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Sample records for fluorescein diacetate staining

  1. NEW METHOD TO DETERMINE 'GIARDIA' CYST VIABILITY: CORRELATION OF FLUORESCEIN DIACETATE AND PROPIDIUM IODIDE STAINING WITH ANIMAL INFECTIVITY

    EPA Science Inventory

    The viability of Giardia muris cysts was studied using the fluorogenic dyes, fluorescein diacetate (FDA) and propidium iodide (PI). Using the mouse model for giardiasis, FDA or PI stained cysts were inoculated into neonatal mice. Feces were examined at days 3, 5, 8, and 11 post-i...

  2. Limitations in the Use of Fluorescein Diacetate/Propidium Iodide (FDA/PI) and Cell Permeable Nucleic Acid Stains for Viability Measurements of Isolated Islets of Langerhans.

    PubMed

    Boyd, Vinc; Cholewa, Olivia Maria; Papas, Klearchos K

    2008-03-01

    BACKGROUND: A review of current literature shows that the combined use of the cell permeable esterase-substrate fluorescein diacetate (FDA) and the cell impermeant nucleic acid stain propidium iodide (PI) to be one of the most common fluorescence-based methods to assess the viability of isolated islets of Langerhans, and it is currently used for islet product release prior to transplantation in humans. However, results from this assay do not correlate with islet viability and function or islet transplantation success in animals or humans (Eckhard et al. 2004; Ricordi et al. 2001). This may be in part attributed to considerable differences as well as discrepancies in the use of these reagents on islets. We critically surveyed the literature and evaluated the impact of a number of variables associated with the use of FDA/PI to determine their reliability in assessing islet cell viability. In addition, we evaluated other fluorescent stains, such as SYTO(R)13, SYTO(R)24 and SYBR(R)14 as possible alternatives to FDA. RESULTS: We found that the stability of stains in storage and stock solutions, the number of islets stained, concentration of stains, staining incubation time, the buffer/media used, and the method of examining islets were significant in the final scoring of viability. For archival file photos, the exposure time and camera/software settings can also impact interpretation of viability. Although our results show that FDA does detect intracellular esterase activity and staining with PI does assess cell membrane integrity, the results obtained from using these stains did not correlate directly with expected islet function and viability per transplantation into diabetic athymic nude mice (Papas et al. 2007). In addition, the use of two nucleic acid stains, such as SYTO(R)13 and PI, for live/dead scoring exhibited staining anomalies which limit their accuracy in assessing islet viability. CONCLUSIONS: From a review of the literature and from our observations on the impact of reagent handling and various staining and imaging parameters used to visually evaluate islets, consistent interpretation of islet cell membrane integrity and viability is dependent upon a number of factors. We discuss the utility and limitations of these reagents in evaluating islet cell membrane integrity and viability. PMID:20814586

  3. Fluorescein eye stain

    MedlinePLUS

    ... eye. The health care provider then shines a blue light at your eye. Any problems on the surface of the cornea will be stained by the dye and appear green under the blue light. The provider can determine the location and ...

  4. Development of an automated ballast water treatment verification system utilizing fluorescein diacetate hydrolysis as a measure of treatment efficacy.

    PubMed

    Akram, A C; Noman, S; Moniri-Javid, R; Gizicki, J P; Reed, E A; Singh, S B; Basu, A S; Banno, F; Fujimoto, M; Ram, J L

    2015-03-01

    Methods for verifying ballast water treatments in foreign vessels are needed to protect the Great Lakes from the discharge of live non-native organisms or pathogens. A prototype automated viability test system using fluorescein diacetate (FDA), a membrane permeable fluorogen, to differentiate live from dead bacteria and algae is described. The automated fluorescence intensity detection device (AFIDD) captures cultured algae or organisms in Detroit River water (simulated ballast water) on 0.2 ?m filters, backwashes them from the filter into a cuvette with buffer and FDA for subsequent fluorescence intensity measurements, and washes the filters with sterile water for serial automated reuse. Preliminary manual versions of these procedures were also tested. Tests of various buffers determined N,N-Bis(2-hydroxyethyl)-2-aminoethanesulfonic acid, N,N-Bis(2-hydroxyethyl)taurine (BES) and 3-(N-morpholino)propanesulfonic acid (MOPS) at pH 7.0 to be the best buffers, causing the least spontaneous FDA breakdown without inhibiting enzymatic activity. Fluorescence in the presence of live organisms increased linearly over time, and the rate of increase was dependent on the sample concentration. Following simulated ballast water treatments with heat or chlorine, the fluorescence produced by Detroit River samples decreased to near control (sterile water) levels. Automated measurements of FDA hydrolysis with a reusable filter backwash system should be applicable to near real-time remote-controlled monitoring of live organisms in ballast water. PMID:25555225

  5. Fluorescent Staining of Tea Pathogenic Fungi in Tea Leaves Using Fluorescein-labeled Lectin

    NASA Astrophysics Data System (ADS)

    Yamada, Kengo; Yoshida, Katsuyuki; Sonoda, Ryoichi

    Fluorochrome-labeled lectin, fluorescein conjugated wheat germ agglutinin (F-WGA) was applied to stain tea pathogenic fungi in tea leaf tissue. Infected leaves were fixed and decolorized with a mixture of ethanol and acetic acid, and cleared with 10% KOH for whole mount before staining with F-WGA. Hyphae of Pestalotiopsis longiseta, Pseudocercospora ocellata, Botrytis cinerea and Colletotrichum theae-sinensis fluoresced brightly in whole mount and sectioned samples of infected leaf tissue. In browned tissue, hyphae did not fluoresce frequently in whole mount sample. Autofluorescence of leaf tissue was strong in browned tissue of sections, it was removed by 10% KOH treatment before staining. Penetration hyphae of C. theae-sinensis in cell wall of trichome and hyphae in basal part of trichome did not fluoresced frequently. In whole mount samples of tea leaf infected with Exobasidium vexans and E. reticulatum, hymenia appeared on leaf surface fluoresced, but hyphae in leaf tissue did not fluoresce. In sectioned samples, hyphae fluoresced brightly when sections were treated with 10% KOH before staining.

  6. The photography of fluorescein

    SciTech Connect

    Welch, J.D.

    1982-06-01

    The last few years have seen a number of new flaps described and a renewed interest in the use of fluorescein, but there have been few photographs of the fluorescein effect, because special light sources were required with the filters that were employed. The realization that fluorescein can be excited by electromagnetic radiation in the visible range allows a simplified technique in which an ordinary electronic flash unit may serve as the only light source. The photography of fluorescein is not difficult to perform, and since minimal additional equipment is required, all workers who use fluorescein should begin to document their work more accurately and dramatically.

  7. New fluorescein precursors for live bacteria detection.

    PubMed

    Guilini, Celia; Baehr, Corinne; Schaeffer, Etienne; Gizzi, Patrick; Rufi, Frédéric; Haiech, Jacques; Weiss, Etienne; Bonnet, Dominique; Galzi, Jean-Luc

    2015-09-01

    Swiftness, reliability, and sensitivity of live bacteria detection in drinking water are key issues for human safety. The most widespread used indicator of live bacteria is a caged form of carboxyfluorescein in which 3' and 6' hydroxyl groups are masked as acetate esters (CFDA). This derivatization altogether abolishes fluorescein fluorescence and renders the molecule prone to passive diffusion through bacterial membranes. Once in the cytoplasm, acetate groups from CFDA are removed by bacterial hydrolases and fluorescence develops, rendering live but not dead cells detectable. Yet the reagent, carboxyfluorescein diacetate, still possesses a free carboxyl group whose ionization constant is such that the majority of the probe is charged at physiological pH. This unfavors probe permeation through membranes. Here, we prepare several chemical modifications of the carboxyl moiety of CFDA, in order to neutralize its charge and improve its passive diffusion through membranes. We show that the ethylamido derivative of the 5-carboxyl group from 5-carboxy-fluorescein diacetate or from Oregon green diacetate or from Oregon green diacetoxymethylester are stable molecules in biological media, penetrate into bacterial cells and are metabolized into fluorescent species. Only live bacteria are revealed since bleached samples are not labeled. Other derivatives with modification of the 5-carboxyl group with an ester group or with a thiourea-based moiety were almost inefficient probes. The most interesting probe, triembarine (5-ethylaminocarboxy-oregon green, 3',6'diacetoxymethyl ester) leads to 6-10 times more sensitive detection of bacteria as compared to CFDA. Addition of contrast agents (trypan blue or brilliant blue R) improve the signal-to-noise ratio by quenching extracellular fluorescence while bromophenol blue quenches both intracellular and extracellular fluorescence, allowing standardization of detections. PMID:26260548

  8. Peptide Targeting of Fluorescein-Based Sensors to Discrete Intracellular Locales

    PubMed Central

    Radford, Robert J.; Chyan, Wen

    2014-01-01

    Fluorescein-based sensors are the most widely applied class of zinc probes but display adventitious localization in live cells. We present here a peptide-based localization strategy that affords precision in targeting of fluorescein-based zinc sensors. By appending the zinc-selective, reaction-based probe Zinpyr-1 diacetate (DA-ZP1) to the N-terminus of two different targeting peptides we achieve programmable localization and avoid unwanted sequestration within acidic vesicles. Furthermore, this approach can be generalized to other fluorescein-based sensors. When appended to a mitochondrial targeting peptide, the esterase-activated profluorophore 2?,7?-dichlorofluorescein diacetate can be used effectively at concentrations four-times lower than previously reported for analogous, non-acetylated derivatives. These results demonstrate on-resin or in-solution esterification of fluorescein to be an effective strategy to facilitate peptide-based targeting in live cells. PMID:25512838

  9. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 2011-04-01 2011-04-01 false Calcium diacetate. 582.6197 Section 582.6197...RECOGNIZED AS SAFE Sequestrants 2 § 582.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

  10. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 2011-04-01 2011-04-01 false Calcium diacetate. 182.6197 Section 182.6197...RECOGNIZED AS SAFE Sequestrants 1 § 182.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

  11. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 2010-04-01 2010-04-01 false Calcium diacetate. 582.6197 Section 582.6197...RECOGNIZED AS SAFE Sequestrants 2 § 582.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

  12. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...3 2010-04-01 2009-04-01 true Calcium diacetate. 182.6197 Section 182.6197...RECOGNIZED AS SAFE Sequestrants 1 § 182.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

  13. Confusion over live/dead stainings for the detection of vital microorganisms in oral biofilms - which stain is suitable?

    PubMed Central

    2014-01-01

    Background There is confusion over the definition of the term “viability state(s)” of microorganisms. “Viability staining” or “vital staining techniques” are used to distinguish live from dead bacteria. These stainings, first established on planctonic bacteria, may have serious shortcomings when applied to multispecies biofilms. Results of staining techniques should be compared with appropriate microbiological data. Discussion Many terms describe “vitality states” of microorganisms, however, several of them are misleading. Authors define “viable” as “capable to grow”. Accordingly, staining methods are substitutes, since no staining can prove viability. The reliability of a commercial “viability” staining assay (Molecular Probes) is discussed based on the corresponding product information sheet: (I) Staining principle; (II) Concentrations of bacteria; (III) Calculation of live/dead proportions in vitro. Results of the “viability” kit are dependent on the stains’ concentration and on their relation to the number of bacteria in the test. Generally this staining system is not suitable for multispecies biofilms, thus incorrect statements have been published by users of this technique. To compare the results of the staining with bacterial parameters appropriate techniques should be selected. The assessment of Colony Forming Units is insufficient, rather the calculation of Plating Efficiency is necessary. Vital fluorescence staining with Fluorescein Diacetate and Ethidium Bromide seems to be the best proven and suitable method in biofilm research. Regarding the mutagenicity of staining components users should be aware that not only Ethidium Bromide might be harmful, but also a variety of other substances of which the toxicity and mutagenicity is not reported. Summary –?The nomenclature regarding “viability” and “vitality” should be used carefully. –?The manual of the commercial “viability” kit itself points out that the kit is not suitable for natural multispecies biofilm research, as supported by an array of literature. –?Results obtained with various stains are influenced by the relationship between bacterial counts and the amount of stain used in the test. Corresponding vitality data are prone to artificial shifting. –?As microbiological parameter the Plating Efficiency should be used for comparison. –?Ethidium Bromide is mutagenic. Researchers should be aware that alternative staining compounds may also be or even are mutagenic. PMID:24410850

  14. 21 CFR 582.6754 - Sodium diacetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Sodium diacetate. 582.6754 Section 582.6754 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium diacetate. (a) Product. Sodium diacetate. (b) Conditions of use. This substance is...

  15. 21 CFR 582.6754 - Sodium diacetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Sodium diacetate. 582.6754 Section 582.6754 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium diacetate. (a) Product. Sodium diacetate. (b) Conditions of use. This substance is...

  16. 21 CFR 582.6754 - Sodium diacetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Sodium diacetate. 582.6754 Section 582.6754 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium diacetate. (a) Product. Sodium diacetate. (b) Conditions of use. This substance is...

  17. 21 CFR 582.6754 - Sodium diacetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Sodium diacetate. 582.6754 Section 582.6754 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium diacetate. (a) Product. Sodium diacetate. (b) Conditions of use. This substance is...

  18. 21 CFR 582.6754 - Sodium diacetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Sodium diacetate. 582.6754 Section 582.6754 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium diacetate. (a) Product. Sodium diacetate. (b) Conditions of use. This substance is...

  19. Anterior segment fluorescein cineangiography.

    PubMed

    Kottow, M H; Jednock, N; Sewell, J H

    1978-03-01

    We have developed a technique for performing anterior segment fluorescein cineangiography. Illumination is obtained with a halogen lamp of a standard slide projector that is fitted with a blue excitation filer. Cinematography occurs with a movie camera fitted with an absorption-type barrier filter and mounted to a photo slit lamp through a cineadapter. The technique has been successfully employed with animals, and it is anticipated that the light levels used are tolerable and safe for application with humans. PMID:306767

  20. Phospha-fluorescein: a red-emissive fluorescein analogue with high photobleaching resistance.

    PubMed

    Fukazawa, Aiko; Suda, Shinji; Taki, Masayasu; Yamaguchi, Eriko; Grzybowski, Marek; Sato, Yoshikatsu; Higashiyama, Tetsuya; Yamaguchi, Shigehiro

    2016-01-01

    Phospha-fluorescein (POF), a phosphine oxide-containing analogue of fluorescein, was synthesized and its photophysical properties were examined. Compared with fluorescein and sila-fluorescein, POF displayed significantly red-shifted absorption and fluorescence as well as superior photobleaching resistance, while retaining the pH-responsive characteristics of fluorescein dyes. PMID:26617335

  1. "Geyser" leakage on fluorescein angiography.

    PubMed

    Levy, Jaime; Fagan, Xavier J; Lifshitz, Tova; Schneck, Marina

    2013-01-01

    An 82-year-old patient with diabetes was followed up due to moderate nonproliferative diabetic retinopathy with macular edema in the right eye. Visual acuity was 6/36. Focal macular laser was conducted (A). Three years later, the patient presented with blurry vision in the right eye. Visual acuity was 3/60. Vitreous hemorrhage was observed (B), and neovascularization of the disc was suspected (C). Fluorescein angiography (D, mid venous phase; E-F, recirculation phase) confirmed neovascularization of the disc and depicted a striking vertical leakage. Panretinal photocoagulation was started. Possible explanations for the "geyser" leakage may be either a partial posterior vitreous detachment allowing the fluorescein to track upwards but not elsewhere or a pocket of syneretic vitreous allowing the fluorescein passage in which to diffuse, much like the passage the blood would have taken. PMID:24548789

  2. 77 FR 23269 - Determination That FUNDUSCEIN-25 (fluorescein sodium injection), 25%, and AK-FLUOR (fluorescein...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-18

    ...Determination That FUNDUSCEIN-25 (fluorescein sodium injection), 25%, and AK-FLUOR (fluorescein sodium injection), 25%, Were Not Withdrawn...determined that FUNDUSCEIN-25 (fluorescein sodium injection), 25%, and AK-FLUOR...

  3. Method and compositions for detecting of bloodstains using fluorescin-fluorescein reaction

    DOEpatents

    Di Benedetto, John; Kyle, Kevin; Boan, Terry; Marie, Charlene

    2004-02-17

    A method, compositions and kit are set forth for detecting blood stains. A reactant solution includes fluorescin solubilized (reduced) in acetic acid in ethanol. The solution may be buffered to a pH of approximately 9. After spraying the reactant solution on the suspected area an oxidizer is applied to promote the fluorescin to fluorescein reaction with the blood. The reacted fluorescein is then detected through luminescence for capture by photography.

  4. STUDIES ON FLUORESCENT ANTIBODY STAINING

    PubMed Central

    Goldstein, Gerald; Slizys, Irene S.; Chase, Merrill W.

    1961-01-01

    1. A study has been made of the non-specific fluorescent staining of splenic imprints treated with fluorescent sheep antibody globulins. 2. In tissue imprints made with the spleens of antigen-stimulated animals, no morphological distinction was evident between areas showing non-specific fluorescence and specific fluorescence. 3. Elimination of non-specific fluorescence was not achieved by any one, or any combination of the following: (a) conjugating only gamma globulins with fluorescein isothiocyanate; (b) removal of dialyzable fluorescent products on sephadex, followed by concentration through the use of pressure dialysis; (c) use of crystalline preparations of fluorescein isothiocyanate. 4. Individual preparations of fluorescent antibodies were separated by gradient elution chromatography on diethylaminoethyl (DEAE) cellulose into fractions possessing different numbers of fluorescein radicals per molecule of globulin. 5. The coupling ratio of 50 mg fluorescein isothiocyanate (FITC) per gm of protein, as commonly advocated, can not be recommended for the precise localization of antibody globulin in tissues owing to the capacity of the coupled products to give non-specific fluorescent staining. When crystalline preparations of FITC are used instead of the amorphous product at 50 mg/gm protein, far too high non-specific fluorescence results. 6. A fraction with bright specific fluorescence and no or negligible nonspecific fluorescence was obtained from each fluorescent antibody that was prepared by using 6 to 8 mg of crystalline fluorescein isothiocyanate per gm of globulin and was then subjected to DEAE-cellulose chromatography and gradient elution to eliminate the most highly coupled molecules. PMID:13706641

  5. 21 CFR 184.1754 - Sodium diacetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium diacetate. 184.1754 Section 184.1754 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS...

  6. 21 CFR 184.1754 - Sodium diacetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Sodium diacetate. 184.1754 Section 184.1754 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS...

  7. Stain Reagent Reversible Stain Kits

    E-print Network

    Lebendiker, Mario

    further diluted serially in 1X SDS-PAGE buffer (50 mM Tris·HCl, pH 6.8, 100 mM dithiothreitol, 2% SDS, 0 during the staining process. 1 Water Wash-Enhanced Protein Staining With GelCode ® Blue Stain Reagent to the nanogram level. The main drawbacks of this method is that it requires long (12-hour) staining times

  8. 77 FR 23269 - Determination That FUNDUSCEIN-25 (fluorescein sodium injection), 25%, and AK-FLUOR (fluorescein...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-18

    ... SERVICES Food and Drug Administration Determination That FUNDUSCEIN-25 (fluorescein sodium injection), 25%, and AK-FLUOR (fluorescein sodium injection), 25%, Were Not Withdrawn From Sale for Reasons of Safety... Drug Administration (FDA) has determined that FUNDUSCEIN-25 (fluorescein sodium injection), 25%, and...

  9. Quantitative studies of immunofluorescent staining*

    PubMed Central

    Beutner, Ernst H.; Sepulveda, Marion R.; Barnett, Eugene V.

    1968-01-01

    Reproducible titres of indirect immunofluorescent (IF) staining with antinuclear factor (ANF)-containing sera could be obtained with different antihuman IgG conjugates by quantitative adjustments of their characteristics. Conversely, one ANF yielded a broad range of ANF titre (80-640) upon appropriate adjustments of the conjugate characteristics. The same and related characteristics of the conjugates also afforded a basis for quantitatively defining the conditions under which non-specific staining (NSS) appeared. The salient characteristics of the anti-IgG conjugates include: (1) their strength of antiglobulin (expressed as units/ml of precipitating antibody or ?g antibody N/ml); (2) their apparent fluorescein concentration (in ?g F/ml); (3) their protein concentration (in mg/ml). Optical and immunologic sensitivity ratios are calculated from these conjugate characteristics. Optical sensitivity (expressed as fluorescein concentration to protein concentration (F/P) ratios), immunological sensitivities (expressed as units/1% protein) and the dilution employed serve to characterize quantitatively anti-IgG conjugates adequately to define their specific and non-specific staining properties. PMID:4179321

  10. Gram Stain

    MedlinePLUS

    ... definitively identify the cause of infection. Fungi , including yeast, may also be detected with a Gram stain. ^ ... white blood cells Fungi (in the form of yeasts or molds) may be seen on a Gram ...

  11. Anthelminthic properties of mangostin and mangostin diacetate.

    PubMed

    Keiser, Jennifer; Vargas, Mireille; Winter, Rolf

    2012-06-01

    Few anthelminthic drugs are available for human use despite the significant burden caused by helminth infections. We studied the activities of mangostin, a major bioactive xanthone isolated from the pericarp and fruit of Garcinia mangostana and of the synthetic derivative mangostin diacetate. Mangostin and mangostin diacetate lacked activity against the nematodes Heligmosomoides polygyrus (third-stage larvae (L3)), Ancylostoma ceylanicum L3, and Trichuris muris adults and showed only low activity against A. ceylanicum adults (IC??s of 91 ?g/ml) in vitro. Mangostin showed promising activities (IC?? of 2.9-15.6 ?g/ml) against the trematodes Schistosoma mansoni, Echinostoma caproni, and Fasciola hepatica in vitro. Single oral doses (400 mg/kg and 800 mg/kg) of the drugs achieved worm burden reductions ranging from 0 to 38% and 11-54% against S. mansoni and E. caproni in vivo, respectively. Pharmacokinetic studies would be helpful to understand the differences observed between in vitro and in vivo activities and lacking dose-response relationships. PMID:22265670

  12. 21 CFR 522.1075 - Gonadorelin diacetate tetrahydrate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS § 522.1075 Gonadorelin diacetate tetrahydrate. (a)...

  13. IRRADIATION (GAMMA) OF FINE EMULSION SAUSAGE THAT CONTAINTED SODIUM DIACETATE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Listeria monocytogenes, a psychrotrophic food-borne pathogen, is a frequent post-process contaminant on ready-to-eat meat (RTE) products including frankfurters and bologna. Ionizing radiation can eliminate L. monocytogenes from ready-to-eat meats. Sodium diacetate (SDA) inhibits the growth of L. mo...

  14. Fluorescent Sensors for Zn2+ Based on a Fluorescein Platform

    E-print Network

    Tsien, Roger Y.

    Fluorescent Sensors for Zn2+ Based on a Fluorescein Platform: Synthesis, Properties fluorescent sensors for Zn2+ that utilize fluorescein as a reporting group, Zinpyr-1 and Zinpyr-2, have been. Both Zinpyr sensors have excitation and emission wavelengths in the visible range (500 nm

  15. Immunofluorescence analysis of cytokeratin 8/18 staining is a sensitive assay for the detection of cell apoptosis

    PubMed Central

    DONG, QIAO-MEI; LING, CHUN; ZHAO, LI

    2015-01-01

    Apoptosis is one of the major types of programmed cell death. During this process, cells experience a series of morphological and biochemical changes. Flow cytometric analysis of Annexin V staining of cell surface phosphatidylserine, in combination with a DNA-staining dye to probe the permeability of the cell membrane, is an established method for detecting apoptosis. The present study aimed to show that the immunofluorescence analysis of cytokeratin (CK) 8/18 staining may provide a new and sensitive assay for the detection of apoptotic cells. Tumor cells were treated with 20 ?M cisplatin to induce apoptosis. Following 12 and 24 h of cisplatin treatment, cells were collected and stained with 4?,6-diamidine-2?-phenylindole dihydrochloride (DAPI) and fluorescein-labeled anti-CK8/18 antibody. The apoptotic cells were subsequently examined by fluorescence microscopy. Annexin V-fluorescein isothiocyanate/propidium iodide staining followed by flow cytometric analysis confirmed that cisplatin was able to induce apoptosis in tumor cells. Immunofluorescence analysis demonstrated that apoptotic cells had a distinct CK8/18 staining pattern. In living cells, CK8/18 was uniformly distributed in the cytoplasm and cytosol; however in the apoptotic cells with a condensed and/or fragmented apoptotic nucleus (as identified by DAPI staining), fluorescein-labeled anti-CK8/18 antibody exhibited unusual punctate and/or bubbly staining in the cytosol. In the apoptotic cells that could not be identified by DAPI staining, fluorescein-labeled CK8/18 displayed polarized aggregated staining in the cytosol. These results indicate that fluorescein-conjugated CK8/18 may be a useful and sensitive indicator of cell apoptosis. PMID:25663887

  16. Port-Wine Stain

    MedlinePLUS

    ... related to port-wine stains are sometimes called salmon patches, which may also be called angel kisses ( ... of the baby's neck). Like port-wine stains, salmon patches start as flat, pink or red patches; ...

  17. Acid-fast stain

    MedlinePLUS

    The acid-fast stain is a laboratory test that determines if a sample of tissue, blood, or other body ... dye. The slide is then washed with an acid solution and a different stain is applied. Bacteria ...

  18. Fluorescein-labeled glutathione to study protein S-glutathionylation.

    PubMed

    Landino, Lisa M; Brown, Carolyn M; Edson, Carolyn A; Gilbert, Laura J; Grega-Larson, Nathan; Wirth, Anna Jean; Lane, Kelly C

    2010-07-01

    Numerous studies of S-glutathionylation of cysteine thiols indicate that this protein modification plays a key role in redox regulation of proteins. To facilitate the study of protein S-glutathionylation, we developed a synthesis and purification to produce milligram quantities of fluorescein-labeled glutathione. The amino terminus of the glutathione tripeptide reacted with fluorescein isothiocyanate readily in ammonium bicarbonate. Purification by solid phase extraction on C8 and C18 columns separated excess reactants from desired products. Both oxidized and reduced fluorescein-labeled glutathione reacted with a variety of thiol-containing proteins to yield fluorescent proteins. PMID:20156418

  19. Port-wine stain

    MedlinePLUS

    A port-wine stain is a birthmark in which swollen blood vessels create a reddish-purplish discoloration of the skin. ... Port-wine stains occur in about 3 out of 1,000 people. In rare cases, port-wine stains are ...

  20. Acute Toxicity of Sodium Fluorescein to Ashy Pebblesnails Fluminicola fuscus

    USGS Publications Warehouse

    Stockton, Kelly A.; Moffitt, Christine M.; Blew, David L.; Farmer, C. Neil

    2011-01-01

    Water resource agencies and groundwater scientists use fluorescein dyes to trace ground water flows that supply surface waters that may contain threatened or endangered mollusk species. Since little is known of the toxicity of sodium fluorescein to mollusks, we tested the toxicity of sodium fluorescein to the ashy pebblesnail Fluminicola fuscus. The pebblesnail was selected as a surrogate test species for the threatened Bliss Rapid snail Taylorcocha serpenticola that is endemic to the Snake River and its tributaries in the Hagerman Valley, Idaho. In laboratory tests, we expose replicated groups of snails to a series of concentrations of fluorescein in a static 24 h exposure at 15 degrees C. Following the exposure, we removed snails, rinsed them, and allowed a 48 h recovery in clean water before recording mortality. We estimated 377 mg/L as the median lethal dose. Mortality to snails occurred at concentrations well above those expected in test wells during the monitoring efforts.

  1. The effect of fluorescein volume on lacrimal outflow transit time.

    PubMed

    Tucker, N A; Codère, F

    1994-12-01

    The Jones primary dye test is a commonly used test of lacrimal outflow. Some clinicians, however, find it of limited practical significance because of variable outcome and relatively low sensitivity in documenting normal lacrimal excretory function. We hypothesized that an important variable affecting the transit time may be the volume of fluorescein used in the primary dye test. To accurately determine the exact time from insertion of dye into the eye to its appearance in the nose, using a rigid nasal endoscope, we directly visualized the dye as it appeared at the ostium of the nasolacrimal duct. Fifty nasolacrimal outflow systems were examined in 25 normal volunteers. The fluorescein dye transit time was determined using a single drop of fluorescein on one side and multiple drops of fluorescein on the other side. Using a single drop of fluorescein, the median dye transit time was 8 min, compared to 1.4 min using multiple drops. These results suggest that the volume of fluorescein used may be an important factor affecting variability in the outcome of the primary dye test. PMID:7865446

  2. Gram stain of tissue biopsy

    MedlinePLUS

    ... stain of tissue biopsy test involves using crystal violet stain to test a sample of tissue taken ... microscope slide. The specimen is stained with crystal violet stain and goes through more processing before it ...

  3. Detection of disseminated peritoneal tumors by fluorescein diacrylate in mice

    NASA Astrophysics Data System (ADS)

    Harada, Yoshinori; Furuta, Hirokazu; Murayama, Yasutoshi; Dai, Ping; Fujikawa, Yuta; Urano, Yasuteru; Nagano, Tetsuo; Morishita, Koki; Hasegawa, Akira; Takamatsu, Tetsuro

    2009-02-01

    Tumor invasion to the peritoneum is a poor prognostic factor in cancer patients. Accurate diagnosis of disseminated peritoneal tumors is essential to accurate cancer staging. To date, peritoneal washing cytology during laparotomy has been used for diagnosis of peritoneal dissemination of gastrointestinal cancer, but its sensitivity has not been satisfactory. Thus, a more direct approach is indispensable to detect peritoneal dissemination in vivo. Fluorescein diacrylate (FDAcr) is an esterase-sensitive fluorescent probe derived from fluorescein. In cancer cells, fluorescent fluorescein generated by exogenous application of FDAcr selectively deposits owing to its stronger hydrolytic enzyme activity and its lower leakage rate. We examined whether FDAcr can specifically detect disseminated peritoneal tumors in athymic nude mouse models. Intraperitoneally administered FDAcr revealed disseminated peritoneal microscopic tumors not readily recognized on white-light imaging. These results suggest that FDAcr is a useful probe for detecting disseminated peritoneal tumors.

  4. Apparatus Would Stain Microscope Slides

    NASA Technical Reports Server (NTRS)

    Breeding, James D.

    1993-01-01

    Proposed apparatus meters specific amounts of fluid out of containers at specific times to stain microscope slides. Intended specifically for semiautomated staining of microbiological and hematological samples in microgravity, leakproof apparatus used in other environments in which technicians have little time to allocate to staining procedures and/or exposure to toxic staining agents or to micro-organisms to be stained hazardous. Apparatus adapted to perform almost any staining procedure and accommodates multiple staining reagents, useful for small or remote clinical laboratories.

  5. Port-Wine Stains

    MedlinePLUS

    ... tend to become darker (usually reddish-purple or dark red) as kids grow. Port-wine stains (also ... upsetting for kids, especially if they're large, dark, or on the face. And any birthmark can ...

  6. Quick Stain Removal Guide 

    E-print Network

    Brown, Pamela J.

    1998-07-29

    . ? Sort the clothes into like stacks of colors, construction, fiber content, surface texture and degree of soil. ? Pretreat stains. ? Measure the amount of detergent recommended on the laundry container. ? Avoid overloading the washer... and dirt stains occur on the lower edges of pants and cuffs. Check for colorfastness. Pretreatments can change a garment?s color. Before using, test it in an inconspicuous area on the garment, such as a hem or seam allowance. Apply the pretreat...

  7. [Clinical and fluorescein angiographic aspects in retinal angiomatosis (von Hippel)].

    PubMed

    Samoil?, O; C?lug?ru, D; C?lug?ru, M

    2007-01-01

    Retinal angiomas represent a rare condition consisting of hamartomas isolated or associated with a systemic disease (Von Hippel-Lindau Disease). We present a patient with multiple retinal angiomas, with atypical fluorescein angiography possible due to vascular thrombotic phenomena. Visual acuity was slightly diminished because of hemorrhagic complications. Systemic involvement was excluded with imaging studies. PMID:17937036

  8. Dodecyl and octyl esters of fluorescein as protonophores and uncouplers of oxidative phosphorylation in mitochondria at submicromolar concentrations.

    PubMed

    Shchepinova, Maria M; Denisov, Stepan S; Kotova, Elena A; Khailova, Ljudmila S; Knorre, Dmitry A; Korshunova, Galina A; Tashlitsky, Vadim N; Severin, Fedor F; Antonenko, Yuri N

    2014-01-01

    In our search for fluorescent uncouplers of oxidative phosphorylation, three esters of fluorescein, n-butyl-, n-octyl-, and n-dodecyl-oxycarbonyl-fluorescein (C4-FL, C8-FL, C12-FL) were synthesized and characterized. With increasing liposomal lipid content, the long-chain alkyl derivatives of fluorescein (C8-FL, C12-FL and commercially available C18-FL), but not C4-FL and unsubstituted fluorescein, exhibited an increase in fluorescence polarization reflecting the dye binding to liposomes. C12-FL induced proton permeability in lipid membranes, while C4-FL was inactive. In contrast to C4-FL and C18-FL, C12-FL and C8-FL increased the respiration rate and decreased the membrane potential of isolated rat liver mitochondria with half-maximal effective concentrations of 700nM and 300nM, respectively. The effect of Cn-FL on the respiration correlated with that on proton permeability of the inner mitochondrial membrane, as measured by induction of mitochondria swelling in the potassium acetate medium. Binding of C8-FL to mitochondria depended on their energization, which was apparently associated with pH gradient generation across the inner mitochondrial membrane in the presence of a respiratory substrate. In wild-type yeast cells, C12-FL localized predominantly in plasma membrane, whereas in AD1-8 mutants lacking MDR pumps, it stained cytoplasmic organelles with some preference for mitochondria. Fluorescent uncouplers can be useful as a tool for determining their localization in a cell or distribution between different tissues in a living animal by fluorescent microscopy. PMID:24076107

  9. Four new compounds, ficusal, ficusesquilignan A, B, and ficusolide diacetate from the heartwood of Ficus microcarpa.

    PubMed

    Li, Y C; Kuo, Y H

    2000-12-01

    Three new lignans, ficusal (1) and ficusesquilignans A (2), B (3) and one new gamma-lactone, ficusolide diacetate (4), were isolated from the wood of Ficus microcarpa L.f. Their structures were determined by spectral evidence. PMID:11145132

  10. Detection of tetracysteine-tagged proteins using a biarsenical fluorescein derivative through dry microplate array gel electrophoresis.

    PubMed

    Feldman, Galia; Bogoev, Roumen; Shevirov, Julia; Sartiel, Adam; Margalit, Ilana

    2004-08-01

    The design of an extended-run 96-well sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) system and the development of protein detection technology based upon fluorescein derivatives that bind to peptide epitope tags, allows the creation of a truly high-throughput analysis of protein expression, where less than 20 min are needed to separate proteins and analyze results. We demonstrate the overall capabilities of such a method combination in a complex cell lysate background, while comparing the specific results obtained using a biarsenical fluorescein-derivative and tetracysteine epitope-tagged proteins with total protein staining using a fluorescent gel stain and with Western blotting where an anti-oligohistidine (His) tag antibody has been employed. When applied on purified target proteins without extraneous protein background, the demonstrated sensitivity of the assay on the extended-run 96-array precast SDS-PAGE system allows detection of quantities of tagged protein as low as 1 pmol per band. PMID:15300761

  11. A Rapid Procedure for Preparing Fluorescein-labeled Specific Antibodies from Whole Antiserum-

    E-print Network

    Goldman, Robert D.

    A Rapid Procedure for Preparing Fluorescein-labeled Specific Antibodies from Whole Antiserum- Its conjugation of affinity-purified antibodies with fluorescein (termed DCAPA) is described. This procedure involves the immobilization of antibodies as antigen- antibody complexes on nitrocellulose blots

  12. THE USE OF FLUORESCEIN DYE TO STUDY THE MOVEMENT OF WATER INTO WHEAT KERNELS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The dispersion of fluorescein dye in water was investigated during the tempering process in wheat kernels. Durum and soft wheat kernels were immersed in fluorescein (free acid crystalline) solution (1%, pH7.54) for 12,24 and 48 hours. The fluorescein pattern showed that the dye solution entered th...

  13. Joint fluid Gram stain

    MedlinePLUS

    Gram stain of joint fluid ... A sample of joint fluid is needed. The fluid sample is sent to a lab where a small drop is placed in a ... on how to prepare for the removal of joint fluid, see joint fluid aspiration .

  14. Shimmering Stained Glass.

    ERIC Educational Resources Information Center

    Simon, Gail Murray

    1998-01-01

    Presents an art lesson for fifth- and sixth-graders where they create a translucent design of colored cellophane on black paper inspired by the stained-glass windows of the Middle Ages and the artwork of Lewis Comfort Tiffany. Enables the students to become crafts people rather than just observers of the past. (CMK)

  15. Stained-Glass Pastels

    ERIC Educational Resources Information Center

    Laird, Shirley

    2009-01-01

    The author has always liked the look of stained-glass windows. Usually the designs are simplified and the shapes are easier for younger students to draw. This technique seemed to be the perfect place for her fifth-graders to try their hand at color mixing. The smaller spaces and simple shapes were just what she needed for this group. Her students…

  16. "Stained Glass" Landscape Windows

    ERIC Educational Resources Information Center

    Vannata, Janine

    2008-01-01

    Both adults and children alike marvel at the grand vivid stained-glass windows created by American artist Louis Comfort Tiffany. Today he is commonly recognized as one of America's most influential designers and artists throughout the last nineteenth and early twentieth century. In the lesson described in this article, students created their own…

  17. Ionic Liquid-Based Fluorescein Colorimetric pH Nanosensors

    PubMed Central

    Das, Susmita; Magut, Paul K. S.; de Rooy, Sergio L.; Hasan, Farhana; Warner, Isiah M.

