldin, P. (1991). The teaching portfolio. Bolton, MA: Anker. Young, C. O., Sr., & Young, L. H. (1999). Assessing Learning in Interactive Courses. Journal on Excellence in College Teaching, 10 (1), 63-76.

  19. Practical hacking techniques and countermeasures

    CERN Document Server

    Spivey, Mark D

    2006-01-01

    Examining computer security from the hacker''s perspective, Practical Hacking Techniques and Countermeasures employs virtual computers to illustrate how an attack is executed, including the script, compilation, and results. It provides detailed screen shots in each lab for the reader to follow along in a step-by-step process in order to duplicate and understand how the attack works. It enables experimenting with hacking techniques without fear of corrupting computers or violating any laws.Written in a lab manual style, the book begins with the installation of the VMware® Workstation product and guides the users through detailed hacking labs enabling them to experience what a hacker actually does during an attack. It covers social engineering techniques, footprinting techniques, and scanning tools. Later chapters examine spoofing techniques, sniffing techniques, password cracking, and attack tools. Identifying wireless attacks, the book also explores Trojans, Man-in-the-Middle (MTM) attacks, and Denial of S...

  20. Ultrasonic techniques for fluids characterization

    CERN Document Server

    Povey, Malcolm J W

    1997-01-01

    This book is a comprehensive and practical guide to the use of ultrasonic techniques for the characterization of fluids. Focusing on ultrasonic velocimetry, the author covers the basic topics and techniques necessaryfor successful ultrasound measurements on emulsions, dispersions, multiphase media, and viscoelastic/viscoplastic materials. Advanced techniques such as scattering, particle sizing, and automation are also presented. As a handbook for industrial and scientific use, Ultrasonic Techniques for Fluids Characterization is an indispensable guide to chemists and chemical engineers using ultrasound for research or process monitoring in the chemical, food processing, pharmaceutical, cosmetic, biotechnology,and fuels industries. Key Features * Appeals to anyone using ultrasound to study fluids * Provides the first detailed description of the ultrasound profiling technique for dispersions * Describes new techniques for measuring phase transitions and nucleation, such as water/ice and oil/fat * Presents the l...

  1. Luminescence techniques: Instrumentation and methods

    DEFF Research Database (Denmark)

    Bøtter-Jensen, L.

    1997-01-01

    This paper describes techniques, instruments and methods used in luminescence dating and environmental dosimetry in many laboratories around the world. These techniques are based on two phenomena - thermally stimulated luminescence and optically stimulated luminescence. The most commonly used...... luminescence stimulation and detection techniques are reviewed and information is given on recent developments in instrument design and on the stale of the art in luminescence measurements and analysis. (C) 1998 Elsevier Science Ltd. All rights reserved....

  2. Optical pulses, lasers, measuring techniques

    CERN Document Server

    Früngel, Frank B A

    1965-01-01

    High Speed Pulse Technology: Volume II: Optical Pulses - Lasers - Measuring Techniques focuses on the theoretical and engineering problems that result from the capacitor discharge technique.This book is organized into three main topics: light flash production from a capacitive energy storage; signal transmission and ranging systems by capacitor discharges and lasers; and impulse measuring technique. This text specifically discusses the air spark under atmospheric conditions, industrial equipment for laser flashing, and claims for light transmitting system. The application of light impulse sign

  3. Correlation of cure monitoring techniques

    Science.gov (United States)

    Chang, S. S.; Mopsik, F. I.; Hunston, D. L.

    Six different composite matrix or neat resin cure-monitoring methods are presently used to follow the cure process in a model epoxy system, and the results obtained are compared. Differential scanning calorimetry, viscosity monitoring, the ultrasonic shear wave propagation technique, dielectric spectrometry, and two different fluorescence intensity techniques are compared with a view to common traits and differences. Dielectric fluorescence and ultrasonic measurement techniques are noted to be applicable to on-line process monitoring.

  4. A summary of virtualization techniques

    OpenAIRE

    Rodríguez Haro, Fernando; Freitag, Fèlix; Navarro Moldes, Leandro; Hernández Sánchez, Efraín; Farías Mendoza, Nicandro; Guerrero Ibáñez, Juan Antonio; González Potes, Apolinar

    2012-01-01

    Nowadays, virtualization is a technology that is applied for sharing the capabilities of physical computers by splitting the resources among OSs. The concept of Virtual Machines (VMs) started back in 1964 with a IBM project called CP/CMS system. Currently, there are several virtualization techniques that can be used for supporting the execution of entire operating systems. We classify the virtualization techniques from the OS view. First, we discuss two techniques that executes modified gu...

  5. Geolocation Techniques Principles and Applications

    CERN Document Server

    Gentile, Camillo; Raulefs, Ronald; Teolis, Carole

    2013-01-01

    Geolocation Techniques: Principles and Applications provides a comprehensive overview of geolocation technologies and techniques, from radio-frequency based to inertial based. The focus of this book is to provide an overview on the different types of infra-structure supported by most commercial localization systems as well as on the most popular computational techniques which these systems employ. This book can serve as a reference for scholarly activities such as teaching, self-learning, or research.

  6. Detection Techniques of Femtosecond Lasers

    Institute of Scientific and Technical Information of China (English)

    LIU Li-peng; ZHOU Ming; DAI Qi-xun; CAI Lan

    2004-01-01

    The measurement techniques of femtosecond spectroscopy are effective method to investigate ultrafast dynamics, they are widely used in the fields of physics, chemistry and biology. In this paper, the principle, experiment setup and the approaches to deal with the experiment data were presented. Then different measurement techniques such as transient absorption spectroscopy, photon echoes, optical Kerr effect and degenerate four-wave mixing were explained with special examples. At last, the application prospect of measurement techniques of femtosecond spectroscopy was forecasted.

  7. Common Tuina Techniques in Children

    Institute of Scientific and Technical Information of China (English)

    Editor

    2004-01-01

    @@ Tuina techniques in children are similar to those in adults. Some are same in name, but different in methods,such as pushing technique. Some techniques are just applied to children rather than to adults, such as pushing method. In clinical practice, such intense Tuina techniques as nailing, grasping, and pinching are practiced as the ending manipulations, so as not to hurt children and affect treatment. Such media as ginger juice,Talcum powder and egg white are commonly used to prevent from skin abrasion and improve therapeutic effects.

  8. Radar interferometry persistent scatterer technique

    CERN Document Server

    Kampes, Bert M

    2006-01-01

    Only book on Permanent Scatterer technique of radar interferometryExplains the Permanent Scatterer technique in detail, possible pitfalls, and details a newly developed stochastic model and estimator algorithm to cope with possible problems for the application of the PS techniqueThe use of Permanent Scatterer allows very precise measurements of the displacement of hundreds of points per square kilometerDescribes the only technique currently able to perform displacement measurements in the past, utilizing the ERS satellite data archive using data acquired from 1992-prese

  9. Helium II level measurement techniques

    Science.gov (United States)

    Celik, D.; Hilton, D. K.; Zhang, T.; Van Sciver, S. W.

    2001-05-01

    In this paper, a survey of cryogenic liquid level measurement techniques applicable to superfluid helium (He II) is given. The survey includes both continuous and discrete measurement techniques. A number of different probes and controlling circuits for this purpose have been described in the literature. They fall into one of the following categories: capacitive liquid level gauges, superconducting wire liquid level gauges, thermodynamic (heat transfer-based) liquid level gauges, resistive gauges, ultrasound and transmission line-based level detectors. The present paper reviews these techniques and their suitability for He II service. In addition to these methods, techniques for measuring the total liquid volume and mass gauging are also discussed.

  10. Optimization techniques for Transportation Problems

    Directory of Open Access Journals (Sweden)

    Gauthaman.P

    2017-06-01

    Full Text Available This paper infers about optimization technique for various problems in transportation engineering. While for pavement engineering, maintenance is priority issue, for traffic it is signalling which is priority issue. Many optimization methods are discussed though given importance of genetic algorithm approach. While optimization techniques nearly approach practicality, research works are on for modern optimization techniques which not only adds ease of structure but also provide compatibility to modern day problems encountered in transportation engineering. Some of the modern tools are discussed to employ optimization techniques which are quite simple to use and implement once it is calibrated to the desired objective.

  11. GPU Pro advanced rendering techniques

    CERN Document Server

    Engel, Wolfgang

    2010-01-01

    This book covers essential tools and techniques for programming the graphics processing unit. Brought to you by Wolfgang Engel and the same team of editors who made the ShaderX series a success, this volume covers advanced rendering techniques, engine design, GPGPU techniques, related mathematical techniques, and game postmortems. A special emphasis is placed on handheld programming to account for the increased importance of graphics on mobile devices, especially the iPhone and iPod touch.Example programs and source code can be downloaded from the book's CRC Press web page. 

  12. Optical techniques in regenerative medicine

    CERN Document Server

    Morgan, Stephen P

    2013-01-01

    In regenerative medicine, tissue engineers largely rely on destructive and time-consuming techniques that do not allow in situ and spatial monitoring of tissue growth. Furthermore, once the therapy is implanted in the patient, clinicians are often unable to monitor what is happening in the body. To tackle these barriers, optical techniques have been developed to image and characterize many tissue properties, fabricate tissue engineering scaffolds, and characterize the properties of the scaffolds. Optical Techniques in Regenerative Medicine illustrates how to use optical imaging techniques and

  13. Developing Fighting Technique Through Visualization

    Directory of Open Access Journals (Sweden)

    Tim Lajcik

    2012-07-01

    Full Text Available Visualization is a training technique that involves creating a detailed mental “movie” of successful performance. This article describes a type of visualization called “mental rehearsal” and explains how it can be used to reinforce the neuromuscular pattern of proper fighting technique. Drawing on his experience as a professional fighter and college coach, his studies in sport psychology as a college student, and his exposure to mental training techniques at the U.S. Olympic Training Center, the author reveals how to use mental imagery to facilitate the mastery of martial art technique.    

  14. Comparison of Composite Restoration Techniques

    Directory of Open Access Journals (Sweden)

    András Katona

    2016-01-01

    Full Text Available This article briefly presents the classification, and possible ways of application of composites. Our objective was to compare three different restoration technique in class II cavity preparations, with the aim to determine which technique is the most advantageous in practical point of view. Artificial teeth fabricated from self-curing acrylic resin were used as models. Comparison of marginal sealing was performed macroscopically via direct visual inspection and via tactile control. Two parameters were examined on esthetic evaluation, which were the anatomical shape and the optical appearance. The time required to prepare the restorations were also measured. After the evaluation of the results, the bulk-fill technique was demonstrated to require the shortest time to be performed. This was followed by the oblique (Z technique, whereas the horizontal incremental technique appeared to be the most time-consuming method. Based on the esthetic evaluation, a remarkable difference can be observed between the bulk-fill technique and the other two incremental techniques, while the mean scores for the oblique and the horizontal techniques were comparable. Based on the results of the visual inspection and instrumental examination, there were no differences in terms of marginal sealing between the three applied methods. Based on these, the use of the appropriate incremental technique can reduce the development of secondary caries and increasing the longevity of composite restorations.

  15. Parachute technique for partial penectomy

    Directory of Open Access Journals (Sweden)

    Fernando Korkes

    2010-04-01

    Full Text Available PURPOSE: Penile carcinoma is a rare but mutilating malignancy. In this context, partial penectomy is the most commonly applied approach for best oncological results. We herein propose a simple modification of the classic technique of partial penectomy, for better cosmetic and functional results. TECHNIQUE: If partial penectomy is indicated, the present technique can bring additional benefits. Different from classical technique, the urethra is spatulated only ventrally. An inverted "V" skin flap with 0.5 cm of extension is sectioned ventrally. The suture is performed with vicryl 4-0 in a "parachute" fashion, beginning from the ventral portion of the urethra and the "V" flap, followed by the "V" flap angles and than by the dorsal portion of the penis. After completion of the suture, a Foley catheter and light dressing are placed for 24 hours. CONCLUSIONS: Several complex reconstructive techniques have been previously proposed, but normally require specific surgical abilities, adequate patient selection and staged procedures. We believe that these reconstructive techniques are very useful in some specific subsets of patients. However, the technique herein proposed is a simple alternative that can be applied to all men after a partial penectomy, and takes the same amount of time as that in the classic technique. In conclusion, the "parachute" technique for penile reconstruction after partial amputation not only improves the appearance of the penis, but also maintains an adequate function.

  16. A numerical grid generation technique

    NARCIS (Netherlands)

    Gilding, B.H.

    1988-01-01

    The paper describes a technique for the generation of boundary-fitted curvilinear coordinate systems for the numerical solution of partial differential equations in two space dimensions. The technique is algebraic, has a transfinite character, and is based on the blending of shearing transformations

  17. Instrumental Techniques in Archeological Research

    Science.gov (United States)

    1988-09-01

    leather, amber, and mummified material. Evidently this is preliminary work. Schwarcz , H. P., 1986, "ESR Dating of Tooth Enamel," Abstracts of the 1984...Radiocarbon dating Contact: H. P. Schwarcz Technique(s): ESR 79 Illinois State Geological Survey McCrone Associates Inc. State Geological Survey

  18. Crack and flip phacoemulsification technique.

    Science.gov (United States)

    Fine, I H; Maloney, W F; Dillman, D M

    1993-11-01

    The crack and flip phacoemulsification technique combines the advantages of circumferential division of the nucleus and nucleofactis techniques. As such, it adds safety and control to the procedure. We describe each of the surgical maneuvers, including machine settings, and explain the rationale for maneuvers and machine settings.

  19. Magnetic characterization techniques for nanomaterials

    CERN Document Server

    2017-01-01

    Sixth volume of a 40 volume series on nanoscience and nanotechnology, edited by the renowned scientist Challa S.S.R. Kumar. This handbook gives a comprehensive overview about Magnetic Characterization Techniques for Nanomaterials. Modern applications and state-of-the-art techniques are covered and make this volume an essential reading for research scientists in academia and industry.

  20. Software Agent Techniques in Design

    DEFF Research Database (Denmark)

    Hartvig, Susanne C

    1998-01-01

    This paper briefly presents studies of software agent techniques and outline aspects of these which can be applied in design agents in integrated civil engineering design environments.......This paper briefly presents studies of software agent techniques and outline aspects of these which can be applied in design agents in integrated civil engineering design environments....

  1. Field techniques for sampling ants

    Science.gov (United States)

    Ants occur in most environments and ecologists ask a diverse array of questions involving ants. Thus, a key consideration in ant studies is to match the environment and question (and associated environmental variables) to the ant sampling technique. Since each technique has distinct limitations, usi...

  2. Passage Retrieval: A Probabilistic Technique.

    Science.gov (United States)

    Melucci, Massimo

    1998-01-01

    Presents a probabilistic technique to retrieve passages from texts having a large size or heterogeneous semantic content. Results of experiments comparing the probabilistic technique to one based on a text segmentation algorithm revealed that the passage size affects passage retrieval performance; text organization and query generality may have an…

  3. A disposition of interpolation techniques

    NARCIS (Netherlands)

    Knotters, M.; Heuvelink, G.B.M.

    2010-01-01

    A large collection of interpolation techniques is available for application in environmental research. To help environmental scientists in choosing an appropriate technique a disposition is made, based on 1) applicability in space, time and space-time, 2) quantification of accuracy of interpolated v

  4. A disposition of interpolation techniques

    NARCIS (Netherlands)

    Knotters, M.; Heuvelink, G.B.M.

    2010-01-01

    A large collection of interpolation techniques is available for application in environmental research. To help environmental scientists in choosing an appropriate technique a disposition is made, based on 1) applicability in space, time and space-time, 2) quantification of accuracy of interpolated v

  5. Photogrammetric techniques for aerospace applications

    Science.gov (United States)

    Liu, Tianshu; Burner, Alpheus W.; Jones, Thomas W.; Barrows, Danny A.

    2012-10-01

    Photogrammetric techniques have been used for measuring the important physical quantities in both ground and flight testing including aeroelastic deformation, attitude, position, shape and dynamics of objects such as wind tunnel models, flight vehicles, rotating blades and large space structures. The distinct advantage of photogrammetric measurement is that it is a non-contact, global measurement technique. Although the general principles of photogrammetry are well known particularly in topographic and aerial survey, photogrammetric techniques require special adaptation for aerospace applications. This review provides a comprehensive and systematic summary of photogrammetric techniques for aerospace applications based on diverse sources. It is useful mainly for aerospace engineers who want to use photogrammetric techniques, but it also gives a general introduction for photogrammetrists and computer vision scientists to new applications.