    2014-01-01

    A novel pH sensitive, colorimetric ionic liquid nanosensor based on phosphonium salts of fluorescein is reported. Herein, fluorescein salts of various stoichiometries were synthesized by use of a trihexyltetradecylphosphonium cation [TTP]+ in combination with dianionic [FL]2? and monoanionic [FL]? fluorescein. Nanomaterials derived from these two compounds yielded contrasting colorimetric responses in neutral and acidic environments. Variations in fluorescence spectra as a function of pH were also observed. Examination of TEM and DLS data revealed significant expansion in the diameter of [TTP]2[FL] nanodroplets in acidic environments of variable pHs. A similar trend was also observed for [TTP][FL] nanoparticles. The pH dependent colorimetric and other optical properties of these nanomaterials are attributed to alterations in molecular orientations and stacking as suggested by measuring the absorption, fluorescence, and zeta potential. Since the pH is an important indicator for many diseases, including cancer, these nanosensors are considered to be potential candidates for biomedical applications. PMID:25264488

  18. Removing Stains from Washable Fabrics. 

    E-print Network

    Beard, Ann Vanderpoorten

    1988-01-01

    to fabrics labeled "dry clean". Fabrics labeled "dry clean" should be taken to the cleaner as soon as possible after being stained. Identify the fiber content and the type of stain for the cleaner. And remember that not all stains can be removed - by you... or by the dry cleaner. *Extension clothing and textiles specialist, TheTexasA&M Uni versity System. ? Heat sets stains. Do not dry a stained fabric in the dryer or press it until you are sure all of the stain has been removed. ? Treat stains quickly. Old...

  19. Ayanin diacetate-induced cell death is amplified by TRAIL in human leukemia cells

    SciTech Connect

    Marrero, Maria Teresa; Estevez, Sara; Negrin, Gledy; Quintana, Jose; Leon, Francisco; Estevez, Francisco

    2012-11-09

    Highlights: Black-Right-Pointing-Pointer Ayanin diacetate as apoptotic inducer in leukemia cells. Black-Right-Pointing-Pointer Cell death was prevented by caspase inhibitors and by the overexpression of Bcl-x{sub L}. Black-Right-Pointing-Pointer The intrinsic and the extrinsic pathways are involved in the mechanism of action. Black-Right-Pointing-Pointer Death receptors are up-regulated and TRAIL enhances apoptotic cell death. -- Abstract: Here we demonstrate that the semi-synthetic flavonoid ayanin diacetate induces cell death selectively in leukemia cells without affecting the proliferation of normal lymphocytes. Incubation of human leukemia cells with ayanin diacetate induced G{sub 2}-M phase cell cycle arrest and apoptosis which was prevented by the non-specific caspase inhibitor z-VAD-fmk and reduced by the overexpression of Bcl-x{sub L}. Ayanin diacetate-induced cell death was found to be associated with: (i) loss of inner mitochondrial membrane potential, (ii) the release of cytochrome c, (iii) the activation of multiple caspases, (iv) cleavage of poly(ADP-ribose) polymerase and (v) the up-regulation of death receptors for TRAIL, DR4 and DR5. Moreover, the combined treatment with ayanin diacetate and TRAIL amplified cell death, compared to single treatments. These results provide a basis for further exploring the potential applications of this combination for the treatment of cancer.

  20. Microscopic oxygen imaging based on fluorescein bleaching efficiency measurements.

    PubMed

    Beutler, Martin; Heisterkamp, Ines M; Piltz, Bastian; Stief, Peter; De Beer, Dirk

    2014-05-01

    Photobleaching of the fluorophore fluorescein in an aqueous solution is dependent on the oxygen concentration. Therefore, the time-dependent bleaching behavior can be used to measure of dissolved oxygen concentrations. The method can be combined with epi-fluorescence microscopy. The molecular states of the fluorophore can be expressed by a three-state energy model. This leads to a set of differential equations which describe the photobleaching behavior of fluorescein. The numerical solution of these equations shows that in a conventional wide-field fluorescence microscope, the fluorescence of fluorescein will fade out faster at low than at high oxygen concentration. Further simulation showed that a simple ratio function of different time-points during a fluorescence decay recorded during photobleaching could be used to describe oxygen concentrations in an aqueous solution. By careful choice of dye concentration and excitation light intensity the sensitivity in the oxygen concentration range of interest can be optimized. In the simulations, the estimation of oxygen concentration by the ratio function was very little affected by the pH value in the range of pH 6.5-8.5. Filming the fluorescence decay by a charge-coupled-device (ccd) camera mounted on a fluorescence microscope allowed a pixelwise estimation of the ratio function in a microscopic image. Use of a microsensor and oxygen-consuming bacteria in a sample chamber enabled the calibration of the system for quantification of absolute oxygen concentrations. The method was demonstrated on nitrifying biofilms growing on snail and mussel shells, showing clear effects of metabolic activity on oxygen concentrations. PMID:24610786

  1. 76 FR 75886 - Determination That DEMULEN 1/50-28 (Ethinyl Estradiol; Ethynodiol Diacetate) Tablet and Four...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-05

    ... SERVICES Food and Drug Administration Determination That DEMULEN 1/50-28 (Ethinyl Estradiol; Ethynodiol... estradiol; 4901 Searle ethynodiol diacetate) Pkwy., Skokie, Tablet, 0.05 mg; 1 mg. IL 60077. NDA 018160 DEMULEN 1/35-28 Do. (ethinyl estradiol; ethynodiol diacetate) Tablet, 0.035 mg; 1 mg. ] NDA 018168...

  2. Coffee stain technique.

    PubMed

    Hodge, Gerald P

    2002-01-01

    Backgrounds prepared with coffee grounds and Conté dust produce many unexpected patterns. The technique adds interest to artwork and is equally good for a carefully prepared drawing as it is for a quick, casual sketch. A materials list is provided which includes a variety of surface choices. The article provides step-by-step directions for the preparation of an attractive coffee-stained background, gives detailed instructions describing how to transfer a sketch, offers tips for drawing with Conté and carbon pencils and tells how to apply highlights. PMID:12199191

  3. Control of Listeria monocytogenes on commercial frankfurters prepared with and without potassium lactate and sodium diacetate and surface treated with lauric arginate....(SLIC®) delivery method

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The viability of Listeria monocytogenes was monitored on commercially-produced frankfurters that were formulated with no, low, or high levels of potassium lactate and sodium diacetate (UltraLac KL6810; low = 0.68% lactate and 0.097% diacetate and high = 1.36% lactate and 0.19% diacetate), and then t...

  4. Flash Pasteurization inactivation of Listeria innocua on frankfurters that contain potassium lactate and sodium diacetate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Listeria monocytogenes, a psychrotrophic food-borne pathogen, is a recurring post-process contaminant on ready-to-eat meat (RTE) products including frankfurters. Potassium lactate (PL) and sodium diacetate (SDA) are FDA approved antimicrobials that inhibit the growth of L. monocytogenes when incorp...

  5. Fluorogenic sequencing using halogen-fluorescein-labeled nucleotides.

    PubMed

    Chen, Zitian; Duan, Haifeng; Qiao, Shuo; Zhou, Wenxiong; Qiu, Haiwei; Kang, Li; Xie, X Sunney; Huang, Yanyi

    2015-05-26

    Fluorogenic sequencing is a sequencing-by-synthesis technology that combines the advantages of pyrosequencing and fluorescence detection. With native duplex DNA as the major product, we employ polymerase to incorporate the complement- arily matched terminal phosphate-labeled fluorogenic nucleotides into the DNA template and release halogen-fluorescein as the reporter. This red-emitting fluorophore successfully avoids spectral overlap with the autofluorescence background of the flow chip. We fully characterized the enzymatic reaction kinetics of the new substrates, and performed a 35-base sequencing experiment with 60 reaction cycles. Our achievement expands the substrate repertoire for fluorogenic sequencing, and extends the spectral range to obtain better signal-to-background performance. PMID:25846104

  6. Registration of Multimodal Fluorescein Images Sequence of the Retina Tae Eun Choe, Isaac Cohen

    E-print Network

    Cohen, Issac

    Registration of Multimodal Fluorescein Images Sequence of the Retina Tae Eun Choe, Isaac Cohen angiograms of the retina. The registration of multimodal fluorescein imagery requires the identification of multimodal retinal images. Several attempts have been made to solve this problem [2]. A common approach

  7. Synthesis of taurine–fluorescein conjugate and evaluation of its retina-targeted efficiency in vitro

    PubMed Central

    Huang, Meihong; Song, Jiaqi; Lu, Bingzheng; Huang, Huizhi; Chen, Yizhen; Yin, Wei; Zhu, Wenbo; Su, Xinwen; Wu, Chuanbin; Hu, Haiyan

    2014-01-01

    In this work, retinal penetration of fluorescein was achieved in vitro by covalent attachment of taurine to fluorescein, yielding the F–Tau conjugate. Nuclear magnetic resonance (NMR) and high resolution mass spectrometry (HRMS) were used to confirm the successful synthesis of F–Tau. The cellular uptake of F–Tau in adult retinal pigment epithelial cells (ARPE-19) and human retinal microvascular endothelial cells (hRMECs) was visualized via confocal scanning microscopy. The results indicated an improvement of solubility and a reduction of logP of F–Tau compared with fluorescein. As compared with fluorescein, F–Tau showed little toxicity, and was retained longer by cells in uptake experiments. F–Tau also displayed higher transepithelial permeabilities than fluorescein in ARPE-19 and hRMECs monolayer cells (P<0.05). These results showed that taurine may be a useful ligand for targeting small-molecule hydrophobic pharmaceuticals into the retina. PMID:26579416

  8. Multispectral Enhancement towards Digital Staining

    PubMed Central

    Bautista, Pinky A.; Yagi, Yukako

    2012-01-01

    Background: Digital staining can be considered as a special form of image enhancement wherein the concern is not only to increase the contrast between the background objects and objects of interest, but to also impart the colors that mark the objects’ unique reactions to a specific stain. In this paper, we extended the previously proposed multispectral enhancement methods such that the colors of the background pixels can also be changed. Methods: In the previous multispectral enhancement methods a shifting factor is provided to the original spectrum. To implement digital staining, a spectral transformation process is introduced prior to spectral shifting. Results: The enhancement method is applied to multispectral images of H&E stained liver tissue. The resulting digitally stained images show good correlation with the serial-section images of the tissue which are physically stained with Masson's trichrome. Conclusion: We have presented a multispectral enhancement method that can be adjusted to produce digitally stained-images. The current experimental results show the viability of the method. However, to achieve robust enhancement performance issues that arise from variations in staining conditions has to be addressed as well. This would be part of our future work. PMID:22002722

  9. Indanones and indenols from 2-alkylcinnamaldehydes via the intramolecular Friedel-Crafts reaction of geminal diacetates.

    PubMed

    Womack, Gary B; Angeles, John G; Fanelli, Vincent E; Indradas, Brinda; Snowden, Roger L; Sonnay, Philippe

    2009-08-01

    When treated with Ac2O at rt in the presence of 4-6 mol % FeCl3, 2-alkylcinnamaldehydes are converted to 2-alkyl-1H-inden-1-yl acetates through the intermediacy of gem-diacetates. Methanolysis of the indenyl acetates yields the corresponding indenols. Saponification yields 2-alkylindanones, providing, in effect, an intramolecular acylation employing catalytic levels of acid. PMID:19572598

  10. Adhesion of a Mycobacterium sp. to cellulose diacetate membranes used in reverse osmosis.

    PubMed Central

    Ridgway, H F; Rigby, M G; Argo, D G

    1984-01-01

    The kinetics of adhesion of a Mycobacterium sp. to cellulose diacetate reverse-osmosis membranes is described. This Mycobacterium sp. (strain BT2-4) was previously implicated in the initial stages of reverse-osmosis membrane biofouling at a wastewater reclamation facility. Adhesion of BT2-4 cells to the cellulose diacetate membrane surfaces occurred within 1 to 2 h at 30 degrees C and exhibited saturation-type kinetics which conformed closely to the Langmuir adsorption isotherm (Pearson r correlation coefficient = 0.977), a mathematical expression describing the partitioning of substances between a solution and solid-liquid interface. This suggests that the cellulose diacetate membrane surfaces may possess a finite number of available binding sites to which the mycobacteria can adhere. Treatment of the attached mycobacteria with different enzymes suggested that cell surface polypeptides, alpha-1, 4- or alpha-1,6-linked glucan polymers, and carboxyl ester bond-containing substances (possibly peptidoglycolipids) may be involved in mycobacterial adhesion. The possible implication of these findings for reverse-osmosis membrane biofouling are discussed. Images PMID:6696424

  11. A tracer test at the Beowawe geothermal field, Nevada, using fluorescein and tinopal CBS

    SciTech Connect

    Rose, P.E.; Adams, M.C.; Benoit, D.

    1995-12-31

    An interwell tracer test using fluorescein and tinopal CBS was performed at the Beowawe geothermal field in north-central Nevada in order to assess the effects of recent changes to the injection strategy. Fluorescein return curves established injection-production flow patterns and verified that produced water is being reinjected into a region of the reservoir that is in excellent communication with the production wells. An analysis of the tinopal CBS return curves indicated that tinopal CBS was apparently strongly adsorbed onto the reservoir rock. The fluorescein return curves were used to estimate the overall (fractures and matrix) reservoir volume.

  12. Hepatic transport mechanisms of cholyl-L-lysyl-fluorescein.

    PubMed

    de Waart, Dirk R; Häusler, Stephanie; Vlaming, Maria L H; Kunne, Cindy; Hänggi, Emanuel; Gruss, Hans-Jurgen; Oude Elferink, Ronald P J; Stieger, Bruno

    2010-07-01

    Cholyl-L-lysyl-fluorescein (CLF) is a fluorescent bile salt derivative that is being developed as an agent for determining in vivo liver function. However, the mechanisms of uptake and excretion by hepatocytes have not been rigorously studied. We have directly assessed the transport capacity of various hepatobiliary transporters for CLF. Uptake experiments were performed in Chinese hamster ovary cells transfected with human NTCP, OATP1B1, OATP1B3, and OATP2B1. Conversely, excretory systems were tested with plasma membrane vesicles from Sf21 insect cells expressing human ABCB11, ABCC2, ABCC3, and ABCG2. In addition, plasma clearance and biliary excretion of CLF were examined in wild-type, Abcc2(-/-), and Abcc3(-/-) mice. Human Na(+)-dependent taurocholic-cotransporting polypeptide (NTCP) and ATP-binding cassette B11 (ABCB11) were incapable of transporting CLF. In contrast, high-affinity transport of CLF was observed for organic anion-transporting polypeptide 1B3 (OATP1B3), ABCC2, and ABCC3 with K(m) values of 4.6 +/- 2.7, 3.3 +/- 2.0, and 3.7 +/- 1.0 microM, respectively. In Abcc2(-/-) mice biliary excretion of CLF was strongly reduced compared with wild-type mice. This resulted in a much higher hepatic retention of CLF in Abcc2(-/-) versus wild-type mice: 64 versus 1% of the administered dose (2 h after administration). In mice intestinal uptake of CLF was negligible compared with that of taurocholate. Our conclusion is that human NTCP and ABCB11 are incapable of transporting CLF, whereas OATP1B3 and ABCC2/Abcc2 most likely mediate hepatic uptake and biliary excretion of CLF, respectively. CLF can be transported back into the blood by ABCC3. Enterohepatic circulation of CLF is minimal. This renders CLF suitable as an agent for assessing in vivo liver function. PMID:20388726

  13. Objective Area Measurement Technique for Choroidal Neovascularization from Fluorescein Angiography

    PubMed Central

    Guthrie, Micah J.; Osswald, Christian R.; Valio, Nicole L.; Mieler, William F.; Kang-Mieler, Jennifer J.

    2014-01-01

    The purpose of this study was to develop a non-biased method of quantitatively measuring choroidal neovascularization (CNV) areas based on late-phase fluorescein angiography (FA) images. Experimental CNV was induced in Long Evans rats by laser disruption of the Bruch’s membrane. FA was performed weekly for 5 weeks. Multi-Otsu thresholding (MOT) was used to quantify CNV in late-phase FA images from both experimental rodent CNV and wet age-related macular degeneration patients (wAMD). Images were automatically thresholded into three levels based on the image histogram, with the highest level containing CNV. To determine the technique’s ability to quantify CNV areas, rats were given either triamcinolone acetonide or dexamethasone sodium phosphate to treat CNV and compared to untreated rats. The rat CNV lesion areas measured from 5-week histology sections from each treatment group were compared to areas measured from the corresponding FA images. MOT was able to detect statistical decreases in rodent CNV area in the treatment groups versus control from weeks 3 through 5. The ratio of CNV area measured from histology to area measured from FA images was not statistically different between groups. Finally, to determine the usefulness of MOT on pathological morphologies of CNV, MOT was performed on late-phase FA images from patients with classic and diffuse CNV. The technique was able to segment classical CNV in wAMD patients, but performed poorly with diffuse CNV. MOT provides a robust, objective, and quantifiable area measurement of CNV lesion area in both experimentally-induced and pathological CNV. The results indicate that MOT could be a useful research tool in helping evaluate the effects of therapeutics on CNV growth. PMID:24316422

  14. Colour properties of fluorescein: an interplay of tautomerism and crystal packing

    E-print Network

    Jackson, Sophie

    the colour properties of this organic pigment system, and in particular the possible role of the zwitterionic solid forms of fluorescein were measured using solid state UV/Vis spectroscopy. The measured band gap

  15. Computer-aided modeling of the binding sites of anti-fluorescein antibodies 

    E-print Network

    Harris, Jonathan S

    1996-01-01

    The variable regions of five anti-fluorescein monoclonal antibodies were sequenced and corresponding binding sites were studied by means of: steady state fluorescence, circular dichroism (CD) and computer-aided modeling. Steady state fluorescence...

  16. New stroboscopic light source and technique for intraoperative retinal fluorescein angiography during penetrating keratoplasty

    NASA Astrophysics Data System (ADS)

    Krueger, Ronald R.; Morales, Ronald B.; Chong, Lawrence P.; Smith, Ronald E.

    1994-06-01

    We report the development of a new stroboscopic light source system and technique for performing intraoperative fluorescein angiography during penetrating keratoplasty for aphakic or pseudophakic bullous keratopathy. A controllable pulse xenon light source system with a fiber optic endoilluminator probe is used to perform high-quality intraoperative fluorescein angiography during penetrating keratoplasty in pigmented rabbits and human subjects. Following corneal trephination and extraction of the intraocular lens, a temporary Cobo keratoprosthesis is secured while a 20-gauge endoilluminator is inserted into the vitreous cavity through a limbal incision. The endoilluminator is advanced to a retinal illumination area of approximately 3 DD and 10% fluorescein is injected intravenously. A microscope camera coupled to a 50:50 beamsplitter photographs the passage of fluorescein dye while the surgeon maintains an unaltered view through the operating microscope. Angiograms through a keratoprosthesis show excellent contrast and resolution, comparable to standard fluorescein angiography. Fine peripapillary vessels are seen reproducibly and with great detail in the rabbits. All the phases of retinal angiography can be seen, including arteriolar constriction and capillary nonperfusion in one of four human subjects examined. High quality intraoperative fluorescein angiography can be performed in patients undergoing penetrating keratoplasty for aphakic/ pseudophakic bullous keratopathy. With this technology, preexisting retinal disorders such as cystoid macular edema might be identified in the perioperative setting allowing for important management decisions to be made intraoperatively.

  17. Heterogeneous catalyst for the production of ethylidene diacetate from acetic anhydride

    DOEpatents

    Ramprasad, D.; Waller, F.J.

    1998-06-16

    This invention relates to a process for producing ethylidene diacetate by the reaction of acetic anhydride, acetic acid, hydrogen and carbon monoxide at elevated temperatures and pressures in the presence of an alkyl halide and a heterogeneous, bifunctional catalyst that is stable to hydrogenation and comprises an insoluble polymer having pendant quaternized heteroatoms, some of which heteroatoms are ionically bonded to anionic Group VIII metal complexes, the remainder of the heteroatoms being bonded to iodide. In contrast to prior art processes, no accelerator (promoter) is necessary to achieve the catalytic reaction and the products are easily separated from the catalyst by filtration. The catalyst can be recycled without loss in activity.

  18. Heterogeneous catalyst for the production of ethylidene diacetate from acetic anhydride

    DOEpatents

    Ramprasad, Dorai (Allentown, PA); Waller, Francis Joseph (Allentown, PA)

    1998-01-01

    This invention relates to a process for producing ethylidene diacetate by the reaction of acetic anhydride, acetic acid, hydrogen and carbon monoxide at elevated temperatures and pressures in the presence of an alkyl halide and a heterogeneous, bifunctional catalyst that is stable to hydrogenation and comprises an insoluble polymer having pendant quaternized heteroatoms, some of which heteroatoms are ionically bonded to anionic Group VIII metal complexes, the remainder of the heteroatoms being bonded to iodide. In contrast to prior art processes, no accelerator (promoter) is necessary to achieve the catalytic reaction and the products are easily separated from the catalyst by filtration. The catalyst can be recycled without loss in activity.

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

    PubMed

    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 ?M, 120 ?M, 160 ?M, 200 ?M, 240 ?M, or 320 ?M) for 45 min to evaluate effects of dye dose. The frozen-thawed sperm were stained with 160 ?M 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 (P<0.05). However, there is no significant difference existing among the Hoechst33342-stained groups with respect to the above evaluated parameters. Additionally, along with the staining durations, the adverse effects of the staining procedure on sperm showed a steady increase. However, Hoechst33342 staining did not damage acrosome and in vitro fertilizing capability of frozen-thawed ram 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 ?M. 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 160?M Hoechst33342. In conclusion, Hoechst33342 staining can detrimentally influence viability of frozen-thawed ram sperm except acrosome and in vitro fertilizing capability. Accordingly, the minimum values of Hoechst33342 concentration and staining duration can be set at 160 ?M and 45 min respectively. However, the maximum values of Hoechst33342 concentration and staining duration were not determined based on the current study. Further research on how to reduce injuries caused by freezing, thawing, and Hoechst33342 staining on frozen-thawed ram sperm is needed. PMID:25481668

  20. Whole Blood Cell Staining Device

    NASA Technical Reports Server (NTRS)

    Sams, Clarence F.; Clift, Vaughan L.; McDonald, Kelly E.

    2000-01-01

    An apparatus and method for staining particular cell markers is disclosed. The apparatus includes a flexible tube that is reversibly pinched into compartments with one or more clamps. Each compartment of the tube contains a separate reagent and is in selective fluid communication with adjoining compartments.

  1. Methods for chromosome-specific staining

    DOEpatents

    Gray, Joe W. (Livermore, CA); Pinkel, Daniel (Walnut Creek, CA)

    1995-01-01

    Methods and compositions for chromosome-specific staining are provided. Compositions comprise heterogenous mixtures of labeled nucleic acid fragments having substantially complementary base sequences to unique sequence regions of the chromosomal DNA for which their associated staining reagent is specific. Methods include methods for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes.

  2. Pro-Q Emerald Glycoprotein Stain Kits

    E-print Network

    Lebendiker, Mario

    Pro-Q Emerald Glycoprotein Stain Kits The most advanced technology for staining glycoproteins CAPABILITIES #12;Molecular Probes' proprietary Pro-Q Emerald 300 and Pro-Q Emerald 488 Glycopro- tein Stain with the Pro-Q Emerald reagent (Figure 1). Blot staining requires extra steps, but also provides excellent

  3. The effect of potassium lactate and sodium diacetate on the microbial, sensory, color and chemical characteristics of vacuum-packaged beef top loin steaks 

    E-print Network

    Anwar, Najia

    2000-01-01

    Beef strip loins were injected with potassium lactate (1.5, 2.0, and 2.5%), sodium diacetate (0.1%), and combination of sodium diacetate (0.1%) with 1.5 or 2.0% potassium lactate. Top loin steaks were vacuum-packaged and stored for up to 49 days...

  4. In vivo imaging of human retinal microvasculature using adaptive optics scanning light ophthalmoscope fluorescein angiography

    PubMed Central

    Pinhas, Alexander; Dubow, Michael; Shah, Nishit; Chui, Toco Y.; Scoles, Drew; Sulai, Yusufu N.; Weitz, Rishard; Walsh, Joseph B.; Carroll, Joseph; Dubra, Alfredo; Rosen, Richard B.

    2013-01-01

    The adaptive optics scanning light ophthalmoscope (AOSLO) allows visualization of microscopic structures of the human retina in vivo. In this work, we demonstrate its application in combination with oral and intravenous (IV) fluorescein angiography (FA) to the in vivo visualization of the human retinal microvasculature. Ten healthy subjects ages 20 to 38 years were imaged using oral (7 and/or 20 mg/kg) and/or IV (500 mg) fluorescein. In agreement with current literature, there were no adverse effects among the patients receiving oral fluorescein while one patient receiving IV fluorescein experienced some nausea and heaving. We determined that all retinal capillary beds can be imaged using clinically accepted fluorescein dosages and safe light levels according to the ANSI Z136.1-2000 maximum permissible exposure. As expected, the 20 mg/kg oral dose showed higher image intensity for a longer period of time than did the 7 mg/kg oral and the 500 mg IV doses. The increased resolution of AOSLO FA, compared to conventional FA, offers great opportunity for studying physiological and pathological vascular processes. PMID:24009994

  5. A double fluorescence staining protocol to determine the cross-sectional area of myofibers using image analysis

    NASA Technical Reports Server (NTRS)

    Mozdziak, P. E.; Fassel, T. A.; Schultz, E.; Greaser, M. L.; Cassens, R. G.

    1996-01-01

    A double fluorescence staining protocol was developed to facilitate computer based image analysis. Myofibers from experimentally treated (irradiated) and control growing turkey skeletal muscle were labeled with the anti-myosin antibody MF-20 and detected using fluorescein-5-isothiocyanate (FITC). Extracellular material was stained with concanavalin A (ConA)-Texas red. The cross-sectional area of the myofibers was determined by calculating the number of pixels (0.83 mu m(2)) overlying each myofiber after subtracting the ConA-Texas red image from the MF-20-FITC image for each region of interest. As expected, myofibers in the irradiated muscle were smaller (P < 0.05) than those in the non-irradiated muscle. This double fluorescence staining protocol combined with image analysis is accurate and less labor-intensive than classical procedures for determining the cross-sectional area of myofibers.

  6. The use of fluorescein sodium in the biopsy and gross-total resection of a tectal plate glioma.

    PubMed

    Ung, Timothy H; Kellner, Christopher; Neira, Justin A; Wang, Shih-Hsiu J; D'Amico, Randy; Faust, Phyllis L; Canoll, Peter; Feldstein, Neil A; Bruce, Jeffrey N

    2015-12-01

    Intravenous administration of fluorescein sodium fluoresces glioma burden tissue and can be visualized using the surgical microscope with a specialized filter. Intraoperative guidance afforded through the use of fluorescein may enhance the fidelity of tissue sampling, and increase the ability to accomplish complete resection of tectal lesions. In this report the authors present the case of a 19-year-old man with a tectal anaplastic pilocytic astrocytoma in which the use of fluorescein sodium and a Zeiss Pentero surgical microscope equipped with a yellow 560 filter enabled safe complete resection. In conjunction with neurosurgical navigation, added intraoperative guidance provided by fluorescein may be beneficial in the resection of brainstem gliomas. PMID:26407010

  7. Rapid detection of herpes simplex virus with fluorescein-labeled Helix pomatia lectin.

    PubMed Central

    Slifkin, M; Cumbie, R

    1989-01-01

    The use of fluorescein-conjugated Helix pomatia lectin was shown to be as effective as fluorescein-conjugated monoclonal antibody reagents for the detection and differentiation of herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2) in MRC-5 cell culture. Cells infected with HSV-1 generally displayed a pattern of nongranular or diffuse fluorescence, while cells infected with HSV-2 were identified by the production of fluorescent grains and flecks. This unique nonimmunological reagent, when used in combination with low-speed centrifugation, provides a remarkably specific, sensitive, rapid, and cost-effective means to detect HSV-infected MRC-5 or BHK-21 cells as early as 20 h postinoculation. In contrast to the immunofluorescence method, the serotypes of HSV can be differentiated with only one fluorescein-H. pomatia reagent in MRC-5 cell cultures. Images PMID:2545739

  8. Spectroscopic studies on the interaction of fluorescein and safranine T in PC liposomes.

    PubMed

    Bozkurt, Ebru; Bayraktutan, Tu?ba; Acar, Murat; Toprak, Mahmut

    2013-01-15

    In this study, the fluorescence quenching of fluorescein by safranine T in liposome media had been investigated systematically by fluorescence spectroscopy, UV-vis absorption spectroscopy and fluorescence decay lifetime measurements. The spectroscopic data were analyzed using a Stern-Volmer equation to determine the quenching process. The experimental results showed that the intrinsic fluorescence of fluorescein was strongly quenched by safranine T, and that the quenching mechanism was considered as static quenching by forming a ground-complex. The Stern-Volmer quenching constant Ksv, and the bimolecular quenching constant Kq were estimated. The distances between the donor (fluorescein) and the acceptor (safranine T) were calculated according to the Förster non-radiation energy transfer theory. In addition, the partition coefficient of the safranine T (Kp) in the L-egg lecithin phosphatidylcholine liposomes was also calculated by utilizing the fluorescence quenching. PMID:23099157

  9. Comparative Efficacy of Potassium Levulinate with/without Potassium Diacetate and Potassium Propionate vs Potassium Lactate and Sodium Diacetate for Control of Listeria monocytogenes on commercially prepared uncured t.breast

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We evaluated the efficacy of potassium levulinate, potassium diacetate, and potassium propionate to inhibit Listeria monocytogenes on commercially-prepared, uncured turkey breast during refrigerated storage. Whole muscle, uncured turkey breast chubs (ca. 5 kg each) were formulated with or without po...

  10. Salt stains from evaporating droplets

    PubMed Central

    Shahidzadeh, Noushine; Schut, Marthe F. L.; Desarnaud, Julie; Prat, Marc; Bonn, Daniel

    2015-01-01

    The study of the behavior of sessile droplets on solid substrates is not only associated with common everyday phenomena, such as the coffee stain effect, limescale deposits on our bathroom walls , but also very important in many applications such as purification of pharmaceuticals, de-icing of airplanes, inkjet printing and coating applications. In many of these processes, a phase change happens within the drop because of solvent evaporation, temperature changes or chemical reactions, which consequently lead to liquid to solid transitions in the droplets. Here we show that crystallization patterns of evaporating of water drops containing dissolved salts are different from the stains reported for evaporating colloidal suspensions. This happens because during the solvent evaporation, the salts crystallize and grow during the drying. Our results show that the patterns of the resulting salt crystal stains are mainly governed by wetting properties of the emerging crystal as well as the pathway of nucleation and growth, and are independent of the evaporation rate and thermal conductivity of the substrates. PMID:26012481

  11. Salt stains from evaporating droplets.

    PubMed

    Shahidzadeh, Noushine; Schut, Marthe F L; Desarnaud, Julie; Prat, Marc; Bonn, Daniel

    2015-01-01

    The study of the behavior of sessile droplets on solid substrates is not only associated with common everyday phenomena, such as the coffee stain effect, limescale deposits on our bathroom walls , but also very important in many applications such as purification of pharmaceuticals, de-icing of airplanes, inkjet printing and coating applications. In many of these processes, a phase change happens within the drop because of solvent evaporation, temperature changes or chemical reactions, which consequently lead to liquid to solid transitions in the droplets. Here we show that crystallization patterns of evaporating of water drops containing dissolved salts are different from the stains reported for evaporating colloidal suspensions. This happens because during the solvent evaporation, the salts crystallize and grow during the drying. Our results show that the patterns of the resulting salt crystal stains are mainly governed by wetting properties of the emerging crystal as well as the pathway of nucleation and growth, and are independent of the evaporation rate and thermal conductivity of the substrates. PMID:26012481

  12. Fluorescence spectrum analysis using Fourier series modeling for Fluorescein solution in Ethanol

    E-print Network

    Hadi, Mahasin F

    2011-01-01

    We have measured the fluorescence spectrum for fluorescein solution in ethanol with concentration 1 {\\times} 10-3 mol/liter at different temperatures from room temperature to freezing point of solvent, (T = 153, 183, 223, 253, and 303 K) using liquid nitrogen. Table curve 2D version 5.01 program has been used to determine the fitting curve and fitting equation for each fluorescence spectrum. Fourier series (3 {\\times} 2) was the most suitable fitting equation for all spectra. Theoretical fluorescence spectrum of fluorescein in ethanol at T = 183K was calculated and compared with experimental fluorescence spectrum at the same temperature. There is a good similarity between them.

  13. Quantitative retinal blood flow mapping from fluorescein videoangiography using tracer kinetic modeling.

    PubMed

    Tichauer, Kenneth M; Guthrie, Micah; Hones, Logan; Sinha, Lagnojita; St Lawrence, Keith; Kang-Mieler, Jennifer J

    2015-05-15

    Abnormal retinal blood flow (RBF) has been associated with numerous retinal pathologies, yet existing methods for measuring RBF predominantly provide only relative measures of blood flow and are unable to quantify volumetric blood flow, which could allow direct patient to patient comparison. This work presents a methodology based on linear systems theory and an image-based arterial input function to quantitatively map volumetric blood flow from standard fluorescein videoangiography data, and is therefore directly translatable to the clinic. Application of the approach to fluorescein retinal videoangiography in rats (4 control, 4 diabetic) demonstrated significantly higher RBF in 4-5 week diabetic rats as expected from the literature. PMID:26393691

  14. A Quantitative and Standardized Method for the Evaluation of Choroidal Neovascularization Using MICRON III Fluorescein Angiograms in Rats

    PubMed Central

    Wigg, Jonathan P.; Zhang, Hong; Yang, Dong

    2015-01-01

    Introduction In-vivo imaging of choroidal neovascularization (CNV) has been increasingly recognized as a valuable tool in the investigation of age-related macular degeneration (AMD) in both clinical and basic research applications. Arguably the most widely utilised model replicating AMD is laser generated CNV by rupture of Bruch’s membrane in rodents. Heretofore CNV evaluation via in-vivo imaging techniques has been hamstrung by a lack of appropriate rodent fundus camera and a non-standardised analysis method. The aim of this study was to establish a simple, quantifiable method of fluorescein fundus angiogram (FFA) image analysis for CNV lesions. Methods Laser was applied to 32 Brown Norway Rats; FFA images were taken using a rodent specific fundus camera (Micron III, Phoenix Laboratories) over 3 weeks and compared to conventional ex-vivo CNV assessment. FFA images acquired with fluorescein administered by intraperitoneal injection and intravenous injection were compared and shown to greatly influence lesion properties. Utilising commonly used software packages, FFA images were assessed for CNV and chorioretinal burns lesion area by manually outlining the maximum border of each lesion and normalising against the optic nerve head. Net fluorescence above background and derived value of area corrected lesion intensity were calculated. Results CNV lesions of rats treated with anti-VEGF antibody were significantly smaller in normalised lesion area (p<0.001) and fluorescent intensity (p<0.001) than the PBS treated control two weeks post laser. The calculated area corrected lesion intensity was significantly smaller (p<0.001) in anti-VEGF treated animals at 2 and 3 weeks post laser. The results obtained using FFA correlated with, and were confirmed by conventional lesion area measurements from isolectin stained choroidal flatmounts, where lesions of anti-VEGF treated rats were significantly smaller at 2 weeks (p = 0.049) and 3 weeks (p<0.001) post laser. Conclusion The presented method of in-vivo FFA quantification of CNV, including acquisition variable corrections, using the Micron III system and common use software establishes a reliable method for detecting and quantifying CNV enabling longitudinal studies and represents an important alternative to conventional CNV quantification methods. PMID:26024231

  15. Sodium lactate, sodium diacetate and pediocin: effects and interactions on the thermal inactivation of Listeria monocytogenes on bologna

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects and interactions of temperature (56.3C-60C), sodium lactate (SL; 0-4.8%), sodium diacetate (SD; 0-0.25%) and pediocin (0-10,000 AU) on Listeria monocytogenes inoculated on bologna were studied and a predictive inactivation model was developed. Bologna was manufactured with different SL ...

  16. Effects on reproductive performance by continuous feeding of dienestrol diacetate and supplementary fat in the breeder ration 

    E-print Network

    Cook, Freeman Waldo

    1958-01-01

    In phase I all-beef and soy-added ground beef patties containing sodium lactate, sodium propionate, and sodium diacetate at various levels and combinations were stored for nine months at -10°C. Upon cooking, the addition of sodium lactate increased...

  17. THE EFFECT OF SODIUM LACTATE AND SODIUM DIACETATE ON THE BEHAVIOR OF LISTERIA MONOCYTOGENES IN HAM STORED AT VARIOUS TEMPERATURES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Refrigerated ready-to-eat (RTE) meats contaminated with L. monocytogenes have been implicated in several listeriosis outbreaks. Lactate and diacetate have been shown to control L. monocytogenes in RTE meats at refrigerated temperature. However, there are no models describing their effects under tem...

  18. Ground beef shelf life assessment as influenced by sodium lactate, sodium propionate, sodium diacetate, and soy protein concentrate 

    E-print Network

    Grones, Kelly Leann

    2000-01-01

    In phase I all-beef and soy-added ground beef patties containing sodium lactate, sodium propionate, and sodium diacetate at various levels and combinations were stored for nine months at -10°C. Upon cooking, the addition of sodium lactate increased...