  6. Single Cell Electrical Characterization Techniques.

    Science.gov (United States)

    Mansor, Muhammad Asraf; Ahmad, Mohd Ridzuan

    2015-06-04

    Electrical properties of living cells have been proven to play significant roles in understanding of various biological activities including disease progression both at the cellular and molecular levels. Since two decades ago, many researchers have developed tools to analyze the cell's electrical states especially in single cell analysis (SCA). In depth analysis and more fully described activities of cell differentiation and cancer can only be accomplished with single cell analysis. This growing interest was supported by the emergence of various microfluidic techniques to fulfill high precisions screening, reduced equipment cost and low analysis time for characterization of the single cell's electrical properties, as compared to classical bulky technique. This paper presents a historical review of single cell electrical properties analysis development from classical techniques to recent advances in microfluidic techniques. Technical details of the different microfluidic techniques are highlighted, and the advantages and limitations of various microfluidic devices are discussed.

  7. Single Cell Electrical Characterization Techniques

    Directory of Open Access Journals (Sweden)

    Muhammad Asraf Mansor

    2015-06-01

    Full Text Available Electrical properties of living cells have been proven to play significant roles in understanding of various biological activities including disease progression both at the cellular and molecular levels. Since two decades ago, many researchers have developed tools to analyze the cell’s electrical states especially in single cell analysis (SCA. In depth analysis and more fully described activities of cell differentiation and cancer can only be accomplished with single cell analysis. This growing interest was supported by the emergence of various microfluidic techniques to fulfill high precisions screening, reduced equipment cost and low analysis time for characterization of the single cell’s electrical properties, as compared to classical bulky technique. This paper presents a historical review of single cell electrical properties analysis development from classical techniques to recent advances in microfluidic techniques. Technical details of the different microfluidic techniques are highlighted, and the advantages and limitations of various microfluidic devices are discussed.

  8. Strongly correlated systems experimental techniques

    CERN Document Server

    Mancini, Ferdinando

    2015-01-01

    The continuous evolution and development of experimental techniques is at the basis of any fundamental achievement in modern physics. Strongly correlated systems (SCS), more than any other, need to be investigated through the greatest variety of experimental techniques in order to unveil and crosscheck the numerous and puzzling anomalous behaviors characterizing them. The study of SCS fostered the improvement of many old experimental techniques, but also the advent of many new ones just invented in order to analyze the complex behaviors of these systems. Many novel materials, with functional properties emerging from macroscopic quantum behaviors at the frontier of modern research in physics, chemistry and materials science, belong to this class of systems. The volume presents a representative collection of the modern experimental techniques specifically tailored for the analysis of strongly correlated systems. Any technique is presented in great detail by its own inventor or by one of the world-wide recognize...

  9. Comparing techniques for pterygium surgery

    Directory of Open Access Journals (Sweden)

    Atilla Alpay

    2008-09-01

    Full Text Available Atilla Alpay, Suat Hayri Ugurbas, Berktug ErdoganDepartment of Ophthalmology, Faculty of Medicine, Zonguldak, Karaelmas University, Zonguldak, TurkeyPurpose: To compare various techniques of pterygium surgery including bare sclera, intraoperative mitomycin C application, conjunctival flap reconstruction, and conjunctival autografting technique.Methods: This study is designed to compare 4 currently used techniques in order to determine the complication and recurrence rates after pterygium exision. Included in the study were 77 eyes from 60 patients. Bare sclera technique was used to treat 21 primary pterygia; mitomycin C was used to treat 20 (16 primary, 4 recurrent pterygia; 18 (17 primary, 1 recurrent pterygia were treated by conjuntival flap reconstruction; and 18 (9 primary, 9 recurrent pterygia were treated by conjunctival autografting technique. All patients who underwent surgery were followed up for between 6 months and 2 years.Results: Eight recurrences (38.09% were observed in the bare sclera group whereas there were 5 (25% recurrences in the mitomycin C group. In the conjunctival flap reconstruction group, 6 (33.33% recurrences were detected. In the conjunctival autografting group, 3 recurrences were  observed. There were no major complications threatening visual ability in the surgical patients.Conclusion: A comparison of the groups demonstrated that the recurrence rate was highest in the bare sclera group, and lowest in conjunctival autografting and mitomycin C treatment groups respectively. Although the conjunctival autografting technique is a more difficult and time consuming technique than the others, cosmetic and surgical results were found to be superior. We advise conjunctival autografting for the treatment of pterygium in view of the high recurrence rates of other techniques, and the possible complications of mitomycin C treatment for benign disease.Keywords: pterygium, comparing, techniques, mitomycin C

  10. Nuclear techniques in analytical chemistry

    CERN Document Server

    Moses, Alfred J; Gordon, L

    1964-01-01

    Nuclear Techniques in Analytical Chemistry discusses highly sensitive nuclear techniques that determine the micro- and macro-amounts or trace elements of materials. With the increasingly frequent demand for the chemical determination of trace amounts of elements in materials, the analytical chemist had to search for more sensitive methods of analysis. This book accustoms analytical chemists with nuclear techniques that possess the desired sensitivity and applicability at trace levels. The topics covered include safe handling of radioactivity; measurement of natural radioactivity; and neutron a

  11. PIGE technique implementation at ININ

    Energy Technology Data Exchange (ETDEWEB)

    Policroniades, R., E-mail: rafael.policroniades@inin.gob.mx; Martínez-Quiroz, E.; Méndez-Garrido, B.; Murillo, G.; Moreno, E.; Villaseñor, P. [Departamento de Aceleradores, Instituto Nacional de Investigaciones Nucleares (ININ), Carr. México-Toluca S/N, La Marquesa, Ocoyoacac, México CP 52750 (Mexico)

    2015-07-23

    In this work, we present a general overview about the implementation at ININ of a Particle Induced Gamma Emission (PIGE) analysis technique, based on the bombardment of samples by protons and deuterons at different energies within our tandem accelerator facility. As it is well known, this technique is based on the detection of γ-rays emitted by nuclei in a target following a charged particle irradiation. The main feature of this technique, apart from being non-destructive and low time consuming, is that it allows a multi-elemental analysis of a sample, permitting an isotopic identification of many nuclides. Advances and some preliminary results are presented.

  12. Low Light Level TV Techniques.

    Science.gov (United States)

    Gildea, J

    1970-10-01

    As the science of low light level sensing becomes better understood, the demand for systems with this capability has increased considerably in recent years. Low light level television systems are part of these low light sensing devices in which interest has grown. Development of low light level TV systems has, in turn, stimulated technical advances in new tube types with improved performance, development of electronic techniques which enhance the over-all performance, and design techniques which make the system more versatile and adaptable. A general look at some of these developments and techniques gives insight into the versatility and adaptability of low light level TV.

  13. A simplified indirect bonding technique

    Directory of Open Access Journals (Sweden)

    Radha Katiyar

    2014-01-01

    Full Text Available With the advent of lingual orthodontics, indirect bonding technique has become an integral part of practice. It involves placement of brackets initially on the models and then their transfer to teeth with the help of transfer trays. Problems encountered with current indirect bonding techniques used are (1 the possibility of adhesive flash remaining around the base of the brackets which requires removal (2 longer time required for the adhesive to gain enough bond strength for secure tray removal. The new simplified indirect bonding technique presented here overcomes both these problems.

  14. PIGE technique implementation at ININ

    Science.gov (United States)

    Policroniades, R.; Martínez-Quiroz, E.; Méndez-Garrido, B.; Murillo, G.; Moreno, E.; Villaseñor, P.

    2015-07-01

    In this work, we present a general overview about the implementation at ININ of a Particle Induced Gamma Emission (PIGE) analysis technique, based on the bombardment of samples by protons and deuterons at different energies within our tandem accelerator facility. As it is well known, this technique is based on the detection of γ-rays emitted by nuclei in a target following a charged particle irradiation. The main feature of this technique, apart from being non-destructive and low time consuming, is that it allows a multi-elemental analysis of a sample, permitting an isotopic identification of many nuclides. Advances and some preliminary results are presented.

  15. Scalable Techniques for Formal Verification

    CERN Document Server

    Ray, Sandip

    2010-01-01

    This book presents state-of-the-art approaches to formal verification techniques to seamlessly integrate different formal verification methods within a single logical foundation. It should benefit researchers and practitioners looking to get a broad overview of the spectrum of formal verification techniques, as well as approaches to combining such techniques within a single framework. Coverage includes a range of case studies showing how such combination is fruitful in developing a scalable verification methodology for industrial designs. This book outlines both theoretical and practical issue

  16. New techniques on embryo manipulation.

    Science.gov (United States)

    Escribá, M J; Valbuena, D; Remohí, J; Pellicer, A; Simón, C

    2002-01-01

    For many years, experience has been accumulated on embryo and gamete manipulation in livestock animals. The present work is a review of these techniques and their possible application in human embryology in specific cases. It is possible to manipulate gametes at different levels, producing paternal or maternal haploid embryos (hemicloning), using different techniques including nuclear transfer. At the embryonic stage, considering practical, ethical and legal issues, techniques will be reviewed that include cloning and embryo splitting at the cleavage stage, morula, or blastocyst stage.

  17. Artificial intelligence techniques in Prolog

    CERN Document Server

    Shoham, Yoav

    1993-01-01

    Artificial Intelligence Techniques in Prolog introduces the reader to the use of well-established algorithmic techniques in the field of artificial intelligence (AI), with Prolog as the implementation language. The techniques considered cover general areas such as search, rule-based systems, and truth maintenance, as well as constraint satisfaction and uncertainty management. Specific application domains such as temporal reasoning, machine learning, and natural language are also discussed.Comprised of 10 chapters, this book begins with an overview of Prolog, paying particular attention to Prol

  18. Statistical Techniques for Project Control

    CERN Document Server

    Badiru, Adedeji B

    2012-01-01

    A project can be simple or complex. In each case, proven project management processes must be followed. In all cases of project management implementation, control must be exercised in order to assure that project objectives are achieved. Statistical Techniques for Project Control seamlessly integrates qualitative and quantitative tools and techniques for project control. It fills the void that exists in the application of statistical techniques to project control. The book begins by defining the fundamentals of project management then explores how to temper quantitative analysis with qualitati

  19. Flow-cytometric method for observing the effects of pulsed electromagnetic fields on the growth of rat bone marrow mesenchymal stem cells%流式细胞仪观察脉冲电磁场干预骨髓间充质干细胞的生长

    Institute of Scientific and Technical Information of China (English)

    黄钊; 苏伟; 崔向荣; 覃万安

    2011-01-01

    BACKGROUND: Compared with the morphology, DNA electrophoresis and other methods, flow-cytometric method has more advantages on the detection of cell phenotype, proliferation rate and cell cycle of rat bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE: To observe and discuss the effect of specific pulsed electromagnetic fields stimulation on the proliferation and cell cycle of rat BMSCs with the flow-cytometric method (FCM). METHODS: Rat BMSCs were exposed to pulsed electromagnetic fields (frequency for 1 kHz, magnetic flux density for 0.05 mT, power density for 5 mW/cm2). BMSCs without exposure to pulsed electromagnetic fields were used as controls. Expression of CD29, CD31, CD44 CD45, CD105 were detect in the control group. The proliferation rate and cell cycle of passage 3 cells were detected at 3, 6, 9, 12 days. RESULTS AND CONCLUSION: CD29, CD44, CD105 in the 3rd passage non-stimulated BMSCs was positively expressed and the CD31, CD45 was negatively expressed (P < 0.05). The survival rate of passage 3 BMSCs and percentage of S phase following intervention of pulsed electromagnetic fields were greater than those in the control group (P < 0.05). These results indicated that the specific pulsed electromagnetic fields can promote the growth and proliferation of BMSCs to some extent.%背景:应用流式细胞仪检测细胞表型、存活或凋亡细胞计数及细胞周期,较形态学观察、DNA电泳等检测方法更具优势.目的:采用流式细胞仪分选检测特定脉冲电磁场干预对大鼠骨髓间充质干细胞生长周期及其增殖效率的影响.方法:使用振荡频率1 kHz,磁感应强度0.05 mT、功率密度5 mW/cm2的脉冲电磁场照射大鼠骨髓间充质干细胞,以未经磁场干预的细胞为对照.采用流式细胞仪检测未经磁场干预细胞表面抗原CD29、CD31、CD44、CD45和CD105表达率;于传第3代后第3,6,9,12天测定不同干预细胞增殖率及生长周期.结果与结论:未经磁场干预的第3代

  20. Analysis of the Fine-Scale Population Structure of “Candidatus Accumulibacter phosphatis” in Enhanced Biological Phosphorus Removal Sludge, Using Fluorescence In Situ Hybridization and Flow Cytometric Sorting▿

    Science.gov (United States)

    Kim, Jeong Myeong; Lee, Hyo Jung; Kim, Sun Young; Song, Jae Jun; Park, Woojun; Jeon, Che Ok

    2010-01-01

    To investigate the fine-scale diversity of the polyphosphate-accumulating organisms (PAO) “Candidatus Accumulibacter phosphatis” (henceforth referred to as “Ca. Accumulibacter”), two laboratory-scale sequencing batch reactors (SBRs) for enhanced biological phosphorus removal (EBPR) were operated with sodium acetate as the sole carbon source. During SBR operations, activated sludge always contained morphologically different “Ca. Accumulibacter” strains showing typical EBPR performances, as confirmed by the combined technique of fluorescence in situ hybridization (FISH) and microautoradiography (MAR). Fragments of “Ca. Accumulibacter” 16S rRNA genes were retrieved from the sludge. Phylogenetic analyses together with sequences from the GenBank database showed that “Ca. Accumulibacter” 16S rRNA genes of the EBPR sludge were clearly differentiated into four “Ca. Accumulibacter” clades, Acc-SG1, Acc-SG2, Acc-SG3, and Acc-SG4. The specific FISH probes Acc444, Acc184, Acc72, and Acc119 targeting these clades and some helpers and competitors were designed by using the ARB program. Microbial characterization by FISH analysis using specific FISH probes also clearly indicated the presence of different “Ca. Accumulibacter” cell morphotypes. Especially, members of Acc-SG3, targeted by probe Acc72, were coccobacillus-shaped cells with a size of approximately 2 to 3 μm, while members of Acc-SG1, Acc-SG2, and Acc-SG4, targeted by Acc444, Acc184, and Acc119, respectively, were coccus-shaped cells approximately 1 μm in size. Subsequently, cells targeted by each FISH probe were sorted by use of a flow cytometer, and their polyphosphate kinase 1 (ppk1) gene homologs were amplified by using a ppk1-specific PCR primer set for “Ca. Accumulibacter.” The phylogenetic tree based on sequences of the ppk1 gene homologs was basically congruent with that of the 16S rRNA genes, but members of Acc-SG3 with a distinct morphology comprised two different ppk1 genes

  1. Evaluation of the probe dihydrocalcein acetoxymethylester as an indicator of reactive oxygen species formation and comparison with oxidative DNA base modification determined by modified alkaline elution technique.

    Science.gov (United States)

    Rohnstock, A; Lehmann, L

    2007-12-01

    Reactive oxygen species (ROS) play a predominant role in various diseases and the development of fast and easy methods for the quantification of intracellular ROS represents an important goal. Therefore, the aim of the present study was the evaluation of the fluorogenic probe dihydrocalcein acetoxymethylester (AM) for the detection of intracellular ROS. A flow cytometric method was developed using MCF-7 cells and the kinetics of ester hydrolysis and the cellular distribution and stability of calcein were characterized using calcein AM. Then, MCF-7 cells were challenged with model agents for the generation of singlet oxygen (illumination with visible light), peroxyl and hydroxyl radicals (tert-butylhydroperoxide, tBHP), superoxide anion radicals (potassium dioxide), and the intracellular formation of superoxide anion radicals by redox cycling (menadione) and the formation of calcein was compared with the induction of oxidative DNA base modifications assessed by modified alkaline elution technique. Every model agent significantly induced formamidopyrimidine-DNA glycosylase-sensitive sites (i.e. oxidative DNA base modifications) and most also induced DNA strand breaks. In contrast, exclusively tBHP and illumination with visible light induced the intracellular formation of calcein. In conclusion, though intracellular oxidation of dihydrocalcein represents a fast screening method, it detects a limited spectrum of ROS.

  2. and use of barrier techniques

    African Journals Online (AJOL)

    Attitudes of Nigerian dentists towards hepatitis B vaccination and use of barrier techniques .... tine screening of only high-risk patients has been recommended.“ .... i337-1342. Sote EO. AIDS and infection Control: experiences, attitudes,.

  3. Modern techniques of surface science

    CERN Document Server

    Woodruff, D Phil

    2016-01-01

    This fully revised, updated and reorganised third edition provides a thorough introduction to the characterisation techniques used in surface science and nanoscience today. Each chapter brings together and compares the different techniques used to address a particular research question, including how to determine the surface composition, surface structure, surface electronic structure, surface microstructure at different length scales (down to sub-molecular), and the molecular character of adsorbates and their adsorption or reaction properties. Readers will easily understand the relative strengths and limitations of the techniques available to them and, ultimately, will be able to select the most suitable techniques for their own particular research purposes. This is an essential resource for researchers and practitioners performing materials analysis, and for senior undergraduate students looking to gain a clear understanding of the underlying principles and applications of the different characterisation tec...

  4. Survey of semantic modeling techniques

    Energy Technology Data Exchange (ETDEWEB)

    Smith, C.L.

    1975-07-01

    The analysis of the semantics of programing languages was attempted with numerous modeling techniques. By providing a brief survey of these techniques together with an analysis of their applicability for answering semantic issues, this report attempts to illuminate the state-of-the-art in this area. The intent is to be illustrative rather than thorough in the coverage of semantic models. A bibliography is included for the reader who is interested in pursuing this area of research in more detail.

  5. Simple techniques can increase motivation

    Energy Technology Data Exchange (ETDEWEB)

    Smith, E.A.

    1982-12-01

    Productivity depends upon strong motivation. This article describes how the symptoms of weak motivation can be detected among engineers and other employees and what to do about it. Fortunately, most people want to be productive and respond to simple motivation building techniques. These techniques include establishing twoway communication between supervisor and subordinate, delegating authority on the basis of responsibility, effective organization of time, and the use of positive reinforcement (or rewards) and, when conditions merit it, negative reinforcement (or sanctions).

  6. Swabs and other sampling techniques.

    Science.gov (United States)

    Lawrence, J C; Ameen, H

    1998-05-01

    This study explores the absorptive capacity of standard cotton wool-tipped swabs in vitro and in vivo. An alternative wound exudate sampling technique using small filter paper discs is also briefly examined. Results showed a poor uniformity of fluid absorption by swabs and a limited correlation of wound exudate quantities in relation to visual estimates and size of wound. It was found, however, that swabs reliably removed material from a wound surface. The filter paper technique appeared to offer no advantages.

  7. Survey of semantic modeling techniques

    Energy Technology Data Exchange (ETDEWEB)

    Smith, C.L.

    1975-07-01

    The analysis of the semantics of programing languages was attempted with numerous modeling techniques. By providing a brief survey of these techniques together with an analysis of their applicability for answering semantic issues, this report attempts to illuminate the state-of-the-art in this area. The intent is to be illustrative rather than thorough in the coverage of semantic models. A bibliography is included for the reader who is interested in pursuing this area of research in more detail.

  8. Comparing techniques for pterygium surgery.

    Science.gov (United States)

    Alpay, Atilla; Uğurbaş, Suat Hayri; Erdoğan, Berktuğ

    2009-01-01

    To compare various techniques of pterygium surgery including bare sclera, intraoperative mitomycin C application, conjunctival flap reconstruction, and conjunctival autografting technique. This study is designed to compare 4 currently used techniques in order to determine the complication and recurrence rates after pterygium exision. Included in the study were 77 eyes from 60 patients. Bare sclera technique was used to treat 21 primary pterygia;mitomycin C was used to treat 20 (16 primary, 4 recurrent) pterygia;18 (17 primary, 1 recurrent) pterygia were treated by conjuntival flap reconstruction;and 18 (9 primary, 9 recurrent) pterygia were treated by conjunctival autografting technique. All patients who underwent surgery were followed up for between 6 months and 2 years. Eight recurrences (38.09%) were observed in the bare sclera group whereas there were 5 (25%) recurrences in the mitomycin C group. In the conjunctival flap reconstruction group, 6 (33.33%) recurrences were detected. In the conjunctival autografting group, 3 recurrences were observed. There were no major complications threatening visual ability in the surgical patients. A comparison of the groups demonstrated that the recurrence rate was highest in the bare sclera group, and lowest in conjunctival autografting and mitomycin C treatment groups respectively. Although the conjunctival autografting technique is a more difficult and time consuming technique than the others, cosmetic and surgical results were found to be superior. We advise conjunctival autografting for the treatment of pterygium in view of the high recurrence rates of other techniques, and the possible complications of mitomycin C treatment for benign disease.

  9. RECAT - Redundant Channel Alignment Technique

    Science.gov (United States)

    2016-06-07

    distribution unlimited 13. SUPPLEMENTARY NOTES NUWC2015 14. ABSTRACT A problem in the analog-to- digital , (A/D), conversion of broadband tape recorded...Alignment Technique, is used to align data taken on one pass with data from any other pass. The accuracy of this alignment is a function of the digital ...Redundant Channel Alignment Technique; analog-to- digital ; A/D; Broadband Bearing Time Processing 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF

  10. Discovering the Botnet Detection Techniques

    Science.gov (United States)

    Rahim, Aneel; Bin Muhaya, Fahad T.

    Botnet is a network of compromised computers. It just fellow the master slave concept. Bots are comprised computers and do the tasks what ever their master orders them. Internet Relay Chat (IRC) is used for the communication between the master and bots. Information is also encrypted to avoid the effect of third party. In this paper we discuss the Botnets detection techniques and comparative analysis of these techniques on the basis of DNS query, History data and group activity.

  11. Teaching Speaking Through Debate Technique

    Directory of Open Access Journals (Sweden)

    . Suranto

    2016-07-01

    Full Text Available Abstract : Teaching Speaking Through Debate Technique. Speaking is one of the basic competence from the other fourth basic competence (listening, speaking, reading and writing. Speaking ability should be mastered by every students, in order to achieve that competence students should be given the right technique to study sepaking. The successfull of the students speaking can be seen from their ability to express idea, thought and feeling through speaking. The objective of this Action Research is to improve students’s oral communication skill through the debate technique. This study was conducted at MA Ma’arif Nu 5 Sekampung Lampung Timur from March to April 2014. The research data were taken from students in the eleventh class, with 28 students and analyzed qualitatively and quantitatively. The research findings indicate that there are improvements in students’ english speaking skill through the debate technique. By analyzing data qualitatively and quantitatively from the end of the first cycle to the second cycle and it was found that the students’ English speaking skill increased 20,9% over the standard that has been determined by the researcher that is 65%. The researcher concludes that the students’ english speaking skill can be improve through the debate technique in learning process.   Key words : action research, debate technique, english speaking skill

  12. Indirect bonding technique in orthodontics

    Directory of Open Access Journals (Sweden)

    Kübra Yıldırım

    2016-08-01

    Full Text Available ‘Direct Bonding Technique’ which allows the fixed orthodontic appliances to be directly bonded to teeth without using bands decreased the clinic time for bracket bonding and increased esthetics and oral hygiene during orthodontic treatment. However, mistakes in bracket positioning were observed due to decreased direct visual sight and access to posterior teeth. ‘Indirect Bonding Technique’ was developed for eliminating these problems. Initially, decreased bond strength, higher bond failure rate, periodontal tissue irritation, compromised oral hygiene and increased laboratory time were the main disadvantages of this technique when compared to direct bonding. The newly developed materials and modified techniques help to eliminate these negative consequences. Today, the brackets bonded with indirect technique have similar bond strength with brackets bonded directly. Moreover, indirect and direct bonding techniques have similar effects on periodontal tissues. However, indirect bonding technique requires more attention and precision in laboratory and clinical stage, and has higher cost. Orthodontist's preference between these two bonding techniques may differ according to time spent in laboratory and clinic, cost, patient comfort and personal opinion.

  13. Review of advanced imaging techniques

    Directory of Open Access Journals (Sweden)

    Yu Chen

    2012-01-01

    Full Text Available Pathology informatics encompasses digital imaging and related applications. Several specialized microscopy techniques have emerged which permit the acquisition of digital images ("optical biopsies" at high resolution. Coupled with fiber-optic and micro-optic components, some of these imaging techniques (e.g., optical coherence tomography are now integrated with a wide range of imaging devices such as endoscopes, laparoscopes, catheters, and needles that enable imaging inside the body. These advanced imaging modalities have exciting diagnostic potential and introduce new opportunities in pathology. Therefore, it is important that pathology informaticists understand these advanced imaging techniques and the impact they have on pathology. This paper reviews several recently developed microscopic techniques, including diffraction-limited methods (e.g., confocal microscopy, 2-photon microscopy, 4Pi microscopy, and spatially modulated illumination microscopy and subdiffraction techniques (e.g., photoactivated localization microscopy, stochastic optical reconstruction microscopy, and stimulated emission depletion microscopy. This article serves as a primer for pathology informaticists, highlighting the fundamentals and applications of advanced optical imaging techniques.

  14. [Current techniques in tonsil surgery].

    Science.gov (United States)

    Coromina Isern, Jordi; Esteller Moré, Eduard

    2010-12-01

    In recent years, consolidation of tonsillar hypertrophy as the principal surgical procedure has led to the emergence of new techniques. Most aim to reduce volume (tonsillectomy or tonsil reduction). These techniques have considerably decreased intra- and postoperative hemorrhages and pain intensity. The present article describes the mechanisms and the advantages and disadvantages of the various techniques, including electro-dissection using electrical scalpels, reduction using a microdebrider, ultrasonic scalpel, radiofrequency (with its different variations) and CO(2) laser. When techniques that reduce tonsil volume are used, the possibility of recurrence of the tonsillar hypertrophy is high if less than 85% of the tonsil is removed. There is also a considerable possibility of infection of the remaining tonsils, whichever technique is used, and therefore these techniques are not valid in the case of repetitive tonsillitis. Recently, alternatives to classical adenoidectomy using adenoid curette have also appeared. Bleeding can be minimized by using a microdebrider, radiofrequency or a blood coagulator. We also discuss the concept of partial adenoidectomy, which is preferred in patients at risk of velopharyngeal insufficiency.

  15. Genetic techniques for the archaea.

    Science.gov (United States)

    Farkas, Joel A; Picking, Jonathan W; Santangelo, Thomas J

    2013-01-01

    Genetic techniques for the Archaea have undergone a rapid expansion in complexity, resulting in increased exploration of the role of Archaea in the environment and detailed analyses of the molecular physiology and information-processing systems in the third domain of life. Complementary gains in describing the ever-increasing diversity of archaeal organisms have allowed these techniques to be leveraged in new and imaginative ways to elucidate shared and unique aspects of archaeal diversity and metabolism. In this review, we introduce the four archaeal clades for which advanced genetic techniques are available--the methanogens, halophiles, Sulfolobales, and Thermococcales--with the aim of providing an overall profile of the advantages and disadvantages of working within each clade, as essentially all of the genetically accessible archaeal organisms require unique culturing techniques that present real challenges. We discuss the full repertoire of techniques possible within these clades while highlighting the recent advances that have been made by taking advantage of the most prominent techniques and approaches.

  16. Microscopy techniques in flavivirus research.

    Science.gov (United States)

    Chong, Mun Keat; Chua, Anthony Jin Shun; Tan, Terence Tze Tong; Tan, Suat Hoon; Ng, Mah Lee

    2014-04-01

    The Flavivirus genus is composed of many medically important viruses that cause high morbidity and mortality, which include Dengue and West Nile viruses. Various molecular and biochemical techniques have been developed in the endeavour to study flaviviruses. However, microscopy techniques still have irreplaceable roles in the identification of novel virus pathogens and characterization of morphological changes in virus-infected cells. Fluorescence microscopy contributes greatly in understanding the fundamental viral protein localizations and virus-host protein interactions during infection. Electron microscopy remains the gold standard for visualizing ultra-structural features of virus particles and infected cells. New imaging techniques and combinatory applications are continuously being developed to push the limit of resolution and extract more quantitative data. Currently, correlative live cell imaging and high resolution three-dimensional imaging have already been achieved through the tandem use of optical and electron microscopy in analyzing biological specimens. Microscopy techniques are also used to measure protein binding affinities and determine the mobility pattern of proteins in cells. This chapter will consolidate on the applications of various well-established microscopy techniques in flavivirus research, and discuss how recently developed microscopy techniques can potentially help advance our understanding in these membrane viruses.

  17. Nontraditional manufacturing technique-Nano machining technique based on SPM

    Institute of Scientific and Technical Information of China (English)

    DONG; Shen; YAN; Yongda; SUN; Tao; LIANG; Yingchun; CHENG

    2004-01-01

    Nano machining based on SPM is a novel, nontraditional advanced manufacturing technique. There are three main machining methods based on SPM, i.e.single atom manipulation, surface modification using physical or chemical actions and mechanical scratching. The current development of this technique is summarized. Based on the analysis of mechanical scratching mechanism, a 5 μm micro inflation hole is fabricated on the surface of inertial confinement fusion (ICF) target. The processing technique is optimized. The machining properties of brittle material, single crystal Ge, are investigated. A micro machining system combining SPM and a high accuracy stage is developed. Some 2D and 3D microstructures are fabricated using the system. This method has broad applications in the field of nano machining.

  18. Modified technique in treating recurrent priapism: a technique report

    Directory of Open Access Journals (Sweden)

    Wei Chen

    2015-04-01

    Full Text Available Recurrent ischemic priapism is a problem in clinical treatment. Most of the cases require more invasive surgery to shunt the blood stasis. We introduce a modified technique in treating recurrent ischemic priapism. The technique described is applied to acute ischaemic priapic episodes in patients with a history of stuttering priapism. It was carried out by a Winter's shunt combined with a continuous cavernosal irrigation system. Priapism was effectively resolved on the patients without recurrence. The four patients who received this treatment recovered most sexual function after 6 months follow-up.

  19. LANDING TECHNIQUES IN BEACH VOLLEYBALL

    Directory of Open Access Journals (Sweden)

    Markus Tilp

    2013-09-01

    Full Text Available The aims of the present study were to establish a detailed and representative record of landing techniques (two-, left-, and right-footed landings in professional beach volleyball and compare the data with those of indoor volleyball. Beach volleyball data was retrieved from videos taken at FIVB World Tour tournaments. Landing techniques were compared in the different beach and indoor volleyball skills serve, set, attack, and block with regard to sex, playing technique, and court position. Significant differences were observed between men and women in landings following block actions (χ²(2 = 18.19, p < 0.01 but not following serve, set, and attack actions. Following blocking, men landed more often on one foot than women. Further differences in landings following serve and attack with regard to playing technique and position were mainly observed in men. The comparison with landing techniques in indoor volleyball revealed overall differences both in men (χ²(2 = 161.4, p < 0.01 and women (χ²(2 = 84.91, p < 0.01. Beach volleyball players land more often on both feet than indoor volleyball players. Besides the softer surface in beach volleyball, and therefore resulting lower loads, these results might be another reason for fewer injuries and overuse conditions compared to indoor volleyball

  20. Landing Techniques in Beach Volleyball

    Science.gov (United States)

    Tilp, Markus; Rindler, Michael

    2013-01-01

    The aims of the present study were to establish a detailed and representative record of landing techniques (two-, left-, and right-footed landings) in professional beach volleyball and compare the data with those of indoor volleyball. Beach volleyball data was retrieved from videos taken at FIVB World Tour tournaments. Landing techniques were compared in the different beach and indoor volleyball skills serve, set, attack, and block with regard to sex, playing technique, and court position. Significant differences were observed between men and women in landings following block actions (χ2(2) = 18.19, p volleyball revealed overall differences both in men (χ2(2) = 161.4, p volleyball players land more often on both feet than indoor volleyball players. Besides the softer surface in beach volleyball, and therefore resulting lower loads, these results might be another reason for fewer injuries and overuse conditions compared to indoor volleyball. Key Points About 1/3 of all jumping actions in beach volleyball result in a landing on one foot. Especially following block situations men land on one foot more often than women. Landing techniques are related to different techniques and positions. Landings on one foot are less common in beach volleyball than indoor volleyball. This could be a reason for fewer injuries and overuse conditions. PMID:24149150

  1. fuzzy control technique fuzzy control technique applied to modified ...