  19. Abstract Poly(acrylic acid) + zinc diacetate hybrid com-posites have been prepared by precipitation from aqueous

    E-print Network

    North Texas, University of

    Abstract Poly(acrylic acid) + zinc diacetate hybrid com- posites have been prepared · Polymer-based composites · Poly(acrylic acid) · Zinc acetate · Polymer electric conductivity · Dynamic chemical structures of PLCs, we have also those based on epoxies [5]. Moreover, blends of the PLC + PI type

  20. EFFECTS AND INTERACTIONS OF TEMPERATURE, SODIUM LACTATE, SODIUM DIACETATE AND PEDIOCIN ON THE STARVED CELLS OF LISTERIA MONOCYTOGENES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects and interactions of temperature (55-60 degrees C), sodium lactate (SL; 0.0-4.8%), sodium diacetate (SDA; 0.0-0.4%) and pediocin (0.0-10000 AU) on the starved cells of L. monocytogenes inoculated on the surface of the frankfurters were investigated, and a predictive model was developed. C...

  1. Ultraviolet Light (254 nm) Inactivation of Listeria monocytogenes on frankfurters that contain potassium lactate and sodium diacetate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Listeria monocytogenes, a psychrotrophic food-borne pathogen, is an occasional post-process contaminant on ready-to-eat meat (RTE) products including frankfurters. Potassium lactate (PL) and sodium diacetate (SDA) are FDA approved antimicrobials that inhibit the growth of L. monocytogenes when incor...

  2. Carboxyfluorescein diacetate succinimidyl ester labeling method to study the interaction between Leptospira and macrophages.

    PubMed

    Liu, Boyu; Wang, Yanchun; Guo, Xiaokui; Zhu, Weinan; Zhang, Yan; He, Ping

    2014-12-01

    Leptospirosis, which is caused by pathogenic species of the genus Leptospira, has emerged as one of the most widespread zoonotic diseases in the world. The exact mechanism of pathogenesis remains unknown, and the interaction between Leptospira and macrophages is not well understood. In this study, we report that carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) can efficiently label different Leptospira interrogans strains without affecting bacterial motility, viability, or virulence. Following co-incubation, CFDA-SE-labeled leptospires associated with macrophages were quantified by flow cytometry or confocal microscopy. In addition, we showed that trypan blue efficiently quenched the extracellular fluorescence from the adherent leptospires, which enabled intracellular and extracellular bacteria to be distinguished. PMID:25455022

  3. Process for the production of ethylidene diacetate from dimethyl ether using a heterogeneous catalyst

    DOEpatents

    Ramprasad, Dorai (Allentown, PA); Waller, Francis Joseph (Allentown, PA)

    1998-01-01

    This invention relates to a process for producing ethylidene diacetate by the reaction of dimethyl ether, acetic acid, hydrogen and carbon monoxide at elevated temperatures and pressures in the presence of an alkyl halide and a heterogeneous, bifunctional catalyst that is stable to hydrogenation and comprises an insoluble polymer having pendant quaternized heteroatoms, some of which heteroatoms are ionically bonded to anionic Group VIII metal complexes, the remainder of the heteroatoms being bonded to iodide. In contrast to prior art processes, no accelerator (promoter) is necessary to achieve the catalytic reaction and the products are easily separated from the catalyst by filtration. The catalyst can be recycled for 3 consecutive runs without loss in activity.

  4. Process for the production of ethylidene diacetate from dimethyl ether using a heterogeneous catalyst

    DOEpatents

    Ramprasad, D.; Waller, F.J.

    1998-04-28

    This invention relates to a process for producing ethylidene diacetate by the reaction of dimethyl ether, acetic acid, hydrogen and carbon monoxide at elevated temperatures and pressures in the presence of an alkyl halide and a heterogeneous, bifunctional catalyst that is stable to hydrogenation and comprises an insoluble polymer having pendant quaternized heteroatoms, some of which heteroatoms are ionically bonded to anionic Group VIII metal complexes, the remainder of the heteroatoms being bonded to iodide. In contrast to prior art processes, no accelerator (promoter) is necessary to achieve the catalytic reaction and the products are easily separated from the catalyst by filtration. The catalyst can be recycled for 3 consecutive runs without loss in activity.

  5. Methods for chromosome-specific staining

    DOEpatents

    Gray, J.W.; Pinkel, D.

    1995-09-05

    Methods and compositions for chromosome-specific staining are provided. Compositions comprise heterogeneous mixtures of labeled nucleic acid fragments having substantially complementary base sequences to unique sequence regions of the chromosomal DNA for which their associated staining reagent is specific. Methods include ways for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes. 3 figs.

  6. Spectroscopic properties and amplified spontaneous emission of fluorescein laser dye in ionic liquids as green media

    NASA Astrophysics Data System (ADS)

    AL-Aqmar, Dalal M.; Abdelkader, H. I.; Abou Kana, Maram T. H.

    2015-09-01

    The use of ionic liquids (ILs) as milieu materials for laser dyes is a promising field and quite competitive with volatile organic solvents and solid state-dye laser systems. This paper investigates some photo-physical parameters of fluorescein dye incorporated into ionic liquids; 1-Butyl-3-methylimidazolium chloride (BMIM Cl), 1-Butyl-3-methylimidazolium tetrachloroaluminate (BMIM AlCl4) and 1-Butyl-3-methylimidazolium tetrafluoroborate (BMIM BF4) as promising host matrix in addition to ethanol as reference. These parameters are: absorption and emission cross-sections, fluorescence lifetime and quantum yield, in addition to the transition dipole moment, the attenuation length and oscillator strength were also investigated. Lasing characteristics such as amplified spontaneous emission (ASE), the gain, and the photostability of fluorescein laser dye dissolved in different host materials were assessed. The composition and properties of the matrix of ILs were found that it has great interest in optimizing the laser performance and photostability of the investigated laser dye. Under transverse pumping of fluorescein dye by blue laser diode (450 nm) of (400 mW), the initial ASE for dye dissolved in BMIM AlCl4 and ethanol were decreased to 39% and 36% respectively as time progressed 132 min. Relatively high efficiency and high fluorescence quantum yield (11.8% and 0.82% respectively) were obtained with good photostability in case of fluorescein in BMIM BF4 that was decreased to ?56% of the initial ASE after continuously pumping with 400 mW for 132 min.

  7. Determination of torasemide by fluorescence quenching method with some dihalogenated fluorescein dyes as probes

    NASA Astrophysics Data System (ADS)

    Cui, Zhiping; Liu, Shaopu; Liu, Zhongfang; Li, Yuanfang; Hu, Xiaoli; Tian, Jing

    2013-10-01

    A novel fluorescence quenching method for the determination of torasemide (TOR) with some dihalogenated fluorescein dyes as fluorescence probes was developed. In acidulous medium, TOR could interact with some dihalogenated fluorescein dyes such as dichlorofluorescein (DCF), dibromofluorescein (DBF) and diiodofluorescein (DIF) to form binary complexes, which could lead to fluorescence quenching of above dihalogenated fluorescein dyes. The maximum fluorescence emission wavelengths were located at 532 nm (TOR-DCF), 535 nm (TOR-DBF) and 554 nm (TOR-DIF). The relative fluorescence intensities (?F = F0 - F) were proportional to the concentration of TOR in certain ranges. The detection limits were 4.8 ng mL-1 for TOR-DCF system, 9.8 ng mL-1 for TOR-DBF system and 35.1 ng mL-1 for TOR-DIF system. The optimum reaction conditions, influencing factors were studied; and the effect of coexisting substances was investigated owing to the highest sensitivity of TOR-DCF system. In addition, the reaction mechanism, composition and structure of the complex were discussed by quantum chemical calculation and Job's method. The fluorescence quenching of dihalogenated fluorescein dyes by TOR was a static quenching process judging from the effect of temperature and the Stern-Volmer plots. The method was satisfactorily applied to the determination of TOR in tablets and human urine samples.

  8. Staining calcified dental tissues with food.

    PubMed

    Chan, K C; Hormati, A A; Kerber, P E

    1981-08-01

    The discoloration of enamel caused by food substances was found to be superficial and for dentin and cementum ingressive. Discoloration of cementum exceeded that of dentin, and dentin stained more than enamel. Coffee and soy sauce stained the calcified dental tissues more than the cola beverage and tea. The longer the staining time, the deeper was the discoloration. PMID:6944481

  9. A novel biosensor for Escherichia coli O157:H7 based on fluorescein-releasable biolabels.

    PubMed

    Hu, Rong-Rong; Yin, Zheng-Zhi; Zeng, Yan-Bo; Zhang, Jian; Liu, Hai-Qing; Shao, Yong; Ren, Shi-Bin; Li, Lei

    2016-04-15

    New techniques are required for the rapid and sensitive detection of Escherichia coli O157:H7 (E. coli O157:H7), a pathogenic bacterium responsible for serious and sometimes life-threatening diseases in humans. In this study, we developed a highly sensitive and efficient biosensor for the quantitative detection of E. coli O157:H7 by integrating fluorescein-releasable biolabels with a magnetism-separable probe. Hollow silica nanospheres with a diameter of approximately 350nm were synthesized, enriched with fluorescein, and surface-protected with macromolecule layers of poly (acrylic acid) and poly (dimethyldiallylammonium chloride). These fluorescein-enriched hollow silica nanospheres were characterized using scanning electron microscopy, transmission electron microscopy, and Fourier transform infrared spectroscopy. They were further functionalized as immune labels of E. coli O157:H7 for a sandwich-type immune reaction between this bacterium and magnetic nanoparticles (Fe3O4@SiO2). Next, the E. coli O157:H7 cells were captured, magnetically separated, and quantified based on the fluorescence intensity of the fluorescein released from the biolabels of the fluorescein-enriched hollow silica nanospheres. This analytic process can be completed within 75min, and the biosensor showed a linear relationship ranging from 4 to 4.0×10(8)cfu/mL with a detection limit of 3cfu/mL. These results show that the developed fluorescent sensor has excellent specificity, and good reproducibility and stability. This study used real spiked samples for detection, indicating that this technique has a wide range of potential applications and may be readily adapted for detecting other pathogens. PMID:26584080

  10. Histological stain evaluation for machine learning applications

    PubMed Central

    Azar, Jimmy C.; Busch, Christer; Carlbom, Ingrid B.

    2013-01-01

    Aims: A methodology for quantitative comparison of histological stains based on their classification and clustering performance, which may facilitate the choice of histological stains for automatic pattern and image analysis. Background: Machine learning and image analysis are becoming increasingly important in pathology applications for automatic analysis of histological tissue samples. Pathologists rely on multiple, contrasting stains to analyze tissue samples, but histological stains are developed for visual analysis and are not always ideal for automatic analysis. Materials and Methods: Thirteen different histological stains were used to stain adjacent prostate tissue sections from radical prostatectomies. We evaluate the stains for both supervised and unsupervised classification of stain/tissue combinations. For supervised classification we measure the error rate of nonlinear support vector machines, and for unsupervised classification we use the Rand index and the F-measure to assess the clustering results of a Gaussian mixture model based on expectation–maximization. Finally, we investigate class separability measures based on scatter criteria. Results: A methodology for quantitative evaluation of histological stains in terms of their classification and clustering efficacy that aims at improving segmentation and color decomposition. We demonstrate that for a specific tissue type, certain stains perform consistently better than others according to objective error criteria. Conclusions: The choice of histological stain for automatic analysis must be based on its classification and clustering performance, which are indicators of the performance of automatic segmentation of tissue into morphological components, which in turn may be the basis for diagnosis. PMID:23766933

  11. DETERMINATION OF 'GIARDIA MURIS' CYST VIABILITY BY DIFFERENTIAL INTERFERENCE CONTRAST, PHASE, OR BRIGHTFIELD MICROSCOPY

    EPA Science Inventory

    Recent experiments have demonstrated that fluorogenic substrates are taken up by Giardia cysts and that an excellent correlation exists between animal infectivity and vital staining with fluorescein diacetate (FDA) for viable cysts and propidium iodide (PI) for non-viable cysts. ...

  12. Phenyliodine(III) Diacetate Mediated Oxidative Cyclization of 1-Alkenoyl-1-carbamoyl Cycloalkanes: Access to Spiro-Fused Dihydrofuran-3(2H)-ones.

    PubMed

    Yuan, Jingwen; Zhang, Qian; Yu, Mangfei; Huang, Peng; Zhang, Rui; Dong, Dewen

    2015-10-16

    Facile and efficient synthesis of spiro-fused dihydrofuran-3(2H)-ones was developed via phenyliodine(III) diacetate (PIDA) mediated oxidative cyclization of 1-alkenoyl-1-carbamoyl cycloalkanes under very mild conditions. PMID:26444337

  13. Analysis of chemical equilibrium of silicon-substituted fluorescein and its application to develop a scaffold for red fluorescent probes.

    PubMed

    Hirabayashi, Kazuhisa; Hanaoka, Kenjiro; Takayanagi, Toshio; Toki, Yuko; Egawa, Takahiro; Kamiya, Mako; Komatsu, Toru; Ueno, Tasuku; Terai, Takuya; Yoshida, Kengo; Uchiyama, Masanobu; Nagano, Tetsuo; Urano, Yasuteru

    2015-09-01

    Fluorescein is a representative green fluorophore that has been widely used as a scaffold of practically useful green fluorescent probes. Here, we report synthesis and characterization of a silicon-substituted fluorescein, i.e., 2-COOH TokyoMagenta (2-COOH TM), which is a fluorescein analogue in which the O atom at the 10' position of the xanthene moiety of fluorescein is replaced with a Si atom. This fluorescein analogue forms a spirolactone ring via intramolecular nucleophilic attack of the carboxylic group in a pH-dependent manner. Consequently, 2-COOH TM exhibits characteristic large pH-dependent absorption and fluorescence spectral changes: (1) 2-COOH TM is colorless at acidic pH, whereas fluorescein retains observable absorption and fluorescence even at acidic pH, and the absorption maximum is also shifted; (2) the absorption spectral change occurs above pH 7.0 for 2-COOH TM and below pH 7.0 for fluorescein; (3) 2-COOH TM shows a much sharper pH response than fluorescein because of its pKa inversion, i.e., pKa1 > pKa2. These features are also different from those of a compound without the carboxylic group, 2-Me TokyoMagenta (2-Me TM). Analysis of the chemical equilibrium between pH 3.0 and 11.0 disclosed that 2-COOH TM favors the colorless and nonfluorescent lactone form, compared with fluorescein. Substitution of Cl atoms at the 4' and 5' positions of the xanthene moiety of 2-COOH TM to obtain 2-COOH DCTM shifted the equilibrium so that the new derivative exists predominantly in the strongly fluorescent open form at physiological pH (pH 7.4). To demonstrate the practical utility of 2-COOH DCTM as a novel scaffold for red fluorescent probes, we employed it to develop a probe for ?-galactosidase. PMID:26237524

  14. Effect of cellulose diacetate films on reactions of oxygen and hydrogen peroxide on platinum and isotropic pyrocarbon

    SciTech Connect

    Zhutaeva, G.V.; Makarova, E.V.; Tkhi Khan, F.

    1995-12-20

    The authors examined how cellulose diacetate (CDA) films on the surface and pyrocarbon electrodes affect electoreduction of oxygen and electrooxidation of hydrogen peroxide in aqueous solutions of electrolytes. The authors show that adsorption interaction of the CDA polymer with the electrode surface is a complex process and is not explained by either only kinetic or only diffusion retardation. The authors determined the permeability of the polymer films to oxygen and hydrogen peroxide.

  15. Evaluation of frankfurters formulated with potassium lactate and sodium diacetate and innocualted with Listeria monocytogenes before and after irradiation treatment 

    E-print Network

    Knight, Timothy David

    2006-08-16

    significant surface bactericidal effect through 12 wk of refrigerated vacuum packaged storage regardless of the inclusion or exclusion of lactate in the frankfurter formulation. Sensory and physiochemical changes of all treatments were minimal... or synergistic bactericidal/bacteriostatic effect while minimally affecting food quality. The combination of lactate/diacetate and pasteurizing doses of irradiation are effective at inhibiting the growth of L. monocytogenes on frankfurters...

  16. Tunneling Spectroscopy Analysis of Hexachloro-Fluorescein Phosphoramidite Fluorescent Dye Attached to Deoxyribonucleic Acid

    NASA Astrophysics Data System (ADS)

    Kawahara, Toshio; Takahashi, Takuya; Tanaka, Hiroyuki; Kawai, Tomoji

    2005-07-01

    Scanning tunneling microscopy (STM) was used to observe hexachloro-fluorescein phosphoramidite (HEX) attached to single-stranded deoxyribonucleic acid (ssDNA) with molecular resolution. Scanning tunneling spectroscopy (STS) was also used to study the electric properties of HEX in single-molecular spectroscopy. In the STM topographic images, the bright HEX molecule and each base subunit of DNA could be clearly observed, just as with fluorescein isothiocyanate (FITC) attached to ssDNA. In contrast to FITC, HEX molecules usually did not show a clear peak in their tunneling spectra. Two types of HEX molecules seemed to have different apparent heights, and only the HEX with the larger height in topographic images showed a peak at +0.6 V. The conformation of the HEX seems to affect the measured spectra. Thus, we obtained another molecule marker in addition to FITC with different spectral features for STM.

  17. Effect of liposomal fluidity on skin permeation of sodium fluorescein entrapped in liposomes

    PubMed Central

    Subongkot, Thirapit; Ngawhirunpat, Tanasait

    2015-01-01

    The purpose of this study was to investigate the effect of ultradeformable liposome components, Tween 20 and terpenes, on vesicle fluidity. The fluidity was evaluated by electron spin resonance spectroscopy using 5-doxyl stearic acid and 16-doxyl stearic acid as spin labels for phospholipid bilayer fluidity at the C5 atom of the acyl chain near the polar head group (hydrophilic region) and the C16 atom of the acyl chain (lipophilic region), respectively. The electron spin resonance study revealed that Tween 20 increased the fluidity at the C5 atom of the acyl chain, whereas terpenes increased the fluidity at the C16 atom of the acyl chain of the phospholipid bilayer. The increase in liposomal fluidity resulted in the increased skin penetration of sodium fluorescein. Confocal laser scanning microscopy showed that ultradeformable liposomes with terpenes increase the skin penetration of sodium fluorescein by enhancing hair follicle penetration. PMID:26229462

  18. Detection of the halogenating activity of heme peroxidases in leukocytes by aminophenyl fluorescein.

    PubMed

    Flemmig, J; Remmler, J; Zschaler, J; Arnhold, J

    2015-06-01

    The formation of hypochlorous and hypobromous acids by heme peroxidases is a key property of certain immune cells. These products are not only involved in defense against pathogenic microorganisms and in regulation of inflammatory processes, but contribute also to tissue damage in certain pathologies. After a short introduction about experimental approaches for the assessment of the halogenating activity in vitro and in cell suspensions, we are focusing on novel applications of fluorescent dye systems to detect the formation of hypochlorous acid (HOCl) in leukocytes. Special attention is directed to properties and applications of the non-fluorescent dye aminophenyl fluorescein that is converted by HOCl, HOBr, and other strong oxidants to fluorescein. This dye allows the detection of the halogenating activity in samples containing free myeloperoxidase and eosinophil peroxidase as well as in intact granulocytes using fluorescence spectroscopy and flow cytometry, respectively. PMID:25536418

  19. Investigation of Fluorescence Resonance Energy Transfer between Fluorescein and Rhodamine 6G

    NASA Astrophysics Data System (ADS)

    Saha, Jaba; Datta Roy, Arpan; Dey, Dibyendu; Chakraborty, Santanu; Bhattacharjee, D.; Paul, P. K.; Hussain, Syed Arshad

    2015-10-01

    Fluorescence Resonance Energy Transfer between two organic dyes Fluorescein and Rhodamine 6G was investigated in aqueous solution in presence and absence of synthetic clay laponite. Spectroscopic studies suggest that both the dyes were present mainly as monomer in solution. Fluorescence Resonance Energy Transfer occurred from Fluorescein to Rhodamine 6G in solutions. Energy transfer efficiency increases in presence of laponite and the maximum efficiency was 72.00% in aqueous laponite dispersion. Energy transfer efficiency was found to be pH sensitive. It has been demonstrated that with proper calibration it is possible to use the present system under investigation to sense pH over a wide range from 1.5 to 8.0.

  20. Fluorescein and Rosamine Derivatives as Donors/Acceptors for "Through-bond" Energy Transfer Cassettes 

    E-print Network

    Castro, Juan C.

    2010-07-14

    .6, 30.5 9 H t-Bu H 518 101.0 10 CN t-Bu H 564, 614 23.2, 47.4 11 H t-Bu I 532 90.8 12 CN t-Bu I 582, 636 33.6, 68.3 In 1871, Adolph Von Baeyer synthesized fluorescein. It was created by the condensation of two equivalents of resorcinol...

  1. Use of indocyanine green and sodium fluorescein for anterior segment angiography in ophthalmologically normal cats.

    PubMed

    Pirie, Chris G; Alario, Anthony

    2015-10-01

    OBJECTIVE To assess and compare results of anterior segment angiography of ophthalmologically normal cats following IV injection with indocyanine green and sodium fluorescein dyes. ANIMALS 10 client-owned cats. PROCEDURES Anterior segment angiography was performed in anesthetized cats following administration of 0.25% indocyanine green (1.0 mg/kg, IV) or 10% sodium fluorescein (20 mg/kg, IV) solution. All cats received both treatments. Imaging (1 eye/cat) was performed with a full-spectrum digital single-lens reflex camera equipped with an adaptor (1 image/s for 30 seconds) immediately following IV dye injection and 1, 2, 3, 4, and 5 minutes after injection. Onset and duration of arterial, capillary, and venous phases of iris vasculature were identified and compared statistically between treatments. Degree of iridal pigmentation, leakage of dye from iris vasculature, and image quality were subjectively assessed. RESULTS No differences were found in onset or duration of vascular phases between treatments. Visibility of the iris vasculature was not impaired by poor or moderate iridal pigmentation with either method. Indocyanine green provided subjectively better vascular detail and image contrast than sodium fluorescein. No vascular dye leakage was observed following indocyanine green administration. Leakage of dye from blood vessels in the stroma (in 10 cats) and presence of dye in the anterior chamber (in 5 cats) were detected after sodium fluorescein administration. CONCLUSIONS AND CLINICAL RELEVANCE Images obtained with either fluorescent dye were considered to be of diagnostic quality. Lack of leakage following indocyanine green administration suggested this treatment may have better diagnostic utility for anterior segment angiography. The photographic equipment used provided a cost-effective alternative to existing imaging systems. PMID:26413828

  2. Fluorescein-N-Methylimidazole Conjugate as Cu(2+) Sensor in Mixed Aqueous Media Through Electron Transfer.

    PubMed

    Helal, Aasif; Kim, Hong-Seok; Yamani, Zain H; Nasiruzzaman Shaikh, M

    2016-01-01

    A new highly selective, chromogenic, and fluorogenic Cu(2+) chemosensor, fluorescein-N-methylimidazole conjugate 1, and another fluorescein-N-imidazole conjugate 2 were synthesized and investigated by UV-visible and fluorescence spectroscopy. The sensing of Cu(2+) quenches the emission band of 1 at ?max = 525 nm, with an association constant (K a = 1.0 x 10(7) M(-1)) and a stoichiometry of 1:1 in a buffered H2O: MeOH solution (4:1, pH = 7.4). The Cu(2+) detection limit for chemosensor 1 is 37 nM. The presence of the N-methyl group in 1 increased the Cu(2+) binding selectivity, resulting in a stronger binding constant and a broader pH working range (pH 5-10) in comparison to 2. The fluorescence in 1 and 2 is caused by electron transfer phenomenon from the imidazole nitrogen to fluorescein, which is readily inhibited by Cu(2+) binding. PMID:26573285

  3. A selectively fluorescein-based colorimetric probe for detecting copper(II) ion

    NASA Astrophysics Data System (ADS)

    Zhang, Li; Zhang, Xianhong

    2014-12-01

    A novel fluorescein derivative 3-bromo-5-methylsalicylaldehyde fluorescein hydrazone (BMSFH) has been synthesized by reacting fluorescein hydrazide with 3-bromo-5-methylsalicylaldehyde and was developed as a new colorimetric probe for detection of Cu2+. In the presence of Cu2+ the BMSFH exhibits a rapid color change from colorless to yellow together with an obvious new band appeared at 502 nm in the UV-vis absorption spectra. However, other common alkali-, alkaline earth-, transition- and rare earth metal ions induced no or minimal spectral changes. This change is attributed to BMSFH via coordination with Cu2+ in a 1:1 stoichiometry and this binding to Cu2+ is reversible, as indicated by the bleaching of the color when the Cu2+ is extracted with EDTA. Experimental results indicate that the BMSFH can provide a rapid, selective and sensitive response to Cu2+ with a linear dynamic range 3.0-330 ?mol/L and can be used as a potential Cu2+ colorimetric probe in aqueous solution.

  4. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...2011-04-01 false Dye and chemical solution stains. 864...DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains...

  5. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 false Dye and chemical solution stains. 864...DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains...

  6. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...2011-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture...Stained Cotton § 28.442 Middling Yellow Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is...

  7. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...2014-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture...Stained Cotton § 28.442 Middling Yellow Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is...

  8. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...2013-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture...Stained Cotton § 28.442 Middling Yellow Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is...

  9. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...2010-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture...Stained Cotton § 28.442 Middling Yellow Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is...

  10. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...2012-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture...Stained Cotton § 28.442 Middling Yellow Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is...

  11. Encapsulation of T4 bacteriophage in electrospun poly(ethylene oxide)/cellulose diacetate fibers.

    PubMed

    Korehei, Reza; Kadla, John F

    2014-01-16

    Phage therapy is a potentially beneficial approach to food preservation and storage. Sustained delivery of bacteriophage can prevent bacterial growth on contaminated food surfaces. Using coaxial electrospinning bacteriophage can be encapsulated in electrospun fibers with high viability. The resulting bio-based electrospun fibers may have potential as a food packaging material. In the present work, T4 bacteriophage (T4 phage) was incorporated into core/shell electrospun fibers made from poly(ethylene oxide) (PEO), cellulose diacetate (CDA), and their blends. Fibers prepared using PEO as the shell polymer showed an immediate burst release of T4 phage upon submersion in buffer. The blending of CDA with PEO significantly decreased the rate of phage release, with no released T4 phage being detected from the solely CDA fibers. Increasing the PEO molecular weight increased the electrospun fiber diameter and viscosity of the releasing medium, which resulted in a relatively slower T4 phage release profile. SEM analyses of the electrospun fiber morphologies were in good agreement with the T4 phage release profiles. Depending on the PEO/CDA ratio, the post-release electrospun fiber morphologies varied from discontinuous fibers to minimally swollen fibers. From these results it is suggested that the T4 phage release mechanism is through solvent activation/polymer dissolution in the case of the PEO fibers and/or by diffusion control from the PEO/CDA blend fibers. PMID:24188849

  12. Degradation of phorbol 12,13-diacetate in aqueous solution by gamma irradiation

    NASA Astrophysics Data System (ADS)

    Kongmany, Santi; Furuta, Masakazu; Matsuura, Hiroto; Okuda, Shuichi; Imamura, Kiyoshi; Maeda, Yasuaki

    2014-12-01

    Phorbol esters (PEs) are highly toxic compounds that cause skin irritation, inflammation, and tumor promotion upon contact with humans or animals. These compounds are naturally present in Jatropha curcas L. To promote the use of J. curcas seed oil in bio-diesel production industries and reduce environmental concerns, it is necessary to find methods of degrading PEs. In this study, the degradation of phorbol 12,13-diacetate (PDA), as a representative PE, in aqueous solution at a concentration of 10 mg/L by 60Co-?-irradiation was investigated. The results demonstrate that PDA was effectively degraded by this treatment and the degradation efficiency increased with the absorbed dose within the range of 0.5-3 kGy. Complete degradation of PDA was achieved at a dose of 3 kGy. In the presence of radical scavengers (i.e., methanol, tert-butanol, 2-propanol), reactive species from water radiolysis were scavenged, and significant inhibition of PDA degradation was observed at absorbed doses less than 1 kGy. In the presence of nitrous oxide, the generation of hydroxyl radicals (rad OH) was promoted during gamma irradiation and PDA degradation was drastically enhanced.

  13. DETERMINATION OF ALIPHATIC AMINES IN WATER USING DERIVATIZATION WITH FLUORESCEIN ISOTHIOCYANATE AND CAPILLARY ELECTROPHORESIS/LASER-INDUCED FLUORESCENCE DETECTION.

    EPA Science Inventory

    Detection-oriented derivatization of aliphatic amines and amine functional groups in coumpounds of environmental interest was studied using fluorescein isothiocyanate (FITC) with separation/determination by capillary electrophoresis/laser-induced fluorescence. Determinative level...

  14. SYPRO Ruby Protein Gel Stain Advanced staining technology for 2-D gels and proteomics

    E-print Network

    Lebendiker, Mario

    for proteomics. It provides the same low nanogram sensitivity as the best silver staining techniques (Figure 1), but stains more proteins and has a much broader linear quantitation range -- extending over three orders

  15. Negative Stain Electron Microscopy of Microtubules Negative staining is a rapid, qualitative method for analyzing

    E-print Network

    Mitchison, Tim

    Negative Stain Electron Microscopy of Microtubules Negative staining is a rapid, qualitative method of microtubules into flat sheets are common. Cryo-electron microscopy, where microtubules are flash frozen

  16. Fluorescein angiography

    MedlinePLUS

    ... bar to keep your head still during the test. The health care provider will take pictures of the inside of ... to stop taking medicines that could affect the test results. Tell your health care provide about any allergies, particularly reactions to iodine. ...

  17. Pro-Q Diamond Phosphoprotein Gel Stain

    E-print Network

    Lebendiker, Mario

    Pro-Q Diamond Phosphoprotein Gel Stain In-gel Detection Technology for Protein Phosphorylation and phosphoproteomics, the Pro-Q Diamond phos- phoprotein gel stain is a breakthrough technology that provides a simple phosphoproteins, the Pro-Q Diamond signal is linear over three orders of magnitude and the strength of the signal

  18. Mineral Stains at the No Name Prospect

    USGS Multimedia Gallery

    USGS scientist Art Bookstrom looks at greenish copper stain and pale pink cobalt bloom on limonite-stained meta-siltite and meta-argillite at the No Name prospect, near Iron Creek, in the southeastern part of the Idaho cobalt belt, in east-central Idaho....

  19. De-staining and re-staining mucins in formalin fixed paraffin sections.

    PubMed

    Smith, A A; Glickfield, I

    2011-04-01

    Re-staining of formalin fixed paraffin sections sometimes is required and this requires prior de-staining. Some simple and effective protocols for de-staining are described. Mucihematoxylin and mucicarmine can be removed with acid alcohol. Zirconyl hematoxylin can be removed with periodic acid or Sinha's fixative. Alcian blue can be removed with 5% trifluoroacetic acid in dichloromethane. Colloidal iron can be bleached in 1% household bleach in alcohol. PAS can be removed with hydrogen peroxide or ammonium hydroxide. With few exceptions, de-stained sections can be re-stained with mucihematoxylin, PAS or Gabe's trichrome. PMID:20001228

  20. Compact, Automated Centrifugal Slide-Staining System

    NASA Technical Reports Server (NTRS)

    Feeback, Daniel L.; Clarke, Mark S. F.

    2004-01-01

    The Directional Acceleration Vector-Driven Displacement of Fluids (DAVD-DOF) system, under development at the time of reporting the information for this article, would be a relatively compact, automated, centrifugally actuated system for staining blood smears and other microbiological samples on glass microscope slides in either a microgravitational or a normal Earth gravitational environment. The DAVD-DOF concept is a successor to the centrifuge-operated slide stainer (COSS) concept, which was reported in Slide-Staining System for Microgravity or Gravity (MSC-22949), NASA Tech Briefs, Vol. 25, No. 1 (January, 2001), page 64. The COSS includes reservoirs and a staining chamber that contains a microscope slide to which a biological sample is affixed. The staining chamber is sequentially filled with and drained of staining and related liquids from the reservoirs by use of a weighted plunger to force liquid from one reservoir to another at a constant level of hypergravity maintained in a standard swing-bucket centrifuge. In the DAVD-DOF system, a staining chamber containing a sample would also be sequentially filled and emptied, but with important differences. Instead of a simple microscope slide, one would use a special microscope slide on which would be fabricated a network of very small reservoirs and narrow channels connected to a staining chamber (see figure). Unlike in the COSS, displacement of liquid would be effected by use of the weight of the liquid itself, rather than the weight of a plunger.

  1. Tooth staining effects of an alexidine mouthwash.

    PubMed

    Formicola, A J; Deasy, M J; Johnson, D H; Howe, E E

    1979-04-01

    The primary purpose of this study was to determine the amount of tooth staining produced by an alexidine mouthrinse. One hundred and eighty subjects rinsed twice daily for 1 month with either 15 ml of alexidine (0.035%) or a placebo solution. Prior to the study, the subjects were classified according to their smoking, coffee and tea drinking habits and these factors were subsequently considered in the analysis of the stain scores. Additionally, the effects on staining of a prior prophylaxis and the use of a fluoridated toothpaste during the study were determined. Upon termination of the study, subjects utilizing the active mouthrinse manifested a greater degree of staining than placebo users. The amount and intensity of the stain due to alexidine were not influenced (increased) by smoking, tea or coffee drinking habits. A prior prophylaxis did not reduce the staining propensity of alexidine users. The method of scoring developed can be used to assess the degree of tooth staining induced by antiplaque agents. PMID:374706

  2. Indicating pressure and environmental effects by means of the spectral shift with rhodamine B and fluorescein

    NASA Astrophysics Data System (ADS)

    Johann, R. M.

    2015-07-01

    Fluorescence absorption and emission wavelengths can be influenced by environmental conditions, such as pressure, temperature and concentration. Here those effects are explored with an emphasis on determining the potential of rhodamine B and fluorescein as high-pressure indicators. The red shift of the emission peak maxima of rhodamine B and fluorescein are investigated in dependence of pressure up to 200 MPa using as the solvents water, ethanol and poly(dimethylsiloxane) (PDMS) with rhodamine B and water, polystyrene beads and melamine resin beads with fluorescein. Emission spectra recording and peak fitting is done automatically at time intervals of down to a second and with 0.3 nm wavelength resolution. The wavenumber-pressure relation for rhodamine B reveals increasing divergence from linear behavior in the sequence of the solvents water, ethanol and silicone rubber. Graphical correlation of the data diverging only slightly from linearity with a selection of polarity functions is enabled using the concept of `deviation from linearity (DL)' plots. Using the example of rhodamine B dissolved in PDMS elastomer it is shown that there is a temperature induced irreversible molecular reordering, when scanning between 3 and ˜50°C, and a polarity change in the proximity of the embedded dye molecule. Swelling studies are performed with PDMS containing rhodamine B, where the elastomer is first put in water, then in ethanol and again in water. There a complex solvent exchange process is revealed in the elastomer demonstrating the feasibility of fluorescence spectroscopy, when observing variations in wavelength, to indicate and enlighten molecular rearrangements and swelling dynamics in the polymer, and polarity changes and solvent exchange processes in the dye solvation shell.

  3. Vessel extraction from non-fluorescein fundus images using orientation-aware detector.

    PubMed

    Yin, Benjun; Li, Huating; Sheng, Bin; Hou, Xuhong; Chen, Yan; Wu, Wen; Li, Ping; Shen, Ruimin; Bao, Yuqian; Jia, Weiping

    2015-12-01

    The automatic extraction of blood vessels in non-fluorescein eye fundus images is a tough task in applications such as diabetic retinopathy screening. However, vessel shapes have complex variations, and accurate modeling of retinal vascular structures is challenging. We have therefore developed a new approach to accurately extract blood vessels in non-fluorescein fundus images using an orientation-aware detector (OAD). The detector was designed according to the intrinsic property of vessels being locally oriented and having linearly elongated structures. We employ the OAD to extract vessel shapes with no assumptions on parametric orientations of vessel shapes. The orientations of vessels can be efficiently modeled by the energy distribution of Fourier transformation. Accordingly, both wide and thin vessels can be extracted with two-scale segmentation in which line operators are applied in large scale and the Gabor filter bank is applied in small scale. A post-processing technique, based on the path opening operation, is applied to eliminate false responses to nonvascular areas, such as retinal structures (optic disc and macula) and pathologies (exudates, hemorrhages,and microaneurysms). This makes the detector robust and structure-aware. By achieving a competitive CAL measurement of 80.82% for the DRIVE database and 68.94% for the STARE, the experimental results demonstrated that the OAD approach outperforms existing segmentation methods. Furthermore, the proposed approach effectively works with non-fluorescein fundus images and proves highly accurate and robust in complicated regions such as the central reflex, close vessels, and crossover points, despite a high level of illumination noise in the original data. PMID:26474120

  4. Fluorescein angiogram findings in a case of cutis marmorata telangiectatica congenita.

    PubMed

    Soohoo, Jeffrey R; McCourt, Emily A; Lenahan, Deborah S; Oliver, Scott C N

    2013-01-01

    Cutis marmorata telangiectatica congenita is a well-characterized cutaneous vascular disorder with variable and rare ocular involvement. It has been reported in association with glaucoma, bilateral congenital retinal detachments, bilateral tractional retinal detachments secondary to proliferative vitreoretinopathy, and retinoblastoma. This case demonstrates novel findings of bilateral peripheral retinal vascular abnormalities and retinal nonperfusion on fluorescein angiography without retinal detachment that have not previously been described in cutis marmorata telangiectatica congenita. Laser photocoagulation was applied to areas of retinal nonperfusion with stability in the retinal pathology at follow-up examination 3 months later. PMID:23758322

  5. [Effect of heat-staining procedure on the gram staining properties of mycobacteria].

    PubMed

    Nakamura, M; Harano, Y; Koga, T

    1991-03-01

    Since the establishment of Gram stain by H.C.Y. Gram in 1884, it has been widely and routinely used as an aid for differentiation of bacteria. The bacteria are divided into three categories by the staining properties; Gram-positive, -negative, and -indefinite. All the text books in the world describe that mycobacteria such as M. tuberculosis are Gram-positive. By the merest chance, however, it was found that M. lepraemurium grown in tissues was not stained by the routinely used Gram staining method. Therefore, we tried to stain some of the mycobacteria by the Gram staining procedure which is widely used at present. The results obtained indicated that the mycobacteria tested were divided into three groups; the unstainable group such as M. leprae and M. lepraemurium, the Gram-positive and difficult-to-stain group which involves such slow growing mycobacteria as M. tuberculosis, M. avium, and M. intracellulare, and the Gram-indefinite group which contains such rapid growing mycobacteria as M. phlei, M. smegmatis, and M. chelonae. However, if Gram stain is carried out by the heating procedure at the first staining step, all the mycobacteria would become Gram-positive. Therefore, we emphasize that Gram staining of mycobacteria should be performed by the heating procedure. PMID:1712050

  6. Formation of coffee stains on porous surfaces.

    PubMed

    Dou, Rui; Derby, Brian

    2012-03-27

    During the drying of drops of nanoparticle suspensions, segregation can occur by internal fluid flows toward the contact line, if the contact line is pinned. This leads to a characteristic ring deposit or coffee stain. On solid substrates coffee staining can be eliminated through the use of solvent mixtures that promote Marangoni flows to oppose these drying-induced flows. Here it is shown that a suspension, optimized to eliminate the formation of coffee stains on a range of solid surfaces, shows coffee staining on a number of porous surfaces. This behavior is shown to be consistent with a mechanism of fluid removal through capillary flow (draining) of the solvent into the porous substrate, combined with filtration of the particles by the small pore size, in addition to the flow from solvent evaporation. The extent of capillary driven coffee staining is a function of substrate pore size: if the pore size is small, capillary flow is slow, reducing the observed coffee staining. However, if the pore size is too large, the nanoparticles are absorbed into the material along with the draining solute and no deposition of particles is observed. PMID:22385387

  7. Wide-Field Fluorescein Angiography in Wet Age-Related Macular Degeneration

    PubMed Central

    Beare, Nicholas

    2014-01-01

    Purpose. The aim of our study was to investigate if peripheral retinal ischaemia contributed to the pathogenesis of neovascular AMD (NvAMD), using wide-field fluorescein angiography (WFFA). Methods. This prospective study included 30 consecutive patients with newly diagnosed NvAMD in the index eye. Wide-field colour fundus images and fluorescein angiograms were obtained using P200C optomap FA and analysed using a grid with three concentric circles of 50°, 100°, and 200° centred on the fovea to define zones Z1, Z2, and Z3. Results. Areas of peripheral retinal nonperfusion were seen in 2 (7%) eyes, peripheral vascular leakage in 5 (17%) eyes, and diffuse dye leakage close to the ora in 5 (17%) eyes. A total of one-third of the study eyes showed changes on WFFA in Z2 and Z3. On comparing index eyes to nonindex eyes in these patients, the presence of NvAMD was associated with peripheral FA changes (P = 0.009, Fisher's test). Conclusion. Frank peripheral retinal non-perfusion does not appear to be associated with NvAMD. In some patients with active NvAMD there is degradation of the peripheral blood-retina barrier. Smoking was also found to be associated with the above-mentioned abnormalities. PMID:25379537

  8. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...2011-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture... § 28.441 Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of...

  9. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...2013-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture... § 28.441 Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of...

  10. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...2010-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture... § 28.441 Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of...

  11. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...2014-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture... § 28.441 Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of...

  12. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...2012-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture... § 28.441 Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of...

  13. Compositions for chromosome-specific staining

    DOEpatents

    Gray, J.W.; Pinkel, D.

    1998-05-26

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. The methods produce staining patterns that can be tailored for specific cytogenetic analyses. The probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. The invention provides for automated means to detect and analyze chromosomal abnormalities. 17 figs.

  14. Compositions for chromosome-specific staining

    DOEpatents

    Gray, Joe W. (Livermore, CA); Pinkel, Daniel (Walnut Creek, CA)

    1998-01-01

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.

  15. Immunoperoxidase staining of cervicovaginal smears after radiotherapy.

    PubMed

    Wachtel, M S; Thaler, H T; Gangi, M D; Hajdu, S I

    1992-01-01

    Cervicovaginal smears from 2 women with postirradiation dysplasia, 4 women with postirradiation squamous cell carcinoma of the cervix, 30 women with irradiation atypia and 5 healthy, nonirradiated women were stained immunohistochemically with six keratin antibodies. For four of the antibodies--CK19 (BA17), EMA, PKK-1 and CAM 5.2--squamous cells showing irradiation atypia, postirradiation dysplasia or postirradiation squamous cell carcinoma were more likely to stain positively than were nonirradiated squamous cells. For three of the antibodies in which multiple squamous cells stained positively, the proportion of squamous cells showing postirradiation dysplasia or postirradiation squamous cell carcinoma staining strongly was equal to or greater than the corresponding overall proportion for squamous cells showing irradiation atypia. This was statistically significant with only one antibody, PKK-1. No statistically significant differences were seen in staining of irradiated and nonirradiated squamous cells by MAK-6 and AE1:AE3. The data show that some keratin antigens are more often expressed in the irradiated groups and that there may be differences in the degree of antigen expression between squamous cells showing postirradiation dysplasia or postirradiation squamous cell carcinoma and squamous cells showing irradiation atypia. PMID:1580112

  16. Histopathological confirmation of similar intramucosal distribution of fluorescein in both intravenous administration and local mucosal application for probe-based confocal laser endomicroscopy of the normal stomach.

    PubMed

    Nonaka, Kouichi; Ohata, Ken; Ban, Shinichi; Ichihara, Shin; Takasugi, Rumi; Minato, Yohei; Tashima, Tomoaki; Matsuyama, Yasushi; Takita, Maiko; Matsuhashi, Nobuyuki; Neumann, Helmut

    2015-12-16

    Probe-based confocal laser endomicroscopy (pCLE) is capable of acquiring in vivo magnified cross-section images of the gastric mucosa. Intravenous injection of fluorescein sodium is used for confocal imaging. However, it is still under debate if local administration of the dye to the mucosa is also effective for confocal imaging as it is not yet clear if topical application also reveals the intramucosal distribution of fluorescein. The objective of this study was to evaluate the intramucosal distribution of fluorescein sodium after topical application and to compare the distribution to the conventional intravenous injection used for confocal imaging. pCLE of the stomach uninfected with Helicobacter pylori was performed in a healthy male employing intravenous administration and local mucosal application of fluorescein. The mucosa of the lower gastric body was biopsied 1 min and 5 min after intravenous administration or local mucosal application of fluorescein, and the distribution of fluorescein in the biopsy samples was examined histologically. Green fluorescence was already observed in the cytoplasm of fundic glandular cells in the biopsied deep mucosa 1 min after local mucosal application of fluorescein. It was also observed in the foveolar lumen and inter-foveolar lamina propria, although it was noted at only a few sites. In the tissue biopsied 5 min after the local mucosal application of fluorescein, green fluorescence was more frequently noted in the cytoplasm of fundic glandular cells than in that 1 min after the local mucosal application of fluorescein, although obvious green fluorescence was not identified in the foveolar lumen or inter-foveolar lamina propria. The distribution of intravenously administered fluorescein in the cytoplasm of fundic glandular cells was also clearly observed similarly to that after local mucosal application of fluorescein. Green fluorescence in more cells was observed in many cells 5 min after intravenous administration compared with that after 1 min. The presence of fluorescein in the mucosa was observed within a short time after local mucosal application of fluorescein, suggesting that pCLE images similarly to those after intravenous fluorescein administration can be acquired by local mucosal application of fluorescein. PMID:26677449

  17. Histopathological confirmation of similar intramucosal distribution of fluorescein in both intravenous administration and local mucosal application for probe-based confocal laser endomicroscopy of the normal stomach

    PubMed Central

    Nonaka, Kouichi; Ohata, Ken; Ban, Shinichi; Ichihara, Shin; Takasugi, Rumi; Minato, Yohei; Tashima, Tomoaki; Matsuyama, Yasushi; Takita, Maiko; Matsuhashi, Nobuyuki; Neumann, Helmut

    2015-01-01

    Probe-based confocal laser endomicroscopy (pCLE) is capable of acquiring in vivo magnified cross-section images of the gastric mucosa. Intravenous injection of fluorescein sodium is used for confocal imaging. However, it is still under debate if local administration of the dye to the mucosa is also effective for confocal imaging as it is not yet clear if topical application also reveals the intramucosal distribution of fluorescein. The objective of this study was to evaluate the intramucosal distribution of fluorescein sodium after topical application and to compare the distribution to the conventional intravenous injection used for confocal imaging. pCLE of the stomach uninfected with Helicobacter pylori was performed in a healthy male employing intravenous administration and local mucosal application of fluorescein. The mucosa of the lower gastric body was biopsied 1 min and 5 min after intravenous administration or local mucosal application of fluorescein, and the distribution of fluorescein in the biopsy samples was examined histologically. Green fluorescence was already observed in the cytoplasm of fundic glandular cells in the biopsied deep mucosa 1 min after local mucosal application of fluorescein. It was also observed in the foveolar lumen and inter-foveolar lamina propria, although it was noted at only a few sites. In the tissue biopsied 5 min after the local mucosal application of fluorescein, green fluorescence was more frequently noted in the cytoplasm of fundic glandular cells than in that 1 min after the local mucosal application of fluorescein, although obvious green fluorescence was not identified in the foveolar lumen or inter-foveolar lamina propria. The distribution of intravenously administered fluorescein in the cytoplasm of fundic glandular cells was also clearly observed similarly to that after local mucosal application of fluorescein. Green fluorescence in more cells was observed in many cells 5 min after intravenous administration compared with that after 1 min. The presence of fluorescein in the mucosa was observed within a short time after local mucosal application of fluorescein, suggesting that pCLE images similarly to those after intravenous fluorescein administration can be acquired by local mucosal application of fluorescein. PMID:26677449

  18. Automated single-slide staining device

    NASA Technical Reports Server (NTRS)

    Wilkins, J. R.; Mills, S. M. (inventors)

    1977-01-01

    A simple apparatus and method is disclosed for making individual single Gram stains on bacteria inoculated slides to assist in classifying bacteria in the laboratory as Gram-positive or Gram-negative. The apparatus involves positioning a single inoculated slide in a stationary position and thereafter automatically and sequentially flooding the slide with increments of a primary stain, a mordant, a decolorizer, a counterstain and a wash solution in a sequential manner without the individual lab technician touching the slide and with minimum danger of contamination thereof from other slides.

  19. Detection Of Concrete Deterioration By Staining

    DOEpatents

    Guthrie, Jr., George D. (Santa Fe, NM); Carey, J. William (Santa Fe, NM)

    1999-09-21

    A method using concentrated aqueous solutions of sodium cobaltinitrite and a rhodamine dye is described which can be used to identify concrete that contains gels formed by the alkali-silica reaction (ASR), and to identify degraded concrete which results in a porous or semi-permeable paste due to carbonation or leaching. These solutions present little health or environmental risk, are readily applied, and rapidly discriminate between two chemically distinct gels; K-rich, Na--K--Ca--Si gels are identified by yellow staining, and alkali-poor, Ca--Si gels are identified by pink staining.

  20. Silver staining DNA in polyacrylamide gels.

    PubMed

    Bassam, Brant J; Gresshoff, Peter M

    2007-01-01

    This protocol describes a simple silver staining method used to visualize DNA fragments and other organic molecules with unsurpassed detail following traditional polyacrylamide gel electrophoresis (PAGE). Sensitivity rivals radioisotopic methods and DNA in the picogram range can be reliably detected. The described protocol is fast (approximately 1 h) and is implemented using readily available chemicals and materials. To achieve the sensitivity and visual clarity expected, quality reagents and clean handling are important. The updated protocol described here is based on the widely used method of Bassam et al. (1991), but provides improved image contrast and less risk of staining artefacts. PMID:18007600

  1. Intracellular release of fluorescein anion from layered double hydroxide nanoparticles indicating endosomal escape

    NASA Astrophysics Data System (ADS)

    Tanaka, M.; Aisawa, S.; Hidetoshi, H.; Narita, E.; Dong, Q.; Yin, S.; Sato, T.

    2013-12-01

    In recent years, layered double hydroxide (LDH) has been attempted to be applied to a molecular container due to their anion exchange ability, low cytotoxicity and good biocompatibility. In this paper, we investigated the intracellular behaviour of LDH particles in mammalian cells after internalization. Nanoparticles of fluorescein (Fluo) intercalated LDH, Fluo/LDH, were prepared by the coprecipitation followed by subsequent hydrothermal treatment. As-prepared Fluo/LDH particles have the LDH structure and morphology of hexagonal sheet of 100 nm on the average. In addition, Fluo/LDH also exhibited high green fluorescence and low cytotoxicity. By a confocal laser scanning microscopy, the dim green fluorescence was observed throughout cells, including the nucleus. This result indicated that Fluo/LDH released guest anion (Fluo) from LDH structure inside cells. Furthermore, because the fluorescence was observed throughout the cell, Fluo was not retained within endosome structure, i.e., Fluo/LDH was dissolved to release Fluo from endosome.

  2. Early events in herpes simplex virus type 1 infection: photosensitivity of fluorescein isothiocyanate-treated virions

    SciTech Connect

    DeLuca, N.; Bzik, D.; Person, S.; Snipes, W.

    1981-02-01

    Herpes simplex virus type 1 is photosensitized by treatment with fluorescein isothiocyanate (FITC). The inactivation of FITC-treated virions upon subsequent exposure to light is inhibited by the presence of sodium azide, suggesting the involvement of singlet oxygen in the process. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis revealed that treatment with FITC plus light induces crosslinks in viral envelope glycoproteins. Treatment of virions with high concentrations of FITC (50 ..mu..g/ml) plus light causes a reduction in the adsorption of the virus to monolayers of human embryonic lung cells. For lower concentrations of FITC (10 ..mu..g/ml) plus light, treated virions adsorb to the host cells, but remain sensitive to light until entry occurs. The loss of light sensitivity coincides with the development of resistance to antibodies. These results are most consistent with a mechanism of entry for herpes simplex virus involving fusion of the viral membrane with the plasma membrane of the host cell.

  3. Automated detection of leakage in fluorescein angiography images with application to malarial retinopathy.

    PubMed

    Zhao, Yitian; MacCormick, Ian J C; Parry, David G; Leach, Sophie; Beare, Nicholas A V; Harding, Simon P; Zheng, Yalin

    2015-01-01

    The detection and assessment of leakage in retinal fluorescein angiogram images is important for the management of a wide range of retinal diseases. We have developed a framework that can automatically detect three types of leakage (large focal, punctate focal, and vessel segment leakage) and validated it on images from patients with malarial retinopathy. This framework comprises three steps: vessel segmentation, saliency feature generation and leakage detection. We tested the effectiveness of this framework by applying it to images from 20 patients with large focal leak, 10 patients with punctate focal leak, and 5,846 vessel segments from 10 patients with vessel leakage. The sensitivity in detecting large focal, punctate focal and vessel segment leakage are 95%, 82% and 81%, respectively, when compared to manual annotation by expert human observers. Our framework has the potential to become a powerful new tool for studying malarial retinopathy, and other conditions involving retinal leakage. PMID:26030010

  4. Fully Automatic Segmentation of Fluorescein Leakage in Subjects With Diabetic Macular Edema

    PubMed Central

    Rabbani, Hossein; Allingham, Michael J.; Mettu, Priyatham S.; Cousins, Scott W.; Farsiu, Sina

    2015-01-01

    Purpose. To create and validate software to automatically segment leakage area in real-world clinical fluorescein angiography (FA) images of subjects with diabetic macular edema (DME). Methods. Fluorescein angiography images obtained from 24 eyes of 24 subjects with DME were retrospectively analyzed. Both video and still-frame images were obtained using a Heidelberg Spectralis 6-mode HRA/OCT unit. We aligned early and late FA frames in the video by a two-step nonrigid registration method. To remove background artifacts, we subtracted early and late FA frames. Finally, after postprocessing steps, including detection and inpainting of the vessels, a robust active contour method was utilized to obtain leakage area in a 1500-?m-radius circular region centered at the fovea. Images were captured at different fields of view (FOVs) and were often contaminated with outliers, as is the case in real-world clinical imaging. Our algorithm was applied to these images with no manual input. Separately, all images were manually segmented by two retina specialists. The sensitivity, specificity, and accuracy of manual interobserver, manual intraobserver, and automatic methods were calculated. Results. The mean accuracy was 0.86 ± 0.08 for automatic versus manual, 0.83 ± 0.16 for manual interobserver, and 0.90 ± 0.08 for manual intraobserver segmentation methods. Conclusions. Our fully automated algorithm can reproducibly and accurately quantify the area of leakage of clinical-grade FA video and is congruent with expert manual segmentation. The performance was reliable for different DME subtypes. This approach has the potential to reduce time and labor costs and may yield objective and reproducible quantitative measurements of DME imaging biomarkers. PMID:25634978

  5. PREDICTIVE MODEL FOR THE COMBINED EFFECT OF TEMPERATURE, SODIUM LACTATE, AND SODIUM DIACETATE ON THE HEAT RESISTANCE OF LISTERIA MONOCYTOGENES IN BEEF

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Listeria monocytogenes continues to be one of the most important foodborne psychrotrophic pathogens of public health significance and a major concern to the food industry and regulatory agencies. Sodium lactate (NaL) and sodium diacetate (SDA) are generally regarded as safe and are used in meat prod...

  6. EFFECTS AND INTERACTIONS OF SODIUM LACTATE, SODIUM DIACETATE, AND PEDIOCIN ON THE THERMAL INACTIVATION OF STARVED CELLS OF LISTERIA MONOCYTOGENES ON THE SURFACE OF BOLOGNA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects and interactions of sodium lactate, sodium diacetate, and pediocin on the thermal inactivation of starved cells of Listeria monocytogenes on the surface of bologna were investigated. The heating temperatures used in the study were 56.3 to 60 degrees C and the antimicrobials were: sodium ...

  7. Effects and interactions of sodium lactate, sodium diacetate, and pediocin on the thermal inactivation of starved cells of Listeria monocytogenes on the surface of bologna

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects and interactions of temperature (56.3-60C) sodium lactate (SL; 0-4.8%), sodium diacetate (SDA; 0-2.5%), and pediocin (0-10,000 AU) on starved Listeria monocytogenes on bologna were investigated. Bologna slices containing SL and SDA in the formulation were dipped in pediocin, surface inoc...

  8. Modeling the Lag Phase and Growth Rate of Listeria monocytogenes in Ground Ham Containing Sodium Lactate and Sodium Diacetate at Various Storage Temperatures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Refrigerated ready-to-eat (RTE) meats contaminated with L. monocytogenes were implicated in several listeriosis outbreaks. Lactate and diacetate have been shown to control L. monocytogenes in RTE meats. The objective of this study was to examine and model the effect of lactate (1.0-4.2%) and diace...

  9. RADIATION (GAMMA) RESISTANCE AND POST-IRRADIATION GROWTH OF LISTERIA MONOCTYTOGENES SUSPENDED IN BEEF BOLOGNA THAT CONTAINED SODIUM DIACETATE AND POTASSIUM LACTATE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Listeria monocytogenes (Lm), a psychrotrophic food-borne pathogen, is a frequent post-process contaminant on ready-to-eat meat (RTE) products including bologna. Ionizing radiation can eliminate Lm from ready-to-eat meats. Sodium diacetate (SDA) and potassium lactate (PL) mixtures inhibit the growt...

  10. Calculation of UV, IR, and NMR Spectra of Diethyl 2,2'-[(1,1'-Biphenyl)-4,4'-Diylbis(Azanediyl)]Diacetate

    NASA Astrophysics Data System (ADS)

    Almodarresiyeh, H. A.; Shahab, S. N.; Zelenkovsky, V. M.; Ariko, N. G.; Filippovich, L. N.; Agabekov, V. E.

    2014-03-01

    The new substance diethyl 2,2'-[(1,1'-biphenyl)-4,4'-diylbis(azanediyl)]diacetate (M13) was modeled using the Hartree-Fock and density functional theory methods and then synthesized. The electronic absorption spectrum of M13 in dimethylformamide solution was calculated. The UV, IR, and NMR spectra of M13 were presented.

  11. Survival and growth of Listeria monocytogenes in broth as a function of temperature, pH, and potassium lactate and sodium diacetate concentrations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to determine the antimicrobial effect of a combination of potassium lactate and sodium diacetate (PURASAL P Opti.Form 4TM, 60% solution) on the survival and growth of Listeria monocytogenes Scott A in pH adjusted broth (5.5, 6.0, 6.5 and 7.0) stored at 4, 10, 17, 24, ...

  12. The Language of Stained-Glass Windows

    ERIC Educational Resources Information Center

    Brew, Charl Anne

    2010-01-01

    The splendor and beauty of stained glass punctuates any room. In this article, the author describes a cross-curriculum project which incorporated the French classes' research and written study of France in the Middle Ages. For the project the author suggested Sainte-Chapelle which is considered a reliquary and was built by Louis IX to house the…

  13. Continuous-flow cytomorphological staining and analysis.

    PubMed

    Tan, Andrew P; Dudani, Jaideep S; Arshi, Armin; Lee, Robert J; Tse, Henry T K; Gossett, Daniel R; Di Carlo, Dino

    2014-02-01

    Cells suspended in bodily fluids are routinely analyzed by cytopathologists as a means of diagnosing malignancies and other diseases. The physical and morphological properties of these suspended cells are evaluated in making diagnostic decisions, which often requires manual concentration, staining, and washing procedures to extract information about intracellular architecture. The need to manually prepare slides for analysis by a cytopathologist is a labor-intensive process, which is ripe for additional automation to reduce costs but also to potentially provide more repeatable and improved accuracy in diagnoses. We have developed a microfluidic system to perform several steps in the preparation of samples for cytopathology that (i) automates colorimetric staining on-chip, and (ii) images cells in flow, as well as provides (iii) additional quantitative analyses of captured images to aid cytopathologists. A flow-through approach provides benefits by allowing staining and imaging to be performed in a continuous, integrated manner, which also overcomes previous challenges with in-suspension colorimetric staining. We envision such a tool may reduce costs and aid cytopathologists in identifying rare or characteristic cells of interest by providing isolated images along with quantitative metrics on single cells from various rotational angles, allowing efficient determination of disease etiology. PMID:24217244

  14. Lighting up Protons with MorphFl, a Fluorescein-Morpholine Dyad: An Experiment for the Organic Laboratory

    ERIC Educational Resources Information Center

    Miller, Tyson A.; Spangler, Michael; Burdette, Shawn C.

    2011-01-01

    A two-period organic laboratory experiment that includes fluorescence sensing is presented. The pH-sensitive sensor MorphFl is prepared using a Mannich reaction between a fluorescein derivative and the iminium ion of morpholine. During the first laboratory, students prepare MorphFl. The second session begins with characterizing the sensor using…

  15. Methods and compositions for chromosome-specific staining

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel

    2003-07-22

    Methods and compositions for chromosome-specific staining are provided. Compositions comprise heterogenous mixtures of labeled nucleic acid fragments having substantially complementary base sequences to unique sequence regions of the chromosomal DNA for which their associated staining reagent is specific. Methods include methods for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes.

  16. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Dye and chemical solution stains. 864.1850 Section... (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains for medical purposes are mixtures...

  17. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Dye and chemical solution stains. 864.1850 Section... (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains for medical purposes are mixtures...

  18. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Dye and chemical solution stains. 864.1850 Section... (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains for medical purposes are mixtures...

  19. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Dye and chemical solution stains. 864.1850 Section... (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains for medical purposes are mixtures...

  20. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Dye and chemical solution stains. 864.1850 Section... (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains for medical purposes are mixtures...

  1. Evaluation of lanthanide salts as alternative stains to uranyl acetate.

    PubMed

    Hosogi, Naoki; Nishioka, Hideo; Nakakoshi, Masamichi

    2015-12-01

    Uranyl acetate (UAc) has been generally used not only as a superb staining reagent for ultrathin sections of plastic-embedded biological materials, but also as high-contrast negative stains for biological macromolecules such as particles of protein or virus. However, the use and purchase of radioactive UAc have been restricted. In this study, we determine the performance of ytterbium triacetate, lutetium triacetate, samarium triacetate and gadolinium triacetate as new staining reagents for biological electron microscopy. We observed chemically fixed spinach (Spinacia oleracea) leaves stained with these reagents. Ultrathin sections were stained with these reagents. Some of them were counterstained with lead citrate. The transmission electron microscopy contrast of spinach organelles was evaluated in sections exposed to the conventional stain and new stains. We show acetate salts of samarium, gadolinium, ytterbium and lutetium could be excellent substitutes for UAc for thin section staining and for negative staining. In addition, each reagent showed appreciable negative-staining effects. PMID:26374081

  2. Staining is complete in less than three hours The procedure includes only three steps --fixation, oxidation and staining

    E-print Network

    Lebendiker, Mario

    lipopolysaccharides in gels Pro-Q Emerald 300 Lipopolysaccharide Stain Kit Pro-Q Emerald 300 Lipopolysaccharide Stain Kit #12;Molecular Probes' proprietary Pro-Q Emerald 300 Lipopolysaccharide Stain Kit provides the most). The Pro-Q Emerald 300 reagent in the kit stains the periodate-oxidized carbohydrates of LPS with a bright

  3. Improved Whole-Blood-Staining Device

    NASA Technical Reports Server (NTRS)

    Sams, Clarence F.; Crucian, Brian; Paul, Bonnie; Melton, Shannon; Guess, Terry

    2012-01-01

    Dramatic improvements have been made in NASA s Whole Blood Staining Device (WBSD) since it was last described in "Whole-Blood-Staining Device," NASA Tech Briefs, Vol. 23, No. 10 (October 1999), page 64. The new system has a longer shelf life, a simpler and more effective operational procedure, improved interface with instrumentation, and shorter processing time. More specifically, the improvements have targeted bag and locking clip materials, sampling ports, and air pocket prevention. The WBSD stains whole blood collected during spaceflight for subsequent flow cytometric analysis. In short, the main device stains white blood cells by use of monoclonal antibodies conjugated to various fluorochromes, followed by lysing and fixing of the cells by use of a commercial reagent that has been diluted according to NASA safety standards. This system is compact, robust, and does not require electric power, precise mixing, or precise incubation times. Figure 1 depicts the present improved version for staining applications, which is a poly(tetrafluoroethylene) bag with a Luer-lock port and plastic locking clips. An InterLink (or equivalent) intravenous- injection port screws into the Luer-lock port. The inflatable/collapsible nature of the bag facilitates loading and helps to minimize the amount of air trapped in the fully loaded bag. Some additional uses have been identified for the device beyond whole blood staining. The WBSD has been configured for functional assays that require culture of live cells by housing sterile culture media, mitogens, and fixatives prior to use [Figure 2(a)]. Simple injection of whole blood allows cell-stimulation culture to be performed in reduced gravity conditions, and product stabilization prior to storage, while protecting astronauts from liquid biohazardous materials. Also, the improved WBSD has reconstituted powdered injectable antibiotics by mixing them with diluent liquids [Figure 2(b)]. Although such mixing can readily be performed on Earth by shaking in glass vials, it cannot readily be performed this way in outer space without entraining air bubbles. The present device can be preloaded with the powder and diluent(s) in separate compartments. The powder and diluent( s) can be mixed, without introducing air bubbles, by removing the clip(s), then shaking. This use of the device could also be advantageous in terrestrial applications because it maintains the isolation of the constituents until the time of use.

  4. Histochemical staining of cadmium with thiazolylazophenol derivatives.

    PubMed

    Sumi, Y

    1980-01-01

    In the search for a more sensitive and specific chelant for cadmium, thiazolylazophenol derivatives were synthesized and their staining properties for cadmium and zinc were examined. The compounds synthesized for this study were identified as being thiazolylazophenol derivatives via analysis of their melting point, absorption maxima and their C, H, N, S content. Thiazolylazo-p-methoxyphenol (TAMe) was found to stain cadmium and zinc differentially and with the highest optical density amongst dyestuffs reported to date. The plots of absorbance vs. metal concentration for thiazolylazophenol chelates gave a straight line obeying the Beer-Lambert law. Only the plot for TAMe-zinc chelates deviated significantly from a straight line above a specific concentration. The change from a 2:1 (chelant:metal) to a 3:1 (chelant:metal) complex is suggested to be the reason for the differential coloring. PMID:6156927

  5. Laser Treatment of Port Wine Stains

    NASA Astrophysics Data System (ADS)

    Majaron, Boris; Nelson, J. Stuart

    Port wine stain (PWS), also called nevus flammeus, is a congenital, cutaneous vascular malformation involving post-capillary venules which produce a light pink to red to dark-red-violet discoloration of human skin [1]. PWS occurs in an estimated 3 children per 1000 live births, affecting males and females and all racial groups equally [2]. There appears to be no hereditary predilection for PWS within families. There are no known risk factors or ways to prevent PWS.

  6. Aptamer Stainings for Super-resolution Microscopy.

    PubMed

    de Castro, Maria Angela Gomes; Rammner, Burkhard; Opazo, Felipe

    2016-01-01

    Fluorescence microscopy is an invaluable tool to visualize molecules in their biological context with ease and flexibility. However, studies using conventional light microscopy have been limited to the resolution that light diffraction allows (i.e., ~200 nm). This limitation has been recently circumvented by several types of advanced fluorescence microscopy techniques, which have achieved resolutions of up to ~10 nm. The resulting enhanced imaging precision has helped to find important cellular details that were not visible using diffraction-limited instruments. However, it has also revealed that conventional stainings using large affinity tags, such as antibodies, are not accurate enough for these imaging techniques. Since aptamers are substantially smaller than antibodies, they could provide a real advantage in super-resolution imaging. Here we compare the live staining of transferrin receptors (TfnR) obtained with different fluorescently labeled affinity probes: aptamers, specific monoclonal antibodies, or the natural receptor ligand transferrin. We observed negligible differences between these staining strategies when imaging is performed with conventional light microscopy (i.e., laser scanning confocal microscopy). However, a clear superiority of the aptamer tag over antibodies became apparent in super-resolved images obtained with stimulated emission depletion (STED) microscopy. PMID:26552828

  7. Fullerol-fluorescein isothiocyanate phosphorescent labeling reagent for the determination of glucose and alkaline phosphatase.

    PubMed

    Liu, Jia-Ming; Wang, Hong-Xin; Zhang, Li-Hong; Zheng, Zhi-Yong; Lin, Shao-Qin; Lin, Li-Ping; Wang, Xin-Xing; Lin, Chang-Qing; Liu, Jian-Qin; Huang, Qi-Tong

    2010-09-15

    The active -OH group in fullerol (F-ol) could react with the dissociated -COOH group in fluorescein isothiocyanate (FITC) to form F-ol-(FITC)(n), which could emit room temperature phosphorescence (RTP) signal of F-ol and FITC on acetate cellulose membrane (ACM), respectively. Their RTP signals were enhanced by N,N-dimethylaniline (DMA). The labeling reaction between the -NCS group of FITC in DMA-F-ol-(FITC)(n) and the -NH2 group in wheat germ agglutinin (WGA) produced DMA-F-ol-(FITC)(n)-WGA, which could further take affinity adsorption (AA) reaction with bioactive substances (BS), such as glucose and alkaline phosphatase (AP), to produce DMA-F-ol-(FITC)(n)-WGA-BS. Both of these two products could maintain the good RTP characteristics of F-ol and FITC. Based on the facts above, a new phosphorescent labeling reagent, DMA-F-ol-FITC, was developed, and a new affinity adsorption solid substrate room temperature phosphorimetry (AASSRTP) for the determination of BS was established. This method was applied to the determination of BS in human serum and the diagnosis of diseases, with the results agreeing very well with those of enzyme-linked immunosorbent assay (ELISA). The mechanism of DMA-F-ol-(FITC)(n) labeling of WGA and AASSRTP for the determination of BS is discussed. PMID:20507821

  8. Estimating retinal vascular permeability using the adiabatic approximation to the tissue homogeneity model with fluorescein videoangiography

    NASA Astrophysics Data System (ADS)

    Tichauer, Kenneth M.; Osswald, Christian R.; Dosmar, Emily; Guthrie, Micah J.; Hones, Logan; Sinha, Lagnojita; Xu, Xiaochun; Mieler, William F.; St. Lawrence, Keith; Kang-Mieler, Jennifer J.

    2015-06-01

    Clinical symptoms of diabetic retinopathy are not detectable until damage to the retina reaches an irreversible stage, at least by today's treatment standards. As a result, there is a push to develop new, "sub-clinical" methods of predicting the onset of diabetic retinopathy before the onset of irreversible damage. With diabetic retinopathy being associated with the accumulation of long-term mild damage to the retinal vasculature, retinal blood vessel permeability has been proposed as a key parameter for detecting preclinical stages of retinopathy. In this study, a kinetic modeling approach used to quantify vascular permeability in dynamic contrast-enhanced medical imaging was evaluated in noise simulations and then applied to retinal videoangiography data in a diabetic rat for the first time to determine the potential for this approach to be employed clinically as an early indicator of diabetic retinopathy. Experimental levels of noise were found to introduce errors of less than 15% in estimates of blood flow and extraction fraction (a marker of vascular permeability), and fitting of rat retinal fluorescein angiography data provided stable maps of both parameters.

  9. Correlation between Fluorescein Angiographic Findings and Visual Acuity in Behçet Retinal Vasculitis

    PubMed Central

    Kim, Min; Kwon, Hee Jung; Choi, Eun Young; Kim, Sung Soo; Koh, Hyoung Jun

    2015-01-01

    Purpose To identify significant fluorescein angiographic (FA) characteristics associated with visual acuity (VA) in Behçet retinal vasculitis. Materials and Methods Retrospective review of 86 eyes of 48 patients (age: 35.6±10.2 years) with Behçet retinal vasculitis were performed. VA and FA findings as well as correlation between them were assessed. Results The mean initial VA of eyes with posterior pole-involved vasculitis (63 eyes; 73.3%) was significantly worse than that of those with peripheral vasculitis (23 eye; 26.7%) (logarithm of the minimum angle of resolution VA: 0.554±0.572 vs. 0.078±0.148; p<0.0001). Subgroup analysis revealed a more severe and diffuse pattern of vascular leakage in posterior pole-involved vasculitis compared to peripheral vasculitis (p<0.0001). Retinal vascular leakage (?=0.345; p<0.0001), optic disc hyperfluorescence (?=0.147; p=0.032), and macular leakage (?=0.107; p=0.047) were significantly associated with worse initial VA. During the follow up (mean: 33.3±17.9 months), the change of leakage showed no significant correlation with change of VA in posterior pole-involved vasculitis (?=0.199, p=0.092). Conclusion Posterior pole involvement, the degree of retinal vascular leakage, optic disc hyperfluorescence, and macular leakage are significantly associated with VA in Behçet retinal vasculitis. PMID:26069134

  10. Synthesis, crystal structure and luminescent properties of a new samarium-fluorescein metal-organic framework

    NASA Astrophysics Data System (ADS)

    Thomas, Jesty; Ambili, K. S.

    2015-10-01

    A new metal-organic framework with empirical formula C43H30NO12Sm was solvothermally synthesized using SmCl3, fluorescein and N, N-Dimethyl formamide (DMF) and characterized by single crystal X-ray diffraction, powder X-ray diffraction, infrared spectroscopy, UV-Visible spectroscopy, scanning electron microscopy, optical microscopy, photoluminescence spectroscopy, CHN elemental analysis and thermogravimetric analysis. Single crystal X-ray diffraction revealed that the crystal structure belongs to the triclinic system, P-1 space group with a = 12.113 (6) Å, b = 12.1734 (7) Å, c = 13.2760(8) Å, ? = 67.930(3)?, ? = 87.779(3)?, ? = 77.603(3)? and V = 1769.71 (17) Å3. The photoluminescence spectrum showed emission peaks at 550 nm, 600 nm and 647 nm due to the characteristic transitions 4G5/2 to 6H5/2, 4G5/2 to 6H7/2 and 4G5/2 to 6H9/2 respectively, when excited at 398 nm.

  11. Retinal vein-to-vein anastomoses in Sturge-Weber syndrome documented by ultra-widefield fluorescein angiography.

    PubMed

    Quan, Ann V; Moore, Grant H; Tsui, Irena

    2015-06-01

    We report the case of a 6-year-old boy with Sturge-Weber syndrome and unilateral glaucoma in his left eye. He was born with a port wine mark involving his upper left eyelid. On ultra-widefield fluorescein angiography, he was found to have several vein-to-vein anastomoses in his left retina. To our knowledge, this is the first documentation of retinal vein-to-vein anastomoses in Sturge-Weber syndrome. PMID:25944745

  12. A Unified Strategy to 6-5-6-5-6-Membered Epipolythiodiketopiperazines: Studies towards the Total Synthesis of Scabrosin Diacetate and Haematocin.