    African Journals Online (AJOL)

    eobe

    ABSTRACT. In this paper, fuzzy control technique is applied to the modified mathematical model for malaria control presented ... be devised for rule-based systems that deals with continuous ... necessary to use fuzzy logic as it is not easy to follow a particular .... point movement and control is realized and designed. (e.g. α1 ...

  2. Twin - Arch technique. Revival of the "edgewise -Technique"?

    Directory of Open Access Journals (Sweden)

    Jacob Karp

    2012-01-01

    The SNB – Bracket brings a new dimension into the orthodontic world which is most apparent in extraction cases. Its Teflon – like material has a very low friction coefficient thus, reducing the treatment time considerably. Through the use of low dimensioned arch wires, the Twin – Arch Technique becomes a Light – wire system and simultaneously provides good anchorage and torque control.

  3. Four Flaps Technique for Neoumbilicoplasty

    Directory of Open Access Journals (Sweden)

    Young Taek Lee

    2015-05-01

    Full Text Available The absence or disfigurement of the umbilicus is both cosmetically and psychologically distressing to patients. The goal of aesthetically pleasing umbilical reconstruction is to create a neoumbilicus with sufficient depth and good morphology, with natural-looking superior hooding and minimal scarring. Although many reports have presented techniques for creating new and attractive umbilici, we developed a technique that we term the "four flaps technique" for creating a neoumbilicus in circumstances such as the congenital absence of the umbilicus or the lack of remaining umbilical tissue following the excision of a hypertrophic or scarred umbilicus. This method uses the neighboring tissue by simply elevating four flaps and can yield sufficient depth and an aesthetically pleasing shape with appropriate superior hooding.

  4. Infrared Devices And Techniques (Revision

    Directory of Open Access Journals (Sweden)

    Rogalski A.

    2014-12-01

    Full Text Available The main objective of this paper is to produce an applications-oriented review covering infrared techniques and devices. At the beginning infrared systems fundamentals are presented with emphasis on thermal emission, scene radiation and contrast, cooling techniques, and optics. Special attention is focused on night vision and thermal imaging concepts. Next section concentrates shortly on selected infrared systems and is arranged in order to increase complexity; from image intensifier systems, thermal imaging systems, to space-based systems. In this section are also described active and passive smart weapon seekers. Finally, other important infrared techniques and devices are shortly described, among them being: non-contact thermometers, radiometers, LIDAR, and infrared gas sensors.

  5. Evaluation of standard watermarking techniques

    Science.gov (United States)

    Marini, Enrico; Autrusseau, Florent; Le Callet, Patrick; Campisi, Patrizio

    2007-02-01

    In the last decade digital watermarking techniques have been devised to answer the ever-growing need to protect the intellectual property of digital still images, video sequences or audio from piracy attacks. Because of the proliferation of watermarking algorithms and their applications some benchmarks have been created in order to help watermarkers comparing their algorithms in terms of robustness against various attacks (i.e. Stirmark, Checkmark). However, no equal attention has been devoted to the proposition of benchmarks tailored to assess the watermark perceptual transparency. In this work, we study several watermarking techniques in terms of the mark invisibility through subjective experiments. Moreover, we test the ability of several objective metrics, used in the literature mainly to evaluate distortions due to the coding process, to be correlated with subjective scores. The conclusions drawn in the paper are supported by extensive experimentations using both several watermarking techniques and objective metrics.

  6. Lead extraction. Indications and techniques.

    Science.gov (United States)

    Byrd, C L; Schwartz, S J; Hedin, N

    1992-11-01

    Each of the extraction techniques and their ancillary tools was reported as used successfully; however, until now, no technique has been successful when used in more than a few isolated instances. The technique for intravascular countertraction and the associated tools described in this paper were devised and selected in an attempt to develop one technique to be used on all patients, with all types of leads, and with a very low complication rate. Its versatility permitted single or multiple lead extractions combined with the precision of selecting and extracting a specific lead. In our experience, as well as the experience of others, the techniques described in this paper have proved to be superior by minimizing the inherent risk and morbidity, allowing us to expand the indications for lead removal beyond septicemia and free-floating leads, to include infection, abandonment of pockets, and replacement of malfunctioning or fractured leads. Intravascular countertraction was a consistently safe and efficacious method of removing transvenous pacemaker leads regardless of the duration of the implant, thus permitting extractions in patients not considered candidates for a more extensive surgical procedure. Intravascular countertraction encompasses surgical and fluoroscopic techniques possessed by most physicians experienced in pacemaker and automatic implantable cardioverter defibrillator implants. However, there is a learning curve, predicating caution for the inexperienced physician. In addition, advanced surgical skills may be needed in handling associated conditions such as debridement and primary closure of chronically inflamed tissues, especially in submuscular pockets and sinus tracts in the neck. Although the potential for a cardiovascular complication is small, it does exist, and cardiovascular surgical backup is a recommended precaution.

  7. Resolution enhancement techniques in microscopy

    Science.gov (United States)

    Cremer, Christoph; Masters, Barry R.

    2013-05-01

    We survey the history of resolution enhancement techniques in microscopy and their impact on current research in biomedicine. Often these techniques are labeled superresolution, or enhanced resolution microscopy, or light-optical nanoscopy. First, we introduce the development of diffraction theory in its relation to enhanced resolution; then we explore the foundations of resolution as expounded by the astronomers and the physicists and describe the conditions for which they apply. Then we elucidate Ernst Abbe's theory of optical formation in the microscope, and its experimental verification and dissemination to the world wide microscope communities. Second, we describe and compare the early techniques that can enhance the resolution of the microscope. Third, we present the historical development of various techniques that substantially enhance the optical resolution of the light microscope. These enhanced resolution techniques in their modern form constitute an active area of research with seminal applications in biology and medicine. Our historical survey of the field of resolution enhancement uncovers many examples of reinvention, rediscovery, and independent invention and development of similar proposals, concepts, techniques, and instruments. Attribution of credit is therefore confounded by the fact that for understandable reasons authors stress the achievements from their own research groups and sometimes obfuscate their contributions and the prior art of others. In some cases, attribution of credit is also made more complex by the fact that long term developments are difficult to allocate to a specific individual because of the many mutual connections often existing between sometimes fiercely competing, sometimes strongly collaborating groups. Since applications in biology and medicine have been a major driving force in the development of resolution enhancing approaches, we focus on the contribution of enhanced resolution to these fields.

  8. CLEAN Technique for Polarimetric ISAR

    Directory of Open Access Journals (Sweden)

    M. Martorella

    2008-01-01

    Full Text Available Inverse synthetic aperture radar (ISAR images are often used for classifying and recognising targets. To reduce the amount of data processed by the classifier, scattering centres are extracted from the ISAR image and used for classifying and recognising targets. This paper addresses the problem of estimating the position and the scattering vector of target scattering centres from polarimetric ISAR images. The proposed technique is obtained by extending the CLEAN technique, which was introduced in radar imaging for extracting scattering centres from single-polarisation ISAR images. The effectiveness of the proposed algorithm, namely, the Polarimetric CLEAN (Pol-CLEAN is tested on simulated and real data.

  9. Computerized proof techniques for undergraduates

    Science.gov (United States)

    Smith, Christopher J.; Tefera, Akalu; Zeleke, Aklilu

    2012-12-01

    The use of computer algebra systems such as Maple and Mathematica is becoming increasingly important and widespread in mathematics learning, teaching and research. In this article, we present computerized proof techniques of Gosper, Wilf-Zeilberger and Zeilberger that can be used for enhancing the teaching and learning of topics in discrete mathematics. We demonstrate by examples how one can use these computerized proof techniques to raise students' interests in the discovery and proof of mathematical identities and enhance their problem-solving skills.

  10. Drilling techniques for osteochondritis dissecans.

    Science.gov (United States)

    Heyworth, Benton E; Edmonds, Eric W; Murnaghan, M Lucas; Kocher, Mininder S

    2014-04-01

    Although the advanced stages of osteochondritis dissecans remain challenging to treat, most early-stage lesions in skeletally immature patients, if managed appropriately, can be stimulated to heal. For stable lesions that do not demonstrate adequate healing with nonoperative measures, such as activity modification, weight-bearing protection, or bracing, drilling of the subchondral bone has emerged as the gold standard of management. Several techniques of drilling exist, including transarticular drilling, retroarticular drilling, and notch drilling. Although each technique has been shown to be effective in small retrospective studies, higher-powered prospective comparative studies are needed to better elucidate their relative advantages and disadvantages.

  11. New Techniques in Neutron Scattering

    DEFF Research Database (Denmark)

    Birk, Jonas Okkels

    . The instrument is ideally suited for solid state experiments with extreme sample environments such as large pressures and strong magnetic fields. CAMEA combines the time-of-flight technique to determine the energy of the incoming neutrons with a complex multiplex backend that will analyse and detect......Neutron scattering is an important experimental technique in amongst others solid state physics, biophysics, and engineering. This year construction of European Spallation Source (ESS) was commenced in Lund, Sweeden. The facility will use a new long pulsed source principle to obtain higher...

  12. Simple technique for facial dimple

    Directory of Open Access Journals (Sweden)

    Ahmed Hassan El-Sabbagh

    2015-01-01

    Full Text Available Background: Subjects seeking aesthetic surgery for facial dimples are increasing in number. Literature on dimple creation surgery are sparse. Various techniques have been used with their own merits and disadvantages. Materials and Methods: Facial dimples were created in 23 cases. All the subjects were females. Five cases were bilateral and the rest were unilateral. Results: Minor complications such as swelling and hematoma were observed in four cases. Infection occurred in two cases. Most of the subjects were satisfied with the results. Conclusions: Suturing technique is safe, reliable and an easily reproducible way to create facial dimple. Level of Evidence: IV: Case series.

  13. Environmental monitoring using optical techniques

    Science.gov (United States)

    Svanberg, Sune

    2003-11-01

    An overview of optical techniques for environmental monitoring is presented. Range-resolved measurements of atmospheric pollutants can be performed using the differential absorption lidar technique. Fluorescence lidar allows assessment of vegetation status and also the conditions of the facades of historical buildings. Diode lasers provide particularly realistic schemes for atmospheric gas analysis, where certain wavelength ranges, which are not easily directly assessed, can be reached by sum- and difference frequency generation. Finally, the gas correlation principle can be used for real-time imaging of hydrocarbons. Several types of such optical environmental monitoring are illustrated with examples from research at the Lund Institute of Technology, Sweden.

  14. Surface analysis the principal techniques

    CERN Document Server

    Vickerman, John C

    2009-01-01

    This completely updated and revised second edition of Surface Analysis: The Principal Techniques, deals with the characterisation and understanding of the outer layers of substrates, how they react, look and function which are all of interest to surface scientists. Within this comprehensive text, experts in each analysis area introduce the theory and practice of the principal techniques that have shown themselves to be effective in both basic research and in applied surface analysis. Examples of analysis are provided to facilitate the understanding of this topic and to show readers how they c

  15. Multiple frequency atmospheric radar techniques

    Science.gov (United States)

    Stitt, Gary Richard

    The use of multiple frequency coding to improve the vertical resolution of pulsed-Doppler very high frequency atmospheric radars, especially with regards to the two-frequency techniques known as frequency domain interferometry (FDI), is presented. This technique consists of transmitting alternate pulses on two distinct carrier frequencies. The two resulting time series are used to evaluate the normalized cross-correlation function, whose magnitude and phase are related to the thickness and position of a scattering layer. These same time series are also used to evaluate cross-spectra, which yield magnitude and phase values for each Doppler frequency component of the return signal.

  16. Development of laser decontamination technique

    Energy Technology Data Exchange (ETDEWEB)

    Ogawa, Ryuichiro; Fukui, Yasutaka; Tanimoto, Kenichi [Japan Nuclear Cycle Development Inst., Oarai, Ibaraki (Japan). Oarai Engineering Center

    2002-06-01

    For decommissioning of nuclear fuel facilities, a laser decontamination technique has been developed. The technique was expected to decontaminate high-level radioactive waste to back ground levels, keep the amount of secondary waste low, and be operated by remote control. In the development, a decontamination experiment was executed. Type and operation mode of the laser oscillator, Type and flow rate of the assist gas, repetition rate of the laser pulse, moving velocity of the laser nozzle and irradiation energy were parameters in the experiment. Hot radioactive waste could be decontaminated to background levels uniformly with optimized parameters, which were determined by comparative evaluation. (author)

  17. High-speed pulse techniques

    CERN Document Server

    Coekin, J A

    1975-01-01

    High-Speed Pulse Techniques covers the many aspects of technique in digital electronics and encompass some of the more fundamental factors that apply to all digital systems. The book describes the nature of pulse signals and their deliberate or inadvertent processing in networks, transmission lines and transformers, and then examines the characteristics and transient performance of semiconductor devices and integrated circuits. Some of the problems associated with the assembly of these into viable systems operating at ultra high speed are also looked at. The book examines the transients and w

  18. Computer animation algorithms and techniques

    CERN Document Server

    Parent, Rick

    2012-01-01

    Driven by the demands of research and the entertainment industry, the techniques of animation are pushed to render increasingly complex objects with ever-greater life-like appearance and motion. This rapid progression of knowledge and technique impacts professional developers, as well as students. Developers must maintain their understanding of conceptual foundations, while their animation tools become ever more complex and specialized. The second edition of Rick Parent's Computer Animation is an excellent resource for the designers who must meet this challenge. The first edition establ

  19. Image Segmentation by Using Threshold Techniques

    CERN Document Server

    Al-amri, Salem Saleh; D., Khamitkar S

    2010-01-01

    This paper attempts to undertake the study of segmentation image techniques by using five threshold methods as Mean method, P-tile method, Histogram Dependent Technique (HDT), Edge Maximization Technique (EMT) and visual Technique and they are compared with one another so as to choose the best technique for threshold segmentation techniques image. These techniques applied on three satellite images to choose base guesses for threshold segmentation image.

  20. EDITORIAL: Imaging Systems and Techniques Imaging Systems and Techniques

    Science.gov (United States)

    Giakos, George; Yang, Wuqiang; Petrou, M.; Nikita, K. S.; Pastorino, M.; Amanatiadis, A.; Zentai, G.

    2011-10-01

    This special feature on Imaging Systems and Techniques comprises 27 technical papers, covering essential facets in imaging systems and techniques both in theory and applications, from research groups spanning three different continents. It mainly contains peer-reviewed articles from the IEEE International Conference on Imaging Systems and Techniques (IST 2011), held in Thessaloniki, Greece, as well a number of articles relevant to the scope of this issue. The multifaceted field of imaging requires drastic adaptation to the rapid changes in our society, economy, environment, and the technological revolution; there is an urgent need to address and propose dynamic and innovative solutions to problems that tend to be either complex and static or rapidly evolving with a lot of unknowns. For instance, exploration of the engineering and physical principles of new imaging systems and techniques for medical applications, remote sensing, monitoring of space resources and enhanced awareness, exploration and management of natural resources, and environmental monitoring, are some of the areas that need to be addressed with urgency. Similarly, the development of efficient medical imaging techniques capable of providing physiological information at the molecular level is another important area of research. Advanced metabolic and functional imaging techniques, operating on multiple physical principles, using high resolution and high selectivity nanoimaging techniques, can play an important role in the diagnosis and treatment of cancer, as well as provide efficient drug-delivery imaging solutions for disease treatment with increased sensitivity and specificity. On the other hand, technical advances in the development of efficient digital imaging systems and techniques and tomographic devices operating on electric impedance tomography, computed tomography, single-photon emission and positron emission tomography detection principles are anticipated to have a significant impact on a

  1. Digital synchronization and communication techniques

    Science.gov (United States)

    Lindsey, William C.

    1992-01-01

    Information on digital synchronization and communication techniques is given in viewgraph form. Topics covered include phase shift keying, modems, characteristics of open loop digital synchronizers, an open loop phase and frequency estimator, and a digital receiver structure using an open loop estimator in a decision directed architecture.

  2. A measurement technique for hydroxyacetone

    Energy Technology Data Exchange (ETDEWEB)

    Klotz, P.J.