    PubMed

    Zipfel, Hannes F; Carreira, Erick M

    2015-08-24

    The family of epipolythiodiketopiperazine (ETP) natural products consists of over 200 members possessing a wide diversity of structures and biological activity. Recently, the subgroup of 6-5-6-5-6-membered ETPs has gained substantial attention, which has resulted in several total syntheses. Despite all the efforts that have been invested into accessing these complex structures, no synthesis of scabrosin diacetate (1?a) and its related esters has been reported. Herein, our attempts towards scabrosin diacetate (1?a) and haematocin (3) starting from diketopiperazine 12?a as a late-stage intermediate are presented. Diketopiperazine 12?a can be conveniently accessed in multigram quantities from aldehyde 18 and diketopiperazine 21 and was envisioned to serve as a general platform for the synthesis of 6-5-6-5-6-membered ETPs. PMID:26179159

  13. Fetal Outcome in Meconium Stained Deliveries

    PubMed Central

    Mundhra, Rajlaxmi; Agarwal, Manika

    2013-01-01

    Objective: To evaluate the foetal outcome in Meconium Stained Amniotic Fluid (MSAF). Material and Methods: This prospective observational study was carried out in the Department of Obstetrics and Gynaecology, North Eastern Indira Gandhi Regional Institute of Health And Medical Sciences, Shillong, India, over a period of eighteen months, from January 2010 to June 2011. A total of 355 pregnant women who had completed more than 37 weeks of gestation, with singleton pregnancies and cephalic presentations were included in this study. One hundred and sixty five cases with MSAF, were thus selected and they were compared with 190 randomly selected controls. Results: Among 165 cases, 27.88 % of the cases had regular visits to the Institute at least 3 times previously, 72.12% cases had no previous visit at all. Primigravidas accounted for a majority of cases and approximately 50% cases had gestational ages of more than 40 weeks Pregnancies complicated with pregnancy induced hypertension had statistically significant higher rates of meconium staining among cases (16.97%), as compared to those among controls (7.89%). 21.81% cases had foetal heart rate abnormalities, as were detected by electronic foetal monitoring and presence of foetal bradycardia was statistically higher in cases compared to that in controls. Casearean section rates were nearly double in cases (49.09%). Neonatal outcome was poor in terms of low Apgar score at birth, birth asphyxia, Meconium Aspiration Syndrome (MAS) and increased neonatal admission among cases as compared to that among controls. Conclusion: Meconium stained amniotic fluid is really worrisome from both, obstetrician’s and paediatrician’s points of view, as it increases the caesarean rates, causes birth asphyxia, MAS and increases neonatal intensive care unit admissions. PMID:24551662

  14. Port wine stain on a child's face (image)

    MedlinePLUS

    Port wine stains are always present at birth. In an infant, they are flat, pink, vascular lesions. Common locations ... may be present anywhere on the body. Port wine stains may appear in association with other syndromes.

  15. Polyvinyl pyrrolidone capped fluorescent anthracene nanoparticles for sensing fluorescein sodium in aqueous solution and analytical application for ophthalmic samples.

    PubMed

    Bhopate, Dhanaji P; Mahajan, Prasad G; Garadkar, Kalyanrao M; Kolekar, Govind B; Patil, Shivajirao R

    2015-11-01

    Based on the known complexation ability between polyvinyl pyrrolidone (PVP) and fluorescein sodium (FL Na(+) ), fluorescent PVP capped anthracene nanoparticles (PVP-ANPs) were prepared using a reprecipitation method for detection of fluorescein in aqueous solution using the fluorescence resonance energy transfer (FRET) approach. A dynamic light scattering histogram of PVP-ANPs showed narrower particle size distribution and the average particle size was 15?nm. The aggregation-induced enhanced emission (AIEE) of PVP-ANPs was red shifted from its monomer by 1087.22?cm(-1) . The maximum emission was seen to occur at 420?nm. The presence of FL Na(+) in the vicinity of PVP-ANPs quenched the fluorescence of PVP-ANPs because of its adsorption on the surface of PVP-ANPs in aqueous suspension. The FL Na(+) and PVP-ANPs were brought close enough, typically to 7.89?nm, which was less than the distance of 10?nm that is required between the energy donor-acceptor molecule for efficient FRET. The quenching results fit into the Stern-Volmer relationship even at temperatures greater than ambient temperatures. The thermodynamic parameters determined from FRET results helped to propose binding mechanisms involving hydrophobic and electrostatic molecular interaction. The fluorescence quenching results were used further to develop an analytical method for estimation of fluorescein sodium from ophthalmic samples available commercially in the market. Copyright © 2015 John Wiley & Sons, Ltd. PMID:25736374

  16. Bleaching of fluorosis stains using sodium hypochlorite.

    PubMed

    Penumatsa, Narendra Varma; Sharanesha, Rajashekhara Bhari

    2015-08-01

    Fluorosis staining is commonly considered an esthetic problem because of the psychological impact of unesthetic maxillary anterior teeth. Numerous treatment approaches have been proposed, ranging from bleaching to enamel reduction to restorative techniques. Bleaching of hypomineralized enamel lesions, using 5% sodium hypochlorite, has been useful clinically. The technique described, in this case, appears to have advantages over other methods for improving the appearance of fluorotic lesions. It is simple, low cost, noninvasive, so the enamel keeps its structure, relatively rapid, and safe; it requires no special materials, and it can be used with safety on young permanent teeth. PMID:26538964

  17. Bleaching of fluorosis stains using sodium hypochlorite

    PubMed Central

    Penumatsa, Narendra Varma; Sharanesha, Rajashekhara Bhari

    2015-01-01

    Fluorosis staining is commonly considered an esthetic problem because of the psychological impact of unesthetic maxillary anterior teeth. Numerous treatment approaches have been proposed, ranging from bleaching to enamel reduction to restorative techniques. Bleaching of hypomineralized enamel lesions, using 5% sodium hypochlorite, has been useful clinically. The technique described, in this case, appears to have advantages over other methods for improving the appearance of fluorotic lesions. It is simple, low cost, noninvasive, so the enamel keeps its structure, relatively rapid, and safe; it requires no special materials, and it can be used with safety on young permanent teeth. PMID:26538964

  18. Several staining techniques to enhance the visibility of Acanthamoeba cysts.

    PubMed

    El-Sayed, Nagwa Mostafa; Hikal, Wafaa Mohamed

    2015-03-01

    Acanthamoeba is one of the most common free-living amoebae. It is widespread in the environment and can infect humans causing keratitis. Delayed diagnosis or misdiagnosis leads to extensive corneal inflammation and profound visual loss. Therefore, accurate and rapid diagnosis of Acanthamoeba keratitis is essential for successful treatment and good prognosis. This study was designed to use different staining techniques to facilitate the identification of Acanthamoeba cysts. Acanthamoeba cysts were isolated by cultivation of either corneal scraping specimens or tap water samples onto non-nutrient agar plates seeded with Escherichia coli. Subcultures were done from positive cultures until unique cysts were isolated. Acanthamoeba cysts were stained temporarily using iodine, eosin, methylene blue, and calcofluor white (CFW) stains and as permanent slides after processing for mounting using modified trichrome, Gimenez and Giemsa staining. These stains were compared on the basis of staining quality including clarity of morphological details, differentiation between cytoplasm and nuclei, color and contrast, and also other characteristics of the staining techniques, including ease of handling, time taken for the procedure, and cost effectiveness. The cysts of Acanthamoeba were recognized in the form of double-walled cysts: the outer wall (ectocyst) that was being differentiated from the variably stained surrounding background and the inner wall (endocyst) that was sometimes stellated, polygonal, round, or oval and visualized as separate from the spherical, sometimes irregular, outline of the ectocyst. Regarding the temporary stains, it was found that they were efficient for visualizing the morphological details of Acanthamoeba cysts. In CFW staining, Acanthamoeba cysts appeared as bluish-white or turquoise oval halos although the internal detail was not evident. On the other hand, the results of permanent-stained slides showed the most consistent stain for identification of Acanthamoeba cysts was modified trichrome followed by Gimenez stain and lastly Giemsa stain that gave poor visibility of Acanthamoeba cysts due to the intense staining background and monochrome staining of parasite. In the present study, multi-attribute ranking of the used staining techniques showed the highest rank for iodine stain (92 %) followed by eosin stain (84 %), Gimenez stain (76 %), methylene blue (72 %), CFW (64 %), modified trichrome (56 %), and the least was Giemsa stain (44 %). In conclusion, the staining techniques enhance the overall visibility of Acanthamoeba cysts. PMID:25346196

  19. PREDICTIVE MODEL FOR THE COMBINED EFFECT OF TEMPERATURE, SODIUM LACTATE, AND SODIUM DIACETATE ON THE HEAT RESISTANCE OF LISTERIA MONOCYTOGENES IN BEEF

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects and interactions of heating temperature (60 - 73.9C), sodium lactate (NaL; 0.0 - 4.8%, w/w) and/or sodium diacetate SDA; 0.0 - 0.25%, w/w) on the heat resistance of a five-strain mixture of Listeria monocytogenes in 75% lean ground beef were examined. Thermal death times were determined...

  20. Catalytic asymmetric synthesis of butane diacetal-protected (4S,5S)-dihydroxycyclohexen-1-one and use in natural product synthesis.

    PubMed

    Burns, David J; Hachisu, Shuji; O'Brien, Peter; Taylor, Richard J K

    2012-10-14

    Due to the lack of availability of unnatural (+)-quinic acid as a starting material, a 6-step synthesis of butane diacetal-protected (4S,5S)-dihydroxycyclohexen-1-one (formally derived from (+)-quinic acid) has been devised. The key catalytic asymmetric step involves a chiral Co-salen-catalysed epoxide ring-opening reaction. (4S,5S)-Dihydroxycyclohexen-1-one was utilised in the synthesis of two cyclohexenone natural products isolated from the mycelia of Lasiodiplodia theobromae. PMID:22930235

  1. Cigarette staining and cleaning of a maxillofacial silicone

    SciTech Connect

    Yu, R.; Koran, A.; Raptis, C.N.; Craig, R.G.

    1983-07-01

    In this study, a maxillofacial silicone elastomer was stained with cigarette smoke. The stain was then removed by solvent extraction using 1,1,1-trichloroethane. The cigarette smoke produced large color changes in the elastomer as measured from spectrophotometric reflectance curves. The solvent was totally effective in removing the cigarette stain without changing the color of the silicone base.

  2. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Middling Yellow Stained Color. 28.441 Section... Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of Strict Middling Tinged Color....

  3. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Strict Middling Yellow Stained Color. 28.441 Section... Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of Strict Middling Tinged Color....

  4. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442... Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is deeper than Middling Tinged Color. below color grade cotton...

  5. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442... Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is deeper than Middling Tinged Color. below color grade cotton...

  6. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Strict Middling Yellow Stained Color. 28.441 Section... Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of Strict Middling Tinged Color....

  7. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442... Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is deeper than Middling Tinged Color. below color grade cotton...

  8. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Strict Middling Yellow Stained Color. 28.441 Section... Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of Strict Middling Tinged Color....

  9. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442... Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is deeper than Middling Tinged Color. below color grade cotton...

  10. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442... Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is deeper than Middling Tinged Color. below color grade cotton...

  11. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Strict Middling Yellow Stained Color. 28.441 Section... Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of Strict Middling Tinged Color....

  12. Non-photorealistic Rendering of Images as Evolutionary Stained Glass

    E-print Network

    Ashlock, Dan

    Non-photorealistic Rendering of Images as Evolutionary Stained Glass Daniel Ashlock Mathematics glass. A collection of points that are the centers of weighted Voronoi tilings are evolved to minimize. A fractal model of stained glass is then run to create a stained glass texture with a similar average color

  13. Spectroscopic characterization of the binding mechanism of fluorescein and carboxyfluorescein in human serum albumin

    NASA Astrophysics Data System (ADS)

    Sulaiman, Saba A. J.; Kulathunga, H. Udani; Abou-Zied, Osama K.

    2015-03-01

    Fluorescein (FL) and some of its precursors have proven to be effective fluorescent tracers in pharmaceutical and medical applications owing to their high quantum yield of fluorescence in physiological conditions and their high membrane permeability. In order to protect FL from metabolic effects during the process of its delivery, human serum albumin (HSA) has been used as a carrier because of its compatibility with the human body. In the present work, we used spectroscopic methods to characterize the binding mechanisms of FL and one of its derivatives, 5(6)- carboxyfluorescein (CFL), in the HSA protein. The absorbance change of the two ligands (FL and CFL) was quantified as a function of the HSA concentration and the results indicate a moderate binding strength for the two ligands inside HSA (1.00 +/- 0.12 x 104 M-1). The quenching effect of FL(CFL) on the fluorescence intensity of W214 (the sole tryptophan in HSA) indicates that FL and CFL occupy Site I in the protein which is known to bind several hydrophobic drugs. By performing site-competitive experiments, the location of the ligands is determined to be similar to that of the anticoagulant drug warfarin. At higher ratios of [ligand]/[HSA], we observed an upward curvature in the Stern-Volmer plots which indicates that the ligands occupy more pockets in Site I, close to W214. Our results indicate that both ligands bind in HSA with a moderate strength that should not affect their release when used as fluorescent reporters. The chemical and physical identities of the two ligands are also preserved inside the HSA binding sites.

  14. Carborhodol: a new hybrid fluorophore obtained by combination of fluorescein and carbopyronine dye cores.

    PubMed

    Sednev, Maksim V; Wurm, Christian A; Belov, Vladimir N; Hell, Stefan W

    2013-04-17

    Asymmetric hybrid fluorophores are built from the structural elements of two (or even more) symmetric dyes and can develop valuable new features which their parents do not possess. A new hybrid carborhodol dye was obtained by the combination of fluorescein and carbopyronine fluorophores. The brightly fluorescent hybrid dye with a linker and reactive group was prepared in 12 steps with overall yield of 1.6%. In aqueous solutions, it has absorption and emission maxima at 586 and 613 nm, respectively. Antibodies labeled with a carborhodol dye possess broad absorption and emission bands so that the effective Stokes shift is increased (compared with small Stokes shifts of the parent dyes) and the fluorescence quantum yield of 39% at a degree of labeling of 5.2. Two samples of secondary antibodies labeled with carborhodol and the benchmark red-emitting rhodamine dye (KK114) were used in two-color imaging experiments with excitation at 514-532 (carborhodol dye) and 633-640 nm (KK114). When emitted light was detected above 650 nm, the novel carborhodol dye provided a lower crosstalk than spectrally similar emitters (e. g., Atto594; crosstalk 40-60% with KK114 under the same conditions). The optical resolution of ca. 80 nm was attained using the new dye in stimulated emission depleted (STED) microscopy. The relatively short fluorescence lifetime in conjugates with antibodies (? = 1.2-1.6 ns) suggests the possibility of dual FLIM with numerous dyes having ? values in the range of 3-5 ns. All of these features make the carborhodol fluorophore a valuable addition to the family of the red-emitting fluorescent dyes. PMID:23517127

  15. Anterior segment angiography of the normal canine eye: a comparison between indocyanine green and sodium fluorescein.

    PubMed

    Pirie, C G; Alario, A

    2014-03-01

    The objective of this study was to assess and compare indocyanine green (IG) and sodium fluorescein (SF) angiographic findings in the normal canine anterior segment using a digital single lens reflex (dSLR) camera adaptor. Images were obtained from 10 brown-eyed Beagles, free of ocular and systemic disease. All animals received butorphanol (0.2 mg/kg IM), maropitant citrate (1.0 mg/kg SC) and diphenhydramine (2.0 mg/kg SC) 20 min prior to propofol (4 mg/kg IV bolus, 0.2 mg/kg/min continuous rate infusion). Standard color imaging was performed prior to the administration of 0.25% IG (1 mg/kg IV). Imaging was performed using a full spectrum dSLR camera, dSLR camera adaptor, camera lens (Canon 60 mm f/2.8 Macro) and an accessory flash. Images were obtained at a rate of 1/s immediately following IG bolus for 30 s, then at 1, 2, 3, 4 and 5 min. Ten minutes later, 10% SF (20 mg/kg IV) was administered. Imaging was repeated using the same adaptor system and imaging sequence protocol. Arterial, capillary and venous phases were identified during anterior segment IG angiography (ASIGA) and their time sequences were recorded. ASIGA offered improved visualization of the iris vasculature in heavily pigmented eyes compared to anterior segment SF angiography (ASSFA), since visualization of the vascular pattern during ASSFA was not possible due to pigment masking. Leakage of SF was noted in a total of six eyes. The use of IG and SF was not associated with any observed adverse events. The adaptor described here provides a cost-effective alternative to existing imaging systems. PMID:24447609

  16. Treatment of port-wine stains: analysis

    SciTech Connect

    van Gemert, M.J.; Welch, A.J.

    1987-08-01

    Port-wine stains (PWS) are bluish red skin stains that are caused by enlarged, ectatic blood vessels in the dermis. Laser treatment of PWS is analyzed from computation of the spatial distribution of heat production by direct absorption of the laser light and subsequent heat conduction. The absorption and scattering caused by oxyhemoglobin, epidermis, and dermis as a function of wavelength are utilized in this analysis. Ideal treatment is defined as coagulating the ectatic blood vessels without irreversible damage to the epidermis and dermis. The analysis shows that a millisecond pulsed, yellow dye laser at 577 nm (one of the large absorption bands in blood) is the laser of choice to treat PWS, offering as close to the ''ideal treatment'' as possible. The blue-green argon laser, which is currently the most frequently used laser for this purpose, is strongly recommended with irradiation times in milliseconds. Other lasers that are in clinical use, such as the red ruby and near-infrared Nd-YAG lasers, can provide selective treatment only when the epidermis is cooled concurrently. The CO/sub 2/ laser, on the other hand, can coagulate the blood vessels only through heat conduction from the hot epidermis; hence, it has neither the treatment selectivity nor any other physical option to force this selectivity.

  17. Mechanism of catalase activity in aqueous solutions of dimanganese(III,IV) ethylenediamine-N,N prime -diacetate

    SciTech Connect

    Rush, J.D.; Maskos, Z. )

    1990-03-07

    Manganous ions, ligated by ethylenediamine-N,N{prime}-diacetate (edda = L) decompose hydrogen peroxide with a rate law {minus}d(H{sub 2}O{sub 2})/dt = k{sub 17}(Mn(edda))(H{sub 2}O{sub 2}) where k{sub 17} = 5.4 M{sup {minus}1} s{sup {minus}1} at pH 7. The reduction of peroxide to water is initiated by the reaction of Mn{sup II}L with a dinuclear Mn{sup III,IV}L{sub 2}. A subsequent fast reaction between the transient product of this reaction and hydrogen peroxide or tert-butyl hydroperoxide effectively oxidizes Mn(II) to Mn(IV) in a concerted step without formation of the hydroxyl radical. The green mixed-valence complex, which is probably a bis({mu}-oxo)-bridged structure, is stable in neutral aqueous solution and exhibits a 16-line ESR signal in frozen solution. The basis of catalase activity is the autocatalytic formation of this complex when hydrogen peroxide is reduced by manganese(II). The catalase cycle is independent of the formation of oxy radicals. Mononuclear Mn{sup III}edda and Mn{sup II}edda react with superoxide radicals, but the decomposition of peroxide is virtually independent of these reactions. In unbuffered solutions, with a moderate excess of hydrogen peroxide, an oscillation in the concentration of the dinuclear complex is detected. 28 refs., 10 figs., 5 tabs.

  18. Phorbol 12,13-diacetate-induced contraction of the canine basilar artery: role of protein kinase C.

    PubMed

    Sugawa, M; Koide, T; Naitoh, S; Takato, M; Matsui, T; Asano, T

    1991-01-01

    The pharmacological and biochemical mechanisms of contractile responses to the protein kinase C (PKC) activator phorbol-12,13-diacetate (PDA) were investigated in canine basilar arteries. In the normal medium, PDA elicited a strong, dose-related, and slow-developing sustained contraction. Among the constrictors examined, including serotonin, prostaglandin F2 alpha, and endothelin, only PDA yielded contractions in a Ca2(+)-free medium. In both media, the PDA-induced contractions were virtually inhibited by either staurosporine, H-7, or quinacrine, while neither neurotransmitter blockades nor R24571 (calmidazolium) exerted significant effects. In addition, it was shown that 8-bromocyclic GMP, but not 8-bromocyclic AMP, markedly curtailed the PDA-induced contractions. Biochemical analysis, furthermore, showed that PDA induced increased phosphorylations of 27- and 96-kDa and proteins other than the myosin light chain (MLC) 20-kDa protein. Thus, the present results open up a novel mechanism of sustained cerebral artery contractions, where PKC activation rather than Ca2+/calmodulin/MLC system plays a key role that is regulated both by phospholipase A2 and by cyclic GMP. PMID:1845765

  19. Effect of alumina on triethylene glycol diacetate-2-propenoic acid butyl ester composite polymer electrolytes for flexible lithium ion batteries

    NASA Astrophysics Data System (ADS)

    Wang, Qiujun; Song, Wei-Li; Fan, Li-Zhen; Shi, Qiao

    2015-04-01

    Triethylene glycol diacetate-2-propenoic acid butyl ester (TEGDA-BA) based composite polymer electrolytes (CPE) are fabricated by incorporating alumina (Al2O3) nanoparticles (average particle size 10-20 nm) as inorganic filler via in situ polymerization. Effects of Al2O3 concentration on ionic conductivities, Li+ transfer numbers and charge/discharge properties are studied in details. Due to the uniformly dispersed Al2O3 nanoparticles, significant improvements in the mechanical flexibility and bendability are presented in the resulting polymer electrolytes. The CPE with 5 wt% Al2O3 nanoparticles exhibits the highest ionic conductivity up to 6.02 × 10-3 S cm-1 at 25 °C and the highest Li+ transference number (0.675), coupled with the most stable electrochemical window (>4.5 V vs. Li/Li+). With the presence of Al2O3, the growth of interface resistance is retarded, which increases the interface stability. The Li|CPE|Li4Ti5O12 and Li|CPE|LiFePO4 cells demonstrate remarkably stable charge/discharge performance and excellent capacity retention during cycling test. The results suggest that the CPE holds great application potential in flexible lithium ion batteries.

  20. Automated single-slide staining device. [in clinical bacteriology

    NASA Technical Reports Server (NTRS)

    Wilkins, J. R.; Mills, S. M.

    1975-01-01

    An automatic single-slide Gram staining device is described. A timer-actuated solenoid controls the dispensing of gentian violet, Gram iodine solution, decolorizer, and 1% aqueous safranin in proper sequence and for the time required for optimum staining. The amount of stain or reagent delivered is controlled by means of stopcocks below each solenoid. Used stains and reagents can be flushed automatically or manually. Smears Gram stained automatically are equal in quality to those prepared manually. The time to complete one Gram cycle is 4.80 min.

  1. Centrifuge-operated specimen staining method and apparatus

    NASA Technical Reports Server (NTRS)

    Clarke, Mark S. F. (Inventor); Feeback, Daniel L. (Inventor)

    1999-01-01

    A method of staining preselected, mounted specimens of either biological or nonbiological material enclosed within a staining chamber where the liquid staining reagents are applied and removed from the staining chamber using hypergravity as the propelling force. In the preferred embodiment, a spacecraft-operated centrifuge and method of diagnosing biological specimens while in orbit, characterized by hermetically sealing a shell assembly. The assembly contains slide stain apparatus with computer control therefor, the operative effect of which is to overcome microgravity, for example on board an International Space Station.

  2. Staining Protocols for Human Pancreatic Islets

    PubMed Central

    Campbell-Thompson, Martha L.; Heiple, Tiffany; Montgomery, Emily; Zhang, Li; Schneider, Lynda

    2012-01-01

    Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and beta mass ranges from 500 to 1500 mg 1-3. With this known heterogeneity, a standard processing and staining procedure was developed so that pancreatic regions were clearly defined and islets characterized using rigorous histopathology and immunolocalization examinations. Standardized procedures for processing human pancreas recovered from organ donors are described in part 1 of this series. The pancreas is processed into 3 main regions (head, body, tail) followed by transverse sections. Transverse sections from the pancreas head are further divided, as indicated based on size, and numbered alphabetically to denote subsections. This standardization allows for a complete cross sectional analysis of the head region including the uncinate region which contains islets composed primarily of pancreatic polypeptide cells to the tail region. The current report comprises part 2 of this series and describes the procedures used for serial sectioning and histopathological characterization of the pancreatic paraffin sections with an emphasis on islet endocrine cells, replication, and T-cell infiltrates. Pathology of pancreatic sections is intended to characterize both exocrine, ductular, and endocrine components. The exocrine compartment is evaluated for the presence of pancreatitis (active or chronic), atrophy, fibrosis, and fat, as well as the duct system, particularly in relationship to the presence of pancreatic intraductal neoplasia4. Islets are evaluated for morphology, size, and density, endocrine cells, inflammation, fibrosis, amyloid, and the presence of replicating or apoptotic cells using H&E and IHC stains. The final component described in part 2 is the provision of the stained slides as digitized whole slide images. The digitized slides are organized by case and pancreas region in an online pathology database creating a virtual biobank. Access to this online collection is currently provided to over 200 clinicians and scientists involved in type 1 diabetes research. The online database provides a means for rapid and complete data sharing and for investigators to select blocks for paraffin or frozen serial sections. PMID:22665223

  3. Hirschsprung's disease diagnosis: Comparison of immunohistochemical, hematoxilin and eosin staining

    PubMed Central

    Memarzadeh, Mehrdad; Talebi, Ardeshir; Edalaty, Masod; Hosseinpour, Mehrdad; Vahidi, Nasrin

    2009-01-01

    Background: The diagnosis of Hirschsprung's disease (HD) is based on the absence of ganglion cells. In hemotoxilin and eosin (H and E) as well as acetylcholine esterase staining there are limitations in the diagnosis of immature ganglion cells in neonates. Methods: In this prospective study, 54 biopsies taken from suspected HD patients (five mucosal specimens and 49 full thickness specimens) were studied. In the laboratory, after preparing sections of paraffin embedded tissues, H and E staining slides were compared with immunohistochemical (IHC) staining including: S100, NSE, CD117, CD56, Cathepsin D, Vimentin, BCL2, GFAP, Synaptophysin and chromogranin. Results: The study revealed 30 negative (absence of ganglion cells) cases (55.5%), 17 positive cases (31.04%) and seven suspected cases (12.9%) of ganglion cells on the H and E staining. On IHC staining with CD56 and Cathepsin D, all of the 17 positive cases detected through H and E, were confirmed for having ganglion cells and out of 30 cases reported negative on H and E staining, 28(93.3%) were reported negative and two (6.7%) positive by IHC staining. Of the seven suspected cases H and E staining), IHC staining detectedganglion cells only in five slides; two remained negative. Conclusions: IHC staining using CD56 and Cathepsin D improved the accuracy of diagnosis in HD when used in addition to H and E staining technique, especially for negative or suspicious slides. PMID:20671847

  4. Factors relating to dental stain formation in the rat.

    PubMed

    McDonald, J L; Schemehorn, B R; Stookey, G K

    1985-05-01

    A series of studies was conducted to investigate the use of the rat as an in vivo model for studies of dental stain and to identify dietary factors which influence stain formation in this model. It was determined that appreciable amounts of stain formed on the molar teeth of rats provided a synthetic diet containing lactalbumin, and the amount of stain increased throughout a four-week test period. Stain formation was also observed when rats were provided their diet by gastric intubation. Topical applications of chlorhexidine generally resulted in an increase in stain formation, as did the presence of tea in the drinking water. These studies support the use of the rat for investigations of dental stain. PMID:3858301

  5. Ultrafast tissue staining with chemical tags

    PubMed Central

    Kohl, Johannes; Ng, Julian; Cachero, Sebastian; Ciabatti, Ernesto; Dolan, Michael-John; Sutcliffe, Ben; Tozer, Adam; Ruehle, Sabine; Krueger, Daniel; Frechter, Shahar; Branco, Tiago; Tripodi, Marco; Jefferis, Gregory S. X. E.

    2014-01-01

    Genetically encoded fluorescent proteins and immunostaining are widely used to detect cellular and subcellular structures in fixed biological samples. However, for thick or whole-mount tissue, each approach suffers from limitations, including limited spectral flexibility and lower signal or slow speed, poor penetration, and high background labeling, respectively. We have overcome these limitations by using transgenically expressed chemical tags for rapid, even, high-signal and low-background labeling of thick biological tissues. We first construct a platform of widely applicable transgenic Drosophila reporter lines, demonstrating that chemical labeling can accelerate staining of whole-mount fly brains by a factor of 100. Using viral vectors to deliver chemical tags into the mouse brain, we then demonstrate that this labeling strategy works well in mice. Thus this tag-based approach drastically improves the speed and specificity of labeling genetically marked cells in intact and/or thick biological samples. PMID:25157152

  6. Fundus fluorescein angiographic findings in patients who underwent ventricular assist device implantation.

    PubMed

    Ozturk, Taylan; Nalcaci, Serhad; Ozturk, Pelin; Engin, Cagatay; Yagdi, Tahir; Akkin, Cezmi; Ozbaran, Mustafa

    2013-09-01

    Disruption of microcirculation in various tissues as a result of deformed blood rheology due to ventricular assist device (VAD) implantation causes novel arteriovenous malformations. Capillary disturbances and related vascular leakage in the retina and choroidea may also be seen in patients supported by VADs. We aimed to evaluate retinal vasculature deteriorations after VAD implantation. The charts of 17 patients who underwent VAD implantation surgery for the treatment of end-stage heart failure were retrospectively reviewed. Eight cases (47.1%) underwent pulsatile pump implantation (Berlin Heart EXCOR, Berlin Heart Mediprodukt GmbH, Berlin, Germany); however, nine cases (52.9%) had continuous-flow pump using centrifugal design (HeartWare, HeartWare Inc., Miramar, FL, USA). Study participants were selected among the patients who had survived with a VAD for at least 6 months, and results of detailed ophthalmologic examinations including optic coherence tomography (OCT) and fundus fluorescein angiography (FA) were documented. All of the 17 patients were male, with a mean age of 48.5 ± 14.8 years (15-67 years). Detailed ophthalmologic examinations including the evaluation of retinal vascular deteriorations via FA were performed at a mean of 11.8 ± 3.7 months of follow-up (6-18 months). Mean best-corrected visual acuity and intraocular pressure were found as logMAR 0.02 ± 0.08 and 14.6 ± 1.9 mm Hg, respectively in the study population. Dilated fundoscopy revealed severe focal arteriolar narrowing in two patients (11.8%), and arteriovenous crossing changes in four patients (23.5%); however, no pathological alteration was present in macular OCT scans. In patients with continuous-flow blood pumps, mean arm-retina circulation time (ARCT) and arteriovenous transit time (AVTT) were found to be 16.8 ± 3.0 and 12.4 ± 6.2 s, respectively; whereas those with pulsatile-flow blood pumps were found to be 17.4 ± 3.6 and 14.0 ± 2.1 s in patients (P=0.526 and P=0.356, respectively). FA also revealed a tendency for increased frequency of dye leakage from the optic disc in our study population. Except for remarkable delays in both ARCT and AVTT as well as a tendency for increased frequency of dye leakage from the optic disc, ophthalmologic evaluations revealed no other significant pathology or vascular deterioration in the retina that could be attributed to artificial heart systems. PMID:23826834

  7. Multi-class stain separation using independent component analysis

    NASA Astrophysics Data System (ADS)

    Trahearn, Nicholas; Snead, David; Cree, Ian; Rajpoot, Nasir

    2015-03-01

    Stain separation is the process whereby a full colour histology section image is transformed into a series of single channel images, each corresponding to a given stain's expression. Many algorithms in the field of digital pathology are concerned with the expression of a single stain, thus stain separation is a key preprocessing step in these situations. We present a new versatile method of stain separation. The method uses Independent Component Analysis (ICA) to determine a set of statistically independent vectors, corresponding to the individual stain expressions. In comparison to other popular approaches, such as PCA and NNMF, we found that ICA gives a superior projection of the data with respect to each stain. In addition, we introduce a correction step to improve the initial results provided by the ICA coefficients. Many existing approaches only consider separation of two stains, with primary emphasis on Haematoxylin and Eosin. We show that our method is capable of making a good separation when there are more than two stains present. We also demonstrate our method's ability to achieve good separation on a variety of different stain types.

  8. Clinical and anatomical approach using Sihler's staining technique (whole mount nerve stain)

    PubMed Central

    Won, Sung-Yoon; Kim, Da-Hye; Yang, Hun-Mu; Park, Jong-Tae; Kwak, Hyun-Ho; Hu, Kyung-Seok

    2011-01-01

    Sihler's staining allows visualization of the nerve distribution within soft tissues without extensive dissection and does not require slide preparation, unlike traditional approaches. This technique can be applied to the mucosa, muscle, and organs that contain myelinated nerve fibers. In particular, Sihler's technique may be considered the best tool for observing nerve distribution within skeletal muscles. The intramuscular distribution pattern of nerves is difficult to observe through manual manipulation due to the gradual tapering of nerves toward the terminal end of muscles, so it should be accompanied by histological studies to establish the finer branches therein. This method provides useful information not only for anatomists but also for physiologists and clinicians. Advanced knowledge of the nerve distribution patterns will be useful for developing guidelines for clinicians who perform operations such as muscle resection, tendon transplantation, and botulinum toxin injection. Furthermore, it is a useful technique to develop neurosurgical techniques and perform electrophysiological experiments. In this review, Sihler's staining technique is described in detail, covering its history, staining protocol, advantages, disadvantages, and possible applications. The application of this technique for determining the arterial distribution pattern is also described additionally in this study. PMID:21519543

  9. Visible luminescence from silicon wafers subjected to stain etches

    NASA Technical Reports Server (NTRS)

    Fathauer, R. W.; George, T.; Ksendzov, A.; Vasquez, R. P.

    1992-01-01

    Etching of Si in a variety of solutions is known to cause staining. These stain layers consist of porous material similar to that produced by anodic etching of Si in HF solutions. In this work, photoluminescence peaked in the red from stain-etched Si wafers of different dopant types, concentrations, and orientations produced in solutions of HF:HNO3:H2O was observed. Luminescence is also observed in stain films produced in solutions of NaNO2 in HF, but not in stain films produced in solutions of CrO3 in HF. The luminescence spectra are similar to those reported recently for porous Si films produced by anodic etching in HF solutions. However, stain films are much easier to produce, requiring no special equipment.

  10. Efficient staining of actinomycetoma and eumycetoma grains using henna extract.

    PubMed

    Batran, S E

    2015-12-01

    The use of natural, nontoxic, convenient and eco-friendly dyes for histopathological diagnosis avoids some of the synthetic dyes' hazards. I used an aqueous extract of henna at a concentration of 20 g/ml and acidified with acetic acid to stain mycetoma grains. Henna stained mycetoma grains orange-red to brown. The engulfed mycetoma grains within inflammatory cells stained well with henna extract compared to hematoxylin and eosin (H & E) and hexamine silver. PMID:26052737

  11. A Chemical Stain for Identifying Arsenic-Treated Wood

    E-print Network

    Florida, University of

    ) and Modified Stannous Chloride Stain (bottom) on New Wood at Reaction Time 5 Hours Figure II.4 Stannous Chloride Stain Progression of Color Change for New 4.0 kg/m3 (top) and 40 kg/m3 (bottom) CCA-Treated Wood.10 Graph of Ash pH vs. Average Percent CCA-Treated Wood Figure II.11 Stannous Chloride Stain Arsenic

  12. Electrostatic control of the coffee stain effect

    NASA Astrophysics Data System (ADS)

    Wray, Alex; Papageorgiou, Demetrios; Sefiane, Khellil; Matar, Omar

    2013-11-01

    The ``coffee stain effect,'' as first explained by Deegan et al. 1997, has received a great deal of attention amongst modellers and experimentalists in recent years, perhaps due in part to its obvious casual familiarity. However, it maintains interest because of its intriguing reliance on an interplay of a trio of effects: contact line pinning, inhomogeneous mass flux, and resulting capillarity-driven flow. What is more, the effect, and especially its suppression or reversal, find applications in fields as diverse as sample recovery, mass spectroscopy and the printing of Organic LEDs. We examine the motion a nanoparticle-laden droplet deposited on a precursor film, incorporating the effects of capillarity, concentration-dependent rheology, together with a heated substrate and resultant mass flux and Marangoni effects. We allow the substrate to act as an electrode and incorporate a second electrode above the droplet. The potential difference together with a disparity in electrical properties between the two regions results in electrical (Maxwell) stresses at the interface. We show via lubrication theory and via direct numerical simulations that the ring effect typically observed may be suppressed or augmented via appropriate use of electric fields. EPSRC DTG

  13. Quantitative chemical analysis of ocular melanosomes in stained and non-stained tissues.

    PubMed

    Biesemeier, Antje; Schraermeyer, Ulrich; Eibl, Oliver

    2011-07-01

    Energy-filtered Analytical Electron Microscopy (AEM) was used to image the ultrastructure and determine quantitatively the chemical composition of rat melanosomes of the choroid and the Retinal Pigment Epithelium (RPE). For the first time, the effect of staining in elemental analysis of melanosomes was investigated. Detection limits and accuracies of the applied methods were determined. Compared to previous work applying only quantitative Energy Dispersive X-ray microanalysis (EDX) in the TEM (Eibl, O., et al., 2006. Micron 37, 262), here we present a combined quantitative EDX and Electron Energy Loss Spectroscopy (EELS) analysis, including N. This yields the fraction of eumelanin and pheomelanin in melanosomes by the S/N mole fraction ratio. Melanosomes of the sepia ink sac, used as eumelanin standard, showed an S/N mole fraction ratio of <0.004. Thus, they consist primarily of eumelanin as reported by degradation analysis. In contrast, melanosomes of the rats contained mixed melanin with significant amounts of pheomelanin (S/N 0.02) in the RPE and the choroid. Consistent with the previous publication, it was shown that oxygen mole fractions are especially large in melanosomes (7-10 at.%) compared to other cell compartments, e.g. 2-4 at.% oxygen in the cytoplasm. In the melanosomes of non-stained tissue, the oxygen mole fraction clearly correlated with the Ca mole fraction. EDX spectra used for quantitative analysis had about 15,000 net counts under the oxygen peak, which is necessary to obtain (i) a small statistical error for oxygen and (ii) optimum minimum detectable mole fractions for S, Ca and transition metals. The precise determination of the oxygen mole fraction in melanosomes is important for understanding metabolism. Therefore, a detailed analysis was carried out on the possible errors affecting quantification. While O, S, and N mole fractions yielded similar results in stained and non-stained ocular melanosomes of rats, transition metals can only be determined reliably in non-stained tissues. High-precision EDX analysis of melanosomes yielded minimum detectable mole fractions of less than 0.04 at.% for Cu and Zn, these elements were present in melanosomes with mole fractions of about 0.3 at.% and 0.1at.%, respectively. Zn is of great importance for metabolism and for age related macular degeneration. Its mole fraction in melanosomes of rats is large enough to be detected and to be quantitatively analyzed by EDX spectroscopy. Ultrastructural information can now be correlated to the elemental composition. This is important to better understand the physical and chemical properties of melanosomal metabolism and turnover. PMID:21330141

  14. Functional expression of the 11 human Organic Anion Transporting Polypeptides in insect cells reveals that sodium fluorescein is a general OATP substrate.

    PubMed

    Patik, Izabel; Kovacsics, Daniella; Német, Orsolya; Gera, Melinda; Várady, György; Stieger, Bruno; Hagenbuch, Bruno; Szakács, Gergely; Özvegy-Laczka, Csilla

    2015-12-15

    Organic Anion Transporting Polypeptides (OATPs), encoded by genes of the Solute Carrier Organic Anion (SLCO) family, are transmembrane proteins involved in the uptake of various compounds of endogenous or exogenous origin. In addition to their physiological roles, OATPs influence the pharmacokinetics and drug-drug interactions of several clinically relevant compounds. To examine the function and molecular interactions of human OATPs, including several poorly characterized family members, we expressed all 11 human OATPs at high levels in the baculovirus-Sf9 cell system. We measured the temperature- and inhibitor-sensitive cellular accumulation of sodium fluorescein and fluorescein-methotrexate, two fluorescent substrates of the OATPs, OATP1B1 and 1B3. OATP1B1 and 1B3 were functional in Sf9 cells, showing rapid uptake (t1/2(fluorescein-methotrexate) 2.64 and 4.16min, and t1/2(fluorescein) 6.71 and 5.58min for OATP1B1 and 1B3, respectively) and high-affinity transport (Km(fluorescein-methotrexate) 0.23 and 0.53?M, and Km(fluorescein) 25.73 and 38.55?M for OATP1B1 and 1B3, respectively) of both substrates. We found that sodium fluorescein is a general substrate of all human OATPs: 1A2, 1B1, 1B3, 1C1, 2A1, 2B1, 3A1, 4A1, 4C1, 5A1 and 6A1, while fluorescein-methotrexate is only transported by 1B1, 1B3, 1A2 and 2B1. Acidic extracellular pH greatly facilitated fluorescein uptake by all OATPs, and new molecular interactions were detected (between OATP2B1 and Imatinib, OATP3A1, 5A1 and 6A1 and estradiol 17-?-d-glucuronide, and OATP1C1 and 4C1 and prostaglandin E2). These studies demonstrate, for the first time, that the insect cell system is suitable for the functional analysis of the entire human OATP family, and for drug-OATP interaction screening. PMID:26415544

  15. Fluorescein and radiolabeled Function-Spacer-Lipid constructs allow for simple in vitro and in vivo bioimaging of enveloped virions.

    PubMed

    Hadac, Elizabeth M; Federspiel, Mark J; Chernyy, Evgeny; Tuzikov, Alexander; Korchagina, Elena; Bovin, Nicolai V; Russell, Stephen; Henry, Stephen M

    2011-09-01

    Tools that can aid in vitro and in vivo imaging and also noninvasively determine half-life and biodistribution are required to advance clinical developments. A Function-Spacer-Lipid construct (FSL) incorporating fluorescein (FSL-FLRO4) was used to label vesicular stomatitis virus (VSV), measles virus MV-NIS (MV) and influenza virus (H1N1). The ability of FSL constructs to label these virions was established directly by FACScan of FSL-FLRO4 labeled VSV and MV, and indirectly following labeled H1N1 and MV binding to a cells. FSL-FLRO4 labeling of H1N1 was shown to maintain higher infectivity of the virus when compared with direct fluorescein virus labeling. A novel tyrosine (125)I radioiodinated FSL construct was synthesized (FSL-(125)I) from FSL-tyrosine. This was used to label VSV (VSV-FSL-(125)I), which was infused into the peritoneal cavity of laboratory mice. Bioscanning showed VSV-FSL-(125)I to localize in the liver, spleen and bloodstream in contrast to the free labels FSL-(125)I or (125)I, which localized predominantly in the liver and thyroid respectively. This is a proof-of-principle novel and rapid method for modifying virions and demonstrates the potential of FSL constructs to improve in vivo imaging of virions and noninvasively observe in vivo biodistribution. PMID:21703308

  16. Quenching of fluorescein-conjugated lipids by antibodies. Quantitative recognition and binding of lipid-bound haptens in biomembrane models, formation of two-dimensional protein domains and molecular dynamics simulations.

    PubMed Central

    Ahlers, M; Grainger, D W; Herron, J N; Lim, K; Ringsdorf, H; Salesse, C

    1992-01-01

    Three model biomembrane systems, monolayers, micelles, and vesicles, have been used to study the influence of chemical and physical variables of hapten presentation at membrane interfaces on antibody binding. Hapten recognition and binding were monitored for the anti-fluorescein monoclonal antibody 4-4-20 generated against the hapten, fluorescein, in these membrane models as a function of fluorescein-conjugated lipid architecture. Specific recognition and binding in this system are conveniently monitored by quenching of fluorescein emission upon penetration of fluorescein into the antibody's active site. Lipid structure was shown to play a large role in affecting antibody quenching. Interestingly, the observed degrees of quenching were nearly independent of the lipid membrane model studied, but directly correlated with the chemical structure of the lipids. In all cases, the antibody recognized and quenched most efficiently a lipid based on dioctadecylamine where fluorescein is attached to the headgroup via a long, flexible hydrophilic spacer. Dipalmitoyl phosphatidylethanolamine containing a fluorescein headgroup demonstrated only partial binding/quenching. Egg phosphatidylethanolamine with a fluorescein headgroup showed no susceptibility to antibody recognition, binding, or quenching. Formation of two-dimensional protein domains upon antibody binding to the fluorescein-lipids in monolayers is also presented. Chemical and physical requirements for these antibody-hapten complexes at membrane surfaces have been discussed in terms of molecular dynamics simulations based on recent crystallographic models for this antibody-hapten complex (Herron et al., 1989. Proteins Struct. Funct. Genet. 5:271-280). Images FIGURE 7 FIGURE 8 PMID:1420916

  17. Klutts 4/2004 Silver Staining of Protein Gels

    E-print Network

    Doering, Tamara

    Klutts 4/2004 1 Silver Staining of Protein Gels Overview: Silver staining is much more sensitive containing: 100 mg of silver nitrate (AgNO3) in 100 ml dH2O E. 100 ml of a solution containing: 3 g sodium

  18. OBSERVATION OF SALMONELLA TYPHIMURIUM FIMBRIAE BY NEGATIVE STAIN

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Washed and unwashed overnight cultures of Salmonella typhimurium were examined for the expression of fimbriae using negative stain. In the course of the evaluation, it was noted that the distribution of bacteria on formvar coated grids was dependent on the negative stain utilized for visualization....

  19. Unusual indelible enamel staining following fixed appliance treatment.

    PubMed

    Hodges, S J; Spencer, R J; Watkins, S J

    2000-12-01

    Two cases are described of indelible enamel staining following fixed appliance therapy. The acquired pigmentation occurred in patients with an identifiable enamel defect prior to treatment. The interaction of factors to cause the staining is discussed and it's prevention in future cases highlighted. Subsequent restoration of the affected teeth is shown. PMID:11099567

  20. 7 CFR 3201.87 - Wood and concrete stains.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...2013-01-01 2013-01-01 false Wood and concrete stains. 3201.87 Section 3201...Designated Items § 3201.87 Wood and concrete stains. (a) Definition. Products...designed to be applied as a finish for concrete and wood surfaces and that...

  1. 7 CFR 3201.87 - Wood and concrete stains.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...2014-01-01 2014-01-01 false Wood and concrete stains. 3201.87 Section 3201...Designated Items § 3201.87 Wood and concrete stains. (a) Definition. Products...designed to be applied as a finish for concrete and wood surfaces and that...

  2. 7 CFR 3201.87 - Wood and concrete stains.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 15 2013-01-01 2013-01-01 false Wood and concrete stains. 3201.87 Section 3201.87... Designated Items § 3201.87 Wood and concrete stains. (a) Definition. Products that are designed to be applied as a finish for concrete and wood surfaces and that contain dyes or pigments to change the...

  3. 7 CFR 3201.87 - Wood and concrete stains.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 15 2014-01-01 2014-01-01 false Wood and concrete stains. 3201.87 Section 3201.87... Designated Items § 3201.87 Wood and concrete stains. (a) Definition. Products that are designed to be applied as a finish for concrete and wood surfaces and that contain dyes or pigments to change the...

  4. In vivo photoacoustic imaging of model of port wine stains.

    PubMed

    Yuan, Kaihua; Yuan, Yi; Gu, Ying; Gao, Jianhua; Xing, Da

    2012-01-01

    Port wine stains are categorized as a benign capillary vascular malformation, which is hard to cure. In this paper, a photoacoustic microscopy system, which integrated a two-dimensional scanning galvanometer, an objective lens and a focused ultrasound transducer, was designed for noninvasive imaging of blood vessels of port wine stains model in vivo. Cock comb was chosen as the port wine stains model in the experiment. The blood vessels in x-y plane and x-z plane were imaged clearly. Experimental results demonstrate that photoacoustic microscopy can image the blood vessels of port wine stains model in vivo with high contrast and high resolution. It has the potential for clinical applications in detecting the blood vessels in port wine stains skin. PMID:22635179

  5. Novel Fluorometric Method for the Determination of Production Rate and Steady-State Concentration of Photochemically Generated Superoxide Radical in Seawater Using 3',6'-(Diphenylphosphinyl)fluorescein.

    PubMed

    Anifowose, Adebanjo Jacob; Takeda, Kazuhiko; Sakugawa, Hiroshi

    2015-12-15

    Superoxide radical (O2(•-)) is an important reactive oxygen species in seawater. Measurements of its production rates and steady-state concentrations generated by photochemical processes have been a Herculean task over the years. In this study, a probe - 3'6'-(diphenylphosphinyl)fluorescein (PF-1) - was used to trap photochemically generated O2(•-) in seawater, thereby yielding fluorescein. The fluorescein produced was measured by an isocratic fluorescence HPLC at excitation/emission wavelengths of 490/513 nm, respectively. The reaction rate constant of PF-1 with O2(•-) (kPF-1) was pH-dependent: (3.2-23.5) × 10(7) M(-1) s(-1) at pHTOT 7.65-8.50. By applying appropriate equations, both the production rate and the steady-state concentration of O2(•-) generated by photochemical reactions in the seawater were quantified. Under the optimized experimental conditions, fluorescein standards (3-50 nM) exhibited linearity in the seawater by HPLC. The photoformation of fluorescein, due to the reaction of PF-1 with the O2(•-) photochemically produced in the seawater, was linear within the 20 min irradiation. The detection limit of the fluorescein photoformation rate was 0.03 pM s(-1), defined as 3? of the lowest standard fluorescein concentration per 20 min irradiation. Using this value, the yield of fluorescein, and the fraction of O2(•-) that reacted with PF-1 in the seawater, the detection limit of the O2(•-) photoformation rate was 1.78 pM s(-1). Superoxide measurements using the proposed method were relatively unaffected by the potential interfering species in seawater. Application of the proposed method to ten (10) seawater samples from the Seto Inland Sea, Japan, resulted in measured O2(•-) photoformation rates of 3.1-8.5 nM s(-1), with steady-state concentrations ranging (0.06-0.3) × 10(-10) M. The method is simple, requires no technical sample preparation, and can be used to analyze a large number of samples. PMID:26569557

  6. Potential lanthanide ion selective reagents. 3. Metal complex formation with 1,7-diaza-4,10-13-trioxacyclopentadecane-N,N'-diacetic acid

    SciTech Connect

    Chang, C.A.; Ochaya, V.O.

    1986-01-29

    Stability constants for the ligand 1,7-diaza-4,10,13-trioxacyclopentadecane-N,N'-diacetic acid (dapda or K21DA) with the lanthanides and several other metal ions have been determined at 25 /sup 0/C in aqueous 0.1 M (CH/sub 3/)/sub 4/NCl medium by a potentiometric method. The results obtained are compared to those obtained for a similar ligand of large cavity size, 1,20-diaza-4,7,13,16-tetraoxacyclooctadecane-N,N'-diacetic acid (dacda or K22DA), which has been previously studied and reported. The stability of dapda is found to reach a peak at Eu(III) with the lanthanide series and is rationalized in terms of the matching of the ligand properties with metal ion characteristics. The transition-metal ions Ni(II), Cu(II), and Zn(II) all form stronger dapda (as compared to dacda) complexes due to a better match of the ligand cavity size and metal ion radius. 18 references, 3 figures, 1 table.

  7. Novel Process for Laser Stain Removal from Archaeological Oil Paintings

    NASA Astrophysics Data System (ADS)

    El-Nadi, Lotfia; El-Feky, Osama; Abdellatif, Galila; Darwish, Sawsan

    2013-03-01

    Some samples of oil paintings (5 × 5 cm) were prepared on wooden panel with four types of fungi commonly encountered on oil paintings were selected for this study. Each of the fungi is associated with different colored stains. Fungus Alternaria tenuis is associated by a dense black stain, Chetomium globosum by a brownish gray stain, Aspergillus flavus by a yellowish stain, and Fusaruim oxysporum by a pinkish stain. Fungi growing on oil paintings affect the surface characteristics by forming a variety of colored patches typically composed of many complex chemical substances that are produced during metabolic processes. These colored stains may be encrusted in spores, present in mycelium or secreted to a substance such as oil paintings surfaces. While the fungal stains can sometimes be extracted with appropriate solvents, there are some stains that resist solvent extraction entirely. Developing new solvent system that might attack the paint structure, and is time consuming and requires a great deal of trial and error. Mechanical stain removal is also problematic in that it often produces abrasion of the surface, markedly deteriorating the artwork, and is extra ordinarily fine and tedious. For these reasons, we decided to examine an alternative physical technique as a new approach to deal with stain removal. Since the stains are due to the existence of fungi, we thought it a good idea to remove them by singlet oxygen. We applied the photo dynamic process through which the fungi stains were covered with organic dye derivatives in solution under controlled illumination in the lab. The samples were then irradiated by low power Laser light from a He-Ne laser, the dye will be photodecomposed and produce singlet oxygen. We report in this work the results obtained as a function of: - The concentration and types of the organic dye in solution, - The presence of certain amounts of liquids added to the solution, - The scanning speed of the laser beam on the sample surface, - The irradiation time. For each case fresh samples were used and photographed before and after the treatment. The results obtained will be speculated and discussed. This procedure was applied to the cleaning of archaeological oil paintings for the first time to our knowledge. The method could well be considered as a new field of combined science and technology applied to laser stain removal and represents a significant addition to the techniques available to art conservation.

  8. Blood–brain barrier transport studies, aggregation, and molecular dynamics simulation of multiwalled carbon nanotube functionalized with fluorescein isothiocyanate

    PubMed Central

    Shityakov, Sergey; Salvador, Ellaine; Pastorin, Giorgia; Förster, Carola

    2015-01-01

    In this study, the ability of a multiwalled carbon nanotube functionalized with fluorescein isothiocyanate (MWCNT–FITC) was assessed as a prospective central nervous system-targeting drug delivery system to permeate the blood–brain barrier. The results indicated that the MWCNT–FITC conjugate is able to penetrate microvascular cerebral endothelial monolayers; its concentrations in the Transwell® system were fully equilibrated after 48 hours. Cell viability test, together with phase-contrast and fluorescence microscopies, did not detect any signs of MWCNT–FITC toxicity on the cerebral endothelial cells. These microscopic techniques also revealed presumably the intracellular localization of fluorescent MWCNT–FITCs apart from their massive nonfluorescent accumulation on the cellular surface due to nanotube lipophilic properties. In addition, the 1,000 ps molecular dynamics simulation in vacuo discovered the phenomenon of carbon nanotube aggregation driven by van der Waals forces via MWCNT–FITC rapid dissociation as an intermediate phase. PMID:25784800

  9. Fluorescein Isothiocyanate-Labeled Lectin Analysis of the Surface of the Nitrogen-Fixing Bacterium Azospirillum brasilense by Flow Cytometry

    PubMed Central

    Yagoda-Shagam, Janet; Barton, Larry L.; Reed, William P.; Chiovetti, Robert

    1988-01-01

    The cell surface of Azospirillum brasilense was probed by using fluorescein isothiocyanate (FITC)-labeled lectins, with binding determined by fluorescence-activated flow cytometry. Cells from nitrogen-fixing or ammonium-assimilating cultures reacted similarly to FITC-labeled lectins, with lectin binding in the following order: Griffonia simplicifolia II agglutinin > Griffonia simplicifolia I agglutinin > Triticum vulgaris agglutinin > Glycine max agglutinin > Canavalia ensiformis agglutinin > Limax flavus agglutinin > Lotus tetragonolobus agglutinin. The fluorescence intensity of cells labeled with FITC-labeled G. simplicifolia I, C. ensiformis, T. vulgaris, and G. max agglutinins was influenced by lectin concentration. Flow cytometry measurements of lectin binding to cells was consistent with measurements of agglutination resulting from lectin-cell interaction. Capsules surrounding nitrogen-fixing and ammonium-assimilating cells were readily demonstrated by light and transmission electron microscopies. Images PMID:16347693

  10. Western Blot of Stained Proteins from Dried Polyacrylamide Gels

    NASA Technical Reports Server (NTRS)

    Gruber, Claudia; Stan-Lotter, Helga

    1996-01-01

    Western blotting of proteins is customarily performed following their separation on polyacrylamide gels, either prior to staining (1) or, as recently reported, following staining (2). We describe here Western blotting with stained gels, which had been dried and some of which had been stored for years. This procedure permits immunological analysis of proteins, to which antisera may have become available only later, or where the application of newly developed sensitive detection methods is desired. Once rehydration of the gels is achieved, proteins can be-transferred to blotting membranes by any appropriate protocol. Proteins stained with Coomassie Blue have to be detected with a non-chromogenic method, such as the film-based enhanced chemiluminescence (ECL)2) procedure (3). Silver stained proteins, which transfer in the colorless form, may be visualized by any detection method, although, because of the usually very low amounts of proteins, detection by ECL is preferable. Blotting of stained proteins from rehydrated gels is as rapid and as quantitative as from freshly prepared gels, in contrast to blotting from wet stained gels, which requires extensive washing and results in low transfer efficiency (2). Together with a photographic record of the gel pattern, unambiguous identification of immunoreactive proteins from complex mixtures is possible. Some further applications of this work are discussed.

  11. Microscopic analysis of MTT stained boar sperm cells

    PubMed Central

    van den Berg, B.M.

    2015-01-01

    The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few min in a diffuse staining of the midpiece of the sperm cells. Upon further incubation the staining of the midpiece became more intense, and gradually the formation of packed formazan granules became more visible. At the same time, a small formazan stained granule appeared medially on the sperm head, which increased in size during further incubation. After incubation for about 1 h the midpiece granules were intensely stained and more clearly distinct as granules, while aggregation of sperm cells occurred. Around 90% of the sperm cells showed these staining events. At the end of the staining the formazan granules have disappeared from both the sperm cells and medium, whereas formazan crystals appeared as thin crystal threads, that became heavily aggregated in the incubation medium. It was concluded that formazan is taken up by lipid droplets in the cytoplasm. Further, the use of the MTT assay to test for sperm viability should be regarded as a qualitative assay, whereas its practical use at artificial insemination (AI) Stations is limited. PMID:26623368

  12. Silver and Cyanine Staining of Oligonucleotides in Polyacrylamide Gel

    PubMed Central

    Li, Wei

    2015-01-01

    To explore why some oligonucleotides in denaturing polyacrylamide gel could not be silver-stained, 134 different oligonucleotides were analyzed using denaturing polyacrylamide gel electrophoresis stained with silver and asymmetric cyanine. As a result, we found that the sensitivity of oligos (dA), (dC), (dG) and (dT) to silver staining could be ranged as (dA) > (dG) > (dC) > (dT) from high to low. It was unexpected that oligo (dT) was hard to be silver-stained. Moreover, the silver staining of an oligonucleotide containing base T could be partially or completely inhibited by base T. The inhibition of silver staining by base T was a competitive inhibition which could be affected by the amounts of the argyrophil nucleobase and base T, the cis-distance between the argyrophil nucleobase and base T, and the gel concentration. The changes of the intensity of an oligonucleotide band caused by the changes of DNA base composition were diverse and interesting. The intensity of some oligonucleotide bands would significantly change when the changes of DNA base composition accumulated to a certain extent (usually ? 4 nt). The sensitivity of cyanine staining of ? 11-nt long oligonucleotides could be enhanced about 250-fold by fixing the gels with methanol fixing solution. PMID:26650843

  13. Evaluation of Common Angling-Induced Sources of Epithelial Damage for Popular Freshwater Sport Fish using Fluorescein

    SciTech Connect

    Colotelo, Alison HA; Cooke, Steven J.

    2011-05-01

    Angling is a popular recreational activity across the globe and a large proportion of fish captured by anglers are released due to voluntary or mandatory catch-and-release practices. The handling associated with hook removal and return of the fish to their environment can cause physical damage to the epidermal layer of the fish which may affect the condition and survival of released fish. This study investigated possible sources of epithelial damage associated with several different handling methods (i.e. landing net types, interactions with different boat floor surfaces, tournament procedures) commonly used in recreational angling for two popular freshwater sport fish species, largemouth bass (Micropterus salmoides) and northern pike (Esox lucius). Epithelial damage was examined using fluorescein, a non-toxic dye, which has been shown to detect latent epithelial damage. Northern pike exhibited extensive epithelial damage after exposure to several of the induced treatments (i.e., interaction with a carpeted surface, knotted nylon net, and line rolling) but relatively little epithelial damage when exposed to others (i.e., knotless rubber nets, smooth boat surfaces, or lip gripping devices). Largemouth bass did not show significant epithelial damage for any of the treatments, with the exception of fish caught in a semi-professional live release tournament. The detection of latent injuries using fluorescein can be an important management tool as it provides visual examples of potential damage that can be caused by different handling methods. Such visualizations can be used to encourage fish friendly angler behaviour and enhance the survival and welfare of released fish. It can also be used to test new products that are intended to or claim to reduce injury to fish that are to be released. Future research should evaluate the relationship between different levels of epithelial damage and mortality across a range of environmental conditions.

  14. 18. INTERIOR DETAIL VIEW OF STAINED GLASS WINDOW LOCATED AT ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    18. INTERIOR DETAIL VIEW OF STAINED GLASS WINDOW LOCATED AT SOUTH SIDE OF ALTAR, NOTE INSCRIPTION DEDICATED IN THE MEMORY OF FATHER DAMIEN - St. Francis Catholic Church, Moloka'i Island, Kalaupapa, Kalawao County, HI

  15. Steinway piano and stained glass clerestory window in lounge area, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Steinway piano and stained glass clerestory window in lounge area, upper deck. Hot water radiators can be seen at base of wall. These run throughout the houseboat. - Houseboat LA DUCHESSE, The Antique Boat Museum, Clayton, Jefferson County, NY

  16. Intravascular staining for discrimination of vascular and tissue leukocytes.

    PubMed

    Anderson, Kristin G; Mayer-Barber, Katrin; Sung, Heungsup; Beura, Lalit; James, Britnie R; Taylor, Justin J; Qunaj, Lindor; Griffith, Thomas S; Vezys, Vaiva; Barber, Daniel L; Masopust, David

    2014-01-01

    Characterization of the cellular participants in tissue immune responses is crucial to understanding infection, cancer, autoimmunity, allergy, graft rejection and other immunological processes. Previous reports indicate that leukocytes in lung vasculature fail to be completely removed by perfusion. Several studies suggest that intravascular staining may discriminate between tissue-localized and blood-borne cells in the mouse lung. Here we outline a protocol for the validation and use of intravascular staining to define innate and adaptive immune cells in mice. We demonstrate application of this protocol to leukocyte analyses in many tissues and we describe its use in the contexts of lymphocytic choriomeningitis virus and Mycobacterium tuberculosis infections or solid tumors. Intravascular staining and organ isolation usually takes 5-30 min per mouse, with additional time required for any subsequent leukocyte isolation, staining and analysis. In summary, this simple protocol should help enable interpretable analyses of tissue immune responses. PMID:24385150

  17. Mallory's Tetrachrome Staining Of Mouse Sections Lise Zakin 2008

    E-print Network

    De Robertis, Eddy M.

    a vacuum oven at 75°C to prevent paraffin from solidifying -Embed embryos in fresh paraffin -Allow is recommended to do a test run to adjust staining times in Groat solution, Acid Fuschine and Aniline solution

  18. Use of carboxyfluorescein diacetate succinimidyl ester (CFSE) dye and fluorescent imaging as an in situ method to visualize lymphoid tissues in egg-layer chickens challenged with Salmonella enterica serovar Enteritidis (SE)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carboxyfluorescein diacetate succinimidyl ester (CFSE) vital dye has been used in leukocyte studies involving mice, rats, sheep, heifers, nonhuman primates, teleost fish and avian embryos. Mice and sheep appear to be the only animals that have received intravenous (IV) CFSE administration, and the ...

  19. Immunohistochemical CD3 staining detects additional patients with celiac disease

    PubMed Central

    Mubarak, Amani; Wolters, Victorien M; Houwen, Roderick HJ; ten Kate, Fiebo JW

    2015-01-01

    AIM: To investigate whether performing immunohistochemical CD3 staining, in order to improve the detection of intra-epithelial lymphocytosis, has an additional value in the histological diagnosis of celiac disease. METHODS: Biopsies obtained from 159 children were stained by hematoxylin and eosin (HE) and evaluated using the Marsh classification. CD3 staining was subsequently evaluated separately and independently. RESULTS: Differences in evaluation between the routine HE sections and CD3 staining were present in 20 (12.6%) cases. In 10 (6.3%) patients the diagnosis of celiac disease (Marsh II and III) changed on examination of CD3 staining: in 9 cases, celiac disease had initially been missed on the HE sections, while 1 patient had been over-diagnosed on the routine sections. In all patients, the final diagnosis based on CD3 staining, was concordant with serological results, which was not found previously. In the other 10 (12.3%) patients, the detection of sole intra-epithelial lymphocytosis (Marsh?I) improved. Nine patients were found to have Marsh?I?on CD3 sections, which had been missed on routine sections. Interestingly, the only patient with negative serology had Giardiasis. Finally, in 1 patient with negative serology, in whom Marsh?I?was suspected on HE sections, this diagnosis was withdrawn after evaluation of the CD3 sections. CONCLUSION: Staining for CD3 has an additional value in the histological detection of celiac disease lesions, and CD3 staining should be performed when there is a discrepancy between serology and the diagnosis made on HE sections. PMID:26140002

  20. Black Stain and Dental Caries: A Review of the Literature

    PubMed Central

    ?y?a, Tomasz; Kawala, Beata; Antoszewska-Smith, Joanna; Kawala, Maciej

    2015-01-01

    Black stain is characterized as a dark line or an incomplete coalescence of dark dots localized on the cervical third of the tooth. Over the last century, the etiology of black stain has been the subject of much debate. Most of the studies concerning this issue were conducted in pediatric population. According to the reviewed articles published between 2001 and 2014, the prevalence of black stain varies from 2.4% to 18% with equal sex distribution. The majority of the authors confirm the correlation between the presence of black stain and lower caries experience. The microflora of this deposit is dominated by Actinomyces spp. and has lower cariogenic potential than nondiscolored dental plaque. Iron/copper and sulfur complexes are thought to be responsible for the dark color. In patients with black stain saliva has higher calcium concentrations and higher buffering capacity. Factors such as dietary habits, socioeconomic status, and iron supplementation may be contributing to the formation of black stain. PMID:25802850

  1. Protocol for Silver Stain, originally from bioprotocol Online modified by JSW 5/5/05 Introduction

    E-print Network

    Mecham, Robert

    Protocol for Silver Stain, originally from bioprotocol Online modified by JSW 5/5/05 Introduction: Silver Staining should be 100-1000 times more sensitive than traditional Coomassie staining. This can. If glass staining jar was previously used for silver staining rinse several times with QH2O. 2. Prepare Fix

  2. Fluorescent multiple staining and CASA system to assess boar sperm viability and membranes integrity in short and long-term extenders

    PubMed Central

    Lange-Consiglio, A.; Meucci, A.; Cremonesi, F.

    2013-01-01

    The aim of this study was to assess the effect on boar spermatozoa quality of in vitro storage in short and long-term extenders by fluorescent multiple staining (FMS) and computer assisted semen analyzer (CASA). Fresh ejaculates from three healthy, sexually mature boars were diluted with equal volumes of six short-term or three long-term commercial extenders and stored at 19°C for 6 days (short-term) or 12 days (long-term). The integrity of spermatozoa membranes was analyzed by FMS using propidium iodide, 5,5’,6,6’-tetrachloro-1,1’,3,3’ tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) and fluorescein isothiocyanate-conjugated peanut agglutinin (PNA). The results obtained from this staining were compared with spermatozoa motility assessed by CASA. Our study showed that the number of viable spermatozoa with non-reacted acrosomes and intact mitochondria was positively correlated with the rate of motile spermatozoa (r2>0.9) irrespective of the extender used. In all extenders the number of motile spermatozoa significantly decreased as preservation period increased (P<0.05). FMS test is a potent indicator of sperm motility because it analyses mitochondrial integrity independently from observable alterations in motility. The best performing extenders were BTS for short-term storage and TRI-x-Cell for long-term storage.

  3. Substituent effect study on experimental ¹³C NMR chemical shifts of (3-(substituted phenyl)-cis-4,5-dihydroisoxazole-4,5-diyl)bis(methylene)diacetate derivatives.

    PubMed

    Kara, Yesim S

    2015-12-01

    Eleven novel (3-(substituted phenyl)-cis-4,5-dihydroisoxazole-4,5-diyl)bis(methylene) diacetate derivatives were synthesized in the present study. These dihydroisoxazole derivatives were characterized by IR, (1)H NMR, (13)C NMR and elemental analyses. Their (13)C NMR spectra were measured in Deuterochloroform (CDCl3). The correlation analysis for the substituent-induced chemical shift (SCS) with Hammett substituent constant (?), inductive substituent constant (?I), different of resonance substituent constants (?R, ?R(o)) and Swain-Lupton substituent parameters (F, R) were performed using SSP (single substituent parameter), and DSP (dual substituent parameter) methods, as well as single and multiple regression analysis. From the result of regression analysis, the effect of substituent on the (13)C NMR chemical shifts was explained. PMID:26172459

  4. Detection of Acid Fast Bacilli in Saliva using Papanicolaou Stain Induced Fluorescence Method Versus Fluorochrome Staining: An Evaluative Study

    PubMed Central

    (Munot), Priya P Lunawat; Mhapuskar, Amit A; Ganvir, S M; Hazarey, Vinay K; Mhapuskar, Madhavi A; Kulkarni, Dinraj

    2015-01-01

    Background: Fifty years after effective chemotherapy, tuberculosis (TB) still remains leading infectious cause of adult mortality. The aim of present study was to evaluate diagnostic utility of papanicolaou (Pap) stain induced fluorescence microscopic examination of salivary smears in the diagnosis of pulmonary TB. Materials and Methods: Cross-sectional study of 100 individuals clinically suspected of suffering from active pulmonary TB. Control group – 50 individuals are suffering from any pulmonary disease other than TB such as pneumonia or bronchiogenic carcinoma. Fluorescence microscopic examination of two salivary smears stained by Pap stain and auramine-rhodamine (A-R) stain respectively for each patient. Ziehl–Neelsen stained sputum smear examined under the light microscope for each patient. Culture was done in all the patients for microbiological confirmation. McNemar's Chi-square analysis, Kappa test, and Z-test. Results: The sensitivities of the three staining methods using culture as a reference method were 93.02%, 88.37% and 87.20% for Pap, A-R and Ziehl–Neelson respectively. Conclusion: Pap-induced fluorescence of salivary smears is a safe, reliable and rapid method, which can prove as a valuable diagnostic tool for diagnosis of TB. PMID:26229384

  5. Reliability of a rapid hematology stain for sputum cytology*

    PubMed Central

    Gonçalves, Jéssica; Pizzichini, Emilio; Pizzichini, Marcia Margaret Menezes; Steidle, Leila John Marques; Rocha, Cristiane Cinara; Ferreira, Samira Cardoso; Zimmermann, Célia Tânia

    2014-01-01

    Objective: To determine the reliability of a rapid hematology stain for the cytological analysis of induced sputum samples. Methods: This was a cross-sectional study comparing the standard technique (May-Grünwald-Giemsa stain) with a rapid hematology stain (Diff-Quik). Of the 50 subjects included in the study, 21 had asthma, 19 had COPD, and 10 were healthy (controls). From the induced sputum samples collected, we prepared four slides: two were stained with May-Grünwald-Giemsa, and two were stained with Diff-Quik. The slides were read independently by two trained researchers blinded to the identification of the slides. The reliability for cell counting using the two techniques was evaluated by determining the intraclass correlation coefficients (ICCs) for intraobserver and interobserver agreement. Agreement in the identification of neutrophilic and eosinophilic sputum between the observers and between the stains was evaluated with kappa statistics. Results: In our comparison of the two staining techniques, the ICCs indicated almost perfect interobserver agreement for neutrophil, eosinophil, and macrophage counts (ICC: 0.98-1.00), as well as substantial agreement for lymphocyte counts (ICC: 0.76-0.83). Intraobserver agreement was almost perfect for neutrophil, eosinophil, and macrophage counts (ICC: 0.96-0.99), whereas it was moderate to substantial for lymphocyte counts (ICC = 0.65 and 0.75 for the two observers, respectively). Interobserver agreement for the identification of eosinophilic and neutrophilic sputum using the two techniques ranged from substantial to almost perfect (kappa range: 0.91-1.00). Conclusions: The use of Diff-Quik can be considered a reliable alternative for the processing of sputum samples. PMID:25029648

  6. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...false Painted, colored or stained glass windows for religious institutions...52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof,...

  7. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...false Painted, colored or stained glass windows for religious institutions...52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof,...

  8. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...false Painted, colored or stained glass windows for religious institutions...52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof,...

  9. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...false Painted, colored or stained glass windows for religious institutions...52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof,...

  10. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...false Painted, colored or stained glass windows for religious institutions...52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof,...

  11. Use of vital dyes to assess embryonic viability in the hamster, Mesocricetus auratus

    SciTech Connect

    Hutz, R.J.; DeMayo, F.J.; Dukelow, W.R.

    1985-05-01

    Experiments were designed to assess the use of the vital dyes trypan blue and fluorescein diacetate as indicators of the viability of hamster ova and embryos. Exclusion of trypan blue and fluorescence with fluorescein diacetate showed high correlations with uptake of (/sup 3/H)uridine by ova and further development of embryos in vitro. Ova killed by freezing and thawing incorporated (/sup 3/H)uridine at background levels. Trypan blue exclusion and fluorescein diacetate uptake were highly correlated with each other (r = 0.99). Trypan blue and fluorescein diacetate serve as excellent indices of viability in ova and early embryos of hamsters.

  12. Methods of biological dosimetry employing chromosome-specific staining

    DOEpatents

    Gray, Joe W. (Livermore, CA); Pinkel, Daniel (Walnut Creek, CA)

    2000-01-01

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.

  13. Methods And Compositions For Chromosome-Specific Staining

    DOEpatents

    Gray, Joe W. (Livermore, CA); Pinkel, Daniel (Walnut Creek, CA)

    2003-08-19

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.

  14. The prediction of human skin responses by using the combined in vitro fluorescein leakage/Alamar Blue (resazurin) assay.

    PubMed

    Clothier, Richard; Starzec, Gemma; Pradel, Lionel; Baxter, Victoria; Jones, Melanie; Cox, Helen; Noble, Linda

    2002-01-01

    A range of cosmetics formulations with human patch-test data were supplied in a coded form, for the examination of the use of a combined in vitro permeability barrier assay and cell viability assay to generate, and then test, a prediction model for assessing potential human skin patch-test results. The target cells employed were of the Madin Darby canine kidney cell line, which establish tight junctions and adherens junctions able to restrict the permeability of sodium fluorescein across the barrier of the confluent cell layer. The prediction model for interpretation of the in vitro assay results included initial effects and the recovery profile over 72 hours. A set of the hand-wash, surfactant-based formulations were tested to generate the prediction model, and then six others were evaluated. The model system was then also evaluated with powder laundry detergents and hand moisturisers: their effects were predicted by the in vitro test system. The model was under-predictive for two of the ten hand-wash products. It was over-predictive for the moisturisers, (two out of six) and eight out of ten laundry powders. However, the in vivo human patch test data were variable, and 19 of the 26 predictions were correct or within 0.5 on the 0-4.0 scale used for the in vivo scores, i.e. within the same variable range reported for the repeat-test hand-wash in vivo data. PMID:12405878

  15. Isolation and characterization of plant growth-promoting rhizobacteria from wheat roots by wheat germ agglutinin labeled with fluorescein isothiocyanate.