    1999-10-04

    Hydroxyacetone (HA) is mainly produced in the atmosphere from oxidation of hydrocarbons of the type, CH{sub 3}(R)C{double{underscore}bond}CH{sub 2}. Tuazon and Atkinson (1990) reported HA yield of 41% from the OH-initiated oxidation of methacrolein in the presence of NOx. Since methacrolein is a major product of isoprene oxidation (Carter and Atkinson, 1996), isoprene, a key biogenic hydrocarbon, is therefore expected to be an important source for HA. Consequently, knowledge of ambient concentration of HA would provide information needed to examine the applicability of isoprene reaction mechanisms developed in laboratory and to assess the contribution of isoprene to photooxidant production. The commonly used GC-FID technique involving cryo-focusing is unsuitable for HA owing to HA's thermal instability. When subjected to a temperature of 100 C for only a few seconds, HA was found to disappear completely. Since HA is highly soluble in water, the authors developed a wet chemical technique similar in principle to the one they reported earlier, namely, derivatization following liquid scrubbing. To increase the sensitivity, they adopted a fluorescence detection scheme based on o-phthaldialdehyde (OPA) chemistry. The technique was deployed in the field during two measurement periods at a NARSTO site located on Long Island, New York. The authors report the principle and the operation of this technique and the results obtained from these field studies.

  3. Biopsy techniques for intraocular tumors

    Directory of Open Access Journals (Sweden)

    Pukhraj Rishi

    2016-01-01

    Full Text Available Biopsy involves the surgical removal of a tissue specimen for histopathologic evaluation. Most intraocular tumors are reliably diagnosed based on the clinical evaluation or with noninvasive diagnostic techniques. However, accurately diagnosing a small percentage of tumors can be challenging. A tissue biopsy is thus needed to establish a definitive diagnosis and plan the requisite treatment. From fine-needle aspiration biopsy (FNAB to surgical excision, all tissue collection techniques have been studied in the literature. Each technique has its indications and limitations. FNAB has been reported to provide for 88-95% reliable and safe ophthalmic tumor diagnosis and has gained popularity for prognostic purposes and providing eye conserving treatment surgeries. The technique and instrumentation for biopsy vary depending upon the tissue involved (retina, choroid, subretinal space, vitreous, and aqueous, suspected diagnosis, size, location, associated retinal detachment, and clarity of the media. The cytopathologist confers a very important role in diagnosis and their assistance plays a key role in managing and planning the treatment for malignancies.

  4. Microprocessor Simulation: A Training Technique.

    Science.gov (United States)

    Oscarson, David J.

    1982-01-01

    Describes the design and application of a microprocessor simulation using BASIC for formal training of technicians and managers and as a management tool. Illustrates the utility of the modular approach for the instruction and practice of decision-making techniques. (SK)

  5. Wavelength conversion techniques and devices

    DEFF Research Database (Denmark)

    Danielsen, Søren Lykke; Mikkelsen, Benny; Hansen, Peter Bukhave

    1997-01-01

    interesting for use in WDM optical fibre networks. However, the perfect converter has probably not yet been fabricated and new techniques such as conversion relying on cross-absorption modulation in electro-absorption modulators might also be considered in pursue of effective conversion devices...

  6. Advances phase-lock techniques

    CERN Document Server

    Crawford, James A

    2008-01-01

    From cellphones to micrprocessors, to GPS navigation, phase-lock techniques are utilized in most all modern electronic devices. This high-level book takes a systems-level perspective, rather than circuit-level, which differentiates it from other books in the field.

  7. Techniques for evaluating cash flow.

    Science.gov (United States)

    Sylvestre, J; Urbancic, F

    1994-01-01

    Jean Sylvestre, Ph.D., and Frank Urbancic, D.B.A., CPA, of the University of South Alabama, write that because of the financial challenges for health care, cash flow problems can arise. They offer techniques for interpreting and evaluating cash flow information as a means to more effective decision making.

  8. Dressing Technique for Intermediate Hierarchies

    OpenAIRE

    Holod, P.; Pakuliak, S.

    1994-01-01

    A generalized AKNS systems introduced and discussed recently in \\cite{dGHM} are considered. It was shown that the dressing technique both in matrix pseudo-differential operators and formal series with respect to the spectral parameter can be developed for these hierarchies.

  9. DEMINERALIZATION AND REMINERALIZATION EVALUATION TECHNIQUES

    NARCIS (Netherlands)

    ARENDS, J; TENBOSCH, JJ

    This paper compares the experimental techniques utilized to assess the de- or remineralization of enamel or dentin in intra-oral studies. In in situ studies, it is important for one to know how much mineral has been lost or gained, and where the loss or gain occurred. The main emphasis in this paper

  10. DEMINERALIZATION AND REMINERALIZATION EVALUATION TECHNIQUES

    NARCIS (Netherlands)

    ARENDS, J; TENBOSCH, JJ

    1992-01-01

    This paper compares the experimental techniques utilized to assess the de- or remineralization of enamel or dentin in intra-oral studies. In in situ studies, it is important for one to know how much mineral has been lost or gained, and where the loss or gain occurred. The main emphasis in this paper

  11. FUTURE ENGINEER TRAINING: MOTIVATING TECHNIQUES

    OpenAIRE

    2016-01-01

    The motivating techniques for training future engineer are proposed. Intrinsic and extrinsic motivations in professional training have been analyzed. The peculiarities of the formation process of students’ motivation in learning language at non-language universities have been systematized. Heuristic training technology based on students’ motivation to cognitive research has been implemented.

  12. Schlenk Techniques for Anionic Polymerization

    KAUST Repository

    Ratkanthwar, Kedar

    2015-09-01

    Anionic polymerization-high vacuum techniques (HVTs) are doubtlessly the most prominent and reliable experimental tools to prepare polymer samples with well-defined and, in many cases, complex macromolecular architectures. Due to the high demands for time and skilled technical personnel, HVTs are currently used in only a few research laboratories worldwide. Instead, most researchers in this filed are attracted to more facile Schlenk techniques. The basic principle of this technique followed in all laboratories is substantially the same, i.e. the use of alternate vacuum and inert gas atmosphere in glass apparatus for the purification/charging of monomer, solvents, additives, and for the manipulation of air-sensitive compounds such as alkyl metal initiators, organometallic or organic catalysts. However, it is executed quite differently in each research group in terms of the structure of Schlenk apparatus (manifolds, connections, purification/storage flasks, reactors, etc.), the use of small supplementary devices (soft tubing, cannulas, stopcocks, etc.) and experimental procedures. The operational methods are partly purpose-oriented while also featured by a high flexibility, which makes it impossible to describe in detail each specific one. In this chapter we will briefly exemplify the application of Schlenk techniques for anionic polymerization by describing the performance of a few experiments from our own work.

  13. Thematic Repetition as Rhetorical Technique.

    Science.gov (United States)

    Allen, Jo

    1991-01-01

    Explores William Harvey's work, "On the Motion of the Heart and Blood in Animals." Highlights Harvey's fear of publication and his strategies for reducing resistance to his ideas. Underscores his use of circular references, metaphors, and organizational techniques that enhance and demonstrate his underlying thesis. (SG)

  14. Generalized fast automatic differentiation technique

    Science.gov (United States)

    Evtushenko, Yu. G.; Zubov, V. I.

    2016-11-01

    A new efficient technique intended for the numerical solution of a broad class of optimal control problems for complicated dynamical systems described by ordinary and/or partial differential equations is investigated. In this approach, canonical formulas are derived to precisely calculate the objective function gradient for a chosen finite-dimensional approximation of the objective functional.

  15. Progress in Exosome Isolation Techniques

    Science.gov (United States)

    Li, Pin; Kaslan, Melisa; Lee, Sze Han; Yao, Justin; Gao, Zhiqiang

    2017-01-01

    Exosomes are one type of membrane vesicles secreted into extracellular space by most types of cells. In addition to performing many biological functions particularly in cell-cell communication, cumulative evidence has suggested that several biological entities in exosomes like proteins and microRNAs are closely associated with the pathogenesis of most human malignancies and they may serve as invaluable biomarkers for disease diagnosis, prognosis, and therapy. This provides a commanding impetus and growing demands for simple, efficient, and affordable techniques to isolate exosomes. Capitalizing on the physicochemical and biochemical properties of exosomes, a number of techniques have been developed for the isolation of exosomes. This article summarizes the advances in exosome isolation techniques with an emphasis on their isolation mechanism, performance, challenges, and prospects. We hope that this article will provide an overview of exosome isolation techniques, opening up new perspectives towards the development more innovative strategies and devices for more time saving, cost effective, and efficient isolations of exosomes from a wide range of biological matrices. PMID:28255367

  16. Simultaneous multislice (SMS) imaging techniques

    NARCIS (Netherlands)

    Barth, M.; Breuer, F.; Koopmans, P.J.; Norris, David Gordon; Poser, B.A.

    2016-01-01

    Simultaneous multislice imaging (SMS) using parallel image reconstruction has rapidly advanced to become a major imaging technique. The primary benefit is an acceleration in data acquisition that is equal to the number of simultaneously excited slices. Unlike in-plane parallel imaging this can have

  17. Biomedical engineering and rehabilitation techniques

    Energy Technology Data Exchange (ETDEWEB)

    Gaussens, G.; Jouve, B.; Laizier, J. (Office des Rayonnements Ionisants, CEN Saclay (France))

    The authors outline the programmes set up by the ORIS for the development of biomaterials and rehabilitation products using the radiochemical techniques. They then discuss the development of instruments designed to make the handling and use of radioactive isotopes easier for users.

  18. Teaching Techniques in Clinical Chemistry.

    Science.gov (United States)

    Wilson, Diane

    This master's thesis presents several instructional methods and techniques developed for each of eleven topics or subject areas in clinical chemistry: carbohydrate metabolism, lipid metabolism, diagnostic enzymology, endocrinology, toxicology, quality control, electrolytes, acid base balance, hepatic function, nonprotein nitrogenous compounds, and…

  19. Advances of the IBIC technique

    Energy Technology Data Exchange (ETDEWEB)

    Breese, M.B.H.; Laird, J.S.; Jamieson, D.N. [Melbourne Univ., Parkville, VIC (Australia). School of Physics

    1993-12-31

    The ion beam induced charge (IBIC) technique has been used for a wide variety of analytical applications in the study of semiconductor materials. This paper briefly reviews these uses and identifies those areas which require further development in order to facilitate the more widespread use of the IBIC method. Progress towards implementing these improvements is discussed. 14 refs., 1 fig.

  20. Practical Techniques for Achieving Consensus.

    Science.gov (United States)

    Kline, John A.

    Consensus is important in the making of a policy decision. If a decision is reached without consensus, morale and unit satisfaction may both suffer. With genuine consensus, a unit tends to willingly support and implement the new policy. After analyzing how observed small groups had actually reached consensus, the following ten techniques were…

  1. Hlaallele Detection Using Molecular Techniques

    Directory of Open Access Journals (Sweden)

    Philip A. Dyer

    1993-01-01

    Full Text Available There are now many molecular biological techniques available to define HLA class I and class II alleles. Some of these are also applicable to other human polymorphic genes, in particular to those non-HLA genes encoded within the Mhc. The range of techniques available allows laboratories to choose those most suited to their purpose. The routine laboratory supporting solid organ transplants will need to type large numbers of potential recipients over a period of time, probably using PCR-SSOP while donors will be typed singly and rapidly using PCR-SSP with HLA allele compatibility determined by heteroduplex analysis. Laboratories supporting bone marrow transplantation, where time is less pressing, can choose from the whole range of techniques to determine accurately donor recipient Mhc compatibility. For disease studies, techniques defining precise HLA allele sequence polymorphisms are needed and high sample numbers have to be accommodated. When an association is established allele sequencing has to be used. In the near future, the precise role of HLA alleles in transplantation and disease susceptibility is likely to be established unambiguously.

  2. Counselling Techniques for Outdoor Leaders.

    Science.gov (United States)

    Chase, Michelle; Chase, Robert

    1992-01-01

    Outdoor leaders need counseling skills to deal with interpersonal conflicts that arise within a group and to facilitate participant growth and change. Person-centered counseling, reality therapy counseling, and behavioral counseling are discussed, as well as how various techniques from each can be used to the benefit of the leader and the group.…

  3. Uranium Detection - Technique Validation Report

    Energy Technology Data Exchange (ETDEWEB)

    Colletti, Lisa Michelle [Los Alamos National Lab. (LANL), Los Alamos, NM (United States). Chemistry Division; Garduno, Katherine [Los Alamos National Lab. (LANL), Los Alamos, NM (United States). Chemistry Division; Lujan, Elmer J. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States). Chemistry Division; Mechler-Hickson, Alexandra Marie [Los Alamos National Lab. (LANL), Los Alamos, NM (United States). Chemistry Division; Univ. of Wisconsin, Madison, WI (United States); May, Iain [Los Alamos National Lab. (LANL), Los Alamos, NM (United States). Chemistry Division; Reilly, Sean Douglas [Los Alamos National Lab. (LANL), Los Alamos, NM (United States). Chemistry Division

    2016-04-14

    As a LANL activity for DOE/NNSA in support of SHINE Medical Technologies™ ‘Accelerator Technology’ we have been investigating the application of UV-vis spectroscopy for uranium analysis in solution. While the technique has been developed specifically for sulfate solutions, the proposed SHINE target solutions, it can be adapted to a range of different solution matrixes. The FY15 work scope incorporated technical development that would improve accuracy, specificity, linearity & range, precision & ruggedness, and comparative analysis. Significant progress was achieved throughout FY 15 addressing these technical challenges, as is summarized in this report. In addition, comparative analysis of unknown samples using the Davies-Gray titration technique highlighted the importance of controlling temperature during analysis (impacting both technique accuracy and linearity/range). To fully understand the impact of temperature, additional experimentation and data analyses were performed during FY16. The results from this FY15/FY16 work were presented in a detailed presentation, LA-UR-16-21310, and an update of this presentation is included with this short report summarizing the key findings. The technique is based on analysis of the most intense U(VI) absorbance band in the visible region of the uranium spectra in 1 M H2SO4, at λmax = 419.5 nm.

  4. New techniques in digital holography

    CERN Document Server

    Picart, Pascal

    2015-01-01

    A state of the art presentation of important advances in the field of digital holography, detailing advances related to fundamentals of digital holography, in-line holography applied to fluid mechanics, digital color holography, digital holographic microscopy, infrared holography, special techniques in full field vibrometry and inverse problems in digital holography

  5. Biopsy techniques for intraocular tumors

    Science.gov (United States)

    Rishi, Pukhraj; Dhami, Abhinav; Biswas, Jyotirmay

    2016-01-01

    Biopsy involves the surgical removal of a tissue specimen for histopathologic evaluation. Most intraocular tumors are reliably diagnosed based on the clinical evaluation or with noninvasive diagnostic techniques. However, accurately diagnosing a small percentage of tumors can be challenging. A tissue biopsy is thus needed to establish a definitive diagnosis and plan the requisite treatment. From fine-needle aspiration biopsy (FNAB) to surgical excision, all tissue collection techniques have been studied in the literature. Each technique has its indications and limitations. FNAB has been reported to provide for 88–95% reliable and safe ophthalmic tumor diagnosis and has gained popularity for prognostic purposes and providing eye conserving treatment surgeries. The technique and instrumentation for biopsy vary depending upon the tissue involved (retina, choroid, subretinal space, vitreous, and aqueous), suspected diagnosis, size, location, associated retinal detachment, and clarity of the media. The cytopathologist confers a very important role in diagnosis and their assistance plays a key role in managing and planning the treatment for malignancies. PMID:27488148

  6. Quantitative Techniques in Volumetric Analysis

    Science.gov (United States)

    Zimmerman, John; Jacobsen, Jerrold J.

    1996-12-01

    Quantitative Techniques in Volumetric Analysis is a visual library of techniques used in making volumetric measurements. This 40-minute VHS videotape is designed as a resource for introducing students to proper volumetric methods and procedures. The entire tape, or relevant segments of the tape, can also be used to review procedures used in subsequent experiments that rely on the traditional art of quantitative analysis laboratory practice. The techniques included are: Quantitative transfer of a solid with a weighing spoon Quantitative transfer of a solid with a finger held weighing bottle Quantitative transfer of a solid with a paper strap held bottle Quantitative transfer of a solid with a spatula Examples of common quantitative weighing errors Quantitative transfer of a solid from dish to beaker to volumetric flask Quantitative transfer of a solid from dish to volumetric flask Volumetric transfer pipet A complete acid-base titration Hand technique variations The conventional view of contemporary quantitative chemical measurement tends to focus on instrumental systems, computers, and robotics. In this view, the analyst is relegated to placing standards and samples on a tray. A robotic arm delivers a sample to the analysis center, while a computer controls the analysis conditions and records the results. In spite of this, it is rare to find an analysis process that does not rely on some aspect of more traditional quantitative analysis techniques, such as careful dilution to the mark of a volumetric flask. Figure 2. Transfer of a solid with a spatula. Clearly, errors in a classical step will affect the quality of the final analysis. Because of this, it is still important for students to master the key elements of the traditional art of quantitative chemical analysis laboratory practice. Some aspects of chemical analysis, like careful rinsing to insure quantitative transfer, are often an automated part of an instrumental process that must be understood by the

  7. New techniques for wine aging

    Directory of Open Access Journals (Sweden)

    m Hatice Kalkan Yıldırı

    2017-01-01

    Full Text Available The aging of wine requires a long time therefore it can cause loss of time and money. Therefore using of new techniques for wine aging shortens the length of aging time and wines may be placed on the market more quickly. Nowadays, gamma irradiation, ultrasonic waves, AC electric field and micro-oxygenation are the new techniques for wine aging. Gamma irradiation (after fermentation is accelerated physical maturation method. Gamma irradiation, in a suitable dosage (200 Gy, is a suitable method for improving some wine defects and producing a higher taste quality in wine. The 20 kHz ultrasonic waves aged wine much more quickly than standard aging, with similar quality. The wine treated by 20 kHz ultrasonic waves had a taste equivalent to 1 year aged wine. Wine maturing with AC electric field promises novel process accelerating aging process of fresh wine when suitable conditions are applied. As a result of research, an optimum treatment (electric field 600 V/cm and duration time 3 min was identified to accelerate wine aging. Harsh and pungent raw wine become harmonious and dainty. This process is equivalent to 6 month aging in oak barrel. Microoxygenation is a very important technique used in aging wines in order to improve their characteristics. The techniques of wine tank aging imply the use of small doses of oxygen (2 ml L−1 month−1 and the addition of wood pieces of oak to the wine. Studies concerning these new techniques demonstrated that maturation of wines become more quickly than standard maturation procedures with keeping and improving the wine quality.

  8. Advanced Tools and Techniques for Formal Techniques in Aerospace Systems

    Science.gov (United States)

    Knight, John C.

    2005-01-01

    This is the final technical report for grant number NAG-1-02101. The title of this grant was "Advanced Tools and Techniques for Formal Techniques In Aerospace Systems". The principal investigator on this grant was Dr. John C. Knight of the Computer Science Department, University of Virginia, Charlottesville, Virginia 22904-4740. This report summarizes activities under the grant during the period 7/01/2002 to 9/30/2004. This report is organized as follows. In section 2, the technical background of the grant is summarized. Section 3 lists accomplishments and section 4 lists students funded under the grant. In section 5, we present a list of presentations given at various academic and research institutions about the research conducted. Finally, a list of publications generated under this grant is included in section 6.

  9. Higher-order techniques in computational electromagnetics

    CERN Document Server

    Graglia, Roberto D

    2016-01-01

    Higher-Order Techniques in Computational Electromagnetics explains 'high-order' techniques that can significantly improve the accuracy, computational cost, and reliability of computational techniques for high-frequency electromagnetics, such as antennas, microwave devices and radar scattering applications.

  10. Flow cytometric assessment of viability of lactic acid bacteria

    NARCIS (Netherlands)

    Bunthof, C.J.; Bloemen, K.; Breeuwer, P.; Rombouts, F.M.; Abee, T.

    2001-01-01

    The viability of lactic acid bacteria is crucial for their applications as dairy starters and as probiotics. We investigated the usefulness of flow cytometry (FCM) for viability assessment of lactic acid bacteria. The esterase substrate carboxyfluorescein diacetate (cFDA) and the dye exclusion DNA b

  11. Flow cytometric detection of viruses in the Zuari estuary, Goa

    Digital Repository Service at National Institute of Oceanography (India)

    Mitbavkar, S.; Rajaneesh, K.M.; SathishKumar, P.

    and microalgae, the most abundant organisms in the ocean and also micro- zooplankton 2 . They have been implicated in phytoplankton mortality and the de- cline of phytoplankton blooms 1 . Marine phytoplankton is responsible for up to half of the total primary...

  12. Flow cytometric immunophenotyping test for staging/monitoring neuroblastoma patients

    National Research Council Canada - National Science Library

    Warzynski, Michael J; Graham, David M; Axtell, Richard A; Higgins, James V; Hammers, Yuki A

    2002-01-01

    .... Following the “rare event” philosophy of selecting one negative and two positive antigens, we initially tried a “cocktail” of CD45 − CD56 very bright+ neuron‐specific enolase (NSE) cytoplasmic...

  13. Flow Cytometric Ploidy Determination of Oral Premalignant and Malignant Lesions

    Science.gov (United States)

    1990-01-01

    However, subjectivity remains an inherent part of the diagnostic process.9 According to Dabelsteen,10 subjectivity is most apparent in the determination...of Silverman 16 and Pindborg43 and Mincer et al.21 that light microscopio features of premalignancy are often not present in original biopsy specimens...was caused either in whole or in part by the presence of doublets was eliminated by syringina,. filtering and visually examining the population. In

  14. IVBT-documented platelet function correlates with flow cytometric data.

    Science.gov (United States)

    Hoffmann, J; Bonacker, G; Kretschmer, V; Schulzki, T; Heimanns, J

    1996-12-01

    Thrombocytopenic patients with identical platelet counts often show different bleeding tendencies owing to significant differences in the platelet function. This could be demonstrated by the in vitro bleeding test (IVBT). Using flow cytometry, we tried to find characteristics of platelet antigen expression in order to explain these differences in function. Thirty patients with bone marrow hypoplasia receiving 65 platelet transfusions (mainly from a cell separator) were observed for 3 to 29 days. Size, granulation and fluorescence of platelet-rich plasma (n = 522 samples) were evaluated using monoclonal antibodies against GP IIIb (collagen receptor), GP IIb/IIIa (fibrinogen receptor) and GP Ib (thrombin receptor). We defined separate gates for each antibody using the results from 50 normals and by laying an orthograde cross over the gate to divide the gate into four equal quadrants. The platelet populations were divided into four different groups according to the occlusion time (OT) of the IVBT and the Simplate time (ST). The thrombocytes with the most impaired function (OT > or = 485 s/ST > 30 min) had significantly less platelet fluorescence when marked with antibodies against GP IIIb and GP Ib than those with short OT and ST (OT platelet fluorescence when marked with anti-GP IIIb and anti-GP Ib than thrombocytopenic patients, who had a spontaneous platelet rise beyond 30,000 platelets/microliters a few days later. One day after platelet transfusion, significantly more platelets with high GP IIIb and Ib expression could be found. We were also able to document better transfusion efficacy of platelet concentrates with high GP IIIb and Ib expression. Finally, patients with high bleeding scores showed less GP Ib expression on the platelets than patients with low bleeding scores. In summary, the IVBT-documented platelet function clearly corresponded to an increased expression of the collagen receptor and the thrombin receptor of platelets.

  15. Flow cytometric DNA ploidy analysis of ovarian granulosa cell tumors

    NARCIS (Netherlands)

    D. Chadha; C.J. Cornelisse; A. Schabert (A.)

    1990-01-01

    textabstractAbstract The nuclear DNA content of 50 ovarian tumors initially diagnosed as granulosa cell tumors was measured by flow cytometry using paraffin-embedded archival material. The follow-up period of the patients ranged from 4 months to 19 years. Thirty-eight tumors were diploid or near-dip

  16. DNA flow cytometric analysis in variable types of hydropic placentas

    Directory of Open Access Journals (Sweden)

    Fatemeh Atabaki pasdar

    2015-05-01

    Full Text Available Background: Differential diagnosis between complete hydatidiform mole, partial hydatidiform mole and hydropic abortion, known as hydropic placentas is still a challenge for pathologists but it is very important for patient management. Objective: We analyzed the nuclear DNA content of various types of hydropic placentas by flowcytometry. Materials and Methods: DNA ploidy analysis was performed in 20 non-molar (hydropic and non-hydropic spontaneous abortions and 20 molar (complete and partial moles, formalin-fixed, paraffin-embedded tissue samples by flow cytometry. The criteria for selection were based on the histopathologic diagnosis. Results: Of 10 cases histologically diagnosed as complete hydatiform mole, 9 cases yielded diploid histograms, and 1 case was tetraploid. Of 10 partial hydatidiform moles, 8 were triploid and 2 were diploid. All of 20 cases diagnosed as spontaneous abortions (hydropic and non-hydropic yielded diploid histograms. Conclusion: These findings signify the importance of the combined use of conventional histology and ploidy analysis in the differential diagnosis of complete hydatidiform mole, partial hydatidiform mole and hydropic abortion.

  17. Flow cytometric data analysis of circulating progenitor cell stability.

    Science.gov (United States)

    Mahar, Ernestine A; Mou, Liping; Hayek, Salim S; Quyyumi, Arshed A; Waller, Edmund K

    2017-02-01

    A recent publication by Mekonnen et al. demonstrated that among women with non-obstructive coronary artery disease, higher levels of circulating progenitor cells in the blood (CPC), were associated with impaired coronary flow reserve [1]. We performed a quality control assessment of the stability of circulating blood progenitor cells in blood samples stored at 4 °C, to determine the time period during which blood samples can be analyzed and yield consistent data for progenitor cell content. Healthy volunteers (n=6) were recruited and underwent phlebotomy, and blood was stored in EDTA tubes at 4 °C. Flow cytometry was performed to quantitate progenitor cell subsets at 0-4 h, 24 h, and 48 h post phlebotomy. All processed samples were fixed with 1% Paraformaldehyde and 1,000,000 total data events were collected. We found no significant differences in PC data for both CD34+ (P=0.68 for one-way ANOVA) and CD34+/CD133+ (P=0.74 for one-way ANOVA).

  18. Flow cytometric data analysis of circulating progenitor cell stability

    Directory of Open Access Journals (Sweden)

    Ernestine A. Mahar

    2017-02-01

    We performed a quality control assessment of the stability of circulating blood progenitor cells in blood samples stored at 4 °C, to determine the time period during which blood samples can be analyzed and yield consistent data for progenitor cell content. Healthy volunteers (n=6 were recruited and underwent phlebotomy, and blood was stored in EDTA tubes at 4 °C. Flow cytometry was performed to quantitate progenitor cell subsets at 0–4 h, 24 h, and 48 h post phlebotomy. All processed samples were fixed with 1% Paraformaldehyde and 1,000,000 total data events were collected. We found no significant differences in PC data for both CD34+ (P=0.68 for one-way ANOVA and CD34+/CD133+ (P=0.74 for one-way ANOVA.

  19. Cytometric analysis of DNA changes induced by sulfur mustard

    Energy Technology Data Exchange (ETDEWEB)

    Smith, W.J.; Sanders, K.M.; Ruddle, S.E.; Gross, C.L.

    1993-05-13

    Sulfur mustard is an alkylating agent which causes severe, potentially debilitating blisters following cutaneous exposure. Its mechanism of pathogenesis is unknown and no antidote exists to prevent its pathology. The biochemical basis of sulfur mustard's vesicating activity has been hypothesized to be a cascade of events beginning with alkylation of DNA. Using human cells in culture, we have assessed the effects of sulfur mustard on cell cycle activity using flow cytometry with propidium iodide. Two distinct patterns emerged, a Gl/S interface block at concentrations equivalent to vesicating doses (>50-micronM) and a G2 block at 10-fold lower concentrations. In addition, noticeable increases in amount of dye uptake were observed at 4 and 24 hours after sulfur mustard exposure. These increases are believed to be related to DNA repair activities and can be prevented by treatment of the cells with niacinamide, which inhibits DNA repair. Other drugs which provide alternate alkylating sites or inhibit cell cycle progression were shown to lower the cytotoxicity of sulfur mustard and to protect against its direct DNA damaging effects.

  20. Flow Cytometric Applicability of Fluorescent Vitality Probes on Phytoplankton

    NARCIS (Netherlands)

    Peperzak, L.; Brussaard, C.P.D.

    2011-01-01

    The applicability of six fluorescent probes (four esterase probes: acetoxymethyl ester of Calcein [Calcein-AM], 5-chloromethylfluorescein diacetate [CMFDA], fluorescein diacetate [FDA], and 2',7'-dichlorofluorescein diacetate [H(2)DCFDA]; and two membrane probes: bis-(1,3-dibutylbarbituric acid) tri

  1. Ultrastructural and flow cytometric analyses of lipid accumulation in microalgae

    Energy Technology Data Exchange (ETDEWEB)

    Solomon, J.A.; Hand, R.E. Jr.; Mann, R.C.

    1986-12-01

    Lipid accumulation in three species of microalgae was investigated with flow cytometry (FCM) and transmission electron microscopy (TEM). Previous studies using batch cultures of a algae have led to the assumption that lipid accumulation in microalgae is a gradual process requiring at least several days for completion. However, FCM reveals, through changes in the chlorophyll:lipid ratio, that the time span required for individual cells to change metabolic state is short. Simultaneous FCM measurements of chlorophyll and nile red (neutral lipid) fluorescence in individual cells of nitrogen-deficient Isochrysis populations revealed a bimodal population distribution as one stage in the lipid accumulation process. The fact that two discrete populations exist, with few cells in an intermediate stage, suggests rapid response to a liqid trigger. Interpretations of light and electron microscopic observations are consistent with this hypothesis. The time required for an entire population to achieve maximum lipid content is considerably longer than that required for a single cell, due to the variation in response time among cells. In this study high lipid cultures were sometimes obtained by using FCM to separate high lipid cells from the remainder of the population. FCM holds much promise for strain enhancement but considerable developmental work, directed at providing more consistent results, remains to be done. 8 refs., 35 figs.

  2. Detachment and flow cytometric quantification of seagrass-associated bacteria.

    Science.gov (United States)

    Trevathan-Tackett, Stacey; Macreadie, Peter; Ralph, Peter; Seymour, Justin

    2014-07-01

    A new protocol was developed to detach bacteria from seagrass tissue and subsequently enumerate cells using flow cytometry (FCM). A method involving addition of the surfactant Tween 80 and vortexing resulted in maximum detachment efficiency of seagrass attached bacteria, providing a robust protocol for precisely enumerating seagrass-associated bacteria with FCM. Using this approach we detected cell concentrations between 2.0×10(5) and 8.0×10(6)cells mg(-1) DW tissue.

  3. Flow cytometric fluorescence lifetime analysis of DNA binding fluorochromes

    Energy Technology Data Exchange (ETDEWEB)

    Crissman, Harry A.; Cui, H. H. (H. Helen); Steinkamp, J. A.

    2002-01-01

    Most flow cytometry (FCM) applications monitor fluorescence intensity to quantitate the various cellular parameters; however, the fluorescence emission also contains information relative to the fluorescence lifetime. Recent developments in FCM (Pinsky et al., 1993; Steinkamp & Crissman, 1993; Steinkamp et al., 1993), provide for the measurement of fluorescence lifetime which is also commonly referred to as fluorescence decay, or the time interval in which a fluorochrome remains in the excited state. Many unbound fluorochromes have characteristic lifetime values that are determined by their molecular structure; however, when the probe becomes bound, the lifetime value is influenced by a number of factors that affect the probe interaction with a target molecule. Monitoring the changes in the lifetime of the probe yields information relating to the molecular conformation, the functional state or activity of the molecular target. In addition, the lifetime values can be used as signatures to resolve the emissions of multiple fluorochrome labels with overlapping emission spectra that cannot be resolved by conventional FCM methodology. Such strategies can increase the number of fluorochrome combinations used in a flow cytometer with a single excitation source. Our studies demonstrate various applications of lifetime measurements for the analysis of the binding of different fluorochromes to DNA in single cells. Data presented in this session will show the utility of lifetime measurements for monitoring changes in chromatin structure associated with cell cycle progression, cellular differentiation, or DNA damage, such as induced during apoptosis. Several studies show that dyes with specificity for nucleic acids display different lifetime values when bound to DNA or to dsRNA. The Phase Sensitive Flow Cytometer is a multiparameter instrument, capable of performing lifetime measurements in conjunction with all the conventional FCM measurements. Future modifications of this instrumentation will provide the capability for simultaneously measuring multiple lifetimes, thereby significantly enhancing the subcellular resolution of the multiple complexes of fluorescent compounds, such as chemotherapeutic agents, in single cells (Sailer et al., 1997b).