    PubMed

    Zhang, Jian; Liu, Jingyang; Meng, Liyuan; Ma, Zhongyou; Tang, Xinyun; Cao, Yuanyuan; Sun, Leni

    2012-04-01

    Thirty-two isolates were obtained from wheat rhizosphere by wheat germ agglutinin (WGA) labeled with fluorescein isothiocyanate (FITC). Most isolates were able to produce indole acetic acid (65.6%) and siderophores (59.3%), as well as exhibited phosphate solubilization (96.8%). Fourteen isolates displayed three plant growth-promoting traits. Among these strains, two phosphate-dissolving ones, WS29 and WS31, were evaluated for their beneficial effects on the early growth of wheat (Triticum aestivum Wan33). Strain WS29 and WS31 significantly promoted the development of lateral roots by 34.9% and 27.6%, as well as increased the root dry weight by 25.0% and 25.6%, respectively, compared to those of the control. Based on 16S rRNA gene sequence comparisons and phylogenetic positions, both isolates were determined to belong to the genus Bacillus. The proportion of isolates showing the properties of plant growth-promoting rhizobacteria (PGPR) was higher than in previous reports. The efficiency of the isolation of PGPR strains was also greatly increased by WGA labeled with FITC. The present study indicated that WGA could be used as an effective tool for isolating PGPR strains with high affinity to host plants from wheat roots. The proposed approach could facilitate research on biofertilizers or biocontrol agents. PMID:22538646

  16. FLUORESCEIN ANGIOGRAPHY VERSUS OPTICAL COHERENCE TOMOGRAPHY-GUIDED PLANNING FOR MACULAR LASER PHOTOCOAGULATION IN DIABETIC MACULAR EDEMA

    PubMed Central

    Kozak, Igor; El-Emam, Sharif Y.; Cheng, Lingyun; Bartsch, Dirk-Uwe; Chhablani, Jay; Freeman, William R.; Arevalo, J. Fernando

    2015-01-01

    Purpose To compare laser photocoagulation plans for diabetic macular edema (DME) using fluorescein angiography (FA) versus optical coherence tomography (OCT) thickness map superimposed on the retina. Methods Fourteen eyes with DME undergoing navigated laser photocoagulation with navigated photocoagulator had FA taken using the same instrument. Optical coherence tomography central retinal thickness map was imported to the photocoagulator and with same magnification aligned onto the retina. Three retina specialists placed laser spot marks separately on FA and OCT image in a masked fashion. The spots placed by each physician were compared between FA and OCT and among physicians. The area of dye leakage on FA and increased central retinal thickness on OCT of the same eye were also compared. Results The average number of spots using FA and OCT template was 36.64 and 40.61, respectively (P = 0.0201). The average area of dye leakage was 7.45 mm2, whereas the average area of increased central retinal thickness on OCT of the same eye was 10.92 mm2 (P = 0.013). Conclusion There is variability in the treatment planning for macular photocoagulation with a tendency to place more spots when guided by OCT than by FA. Integration of OCT map aligned to the retina may have an impact on treatment plan once such information is available. PMID:24695064

  17. Lack of Impact of High Dietary Vitamin A on T Helper 2-Dependent Contact Hypersensitivity to Fluorescein Isothiocyanate in Mice.

    PubMed

    Kobayashi, Chie; Kurohane, Kohta; Imai, Yasuyuki

    2015-11-01

    Overuse of vitamin A as a dietary supplement is a concern in industrialized countries. High-level dietary vitamin A is thought to shift immunity to a T helper 2 (Th2)-dominant one, resulting in the promotion of allergies. We have been studying a fluorescein isothiocyanate (FITC)-induced contact hypersensitivity (CHS) mouse model that involves Th2-type immunity. We fed a diet with a high retinyl palmitate content (250 international units (IU)/g diet) or a control diet (4?IU/g diet) to BALB/c mice for three weeks. No augmentation of FITC-induced CHS was found in mice fed the diet with a high vitamin A content, although accumulation of the vitamin was confirmed in the livers of these animals. The results indicated that relatively short-term feeding of the high-level vitamin A diet did not influence the Th2-driven response at a stage with significant retinol accumulation in the liver. The results were in contrast to the high-dose pyridoxine diets that produced a reduced response in FITC-induced CHS. PMID:26299258

  18. Simultaneous optical coherence tomography angiography and fluorescein angiography in rodents with normal retina and laser-induced choroidal neovascularization.

    PubMed

    Liu, Wenzhong; Li, Hao; Shah, Ronil S; Shu, Xiao; Linsenmeier, Robert A; Fawzi, Amani A; Zhang, Hao F

    2015-12-15

    Fluorescein angiography (FA) is the current clinical imaging standard for vascular related retinal diseases such as macular degeneration and diabetic retinopathy. However, FA is considered invasive and can provide only two-dimensional imaging. In comparison, optical coherence tomography angiography (OCTA) is noninvasive and can generate three-dimensional imaging; investigations of OCTA already demonstrated great promise in retinal vascular imaging. Yet, to further develop and apply OCTA, strengths and weaknesses between OCTA and FA need to be thoroughly compared. To avoid complications in image registration, an ideal comparison requires co-registered and simultaneous imaging by both FA and OCTA. In this Letter, we developed a system with integrated laser-scanning ophthalmoscope FA (SLO-FA) and OCTA, and conducted simultaneous dual-modality retinal vascular imaging in rodents. In imaging healthy rodent eyes, OCTA can resolve retinal capillaries better than SLO-FA does, particularly deep capillaries. In imaging rodent eyes with laser-induced choroidal neovascularization (CNV), OCTA can identify CNV that eludes SLO-FA detection. PMID:26670511

  19. Chromosome-specific staining to detect genetic rearrangements

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel; Tkachuk, Douglas; Westbrook, Carol

    2013-04-09

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyzes. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML) and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar but genetically different diseases, and for many prognostic and diagnostic applications.

  20. Wintergreen oil: a novel method in Wheatley's trichrome staining technique.

    PubMed

    Salleh, Fatmah Md; Anuar, Tengku Shahrul; Yasin, Azlin Mohd; Moktar, Norhayati

    2012-10-01

    Permanent staining of faecal smears by Wheatley's trichrome technique has been used by many scientists for the detection of parasites in the past and it was found to be highly sensitive. This study was conducted to evaluate the use of Wintergreen oil in comparison with xylene in Wheatley's trichrome staining technique, as the reference technique. In a blind comparison study, 500 collected faecal samples from aboriginal communities were examined. Wintergreen oil was found to be more superior than xylene as a clearing agent in the Wheatley's trichrome staining of polyvinyl alcohol-fixed faecal smears for the identification of intestinal protozoa. Elimination of toxic, carcinogenic, and fire hazards makes Wintergreen oil the preferred choice in routine parasitology examinations. PMID:22986100

  1. The role of the Giemsa stain in cytogenetics.

    PubMed

    Dolan, M

    2011-04-01

    In just half a century since the human diploid chromosome number was correctly identified as 46, there has been a rapid expansion in our understanding of both the genetic foundation of normal human development and the development of various constitutional and acquired abnormalities. The ability to detect numerical and structural chromosomal abnormalities was made possible by the Giemsa stain. Despite the recent advent of powerful molecular-based cytogenetic techniques (e.g., fluorescence in situ hybridization, array-based comparative genomic hybridization), Giemsa-based chromosomal banding and staining techniques retain their crucial role in cytogenetics. PMID:21395494

  2. Interfacing Microfluidics with Negative Stain Transmission Electron Microscopy.

    PubMed

    Mukhitov, Nikita; Spear, John M; Stagg, Scott M; Roper, Michael G

    2016-01-01

    A microfluidic platform is presented for preparing negatively stained grids for use in transmission electron microscopy (EM). The microfluidic device is composed of glass etched with readily fabricated features that facilitate the extraction of the grid poststaining and maintains the integrity of the sample. Utilization of this device simultaneously reduced environmental contamination on the grids and improved the homogeneity of the heavy metal stain needed to enhance visualization of biological specimens as compared to conventionally prepared EM grids. This easy-to-use EM grid preparation device provides the basis for future developments of systems with more integrated features, which will allow for high-throughput and dynamic structural biology studies. PMID:26642355

  3. Anisotropic staining of apurinic acid with toluidine blue.

    PubMed

    Erenpreisa, J; Freivalds, T

    1979-04-12

    Using normal rat liver imprints, studies were carried out on the effects of histone extraction and the formation of aldehyde groups from deoxyribose on anisotropic toluidine blue staining of depurinized DNA after sodium bisulfite treatment. The anisotropic effect of bisulfite was found to be determined by binding of bisulfite ions to the aldehyde groups of apurinic acid which, together with free phosphate groups of DNA ensure coparallel attachment of the dye molecules. It was also shown that at pH 5.0 toluidine blue binds with both the phosphate and aldehyde groups of apurinic acid, to give anisotropic staining. PMID:89109

  4. Analysis of Oxiclean: An Interesting Comparison of Percarbonate Stain Removers

    ERIC Educational Resources Information Center

    Bracken, Jeffrey D.; Tietz, David

    2005-01-01

    The study focuses on percarbonate-based stain removers since the percarbonate can be heated to produce additional sodium carbonate. An experiment that provides general chemistry students an opportunity to apply their knowledge of basic stoichiometry to solve a relevant, real-world problem is described.

  5. 5. Downstream elevation, view to southeast. Dark stains on side ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    5. Downstream elevation, view to southeast. Dark stains on side of main girder are from deck drain scuppers, marking deck level within the girders. Compare this view and CA-126-7 to CA-126-19 for indication of severity of siltation of Salt River channel has silted. - Salt River Bridge, Spanning Salt River at Dillon Road, Ferndale, Humboldt County, CA

  6. MP 33354 Pro-Q Sapphire 532 Oligohistidine Gel Stain

    E-print Network

    Lebendiker, Mario

    MP 33354 Pro-Q® Sapphire 532 Oligohistidine Gel Stain Product Information Storage upon receipt: · 6­ polyacrylamide gel electrophoresis (PAGE) and Western blot analysis. With Molecular Probes Pro-Q® Sapphire 532­polyacrylamide gel, eliminating the need to blot the protein to a membrane (Figure 1, top). Pro-Q Sapphire 532

  7. Staining potential of sealants in/on exterior wall substrates

    SciTech Connect

    Chin, I.R.; Gorrell, T.A.; Scheffler, M.J.

    1996-12-31

    The investigation of dozens of exterior walls on buildings by the authors have revealed that certain sealants used to seal joints in exterior walls have caused the following conditions: 1. Local staining and discoloration of masonry wall substrates due to penetration of sealant into substrates. 2. Local change in absorption of masonry wall substrates due to penetration of sealant into the substrates which results in local discoloration of wall substrates after rains. 3. Surface discoloration of sealant due to dirt accumulation on sealant. 4. Stains in the form of streaks of direct on masonry and on metal wall substrates due to transfer by rain water of some of the dirt on the sealant surface onto the adjacent surface of the substrate. Laboratory and field testing of the major commercially available sealants by the authors to date have revealed that silicone sealants have the propensity to stain certain exterior wall substrates while polyurethane sealants do not; that silicone sealants in service discolor more significantly and faster due to dirt accumulation than polyurethane sealants; that sealant that has penetrated into masonry substrates cannot be entirely removed from the substrate by cleaning; and that preconstruction testing of sealants and proper material selection can help to avoid staining of sealant.

  8. Image analysis of dye stained patterns in soils

    NASA Astrophysics Data System (ADS)

    Bogner, Christina; Trancón y Widemann, Baltasar; Lange, Holger

    2013-04-01

    Quality of surface water and groundwater is directly affected by flow processes in the unsaturated zone. In general, it is difficult to measure or model water flow. Indeed, parametrization of hydrological models is problematic and often no unique solution exists. To visualise flow patterns in soils directly dye tracer studies can be done. These experiments provide images of stained soil profiles and their evaluation demands knowledge in hydrology as well as in image analysis and statistics. First, these photographs are converted to binary images classifying the pixels in dye stained and non-stained ones. Then, some feature extraction is necessary to discern relevant hydrological information. In our study we propose to use several index functions to extract different (ideally complementary) features. We associate each image row with a feature vector (i.e. a certain number of image function values) and use these features to cluster the image rows to identify similar image areas. Because images of stained profiles might have different reasonable clusterings, we calculate multiple consensus clusterings. An expert can explore these different solutions and base his/her interpretation of predominant flow mechanisms on quantitative (objective) criteria. The complete workflow from reading-in binary images to final clusterings has been implemented in the free R system, a language and environment for statistical computing. The calculation of image indices is part of our own package Indigo, manipulation of binary images, clustering and visualization of results are done using either build-in facilities in R, additional R packages or the LATEX system.

  9. ANEUPLOIDY TEST DEVELOPMENT: KINETOCHORE STAINING IN MAMMALIAN SYSTEMS

    EPA Science Inventory

    The purpose of the project was to determine the feasibility of using human-derived antibodies against the chromosomal kinetochore region coupled with immunofluorescence staining as a method for evaluating the induction of aneuploidy in mammalian cells in vitro and in vivo. The te...

  10. ENHANCED DAPI STAINING FOR CRYPTOSPORIDIUM IN WATER SAMPLES

    EPA Science Inventory

    The U.S. Environmental Protection Agency's Method 1623 is used to detect and quantify the presence of {ital Cryptosporidium} spp. oocysts in water. The protocol consists of concentrating a sample, staining this concentrate with a fluorescent antibody, and examining the sample mi...

  11. Surface hardness of resin composites after staining and bleaching.

    PubMed

    Okte, Zeynep; Villalta, Patricia; García-Godoy, Franklin; Lu, Huan; Powers, John M

    2006-01-01

    This study investigated the effect of 3 staining solutions and 3 over-the-counter tooth-bleaching systems on the microhardness of 2 dental resin composites. Forty-five specimens of Filtek Supreme and Esthet-X were randomly assigned to 3 groups. Over a 40-day test period, the specimens in each group (n=15) were immersed in 1 of the 2 staining solutions (coffee and red wine) or distilled water as the control for 3 hours a day at room temperature. The 15 specimens in each staining group were further randomly divided into 3 subgroups, and the specimens in each subgroup (n=5) were bleached using one of the bleaching agents (Night Effects, Simply White Night and Opalescence Quick). Surface hardness was measured at 24 hours after polymerization (baseline), after staining and after bleaching. Means and standard deviations were calculated, and the data were analyzed using repeated-measures analysis of variance and Duncan's Test. The microhardness of Esthet-X was significantly higher than Filtek Supreme at baseline (p<0.01). All specimens of both materials immersed in coffee and wine revealed a significant hardness decrease compared to baseline values (p<0.05). In the control group, microhardness was increased, and this increase was statistically significant for Filtek Supreme (p<0.05). After bleaching, there was a significant decrease in mean microhardness for all groups tested (p<0.05). No significant difference was found among bleaching agents. PMID:17024953

  12. MODELING OF ALKANE EMISSIONS FROM A WOOD STAIN

    EPA Science Inventory

    The article discusses full-scale residential house tests to evaluate the effects of organic emissions from a wood finishing product--wood stain--on indoor air quality (IAQ). The test house concentrations of three alkane species, nonane, decane, and undecane, were measured as a fu...

  13. SYPRO Orange and Red protein gel stains provide the following advantages over

    E-print Network

    Lebendiker, Mario

    SYPRO Orange and Red protein gel stains provide the following advantages over conventional and Orange dyes interact with the SDS coat around proteins in the gel they give more consistent staining. SYPRO Orange and Red protein gel stains SYPROTM Orange and Red protein gel stains enabling fast, simple

  14. Fundus autofluorescence in central serous chorioretinopathy: association with spectral-domain optical coherence tomography and fluorescein angiography

    PubMed Central

    Zhang, Peng; Wang, Hai-Yan; Zhang, Zi-Feng; Sun, Dong-Jie; Zhu, Jin-Ting; Li, Juan; Wang, Yu-Sheng

    2015-01-01

    AIM To evaluate the correlation among changes in fundus autofluorescence (AF) measured using infrared fundus AF (IR-AF) and short-wave length fundus AF (SW-AF) with changes in spectral-domain optical coherence tomography (SD-OCT) and fluorescein angiography (FA) in central serous chorioretinopathy (CSC). METHODS Two hundred and twenty consecutive patients with CSC were included. In addition to AF, patients were assessed by means of SD-OCT and FA. Abnormalities in images of IR-AF, SW-AF, FA were analyzed and correlated with the corresponding outer retinal alterations in SD-OCT findings. RESULTS Eyes with abnormalities on either IR-AF or SW-AF were found in 256 eyes (58.18%), among them 256 eyes (100%) showed abnormal IR-AF, but SW-AF abnormalities were present only in 213 eyes (83.20%). The hypo-IR-AF corresponded to accumulation of sub-retinal liquid, collapse of retinal pigment epithelium (RPE) or detachment of RPE with or without RPE leakage point in the corresponding area. The hyper-IR-AF corresponded to the area with loss of the ellipsoid portion of the inner segments and sub-sensory retinal deposits or focal melanogenesis under sensory retina. The hypo-SW-AF corresponded to accumulation of sub-retinal liquid or atrophy of RPE. The hyper-SW-AF associated with sub-sensory retinal deposits, detachment of RPE and focal melanogenesis. CONCLUSION IR-AF was more sensitive than SW-AF and FA for identifying pathological abnormalities in CSC. The characteristics of IR-AF in CSC were attributable to the modification of melanin in the RPE. IR-AF should be used as a common diagnostic tool for identifying pathological lesion in CSC. PMID:26558217

  15. The development of a standardised protocol to measure squamous differentiation in stratified epithelia, by using the fluorescein cadaverine incorporation technique.

    PubMed

    Gray, Alison C; Malton, Joanne; Clothier, Richard H

    2004-06-01

    Fluorescein cadaverine (FC) incorporation into cornified envelopes during squamous differentiation in stratified epithelia acts as a fluorescent substitute for endogenous transglutaminase substrates that can be visualised and quantified. The FC incorporation technique has been used to evaluate squamous differentiation in keratinocytes cultured in a medium that stimulates differentiation and in response to modulation by chemicals. A Standard Operating Procedure for the measurement of squamous differentiation is required as part of the prevalidation procedure for in vitro assays. In the present study, keratinocytes were isolated from the epidermis of 34 human donors. Cellular metabolic activity (resorufin production), total protein (kenacid blue uptake) and transglutaminase activity (FC incorporation) were measured in 87 and 21 independent runs at 6 and 12 days, respectively. Analysis of the control data showed that the cultures had a mean resorufin production that decreased over 12 days. This was inversely related to FC incorporation, which increased over 12 days. Mean protein concentration was reduced over the 12 days, but not in analyses that used donors for whom both 6-day and 12-day data were available. This information was used to define the normal limits within which the data should fall (mean +/- 1 SD). Data sets falling outside the normal limits performed statistically no differently from the normal responders, in experiments conducted to investigate the effects of chemicals on the modulation of squamous differentiation in keratinocytes. This was demonstrated by using compounds that modify transglutaminase expression and/or activity. All-trans retinoic acid significantly inhibited FC incorporation, but stimulated resorufin production at 1 x 10(-7)M and above. Nicotine significantly up-regulated both FC incorporation and resorufin production at 125 microg/ml. Hence, it was concluded that this robust assay approach, in which keratinocytes from a range of donors respond predictably to the test chemicals employed, did not justify the limitations that would be imposed by setting criteria that eliminated all data lying outside the normal range. PMID:15601237

  16. Multiple Function Fluorescein Probe Performs Metal Chelation, Disaggregation, and Modulation of Aggregated A? and A?-Cu Complex.

    PubMed

    Muthuraj, B; Layek, Sourav; Balaji, S N; Trivedi, Vishal; Iyer, Parameswar Krishnan

    2015-11-18

    An exceptional probe comprising indole-3-carboxaldehyde fluorescein hydrazone (FI) performs multiple tasks, namely, disaggregating amyloid ? (A?) aggregates in different biomarker environments such as cerebrospinal fluid (CSF), A?1-40 fibrils, ?-amyloid lysozyme aggregates (LA), and U87 MG human astrocyte cells. Additionally, the probe FI binds with Cu(2+) ions selectively, disrupts the A? aggregates that vary from few nanometers to micrometers, and prevents their reaggregation, thereby performing disaggregation and modulation of amyloid-? in the presence as well as absence of Cu(2+) ion. The excellent selectivity of probe FI for Cu(2+) was effectively utilized to modulate the assembly of metal-induced A? aggregates by metal chelation with the "turn-on" fluorescence via spirolactam ring opening of FI as well as the metal-free A? fibrils by noncovalent interactions. These results confirm that FI has exceptional ability to perform multifaceted tasks such as metal chelation in intracellular conditions using A? lysozyme aggregates in cellular environments by the disruption of ?-sheet rich A? fibrils into disaggregated forms. Subsequently, it was confirmed that FI had the ability to cross the blood-brain barrier and it also modulated the metal induced A? fibrils in cellular environments by "turn-on" fluorescence, which are the most vital properties of a probe or a therapeutic agent. Furthermore, the morphology changes were examined by atomic force microscopy (AFM), polarizable optical microscopy (POM), fluorescence microscopy, and dynamic light scattering (DLS) studies. These results provide very valuable clues on the A? (CSF A? fibrils, A?1-40 fibrils, ?-amyloid lysozyme aggregates) disaggregation behavior via in vitro studies, which constitute the first insights into intracellular disaggregation of A? by "turn-on" method thereby influencing amyloidogenesis. PMID:26332658

  17. Efficacy of chewing gum in preventing extrinsic tooth staining.

    PubMed

    Yankell, S L; Emling, R C

    1997-01-01

    The purpose of this six-week clinical study was to determine the efficacy of sugar-free chewing gum versus no chewing on preventing Peridex (0.12% chlorhexidine)-associated stain. One-hundred and fifty healthy adult subjects, categorized by tea or coffee intake and smoking, were randomly assigned to a chewing or no chewing gum group. All subjects were given Peridex and an ADA-approved toothbrush and fluoride toothpaste to use twice a day. Gum was chewed for 20 minutes five times each day, after toothbrushing and Peridex rinse in the morning and evening, and after each meal. At baseline, all subjects received a professional cleaning to remove all supragingival deposits and extrinsic strain. At three and six weeks, safety and stain intensity and area were monitored on the anterior teeth and posterior Ramfjord teeth using the Lobene stain scoring method. Seventy-two subjects in each group completed the study. Attrition was unrelated to product use. No untoward reactions were reported or observed at any time in the study. At the six-week evaluations, the chewing gum group exhibited significantly lower (p < 0.05-0.001) total stain scores on both anterior and posterior areas evaluated compared to the no chewing group scores. In addition to the stain evaluations, a randomly selected subset of 60 subjects was evaluated for gingivitis at baseline prior to cleaning, and at three and six weeks, on the buccal and lingual surfaces of the Ramfjord teeth. Both the chewing gum and no chewing gum subset subjects had a significant decrease in gingivitis scores from baseline to three weeks (p < 0.001) and from baseline to six weeks (p < 0.05-0.001). There were no significant statistical differences between the two groups at anytime during the study on gingivitis levels. Chewing gum, after product use, did not reduce the efficacy of chlorhexidine on gingivitis scores. PMID:9586534

  18. Uniform staining of Cyclospora oocysts in fecal smears by a modified safranin technique with microwave heating.

    PubMed Central

    Visvesvara, G S; Moura, H; Kovacs-Nace, E; Wallace, S; Eberhard, M L

    1997-01-01

    Cyclospora, a coccidian protist, is increasingly being identified as an important, newly emerging parasite that causes diarrhea, flatulence, fatigue, and abdominal pain leading to weight loss in immunocompetent persons with or without a recent travel history as well as in patients with AIDS. Modified Kinyoun's acid-fast stain is the most commonly used stain to identify the oocyst of this parasite in fecal smears. Oocysts of Cyclospora stain variably by the modified acid-fast procedure, resulting in the possible misidentification of this parasite. We examined fecal smears stained by six different procedures that included Giemsa, trichrome, chromotrope, Gram-chromotrope, acid-fast, and safranin stains. We report on safranin-based stain that uniformly stains oocysts of Cyclospora a brilliant reddish orange, provided that the fecal smears are heated in a microwave oven prior to staining. This staining procedure, besides being superior to acid-fast staining, is fast, reliable, and easy to perform in most clinical laboratories. PMID:9041421

  19. Nuclear Quadrupole Double Resonance Investigation of the Anomalous Temperature Coefficients of the Strong Hydrogen Bonds in Sodium and Potassium Deuterium Diacetate.

    NASA Astrophysics Data System (ADS)

    Shaw, Eric Max

    This thesis was directed at learning more about the unusual electronic environment near hydrogen within strong hydrogen bonds. "Strong" hydrogen bonds are unique in that the hydrogen atom is symmetrically located, or nearly so, between two electronegative atoms; the bond energies are relatively large. In a "normal" hydrogen bond the hydrogen atom is bonded to, and thus physically closer to, a parent atom, and only weakly attracted to another electronegative atom; bond energies are typically small. To examine these bonds, deuterium was substituted for hydrogen and the electric quadrupole coupling constant (QCC) of deuterium was measured using field cycling nuclear magnetic resonance. The electric quadrupole moment of deuterium is sensitive to changes in the surrounding electric field gradient, and is thus a good probe of the immediate electronic structure. The results show that the temperature dependence of the QCC is opposite to, and much larger than, what one would normally expect to observe for deuterium. The QCC is found to decrease strongly with decreasing temperature. This project was the first to study in detail the temperature dependence of deuterium QCCs in strong hydrogen bonds. The magnitude of the deuterium QCCs for the diacetates was found to be strongly depressed relative to typical values for deuterium. These results parallel large shifts in the infrared vibrational frequencies observed in many molecules which contain strong hydrogen bonds. The asymmetry parameter, which is a measure of the departure from axial symmetry of the electric field gradient (EFG) at deuterium, was found to be unusually large for what are known to be linear, or nearly linear, three-center bonds. Based on ab initio Hartree-Fock calculations aimed at determining the EFG at H in the archetypal bifluoride ion, F-H-F^-, the electronic charge density is drastically depleted at H. It is believed that the large reduction in the charge density allows the deuterium EFG to be highly sensitive to the shape of the charge distribution on the atoms to which deuterium is bonded. If these atoms are at points of low crystallographic symmetry, the polarization of these adjacent atoms by other nearby atoms may cause the EFG to depart substantially from being axially symmetric. Also obtained from the molecular orbital calculations for bifluoride ion were the total electronic energy and the electric field gradient at H. From these calculations potential function models for the asymmetric stretch and the bend were constructed. An attempt was made to correlate the predictions made by these models for the temperature dependence of the deuteron quadrupole coupling constant in bifluoride ion with the experimentally observed results for the diacetates.

  20. Widespread Microbial Adaptation to l-Glutamate-N,N-diacetate (L-GLDA) Following Its Market Introduction in a Consumer Cleaning Product.

    PubMed

    Itrich, Nina R; McDonough, Kathleen M; van Ginkel, Cornelis G; Bisinger, Ed C; LePage, Jim N; Schaefer, Edward C; Menzies, Jennifer Z; Casteel, Kenneth D; Federle, Thomas W

    2015-11-17

    l-Glutamate-N,N-diacetate (L-GLDA) was recently introduced in the United States (U.S.) market as a phosphate replacement in automatic dishwashing detergents (ADW). Prior to introduction, L-GLDA exhibited poor biodegradation in OECD 301B Ready Biodegradation Tests inoculated with sludge from U.S. wastewater treatment plants (WWTPs). However, OECD 303A Activated Sludge WWTP Simulation studies showed that with a lag period to allow for growth (40-50 days) and a solids retention time (SRT) that allows establishment of L-GLDA degraders (>15 days), significant biodegradation (>80% dissolved organic carbon removal) would occur. Corresponding to the ADW market launch, a study was undertaken to monitor changes in the ready biodegradability of L-GLDA using activated sludge samples from various U.S. WWTPs. Initially all sludge inocula showed limited biodegradation ability, but as market introduction progressed, both the rate and extent of degradation increased significantly. Within 22 months, L-GLDA was ready biodegradable using inocula from 12 WWTPs. In an OECD 303A study repeated 18 months post launch, significant and sustained carbon removal (>94%) was observed after a 29-day acclimation period. This study systematically documented field adaptation of a new consumer product chemical across a large geographic region and confirmed the ability of laboratory simulation studies to predict field adaptation. PMID:26465169

  1. Synthesis and characterization of polycarbonates by melt phase interchange reactions of alkylene and arylene diacetates with alkylene and arylene diphenyl dicarbonates.

    PubMed

    Sweileh, Bassam A; Al-Hiari, Yusuf M; Kailani, Mohammad H; Mohammad, Hani A

    2010-05-01

    This work presents a new synthetic approach to aromatic and aliphatic polycarbonates by melt polycondensation of bisphenol A diacetates with alkylene- and arylenediphenyl dicarbonates. The diphenyl dicarbonates were prepared from phenyl chloroformate and the corresponding dihydroxy compounds. The process involved a precondensation step under a slow stream of dry argon with the elimination of phenyl acetate, followed by melt polycondensation at high temperature and under vacuum. The potential of this reaction is demonstrated by the successful synthesis of a series of aromatic-aromatic and aromatic-aliphatic polycarbonates having inherent viscosities from 0.19 to 0.43 dL/g. Thus low to intermediate molecular mass polymers were obtained. The (13)C-NMR spectra of the carbon of the carbonate group showed that the formed polycarbonates contain partial random sequence distribution of monomer residues in their chains. The polycarbonates were characterized by inherent viscosity, FTIR, (1)H-NMR and (13)C-NMR spectroscopy. The glass transition temperatures, measured by DSC, of the polycarbonates were in the range 13-108 degrees C. The thermogravimetric curves of showed that these polymers have good thermal stability up to 250 degrees C. The present approach may open the door for novel polycarbonates containing other organic functional groups. PMID:20657506

  2. Lectins stain cells differentially in the coral, Montipora capitata

    USGS Publications Warehouse

    Work, Thierry M.; Farah, Yael

    2014-01-01

    A limitation in our understanding of coral disease pathology and cellular pathogenesis is a lack of reagents to characterize coral cells. We evaluated the utility of plant lectins to stain tissues of a dominant coral, Montipora capitata, from Hawaii. Of 22 lectins evaluated, nine of these stained structures in the upper or basal body wall of corals. Specific structures revealed by lectins that were not considered distinct or evident on routine hematoxylin and eosin sections of coral tissues included apical and basal granules in gastrodermis and epidermis, cnidoglandular tract and actinopharynx cell surface membranes, capsules of mature holotrichous isorhizas, and perivitelline and periseminal cells. Plant lectins could prove useful to further our understanding of coral physiology, anatomy, cell biology, and disease pathogenesis.

  3. Cement line staining in undecalcified thin sections of cortical bone

    NASA Technical Reports Server (NTRS)

    Bain, S. D.; Impeduglia, T. M.; Rubin, C. T.

    1990-01-01

    A technique for demonstrating cement lines in thin, undecalcified, transverse sections of cortical bone has been developed. Cortical bone samples are processed and embedded undecalcified in methyl methacrylate plastic. After sectioning at 3-5 microns, cross-sections are transferred to a glass slide and flattened for 10 min. Sections of cortical bone are stained for 20 sec free-floating in a fresh solution of 1% toluidine blue dissolved in 0.1% formic acid. The section is dehydrated in t-butyl alcohol, cleared in xylene, and mounted with Eukitt's medium. Reversal lines appear as thin, scalloped, dark blue lines against a light blue matrix, whereas bone formation arrest lines are thicker with a smooth contour. With this technique cellular detail, osteoid differentiation, and fluorochrome labels are retained. Results demonstrate the applicability of a one-step staining method for cement lines which will facilitate the assessment of bone remodeling activity in thin sections of undecalcified cortical bone.

  4. Development of Cell Staining Technique for X-Ray Microscopy

    SciTech Connect

    Tseng, P. Y.; Shih, Y. T.; Liu, C. J.; Hsu, T.; Chien, C. C.; Leng, W. H.; Liang, K. S.; Yin, G. C.; Chen, F. R.; Je, J. H.; Margaritondo, G.; Hwu, Y.

    2007-01-19

    We report a technique for detection of sub-cellular organelles and proteins with hard x-ray microscopy. Several metals were used for enhancing contrast for x-ray microscopy. Osmium tetroxide provides an excellent stain for lipid and can delineate cell membrane. Uranyl acetate has high affinity for nucleotide and can stain nucleus. Immunolocalization of specific proteins and sub-cellular organelles was achieved by 3'3 diaminobenzidine (DAB) with nickel enhancement and nanogold-conjugated secondary antibody with silver enhancement. The x-rays emitted from synchrotron source was monochromatized by double crystal monochromator, the photon energy was fixed at 8 keV to optimize the focusing efficiency of the zone plates. The estimated resolution is about 60 nm. When compared with visible light and conventional confocal microscopy, the X-ray microscopy provides a superior resolution to both conventional optical microscopes.

  5. Genetic Variants Associated with Port-Wine Stains

    PubMed Central

    Wooderchak-Donahue, Whitney; Tan, Oon T.; Margraf, Rebecca; Stevenson, David A.; Grimmer, J. Fredrik; Bayrak-Toydemir, Pinar

    2015-01-01

    Background Port-wine stains (PWS) are capillary malformations, typically located in the dermis of the head and neck, affecting 0.3% of the population. Current theories suggest that port-wine stains are caused by somatic mutations that disrupt vascular development. Objectives Understanding PWS genetic determinants could provide insight into new treatments. Methods Our study used a custom next generation sequencing (NGS) panel and digital polymerase chain reaction to investigate genetic variants in 12 individuals with isolated port-wine stains. Importantly, affected and healthy skin tissue from the same individual were compared. A subtractive correction method was developed to eliminate background noise from NGS data. This allowed the detection of a very low level of mosaicism. Results A novel somatic variant GNAQ, c.547C>G, p.Arg183Gly was found in one case with 4% allele frequency. The previously reported GNAQ c.548G>A, p.Arg183Gln was confirmed in 9 of 12 cases with an allele frequency ranging from 1.73 to 7.42%. Digital polymerase chain reaction confirmed novel variants detected by next generation sequencing. Two novel somatic variants were also found in RASA1, although neither was predicted to be deleterious. Conclusions This is the second largest study on isolated, non-syndromic PWS. Our data suggest that GNAQ is the main genetic determinant in this condition. Moreover, isolated port-wine stains are distinct from capillary malformations seen in RASA1 disorders, which will be helpful in clinical evaluation. PMID:26192947

  6. Method and apparatus for staining immobilized nucleic acids

    DOEpatents

    Ramsey, J. Michael (Knoxville, TN); Foote, Robert S. (Oak Ridge, TN); Jacobson, Stephen C. (Knoxville, TN)

    2000-01-01

    A method for staining immobilized nucleic acids includes the steps of affixing DNA probes to a solid substrate, moving target DNA material into proximity with the DNA probes, whereby the target DNA hybridized with specific ones of the DNA probes, and moving a fluorescent dye into proximity with the hybridized target DNA, whereby the fluorescent dye binds to the hybridized DNA to enable subsequent detection of fluorescence.

  7. Coffee Stains from Drops with Receding Contact Lines

    NASA Astrophysics Data System (ADS)

    Freed-Brown, Julian

    2015-03-01

    We present a framework for calculating the surface density profile of a coffee stain deposited by a drying drop with a receding contact line. For standard coffee stains, the fluid pins to the substrate, forces flow towards the exterior of the drop and deposits a thin, concentrated ring of particles. Unlike a pinned drop, a receding drop pushes fluid towards its interior and continuously deposits mass across its substrate as it evaporates. This gives rise to a new class of mountain-like morphologies that are not seen in the standard coffee ring effect but are reminiscent of recent experimental results. For a thin, circular drop with uniform evaporation, we calculate the surface density profile analytically and find that it diverges towards the center of the drop as ? ~r - 1 / 2 , where r is the distance from the center. We estimate how this divergence is softened due to solute interactions at the final stage of drying. Our framework can easily be extended numerically or analytically to investigate novel stain morphologies left by drying drops of different shapes and evaporation profiles. This work is part of a thesis project advised by Tom Witten. It was supported in part by the National Science Foundation's MRSEC Program under Award Number DMR 0820054.

  8. Immunocytochemical staining of T and B lymphocytes in serous effusions.

    PubMed Central

    Ghosh, A K; Spriggs, A I; Mason, D Y

    1985-01-01

    Cell smears from serous effusions containing large numbers of lymphoid cells were stained by the alkaline phosphatase-anti-alkaline phosphatase technique with a panel of monoclonal antibodies, including anti-B and anti-T cell antibodies and anti-HLA-DR. Samples from 17 patients with lymphoproliferative disorders--such as chronic lymphocytic leukaemia and non-Hodgkin's lymphoma--and from 19 patients who had no evidence of lymphoid neoplasia--for example, cases of carcinoma, cardiac failure--were investigated. The majority of lymphoid cells in reactive effusions were T cells, which lacked HLA-DR and showed a marked excess of helper/inducer cells (mean helper to suppressor ratio of 3 X 5). In contrast, lymphoid cells in samples from nine cases of B cell neoplasia were positive for B cell antigen and HLA-DR. In a further four B cell neoplasms most lymphoid cells were reactive T cells. Two cases of T cell lymphoid leukaemia could also be characterised by immunocytochemical staining, both being classified as T helper cell neoplasms. Labelling was performed on routinely prepared, air dried cell smears, which could be stored in the unfixed state for long periods before staining. The technique may therefore be of use in many clinical cytology laboratories for the diagnosis of effusions containing numerous lymphoid cells. Images PMID:3891789

  9. [A duplicate staining method for permanent specimen of Trichinella spiralis encapsulated larvae].