  4. Validation of merocyanine 540 staining as a technique for assessing capacitation-related membrane destabilization of fresh dog sperm

    NARCIS (Netherlands)

    Steckler, D; Stout, T A E; Durandt, C; Nöthling, J O

    2015-01-01

    The aim of this study was to determine whether flow cytometric evaluation of combined merocyanine 540 and Yo-Pro 1 (M540-YP) staining would identify viable dog sperm that had undergone membrane stabilization known to be associated with capacitation in other species, and whether such destabilization

  5. Seismic techniques in coal mining

    Energy Technology Data Exchange (ETDEWEB)

    Bhattacharyya, A.K.; Belleza, G.V.

    1983-01-01

    The aim of this study is to investigate the peripheral fracture zones in coal pillars left for support in underground mines. The fracture zones are caused by the redistribution of stresses in strata resulting from the process of excavation and blasting if it is used. The extent and degree of these fracture zones, in turn, have a direct influence on the ability of pillars to provide stable support to the overlying strata. Seismic methods such as refraction, uphole, and collinear techniques outlined in previous reports are being used to investigate the extent and degree of the peripheral fracture zones. Some of the work that has been carried out and is described in this report, relates to the study of peripheral fracture zones in coal pillars using seismic techniques.

  6. Embodying embodied design research techniques

    DEFF Research Database (Denmark)

    Wilde, Danielle; Tomico, Oscar; Lucero, Andrés

    2015-01-01

    ideation, communication and collaboration techniques enable enhanced creative engagement and assist creativity [2]. By applying such methods to the problem of their reporting, we hope to deepen understanding of how to move towards enriched, nuanced and repeatable methods for embodied design and knowledge...... propose to investigate how embodied research techniques may be used as direct and unmediated vehicles for their own reporting. Rather than engaging in oral presentations, participants will lead other workshop participants through a proven embodied method or approach. Small groups will create mash...... of reporting embodied experiments, so that intangible elements are not lost. Participants will be supported to reflect on unfolding discoveries, to share impressions, as well as outcomes, including documentation experiments that aim to tangibly capture and communicate the processes undertaken. Embodied...

  7. Historical Techniques of Lie Detection

    Directory of Open Access Journals (Sweden)

    Martina Vicianova

    2015-08-01

    Full Text Available Since time immemorial, lying has been a part of everyday life. For this reason, it has become a subject of interest in several disciplines, including psychology. The purpose of this article is to provide a general overview of the literature and thinking to date about the evolution of lie detection techniques. The first part explores ancient methods recorded circa 1000 B.C. (e.g., God’s judgment in Europe. The second part describes technical methods based on sciences such as phrenology, polygraph and graphology. This is followed by an outline of more modern-day approaches such as FACS (Facial Action Coding System, functional MRI, and Brain Fingerprinting. Finally, after the familiarization with the historical development of techniques for lie detection, we discuss the scope for new initiatives not only in the area of designing new methods, but also for the research into lie detection itself, such as its motives and regulatory issues related to deception.

  8. Atmospheric Ice Accretion Measurement Techniques

    Directory of Open Access Journals (Sweden)

    M Virk

    2016-09-01

    Full Text Available Atmospheric icing on structures has proven to be an area of concern in many cold climate geographical regions like arctic and alpine. Difficulties encountered by the communication, construction and power industries in these areas are the subject of intense investigations for researchers from decades. Three main methods of investigation are generally employed by researchers to study atmospheric ice accretion on structures: a continuous field measurements, b lab based simulations using icing wind tunnel & c numerical modelling. This paper presents a brief review study of various techniques to understand and measure the atmospheric ice accretion on structures, anti/de icing techniques and important parameters for numerical modelling of atmospheric ice accretion.

  9. Data mining concepts and techniques

    CERN Document Server

    Han, Jiawei

    2005-01-01

    Our ability to generate and collect data has been increasing rapidly. Not only are all of our business, scientific, and government transactions now computerized, but the widespread use of digital cameras, publication tools, and bar codes also generate data. On the collection side, scanned text and image platforms, satellite remote sensing systems, and the World Wide Web have flooded us with a tremendous amount of data. This explosive growth has generated an even more urgent need for new techniques and automated tools that can help us transform this data into useful information and knowledge.Like the first edition, voted the most popular data mining book by KD Nuggets readers, this book explores concepts and techniques for the discovery of patterns hidden in large data sets, focusing on issues relating to their feasibility, usefulness, effectiveness, and scalability. However, since the publication of the first edition, great progress has been made in the development of new data mining methods, systems, and app...

  10. The ancient Chinese casting techniques

    Directory of Open Access Journals (Sweden)

    Tan Derui

    2011-02-01

    Full Text Available In the course of Chinese civilization, which lasted more than 5,000 years, casting production has made a huge contribution. In this paper, some representative metal castings were presented. According to their forming techniques, they can be grouped into stone mould casting, clay mould casting, ablation casting, lost wax casting, stack casting, permanent mould casting, sand casting, etc. According to their materials, they can be categorized into tin bronze, bimetallic bronze, malleable cast iron, ductile cast iron, brass, cupronickel alloy (Packtong, etc. According to their surface decorative techniques they can be devided into gem inlay, gilding, gold and silver inlay, copper inlay, engraved decoration, surface tin-enrichment, mother-of-pearl inlay, burnished works with gold or silver inlay, surface coloring and cloisonné enamel, etc.

  11. Multilevel techniques for Reservoir Simulation

    DEFF Research Database (Denmark)

    Christensen, Max la Cour

    The subject of this thesis is the development, application and study of novel multilevel methods for the acceleration and improvement of reservoir simulation techniques. The motivation for addressing this topic is a need for more accurate predictions of porous media flow and the ability to carry...... Full Approximation Scheme) • Variational (Galerkin) upscaling • Linear solvers and preconditioners First, a nonlinear multigrid scheme in the form of the Full Approximation Scheme (FAS) is implemented and studied for a 3D three-phase compressible rock/fluids immiscible reservoir simulator...... based on element-based Algebraic Multigrid (AMGe). In particular, an advanced AMGe technique with guaranteed approximation properties is used to construct a coarse multilevel hierarchy of Raviart-Thomas and L2 spaces for the Galerkin coarsening of a mixed formulation of the reservoir simulation...

  12. Alternative Techniques for Teaching Writing

    Directory of Open Access Journals (Sweden)

    Sihindun arumi

    2015-12-01

    Full Text Available Writing as one of language skill is often considered very difficult. It is due to the fact that writing needs to produce and organize ideas using appropriate vocabulary, language use, paragraph organization, and mechanism. It also needs to turn the ideas into a readable text and for foreign language learners, they should also transfer ideas from their native language into target language (foreign language. It raises any problems for them to create a good text. Moreover, the situation in the class does not always supportthem in which the techniques of the teacher in teaching writing is boring and monotonous, do not give enough attention to help students explore their writing skills. So that they attend the writing class only for procedural formality.Thus, it is considered important to elaborate various techniques to build nice classroom atmosphere as well as to improve students’ writing skills.

  13. Apoptosis Evaluation by Electrochemical Techniques.

    Science.gov (United States)

    Yin, Jian; Miao, Peng

    2016-03-01

    Apoptosis has close relevance to pathology, pharmacology, and toxicology. Accurate and convenient detection of apoptosis would be beneficial for biological study, clinical diagnosis, and drug development. Based on distinct features of apoptotic cells, a diversity of analytical techniques have been exploited for sensitive analysis of apoptosis, such as surface plasmon resonance, electrochemical methods, flow cytometry, and some imaging assays. Among them, the features of simplicity, easy operation, low cost, and high sensitivity make electrochemical techniques powerful tools to investigate electron-transfer processes of in vitro biological systems. In this contribution, a general overview of current knowledge on various technical approaches for apoptosis evaluation is provided. Furthermore, recently developed electrochemical biosensors for detecting apoptotic cells and their advantages over traditional methods are summarized. One of the main considerations focuses on designing the recognition elements based on various biochemical events during apoptosis.

  14. Pinch Technique: Theory and Applications

    CERN Document Server

    Binosi, D; 10.1016/j.physrep.2009.05.001

    2009-01-01

    We review the theoretical foundations and most important physical applications of the Pinch Technique (PT). This method allows the construction of off-shell Green's functions in non-Abelian gauge theories that are independent of the gauge-fixing parameter and satisfy ghost-free Ward identities. We first present the diagrammatic formulation of the technique in QCD, deriving at one loop the gauge independent gluon self-energy, quark-gluon vertex, and three-gluon vertex, together with their Abelian Ward identities. The generalization to theories with spontaneous symmetry breaking is carried out in detail, and the connection with the optical theorem and the dispersion relations are explained within the electroweak sector of the Standard Model. The equivalence between the PT and the Feynman gauge of the Background Field Method (BFM) is elaborated, and the crucial differences between the two methods are critically scrutinized. The Batalin-Vilkovisky quantization method and the general formalism of algebraic renorma...

  15. Nanorobotics current approaches and techniques

    CERN Document Server

    Ferreira, Antoine

    2013-01-01

    Nanorobot devices now perform a wide variety of tasks at the nanoscale in a wide variety of fields including but not limited to fields such as manufacturing, medicine, supply chain, biology, and outer space. Nanorobotics: Current Approaches and Techniques is a comprehensive overview of this interdisciplinary field with a wide ranging discussion that includes nano-manipulation and industrial nanorobotics, nanorobotics in biology and medicine, nanorobotic sensing, navigation and swarm behavior, and protein and DNA-based nanorobotics. Also included is the latest on topics such as bio-nano-actuators and propulsion and navigation of nanorobotic systems using magnetic fields. Nanorobotics: Current Approaches and Techniques is an ideal book for scientists, researchers, and engineers actively involved in applied and robotic research and development.

  16. UML-ising formal techniques

    DEFF Research Database (Denmark)

    Bjørner, Dines; George, Chris W.; Haxthausen, Anne Elisabeth

    2004-01-01

    these different UML views are unified, integrated, correlated or merely co-located is for others to dispute. We also seek to support multiple views, but are also in no doubt that there must be sound, well defined relations between such views. We thus report on ways and means of integrating formal techniques...... Modeling Language [10,67,36,20] has caused, for one reason or another, the research community to try formalise one or another facet of UML. In this paper we report on another way to achieve what UML attempts to achieve: Broadness of application, convenience of notation, and multiplicity of views. Whether...... techniques, one that can be believably deployed for as wide a spectrum, or even a wider spectrum of software (and hardware) development, as, respectively than UML....

  17. Digital Fourier analysis advanced techniques

    CERN Document Server

    Kido, Ken'iti

    2015-01-01

    This textbook is a thorough, accessible introduction to advanced digital Fourier analysis for advanced undergraduate and graduate students. Assuming knowledge of the Fast Fourier Transform, this book covers advanced topics including the Hilbert transform, cepstrum analysis, and the two-dimensional Fourier transform. Saturated with clear, coherent illustrations, "Digital Fourier Analysis - Advanced Techniques" includes practice problems and thorough Appendices. As a central feature, the book includes interactive applets (available online) that mirror the illustrations. These user-friendly applets animate concepts interactively, allowing the user to experiment with the underlying mathematics. The applet source code in Visual Basic is provided online, enabling advanced students to tweak and change the programs for more sophisticated results. A complete, intuitive guide, "Digital Fourier Analysis - Advanced Techniques" is an essential reference for students in science and engineering.

  18. Barley Transformation Using Biolistic Techniques

    Science.gov (United States)

    Harwood, Wendy A.; Smedley, Mark A.

    Microprojectile bombardment or biolistic techniques have been widely used for cereal transformation. These methods rely on the acceleration of gold particles, coated with plasmid DNA, into plant cells as a method of directly introducing the DNA. The first report of the generation of fertile, transgenic barley plants used biolistic techniques. However, more recently Agrobacterium-mediated transformation has been adopted as the method of choice for most cereals including barley. Biolistic procedures are still important for some barley transformation applications and also provide transient test systems for the rapid checking of constructs. This chapter describes methods for the transformation of barley using biolistic procedures and also highlights the use of the technology in transient assays.

  19. Full scale lightning test technique

    Science.gov (United States)

    Walko, L. C.; Schneider, J. G.

    1980-01-01

    A test technique was developed for applying a full scale mean value (30 kiloampere peak) simulated lightning return stroke current on a complete flight ready aircraft to assess the threat of lightning to aircraft electrical circuits. A computer-aided generator design was used to establish the parameters of the test system. Data from previous work done on development of low inductance current paths determined the basic system configuration.

  20. A MEASUREMENT TECHNIQUE FOR HYDROXYACETONE.

    Energy Technology Data Exchange (ETDEWEB)

    KLOTZ,P.J.

    1999-10-04

    Hydroxyacetone (HA) is mainly produced in the atmosphere from oxidation of hydrocarbons of the type, CH{sub 3}(R)C=CH{sub 2}. Tuazon and Atkinson (1990) reported HA yield of 41% from the OH-initiated oxidation of methacrolein in the presence of NO{sub x}. Since methacrolein is a major product of isoprene oxidation (Carter and Atkinson, 1996), isoprene, a key biogenic hydrocarbon, is therefore expected to be an important source for HA. Consequently, knowledge of ambient concentration of HA would provide information needed to examine the applicability of isoprene reaction mechanisms developed in laboratory and to assess the contribution of isoprene to photooxidant production. The commonly used GC-FID technique involving cryo-focusing is unsuitable for HA owing to HA's thermal instability. When subjected to a temperature of 100 C for only a few seconds, HA was found to disappear completely. Since HA is highly soluble in water (it's Henry's law constant being {approx}2 x 10{sup 4} M atm{sup -1} at 20 C, Zhou and Lee, unpublished data), we developed a wet chemical technique similar in principle to the one we reported earlier (Lee and Zhou, 1993), namely, based on derivatization following liquid scrubbing. To increase the sensitivity, we adopted a fluorescence detection scheme based on o-phthaldialdehyde (OPA) chemistry. The technique was deployed in the field during two measurement periods at a NARSTO site located on Long Island (LI), New York. We report the principle and the operation of this technique and the results obtained from these field studies.

  1. Hip: Anatomy and US technique

    OpenAIRE

    2011-01-01

    Ultrasound (US) has always had a relatively limited role in the evaluation of the hip due to the deep location of this joint. However, many hip diseases are well detectable at US, but before approaching such a study it is necessary to be thoroughly familiar with the normal anatomy and related US images. The study technique is particularly important as optimization of various parameters is required, such as probe frequency, focalization, positioning of the probe, etc. Also the patient’s positi...

  2. Microcontact Printing Techniques in Bioscience

    Institute of Scientific and Technical Information of China (English)

    FENG Xi-zeng; HOU Sun; CHAN Qi-lin; WANG Li-kai; QIN Ming; HAN Pei-dong

    2004-01-01

    The microcontact printing(μCP) technology available for patterning protein, DNA hybridization, immunoassay and cellular cocultures onto solid surface are reviewed. This review describes some of the techniques currently employed for creating two-dimensional biomolecular microarray, and the research results regarding their effectiveness. In addition, the applications of the impact of μCP technology in the field of biosciences are also presented.

  3. Thermal measurements and inverse techniques

    CERN Document Server

    Orlande, Helcio RB; Maillet, Denis; Cotta, Renato M

    2011-01-01

    With its uncommon presentation of instructional material regarding mathematical modeling, measurements, and solution of inverse problems, Thermal Measurements and Inverse Techniques is a one-stop reference for those dealing with various aspects of heat transfer. Progress in mathematical modeling of complex industrial and environmental systems has enabled numerical simulations of most physical phenomena. In addition, recent advances in thermal instrumentation and heat transfer modeling have improved experimental procedures and indirect measurements for heat transfer research of both natural phe

  4. The ancient Chinese casting techniques

    OpenAIRE

    2011-01-01

    In the course of Chinese civilization, which lasted more than 5,000 years, casting production has made a huge contribution. In this paper, some representative metal castings were presented. According to their forming techniques, they can be grouped into stone mould casting, clay mould casting, ablation casting, lost wax casting, stack casting, permanent mould casting, sand casting, etc. According to their materials, they can be categorized into tin bronze, bimetallic bronze, malleable cast ir...

  5. Electromigration techniques theory and practice

    CERN Document Server

    Dziubakiewicz, Ewelina; Szumski, Michal

    2013-01-01

    The book provides the broad knowledge on electromigration techniques including: theory of CE, description of instrumentation, theory and practice in micellar electrokinetic chromatography, isotachophoresis, capillary isoelectric focusing, capillary and planar electrochromatography (including description of instrumentation and packed and monolithic column preparation), 2D-gel electrophoresis (including sample preparation) and lab-on-a-chip systems. The book also provides the most recent examples of applications including food, environmental, pharmaceutical analysis as well as proteomics.

  6. AINV AND BILUM PRECONDITIONING TECHNIQUES

    Institute of Scientific and Technical Information of China (English)

    谷同祥; 迟学斌; 刘兴平

    2004-01-01

    It was proposed that a robust and efficient parallelizable preconditioner for solving general sparse linear systems of equations, in which the use of sparse approximate inverse (AINV) techniques in a multi-level block ILU (BILUM) preconditioner were investigated. The resulting preconditioner retains robustness of BILUM preconditioner and has two advantages over the standard BILUM preconditioner: the ability to control sparsity and increased parallelism. Numerical experiments are used to show the effectiveness and efficiency of the new preconditioner.

  7. Radar techniques using array antennas

    CERN Document Server

    Wirth, Wulf-Dieter

    2013-01-01

    Radar Techniques Using Array Antennas is a thorough introduction to the possibilities of radar technology based on electronic steerable and active array antennas. Topics covered include array signal processing, array calibration, adaptive digital beamforming, adaptive monopulse, superresolution, pulse compression, sequential detection, target detection with long pulse series, space-time adaptive processing (STAP), moving target detection using synthetic aperture radar (SAR), target imaging, energy management and system parameter relations. The discussed methods are confirmed by simulation stud

  8. Submucosal tunneling techniques: current perspectives

    Directory of Open Access Journals (Sweden)

    Kobara H

    2014-04-01

    Full Text Available Hideki Kobara,1 Hirohito Mori,1 Kazi Rafiq,2 Shintaro Fujihara,1 Noriko Nishiyama,1 Maki Ayaki,1 Tatsuo Yachida,1 Tae Matsunaga,1 Johji Tani,1 Hisaaki Miyoshi,1 Hirohito Yoneyama,1 Asahiro Morishita,1 Makoto Oryu,1 Hisakazu Iwama,3 Tsutomu Masaki1 1Department of Gastroenterology and Neurology, 2Department of Pharmacology, 3Life Science Research Center, Faculty of Medicine, Kagawa University, Miki-cho, Kita-Gun, Kagawa, Japan Abstract: Advances in endoscopic submucosal dissection include a submucosal tunneling technique, involving the introduction of tunnels into the submucosa. These tunnels permit safer offset entry into the peritoneal cavity for natural orifice transluminal endoscopic surgery. Technical advantages include the visual identification of the layers of the gut, blood vessels, and subepithelial tumors. The creation of a mucosal flap that minimizes air and fluid leakage into the extraluminal cavity can enhance the safety and efficacy of surgery. This submucosal tunneling technique was adapted for esophageal myotomy, culminating in its application to patients with achalasia. This method, known as per oral endoscopic myotomy, has opened up the new discipline of submucosal endoscopic surgery. Other clinical applications of the submucosal tunneling technique include its use in the removal of gastrointestinal subepithelial tumors and endomicroscopy for the diagnosis of functional and motility disorders. This review suggests that the submucosal tunneling technique, involving a mucosal safety flap, can have potential values for future endoscopic developments. Keywords: submucosal endoscopy, submucosal tunneling method, natural orifice transluminal endoscopic surgery, peroral endoscopic myotomy, gastrointestinal subepithelial tumor, functional and motility disorders

  9. Cobbler's technique for Iridodialysis repair

    Directory of Open Access Journals (Sweden)

    Surinder Singh Pandav

    2016-01-01

    Full Text Available We describe a novel “Cobbler's technique” for iridodialysis repair in the right eye of a patient aged 18 years, with a traumatic iridodialysis secondary to open globe injury with an iron rod. Our technique is simple with easy surgical maneuvers, that is, effective for repairing iridodialysis. The “Cobbler's technique” allows a maximally functional and cosmetic result for iridodialysis.

  10. Enteral Feeding Set Handling Techniques.

    Science.gov (United States)

    Lyman, Beth; Williams, Maria; Sollazzo, Janet; Hayden, Ashley; Hensley, Pam; Dai, Hongying; Roberts, Cristine

    2017-04-01

    Enteral nutrition therapy is common practice in pediatric clinical settings. Often patients will receive a pump-assisted bolus feeding over 30 minutes several times per day using the same enteral feeding set (EFS). This study aims to determine the safest and most efficacious way to handle the EFS between feedings. Three EFS handling techniques were compared through simulation for bacterial growth, nursing time, and supply costs: (1) rinsing the EFS with sterile water after each feeding, (2) refrigerating the EFS between feedings, and (3) using a ready-to-hang (RTH) product maintained at room temperature. Cultures were obtained at baseline, hour 12, and hour 21 of the 24-hour cycle. A time-in-motion analysis was conducted and reported in average number of seconds to complete each procedure. Supply costs were inventoried for 1 month comparing the actual usage to our estimated usage. Of 1080 cultures obtained, the overall bacterial growth rate was 8.7%. The rinse and refrigeration techniques displayed similar bacterial growth (11.4% vs 10.3%, P = .63). The RTH technique displayed the least bacterial growth of any method (4.4%, P = .002). The time analysis in minutes showed the rinse method was the most time-consuming (44.8 ± 2.7) vs refrigeration (35.8 ± 2.6) and RTH (31.08 ± 0.6) ( P < .0001). All 3 EFS handling techniques displayed low bacterial growth. RTH was superior in bacterial growth, nursing time, and supply costs. Since not all pediatric formulas are available in RTH, we conclude that refrigerating the EFS between uses is the next most efficacious method for handling the EFS between bolus feeds.

  11. Believing in the buttonhole technique.

    Science.gov (United States)

    Kregness, Allison

    2008-11-01

    Vascular access cannulation for dialysis staff is the most difficult obstacle to overcome during the orientation process and can remain a challenge throughout one's career in caring for patients with renal disease. Learning the "buttonhole technique" can make venipuncture more comfortable for the patient and can eliminate the fear of failure for the cannulator. Not only does it preserve the timeframe of the patient's access, but establishes a routine process for the dialysis facility to follow for successful cannulation.

  12. Triangulation of Data Analysis Techniques

    Directory of Open Access Journals (Sweden)

    Lauri, M

    2011-10-01

    Full Text Available In psychology, as in other disciplines, the concepts of validity and reliability are considered essential to give an accurate interpretation of results. While in quantitative research the idea is well established, in qualitative research, validity and reliability take on a different dimension. Researchers like Miles and Huberman (1994 and Silverman (2000, 2001, have shown how these issues are addressed in qualitative research. In this paper I am proposing that the same corpus of data, in this case the transcripts of focus group discussions, can be analysed using more than one data analysis technique. I refer to this idea as ‘triangulation of data analysis techniques’ and argue that such triangulation increases the reliability of the results. If the results obtained through a particular data analysis technique, for example thematic analysis, are congruent with the results obtained by analysing the same transcripts using a different technique, for example correspondence analysis, it is reasonable to argue that the analysis and interpretation of the data is valid.

  13. Techniques for Specifying Bug Patterns

    Energy Technology Data Exchange (ETDEWEB)

    Quinlan, D J; Vuduc, R W; Misherghi, G

    2007-04-30

    We present our on-going work to develop techniques for specifying source code signatures of bug patterns. Specifically, we discuss two approaches. The first approach directly analyzes a program in the intermediate representation (IR) of the ROSE compiler infrastructure using ROSE's API. The second analyzes the program using the bddbddb system of Lam, Whaley, et al.. In this approach, we store the IR produced by ROSE as a relational database, express patterns as declarative inference rules on relations in the language Datalog, and bddbddb implements the Datalog programs using binary decision diagram (BDD) techniques. Both approaches readily apply to large-scale applications, since ROSE provides full type analysis, control flow, and other available analysis information. In this paper, we primarily consider bug patterns expressed with respect to the structure of the source code or the control flow, or both. More complex techniques to specify patterns that are functions of data flow properties may be addressed by either of the above approaches, but are not directly treated here. Our Datalog-based work includes explicit support for expressing patterns on the use of the Message Passing Interface (MPI) in parallel distributed memory programs. We show examples of this on-going work as well.

  14. Microirradiation techniques in radiobiological research

    Indian Academy of Sciences (India)

    Guido A Drexler; Miguel J Ruiz-Gómez

    2015-09-01

    The aim of this work is to review the uses of laser microirradiation and ion microbeam techniques within the scope of radiobiological research. Laser microirradiation techniques can be used for many different purposes. In a specific condition, through the use of pulsed lasers, cell lysis can be produced for subsequent separation of different analytes. Microsurgery allows for the identification and isolation of tissue sections, single cells and subcellular components, using different types of lasers. The generation of different types of DNA damage, via this type of microirradiation, allows for the investigation of DNA dynamics. Ion microbeams are important tools in radiobiological research. There are only a limited number of facilities worldwide where radiobiological experiments can be performed. In the beginning, research was mostly focused on the bystander effect. Nowadays, with more sophisticated molecular and cellular biological techniques, ion microirradiation is used to unravel molecular processes in the field of radiobiology. These include DNA repair protein kinetics or chromatin modifications at the site of DNA damage. With the increasing relevance of charged particles in tumour therapy and new concepts on how to generate them, ion microbeam facilities are able to address unresolved questions concerning particle tumour therapy.

  15. [Our technique in orthotopic neobladder].

    Science.gov (United States)

    Del Boca, C; Colloi, D; Guardamagna, A; Giuberti, A C; Ferrari, C

    1995-09-01

    The strive in the field of orthotopic neobladders derives from the need to improve their morphofunctional aspects and to simplify the surgical procedures. The Authors propose their experience on a new method of orthotopic neobladder in 8 patients submitted to radical cystectomy for advanced bladder neoplasia from march 91 until june 93. The surgical technique was to prepare a reservoir with a simile Camey 2 type procedure modified by them using 50 cm of ileus, 30 of which detubularized and reconfigured into a simile spheric shape with Polygia 75 staplers. The remaining length was left intact for the ureteral anastomosis performing the Wallace 1 type procedure. The advantages of this technique are that: the neobladder is prepared rapidly using staplers, thus reducing operating time the presence of an isoperistaltic segment of ileus for ureteral anastomosis permits an reduced ureteral mobilization with a low probability of reflux a simple reconversion in ileal conduit in case of reservoir failure or neoplastic urethral recurrence is possible. The criteria for selecting the patients are reported and the diagnostic algorithm regarding the follow-up presented. The latter is done with biochemical, echographic, radiological and pressure studies, every 4-6 or 12 months. Particular attention has been focused on the quality of life in relation to the diurnal/nocturnal continence and micturation interval. They conclude that this technique is surgically simple and rapid with satisfactory clinical and urodynamic results.

  16. Survey of Cataract Surgical Techniques in Nigeria

    African Journals Online (AJOL)

    PROF SABE NWOSU

    Objective: To determine the techniques of cataract surgery as currently being ... include phacoemulsification, manual small incision sutureless. 3 cataract surgery (SICS) ... technology automated small incision phacoemulsification technique.5.

  17. EDITORIAL: Imaging systems and techniques Imaging systems and techniques

    Science.gov (United States)

    Yang, Wuqiang; Giakos, George; Nikita, Konstantina; Pastorino, Matteo; Karras, Dimitrios

    2009-10-01

    The papers in this special issue focus on providing the state-of-the-art approaches and solutions to some of the most challenging imaging areas, such as the design, development, evaluation and applications of imaging systems, measuring techniques, image processing algorithms and instrumentation, with an ultimate aim of enhancing the measurement accuracy and image quality. This special issue explores the principles, engineering developments and applications of new imaging systems and techniques, and encourages broad discussion of imaging methodologies, shaping the future and identifying emerging trends. The multi-faceted field of imaging requires drastic adaptation to the rapid changes in our society, economy, environment and technological evolution. There is an urgent need to address new problems, which tend to be either static but complex, or dynamic, e.g. rapidly evolving with time, with many unknowns, and to propose innovative solutions. For instance, the battles against cancer and terror, monitoring of space resources and enhanced awareness, management of natural resources and environmental monitoring are some of the areas that need to be addressed. The complexity of the involved imaging scenarios and demanding design parameters, e.g. speed, signal-to-noise ratio (SNR), specificity, contrast, spatial resolution, scatter rejection, complex background and harsh environments, necessitate the development of a multi-functional, scalable and efficient imaging suite of sensors, solutions driven by innovation, and operation on diverse detection and imaging principles. Efficient medical imaging techniques capable of providing physiological information at the molecular level present another important research area. Advanced metabolic and functional imaging techniques, operating on multiple physical principles, and using high-resolution, high-selectivity nano-imaging methods, quantum dots, nanoparticles, biomarkers, nanostructures, nanosensors, micro-array imaging chips

  18. Swept Impact Seismic Technique (SIST)

    Science.gov (United States)

    Park, C.B.; Miller, R.D.; Steeples, D.W.; Black, R.A.

    1996-01-01

    A coded seismic technique is developed that can result in a higher signal-to-noise ratio than a conventional single-pulse method does. The technique is cost-effective and time-efficient and therefore well suited for shallow-reflection surveys where high resolution and cost-effectiveness are critical. A low-power impact source transmits a few to several hundred high-frequency broad-band seismic pulses during several seconds of recording time according to a deterministic coding scheme. The coding scheme consists of a time-encoded impact sequence in which the rate of impact (cycles/s) changes linearly with time providing a broad range of impact rates. Impact times used during the decoding process are recorded on one channel of the seismograph. The coding concept combines the vibroseis swept-frequency and the Mini-Sosie random impact concepts. The swept-frequency concept greatly improves the suppression of correlation noise with much fewer impacts than normally used in the Mini-Sosie technique. The impact concept makes the technique simple and efficient in generating high-resolution seismic data especially in the presence of noise. The transfer function of the impact sequence simulates a low-cut filter with the cutoff frequency the same as the lowest impact rate. This property can be used to attenuate low-frequency ground-roll noise without using an analog low-cut filter or a spatial source (or receiver) array as is necessary with a conventional single-pulse method. Because of the discontinuous coding scheme, the decoding process is accomplished by a "shift-and-stacking" method that is much simpler and quicker than cross-correlation. The simplicity of the coding allows the mechanical design of the source to remain simple. Several different types of mechanical systems could be adapted to generate a linear impact sweep. In addition, the simplicity of the coding also allows the technique to be used with conventional acquisition systems, with only minor modifications.

  19. Enseignement technique : Séminaire de l'enseignement technique

    CERN Document Server

    2004-01-01

    Mardi 4 mai 2004 SEMINAIRE DE L'ENSEIGNEMENT TECHNIQUE de 14:00 à 17:00 - Training Centre Auditorium - bât. 593 Le SPC - Statistical Process Control - dans une démarche de qualité totale Hakim Bourahla, Maurice Arrius, Charbel Tannous - ABW Concept - F-74950 SCIONZIER, France Ce nouveau séminaire de l'Enseignement Technique sera consacré à la présentation du SPC, le Statistical Process Control. Le SPC apporte une grande efficacité dans l'amélioration de la qualité des produits. Cette méthode, permettant d'assurer une qualité optimum à l'aide d'un outil statistique, est fondée sur deux concepts de base : le suivi et le pilotage des procédés industriels par cartes de contrôle, et l'étude des capabilités des systèmes de production. - Origine et objectifs du SPC - Concepts du ...

  20. Enseignement technique : Séminaire de l'enseigement technique

    CERN Document Server

    2004-01-01

    Mardi 4 mai 2004 SEMINAIRE DE L'ENSEIGNEMENT TECHNIQUE de 14:00 à 17:00 - Training Centre Auditorium - bât. 593 Le SPC - Statistical Process Control - dans une démarche de qualité totale Hakim Bourahla, Maurice Arrius, Charbel Tannous - ABW Concept - F-74950 SCIONZIER, France Ce nouveau séminaire de l'Enseignement Technique sera consacré à la présentation du SPC, le Statistical Process Control. Le SPC apporte une grande efficacité dans l'amélioration de la qualité des produits. Cette méthode, permettant d'assurer une qualité optimum à l'aide d'un outil statistique, est fondée sur deux concepts de base : le suivi et le pilotage des procédés industriels par cartes de contrôle, et l'étude des capabilités des systèmes de production. - Origine et objectifs du SPC - Concepts du ...