    PubMed

    Li, Dan; Yang, Ding; Pi, Ben-Wei; Niu, Li-Na; Zhang, Ying; Wang, Guo-Ying

    2012-04-30

    With single staining method, Trichinella spiralis encapsulated larvae specimens were fixed with formaldehyde alcohol acetic acid fixative solution, and stained with alcohol borax-carmine staining solution (4% borax solution 100 ml, carmine 1 g, and 70% alcohol 100 ml). With duplicate staining, the encapsulated larvae specimens were fixed with formaldehyde alcohol acetic acid fixative solution, and double stained with alcohol borax carmine staining solution and fast green staining solution (fast green 0.1 g, 95% alcohol 100 ml). The results showed that with single staining, it was not clear-cut between the cyst and muscle cells although the larva was differentiable, while with duplicate staining, the larva, cyst and muscle cells were distinguished more clearly. PMID:22908823

  10. En bloc staining with hydroquinone treatment for block face imaging.

    PubMed

    Togo, Akinobu; Ohta, Keisuke; Higashi, Ryuhei; Nakamura, Kei-Ichiro

    2014-11-01

    IntroductionBecause recent three-dimensional (3D) ultrastructural reconstruction techniques such as serial block face scanning electron microscopy (SBFSEM), obtain their images directly from the flat surface of specimens via material contrast[1], specimens should be strongly stained with heavy metals prior to resin embedding in order to obtain higher material contrast using backscattered electrons (BSEs). To enhance membrane contrast for block face imaging (BFI), we usually stain specimens using the method published by Deerinck[2], and the images obtained show TEM-like contrast.However, recently, our research subjects have required reconstruction of a much larger volume, increasing the total image acquisition time. To reduce the total acquisition time, both high sensitivity detectors and a new specimen preparation method that provides much higher contrast are required. Takahashi et al.[3] have reported that hydroquinone (HQ) treatment during traditional electro-conductive staining increases specimen conductivity and drastically reduces the charge problem for SEM observation. They concluded that HQ treatment might increase the efficiency of secondary electron (SE) generation. Because BFI can be performed using SE as well as BSE, we examined whether addition of HQ treatment to en bloc staining protocols increased the contrast for BFI using SE. Materials & methodsMouse liver tissue was used. Mice were deeply anesthetized by diethyl ether and sodium pentobarbital, and tissues were fixed by transcardial perfusion of 2% paraformaldehyde and 2.5% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4) through the left ventricle, followed by heparin-containing saline. After perfusion, liver tissues were removed and cut into small cubes approximately 1 mm(3) in the fixative, and were further fixed in the same fixative for 2 h at 4°C. Subsequently, en blocstaining was performed as follows: the specimens were treated using a reduced-OTO staining method (1.5% potassium ferrocyanide-2% OsO4, 1% thiocarbohydrazide, and then 2% OsO4). Subsequently, specimens were treated with 1% HQ solution. Some specimens were exempted from this step and used as controls. Specimens were further stained with 4% uranyl acetate and Walton's lead aspartate solution.After staining, specimens were dehydrated using an ethanol series and embedded in epoxy resin (EPON812, TAAB). Surface of specimens block were cut with a diamond knife, and the newly created flat surfaces of the specimens were coated with evaporated carbon (50 Å) and observed using a SEM (Quanta 3D FEG, FEI).ResultsThe HQ-treated specimens generated a larger amount of SEs than control specimens when subjected to irradiation with the same beam, although BSE numbers were not evidently increased by the treatment. The present results suggest that HQ treatment increases SE generation efficiency, but does not enhance the recruitment of heavy metals into specimens. HQ treatment increased the contrast-to-noise ratio of BFI for images obtained using SEs, and may reduce the total image acquisition time of recently developed 3D reconstruction methods based on SEM. PMID:25359840

  11. Detailed interrogation of trypanosome cell biology via differential organelle staining and

    E-print Network

    Schnaufer, Achim

    Detailed interrogation of trypanosome cell biology via differential organelle staining of trypanosome cell biology via differential organelle staining and automated image analysis Richard J Wheeler DNA-containing organelles, the kinetoplast (mitochondrial DNA) and nucleus, which provide useful

  12. Colorimetry for the stain technologist. III. The specification of color difference.

    PubMed

    Marshall, P N; Galbraith, W

    1984-11-01

    This paper illustrates the calculation of color differences, involving luminance as well as chromaticity components. Color differences have been calculated for a large number of stained histological objects. Four different color difference formulae have been used, namely, those associated with the FMC 2, U*V*W*, L*u*v* and L*a*b* systems. Comparison has first been made between various hematological substrates after staining with two different azure B-eosin Y stains. Next, comparison is made for the same substrates after staining with one of the azure B stains and a methylene blue-eosin Y stain. Pairwise comparison is also made of various substrates from the epithelium of the uterine cervix after Papanicolaou staining. Finally, pairwise comparison documents color differences accompanying maturation for the erythroid and myeloid cell lines in azure B-eosin Y stained bone marrow. The limitations of current color difference formulae are discussed. PMID:6084879

  13. Machine vision system for automated detection of stained pistachio nuts

    NASA Astrophysics Data System (ADS)

    Pearson, Tom C.

    1995-01-01

    A machine vision system was developed to separate stained pistachio nuts, which comprise of about 5% of the California crop, from unstained nuts. The system may be used to reduce labor involved with manual grading or to remove aflatoxin contaminated product from low grade process streams. The system was tested on two different pistachio process streams: the bi- chromatic color sorter reject stream and the small nut shelling stock stream. The system had a minimum overall error rate of 14% for the bi-chromatic sorter reject stream and 15% for the small shelling stock stream.

  14. Restoration of Fluorosis Stained Teeth: A Case Study.

    PubMed

    Slaska, Barbara; Liebman, Arnold I; Kukleris, Diana

    2015-07-01

    Dental fluorosis manifests by too much ingestion of fluoride resulting in disturbances in enamel mineralization. The result is intrinsic discolorations in the maxillary and mandibular teeth with a poor esthetic appearance. In challenging cases, an esthetic result may be achieved only by a combination of techniques. This case report demonstrates a combination of modalities used to treat a patient presenting with atypical staining as a result of high-level exposure to ingested fluoride present in the drinking water as a child. Conservative treatment consisted of a combination of in-office bleaching to reduce the discoloration and porcelain veneers to create an esthetic result. PMID:26140966

  15. 10. Photocopy of an engraving of a stained glass window ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    10. Photocopy of an engraving of a stained glass window design by Johann Friedrich Overbeck (1789-1869) on which two of the chancel windows in the Church of the Holy Cross are thought to have been based. This copy is of a photocopy obtained from the Treasury of Notre Dame de Paris, Paris, France, by the late Mrs. Walter C. White of Stateburg, South Carolina. Mrs. White's photocopy is in the possession of Mrs. Richard K. Anderson of the Borough House at Stateburg. - Church of the Holy Cross, State Route 261, Stateburg, Sumter County, SC

  16. Detection of reactive oxygen species (ROS) by the oxidant-sensing probe 2',7'-dichlorodihydrofluorescein diacetate in the cyanobacterium Anabaena variabilis PCC 7937

    SciTech Connect

    Rastogi, Rajesh P.; Laboratory of Photobiology and Molecular Microbiology, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005 ; Singh, Shailendra P.; Haeder, Donat-P.; Sinha, Rajeshwar P.

    2010-07-02

    The generation of reactive oxygen species (ROS) under simulated solar radiation (UV-B: 0.30 Wm{sup -2}, UV-A: 25.70 Wm{sup -2} and PAR: 118.06 Wm{sup -2}) was studied in the cyanobacterium Anabaena variabilis PCC 7937 using the oxidant-sensing fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). DCFH-DA is a nonpolar dye, converted into the polar derivative DCFH by cellular esterases that are nonfluorescent but switched to highly fluorescent DCF when oxidized by intracellular ROS and other peroxides. The images obtained from the fluorescence microscope after 12 h of irradiation showed green fluorescence from cells covered with 295, 320 or 395 nm cut-off filters, indicating the generation of ROS in all treatments. However, the green/red fluorescence ratio obtained from fluorescence microscopic analysis showed the highest generation of ROS after UV-B radiation in comparison to PAR or UV-A radiation. Production of ROS was also measured by a spectrofluorophotometer and results obtained supported the results of fluorescence microscopy. Low levels of ROS were detected at the start (0 h) of the experiment showing that they are generated even during normal metabolism. This study also showed that UV-B radiation causes the fragmentation of the cyanobacterial filaments which could be due to the observed oxidative stress. This is the first report for the detection of intracellular ROS in a cyanobacterium by fluorescence microscopy using DCFH-DA and thereby suggesting the applicability of this method in the study of in vivo generation of ROS.

  17. A new staining method for identification of Helicobacter pylori and simultaneous visualization of gastric morphologic features.

    PubMed

    Cohen, L F; Sayeeduddin, M; Phillips, C; Shahab, I

    1997-11-01

    Helicobacter pylori (HP) is prevalent in the general population and is associated with chronic active gastritis, peptic ulcers, gastric carcinoma, and lymphoma. Different methods to diagnose HP colonization include the urea breath test, serologic analysis, and gastric biopsy. Many different staining methods, including silver and Giemsa-based stains, have been used to demonstrate these organisms in gastric biopsy specimens. The tissue morphologic features, however, are often obscured, thus mandating evaluation of both hematoxylin and eosin (H&E) and special stains. This study evaluates the ability of a new staining method that allows simultaneous identification of HP and visualization of tissue morphologic features. We examined formalin-fixed, paraffin-embedded archival tissues from 184 consecutive gastric biopsy specimens using our new staining method, and we compared our results with those previously obtained from H&E and Warthin-Starry (WS) stains. Our new stain uses periodic acid-Schiff, Coleman's Feulgen solution, Mayer's hematoxylin, and methylene blue. Our method is technically simpler than the WS stain and can be completed in approximately 9 minutes. We found HP organisms in 76 (41%) of 184 biopsy specimens by H&E and/or WS staining. Our new staining method identified HP in 83 specimens (45%). The organisms stained bright blue and were easily visualized (compared with the H&E-stained specimens) because of the contrasting magenta mucin. Tissue morphologic features consisted of well-defined cells with dark blue nuclei and pale cytoplasm. Intestinal metaplasia was highlighted magenta Our new staining method revealed chronic active gastritis in 71 cases (39%), intestinal metaplasia in 18 cases (10%), and lymphoid aggregates in 35 cases (19%). These findings were similar to the results obtained with H&E-stained sections. We conclude that our new staining method is inexpensive, quick, and easy to perform and interpret. It has a sensitivity comparable to that of conventional staining methods and simultaneously demonstrates both tissue morphologic features and the presence of HP. PMID:9388068

  18. Determination of total antioxidant capacity by oxygen radical absorbance capacity (ORAC) using fluorescein as the fluorescence probe: First Action 2012.23.

    PubMed

    Ou, Boxin; Chang, Tony; Huang, Dejian; Prior, Ronald L

    2013-01-01

    An improved method for the measurement of oxygen radical absorbance capacity (ORAC) was developed and validated using fluorescein (3',6'-dihydroxyspiro[isobenzofuran-1[3H], 9'[9H]-xanthen]-3-one) as a new fluorescence probe (ORAC(FL)). Randomly methylated beta-cyclodextrin (RMCD) was introduced as the water-solubility enhancer for lipophilic antioxidants. 7% RMCD (w/v) in 50% acetone-H2O mixture sufficiently solubilized vitamin E compounds and other lipophilic phenolic antioxidants in 75 mM phosphate buffer (pH 7.4). Results indicated that fluorescein shows excellent photostability under the plate reader conditions. This ORAC(FL) was validated through linearity, precision, accuracy, and ruggedness. The validation results demonstrated that the ORACFL assay is reliable and robust. The mean of intraday and interday CVs were <15%; for hydrophilic ORAC, LOD and LOQ are 5 and 6.25 microM, respectively; for lipophilic ORAC, LOD and LOQ are 6.25 and 12.5 microM, respectively. It is concluded that unlike other popular methods, the ORAC(FL) assay provides a direct measure of total antioxidant capacity against the peroxyl radicals. PMID:24645517

  19. Evaporation of a sessile droplet: Inside the coffee stain Guillaume Berteloot a,

    E-print Network

    Daerr, Adrian - Laboratoire Matière et Systèmes Complexes, Université Paris 7

    Evaporation of a sessile droplet: Inside the coffee stain Guillaume Berteloot a, , Anna Hoang Evaporation Deposition Coffee stain a b s t r a c t We have investigated experimentally, for the first time (``coffee stain effect''). Direct observations show that there are several distinct phases of growth

  20. LaTeX Coffee Stains http://hanno-rein.de

    E-print Network

    Gerhardy, Philipp

    LaTeX Coffee Stains Hanno Rein http://hanno-rein.de Cambridge University April 3, 2009 1 a coffee stain to your documents. A lot of time can be saved by printing stains directly on the page rather Usage To use the package, simply place the coffee.sty file in the directory with all of your other .tex

  1. Pro-Q Sapphire 365 Oligohistidine Gel StainMP 21876 Revised: 08.ebruary2002

    E-print Network

    Lebendiker, Mario

    Pro-Q Sapphire 365 Oligohistidine Gel StainMP 21876 Revised: 08.ebruary2002 Product Information-Q Sapphire 365 Oligohistidine Gel Stain (P-21876) Introduction The oligohistidine domain is a Ni2+ -binding protein can be easily purified using a nickel-chelating resin. Pro-Q Sapphire 365 oligohistidine gel stain

  2. Pro-Q Sapphire 488 Oligohistidine Gel StainMP 21877 Revised: 08.ebruary2002

    E-print Network

    Lebendiker, Mario

    Pro-Q Sapphire 488 Oligohistidine Gel StainMP 21877 Revised: 08.ebruary2002 Product Information-Q Sapphire 488 Oligohistidine Gel Stain (P-21877) Introduction The oligohistidine domain is a Ni2+ -binding protein can be easily purified using a nickel-chelating resin. Pro-Q Sapphire 488 oligohistidine gel stain

  3. SYPRO Orange and SYPRO Red Protein Gel StainsMP 06650 Revised: 17-January-2003

    E-print Network

    Lebendiker, Mario

    SYPRO® Orange and SYPRO® Red Protein Gel StainsMP 06650 Revised: 17-January-2003 Product Information Introduction Molecular Probes proprietary SYPRO® Orange and SYPRO® Red protein gel stains provide over conventional colorimetric stains: $ High sensitivity. SYPRO Orange and SYPRO Red protein gel

  4. A Comparison of Heat versus Methanol Fixation for Gram Staining Bacteria

    ERIC Educational Resources Information Center

    Minnerath, Jeanne M.; Roland, Jenna M.; Rossi, Lucas C.; Weishalla, Steven R.; Wolf, Melissa M.

    2009-01-01

    Gram staining bacteria is a fundamental technique introduced in general biology and microbiology laboratory courses. Two common problems students encounter when Gram staining bacteria are (1) having a difficult time locating bacterial cells on the microscope slide and (2) over-decolorizing bacterial cells during the staining procedure such that…

  5. IMAGING OF PORT WINE STAIN LESIONS USING A MULTI-SENSOR PHOTOACOUSTIC PROBE

    E-print Network

    Aguilar, Guillermo

    IMAGING OF PORT WINE STAIN LESIONS USING A MULTI-SENSOR PHOTOACOUSTIC PROBE John A. Viator, Steven, California, 92521 gaguilar@engr.ucr.edu ABSTRACT Successful treatment of port wine stain (PWS) birthmarks and distribution of melanin are required to optimize a laser procedure, such as laser therapy of port wine stain

  6. X-ray Microtomography Developed high Z element staining to enhance the visualization of

    E-print Network

    Illustrate how biofilms interact with liquids or substrates and reveal how microbe-scale interactions Stained Unstained Research Accomplishments Biofilm Cryogenic Preparation and Chemical Imaging Cryogenic Preparation Cryogenic preparation STXM spectroscopy cells EPS cells EPS Unstained Stained Stained/hydrated 0

  7. OIL RED AS A HISTOCHEMICAL STAIN FOR NATURAL FIBERS AND PLANT CUTICLE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Oil Red was evaluated as a histochemical stain for the cuticle of plants and for components in cotton and flax fibers. A positive reaction for arachidyl stearate and as well as differential staining of plants after sequential extraction of fatty acids and alcohols confirmed that Oil Red stained wa...

  8. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  9. IEEE TRANSACTIONS ON VISUALIZATION AND COMPUTER GRAPHICS 1 Image-Based Stained Glass

    E-print Network

    Brooks, Stephen

    IEEE TRANSACTIONS ON VISUALIZATION AND COMPUTER GRAPHICS 1 Image-Based Stained Glass Stephen Brooks of a work of stained glass. To this end, we develop a novel approach which involves image warping is first segmented. Each segment is subsequently transformed to match real segments of stained glass

  10. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 19 Customs Duties 1 2013-04-01 2013-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  11. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 19 Customs Duties 1 2011-04-01 2011-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  12. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 19 Customs Duties 1 2014-04-01 2014-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  13. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 19 Customs Duties 1 2012-04-01 2012-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  14. Digital simulation of staining in histopathology multispectral images: enhancement and linear transformation of spectral transmittance.

    PubMed

    Bautista, Pinky A; Yagi, Yukako

    2012-05-01

    Hematoxylin and eosin (H&E) stain is currently the most popular for routine histopathology staining. Special and/or immuno-histochemical (IHC) staining is often requested to further corroborate the initial diagnosis on H&E stained tissue sections. Digital simulation of staining (or digital staining) can be a very valuable tool to produce the desired stained images from the H&E stained tissue sections instantaneously. We present an approach to digital staining of histopathology multispectral images by combining the effects of spectral enhancement and spectral transformation. Spectral enhancement is accomplished by shifting the N-band original spectrum of the multispectral pixel with the weighted difference between the pixel's original and estimated spectrum; the spectrum is estimated using M < N principal component (PC) vectors. The pixel's enhanced spectrum is transformed to the spectral configuration associated to its reaction to a specific stain by utilizing an N × N transformation matrix, which is derived through application of least mean squares method to the enhanced and target spectral transmittance samples of the different tissue components found in the image. Results of our experiments on the digital conversion of an H&E stained multispectral image to its Masson's trichrome stained equivalent show the viability of the method. PMID:22612136

  15. Coproduction of detergent compatible bacterial enzymes and stain removal evaluation.

    PubMed

    Niyonzima, Francois N; More, Sunil S

    2015-10-01

    Most of the detergents that are presently produced contain the detergent compatible enzymes to improve and accelerate the washing performance by removing tough stains. The process is environment friendly as the use of enzymes in the detergent formulation reduces the utilization of toxic detergent constituents. The current trend is to use the detergent compatible enzymes that are active at low and ambient temperature in order to save energy and maintain fabric quality. As the detergent compatible bacterial enzymes are used together in the detergent formulation, it is important to co-produce the detergent enzymes in a single fermentation medium as the enzyme stability is assured, and production cost gets reduced enormously. The review reports on the production, purification, characterization and application of detergent compatible amylases, lipases, and proteases are available. However, there is no specific review or minireview on the concomitant production of detergent compatible amylases, lipases, and proteases. In this minireview, the coproduction of detergent compatible enzymes by bacterial species, enzyme stability towards detergents and detergent components, and stain release analysis were discussed. PMID:26011283

  16. Antibody Staining in C. Elegans Using "Freeze-Cracking"

    PubMed Central

    Duerr, Janet S.

    2013-01-01

    To stain C. elegans with antibodies, the relatively impermeable cuticle must be bypassed by chemical or mechanical methods. "Freeze-cracking" is one method used to physically pull the cuticle from nematodes by compressing nematodes between two adherent slides, freezing them, and pulling the slides apart. Freeze-cracking provides a simple and rapid way to gain access to the tissues without chemical treatment and can be used with a variety of fixatives. However, it leads to the loss of many of the specimens and the required compression mechanically distorts the sample. Practice is required to maximize recovery of samples with good morphology. Freeze-cracking can be optimized for specific fixation conditions, recovery of samples, or low non-specific staining, but not for all parameters at once. For antibodies that require very hard fixation conditions and tolerate the chemical treatments needed to chemically permeabilize the cuticle, treatment of intact nematodes in solution may be preferred. If the antibody requires a lighter fix or if the optimum fixation conditions are unknown, freeze-cracking provides a very useful way to rapidly assay the antibody and can yield specific subcellular and cellular localization information for the antigen of interest. PMID:24145964

  17. Analysis of surface stains on modern gold coins

    NASA Astrophysics Data System (ADS)

    Corregidor, V.; Alves, L. C.; Cruz, J.

    2013-07-01

    It is a mandatory practice in the European Mint Houses to provide a certificate of guarantee of their products specially when issuing commemorative gold or silver coins. This practise should assure satisfaction and trust both for the mint house and for the demanding numismatic collector. For these reasons the Mint Houses follow a strict quality control in all the production steps in order to ensure a no-defect, fully supervised output. In spite of all the undertaken precautions, different surface stains with diverse origin on gold coins recently minted in Europe were observed. Those were compositionally studied by means of IBA techniques at the end-stage nuclear microprobe installed at IST/ITN. From this study it was possible to identify several possible sources for these stains. The presence of defects at the surface of these commemorative coins address the need of improving the quality control system and the results here presented point out where these improvements should occur, in order to reduce/eliminate them and give the customer a product that with time probably will be revalued.

  18. Sperm DNA fragmentation is related to sperm morphological staining patterns.

    PubMed

    Sá, Rosália; Cunha, Mariana; Rocha, Eduardo; Barros, Alberto; Sousa, Mário

    2015-10-01

    In this prospective comparative study, sperm DNA fragmentation (sDNAfrag) was compared at each step of a sequential semen preparation, with semen parameters according to their degree of severity. At each step (fractions) of the sequential procedure, sDNAfrag was determined: fresh (Raw), after gradient centrifugation, washing, and swim-up (SU) for 70 infertile men enrolled in intracytoplasmic sperm injection cycles. sDNAfrag significantly (P = 0.04; P < 0.0001) decreased throughout the steps of semen preparation, with centrifugation and washing not increasing it. A negative correlation to sperm motility was observed in Raw and SU fractions, and a higher sDNAfrag was observed in samples with lower semen quality. Our results confirm that the steps of the sequential procedure do not compromise sperm DNA integrity and progressively decreased sDNAfrag regardless of the sperm abnormality and that semen parameters with lower quality present higher sDNAfrag. Four distinct patterns were observed, of which the entire sperm head staining was the pattern most expressed in all studied fractions. Additionally, the sperm head gene-rich region staining pattern was reduced by the procedure. This suggests that pattern quantification might be a useful adjunct when performing sDNAfrag testing for male infertility. PMID:26278809

  19. Color stability of ceramic brackets immersed in potentially staining solutions

    PubMed Central

    Guignone, Bruna Coser; Silva, Ludimila Karsbergen; Soares, Rodrigo Villamarim; Akaki, Emilio; Goiato, Marcelo Coelho; Pithon, Matheus Melo; Oliveira, Dauro Douglas

    2015-01-01

    OBJECTIVE: To assess the color stability of five types of ceramic brackets after immersion in potentially staining solutions. METHODS: Ninety brackets were divided into 5 groups (n = 18) according to brackets commercial brands and the solutions in which they were immersed (coffee, red wine, coke and artificial saliva). The brackets assessed were Transcend (3M/Unitek, Monrovia, CA, USA), Radiance (American Orthodontics, Sheboygan, WI, USA), Mystique (GAC International Inc., Bohemia, NY, USA) and Luxi II (Rocky Mountain Orthodontics, Denver, CO, USA). Chromatic changes were analyzed with the aid of a reflectance spectrophotometer and by visual inspection at five specific time intervals. Assessment periods were as received from the manufacturer (T0), 24 hours (T1), 72 hours (T2), as well as 7 days (T3) and 14 days (T4) of immersion in the aforementioned solutions. Results were submitted to statistical analysis with ANOVA and Bonferroni correction, as well as to a multivariate profile analysis for independent and paired samples with significance level set at 5%. RESULTS: The duration of the immersion period influenced color alteration of all tested brackets, even though these changes could not always be visually observed. Different behaviors were observed for each immersion solution; however, brackets immersed in one solution progressed similarly despite minor variations. CONCLUSIONS: Staining became more intense over time and all brackets underwent color alterations when immersed in the aforementioned solutions. PMID:26352842

  20. Comparison of animal infectivity, excystation, and fluorogenic dye as measures of Giardia muris cyst inactivation by ozone

    SciTech Connect

    Labatiuk, C.W.; Finch, G.R.; Belosevic, M. ); Schaefer, F.W. III )

    1991-11-01

    Giardia muris cyst viability after ozonation was compared by using fluorescein diacetate-ethidium bromide staining, the C3H/HeN mouse-G. muris model, and in vitro excystation. Bench-scale batch experiments were conducted under laboratory conditions (pH 6.7, 22C) in ozone-demand-free phosphate buffer. There was a significant difference between fluorogenic staining and infectivity with fluorogenic staining overestimating viability compared with infectivity estimates of viability. This suggests that viable cysts as indicated by fluorogenic dyes may not be able to complete the life cycle and produce an infection. No significant differences between infectivity and excystation and between fluorogenic staining and excystation were detected for inactivations up to 99.9%. Only animal infectivity had the sensitivity to detect inactivations greater than 99.9%. Therefore, the animal model is the best method currently available for detecting high levels of G. muris cyst inactivation.

  1. Combined methenamine-silver nitrate and hematoxylin & eosin stain for fungi in tissues.

    PubMed Central

    Huppert, M; Oliver, D J; Sun, S H

    1978-01-01

    Initial examination of hematoxylin & eosin-stained tissue from a human brain specimen did not reveal the fungi which were seen in subsequent tissue sections stained with methenamine-silver nitrate. Microabscesses seen in the hematoxylin & eosin-stained sections were not apparent in the methenamine--silver nitrate-stained tissue. Staining with methenamine--silver nitrate and counterstaining with hematoxylin & eosin proved excellent not only for detecting fungus cells, but also for revealing their relationship to the host cellular response in this case and in examples of experimental murine coccidioidomycosis and histoplasmosis. Images PMID:83328

  2. Posterior capsule staining and posterior continuous curvilinear capsulorhexis in congenital cataract.

    PubMed

    Wakabayashi, Taketoshi; Yamamoto, Narumichi

    2002-11-01

    We report 2 cases of indocyanine green (ICG) staining used for posterior continuous curvilinear capsulorhexis (PCCC) in congenital cataract combined with anterior vitrectomy. In the first case, because of corneal opacity, the visibility of the posterior capsule was poor without staining. After the extraction of the cataract, a PCCC was performed after ICG staining of the posterior capsule. In the second case, after cataract removal, ICG staining was used to better visualize the posterior capsule. In both cases, the PCCC was successfully completed because of better visualization of the stained posterior capsule flap against the transparent anterior hyaloid face of the vitreous. Clear visual axes have been maintained. PMID:12457683

  3. Accomplishments of the Trustees and laboratory staff of the Biological Stain Commission, 2002-2013.

    PubMed

    Dapson, R W

    2014-08-01

    During the 12 years from 2002 to 2013, the Trustees and laboratory personnel of the Biological Stain Commission (BSC) can claim many accomplishments. These accomplishments are itemized under 11 categories: continuous publication of the official journal, Biotechnic & Histochemistry; production of four special issues of Biotechnic & Histochemistry devoted to specific dyes or stains; standardization of staining and dye purity; mechanisms of staining and prediction of dye behavior; publication of books or book chapters; effects of fixation and processing on staining; cancer research; immunohistochemistry; BSC Laboratory activities; miscellaneous publications; and administrative accomplishments. PMID:24665939

  4. Targeted alteration of real and imaginary refractive index of biological cells by histological staining.

    PubMed

    Cherkezyan, L; Subramanian, H; Stoyneva, V; Rogers, J D; Yang, S; Damania, D; Taflove, A; Backman, V

    2012-05-15

    Various staining techniques are commonly used in biomedical research to investigate cellular morphology. By inducing absorption of light, staining dyes change the intracellular refractive index due to the Kramers-Kronig relationship. We present a method for creating 2D maps of real and imaginary refractive indices of stained biological cells using their thickness and absorptance. We validate our technique on dyed polystyrene microspheres and quantify the alteration in refractive index of stained biological cells. We reveal that specific staining of individual organelles can increase their scattering cross-section by orders of magnitudes, implying a major impact in the field of biophotonics. PMID:22627509

  5. Staining with 0.05% neutral red reduces nutrient uptake by wheat roots.

    PubMed

    Trolove, Stephen; Tan, Yong; Reid, Jeff

    2015-11-01

    A number of studies have used a 0.05% solution of neutral red to stain live roots so that short term root growth could be measured. These studies, which used a 5 or 10 min staining time, report no effects of the stain on plant properties such as growth, respiration, or nitrate uptake. This paper reports on two experiments conducted to determine whether this staining technique, with a 15 min stain time, affected macronutrient uptake of 6- and 7-week-old wheat (Triticum aestivum L.) plants grown in solution culture. The results showed that, compared with unstained controls, staining plants with 0.05% neutral red halted or halved nitrate uptake measured over a 4 h period the following day. Potassium uptake was also significantly reduced by staining. In the experiment with smaller plants nutrient uptake rate recovered 5 days after staining, but not in the second experiment with larger plants. Stained roots were 19% narrower than unstained roots, suggesting that the stain affected the root structure. We do not recommend the use of 0.05% neutral red staining, for wheat at least, in experiments where accurate measurement of nutrient uptake rate is important. PMID:26379198

  6. Sizing of single fluorescently stained DNA fragments by scanning microscopy

    PubMed Central

    Laib, Stephan; Rankl, Michael; Ruckstuhl, Thomas; Seeger, Stefan

    2003-01-01

    We describe an approach to determine DNA fragment sizes based on the fluorescence detection of single adsorbed fragments on specifically coated glass cover slips. The brightness of single fragments stained with the DNA bisintercalation dye TOTO-1 is determined by scanning the surface with a confocal microscope. The brightness of adsorbed fragments is found to be proportional to the fragment length. The method needs only minute amount of DNA, beyond inexpensive and easily available surface coatings, like poly-l-lysine, 3-aminoproyltriethoxysilane and polyornithine, are utilizable. We performed DNA-sizing of fragment lengths between 2 and 14 kb. Further, we resolved the size distribution before and after an enzymatic restriction digest. At this a separation of buffers or enzymes was unnecessary. DNA sizes were determined within an uncertainty of 7–14%. The proposed method is straightforward and can be applied to standardized microtiter plates. PMID:14602931

  7. Fat tissue staining and photodynamic/photothermal effects

    NASA Astrophysics Data System (ADS)

    Tuchin, Valery V.; Altshuler, Gregory B.; Yanina, Irina Yu.; Kochubey, Vyacheslav I.; Simonenko, Georgy V.

    2010-02-01

    Cellulite is considered as a disease of the subcutaneous fat layer that appears mostly in women and consists of changes in fat cell accumulation together with disturbed lymphatic drainage, affecting the external appearance of the skin. The photodynamic and selective photothermal treatments may provide reduction the volume of regional or sitespecific accumulations of subcutaneous adipose tissue on the cellular level. We hypothesize that light irradiation of stained fat tissue at selected temperature leads to fat cell lypolytic activity (the enhancement of lipolysis of cell triglycerides due to expression of lipase activity and cell release of free fat acids (FFAs) due to temporal cell membrane porosity), and cell killing due to apoptosis caused by the induced fat cell stress and/or limited cell necrosis.

  8. Disseminated zygomycosis associated with erythroleukaemia: confirmation by lectin stains.

    PubMed Central

    Benbow, E W; Delamore, I W; Stoddart, R W; Reid, H

    1985-01-01

    Zygomycosis is not often diagnosed in the United Kingdom, and so the possible importance of the findings in a patient with disseminated zygomycosis who had been treated with chemotherapy for erythroleukaemia was not appreciated until histological examination of specimens obtained at necropsy provided a presumptive diagnosis. No attempt had therefore been made to identify the organism by culture, and lectin binding methods were used to try to compensate for this. The characteristics of the hyphae on staining with lectins were similar to those previously shown in Rhizopus oryzae and were unlike those of a wide range of other hyphal fungi. Although definite speciation of the fungus was not achieved, these findings confirm that this was a case of zygomycosis and would seem to represent the first such reported confirmation in the absence of culture. Images PMID:2413080

  9. Pulsed photothermal radiometry of port-wine stains

    NASA Astrophysics Data System (ADS)

    Milner, Thomas E.; Nelson, J. Stuart; Tran, N. Q.; Katzir, Abraham; Svaasand, Lars O.; Jacques, Steven L.

    1993-07-01

    The application of pulsed photothermal radiometry (PPTR) diagnostics to characterize port wine stain (PWS) lesions is discussed. Influence of epidermal absorbance and PWS depth on the PPTR signal is analyzed using an in-vitro model of PWS skin consisting of multilayered collagen films. An instrument is constructed and used for patient and animal model PPTR measurements. When an infrared fiber is incorporated, the instrumentation facilitates convenient skin-site accessibility. Modulation of the PPTR signal and homodyne detection substantially improve the system signal-to-noise ratio over previous methods. Given the measured PPTR signal, an algorithm computes the initial temperature distribution in the PWS skin immediately after the laser pulse. The use and limitations of the algorithm are presented and discussed.

  10. Photonic Crystal Hydrogel Enhanced Plasmonic Staining for Multiplexed Protein Analysis.

    PubMed

    Mu, Zhongde; Zhao, Xiangwei; Huang, Yin; Lu, Meng; Gu, Zhongze

    2015-12-01

    Plasmonic nanoparticles are commonly used as optical transducers in sensing applications. The optical signals resulting from the interaction of analytes and plamsonic nanoparticles are influenced by surrounding physical structures where the nanoparticles are located. This paper proposes inverse opal photonic crystal hydrogel as 3D structure to improve Raman signals from plasmonic staining. By hybridization of the plasmonic nanoparticles and photonic crystal, surface-enhanced Raman spectroscopy (SERS) analysis of multiplexed protein is realized. It benefits the Raman analysis by providing high-density "hot spots" in 3D and extra enhancement of local electromagnetic field at the band edge of PhC with periodic refractive index distribution. The strong interaction of light and the hybrid 3D nanostructure offers new insights into plasmonic nanoparticle applications and biosensor design. PMID:26436833

  11. Discoloration of coating resins exposed to staining solutions in vitro.

    PubMed

    Manabe, Atsufumi; Kato, Yukiyo; Finger, Werner J; Kanehira, Masafumi; Komatsu, Masashi

    2009-05-01

    The purpose of this in vitro investigation was to study the effects of coffee and tea immersion on surface discoloration of one commercial temporary resin coating material, White Coat (WHC; Kuraray Medical Inc., Tokyo, Japan), and an experimental one, SI-R20209 (SIR; Shofu Inc., Kyoto, Japan). Disk-shaped specimens were prepared, their colors were determined at baseline, and after immersion in water (control), tea and coffee solutions for 24, 48, and 72 hours. Very little discoloration was found with the water-stored specimens. Staining response was most pronounced after coffee immersion for White Coat and after tea immersion for the experimental material, exceeding the clinically acceptable discoloration threshold value of deltaE=3.3. However, most of the resin shades tested are likely to be sufficiently safe against heavy discoloration when used for short-term restoration only. PMID:19662733

  12. Multi-stained whole slide image alignment in digital pathology

    NASA Astrophysics Data System (ADS)

    Déniz, Oscar; Toomey, David; Conway, Catherine; Bueno, Gloria

    2015-03-01

    In Digital Pathology, one of the most simple and yet most useful feature is the ability to view serial sections of tissue simultaneously on a computer monitor. This enables the pathologist to evaluate the histology and expression of multiple markers for a patient in a single review. However, the rate limiting step in this process is the time taken for the pathologist to open each individual image, align the sections within the viewer, with a maximum of four slides at a time, and then manually move around the section. In addition, due to tissue processing and pre-analytical steps, sections with different stains have non-linear variations between the two acquisitions, that is, they will stretch and change shape from section to section. To date, no solution has come close to a workable solution to automatically align the serial sections into one composite image. This research work address this problem to obtain an automated serial section alignment tool enabling the pathologists to simply scroll through the various sections in a single viewer. To this aim a multi-resolution intensity-based registration method using mutual information as a similarity metric, an optimizer based on an evolutionary process and a bilinear transformation has been used. To characterize the performance of the algorithm 40 cases x 5 different serial sections stained with hematoxiline-eosine (HE), estrogen receptor (ER), progesterone receptor (PR), Ki67 and human epidermal growth factor receptor 2 (Her2), have been considered. The qualitative results obtained are promising, with average computation time of 26.4s for up to 14660x5799 images running interpreted code.

  13. Surface discoloration of composite resins: Effects of staining and bleaching

    PubMed Central

    Poggio, Claudio; Beltrami, Riccardo; Scribante, Andrea; Colombo, Marco; Chiesa, Marco

    2012-01-01

    Background: The purpose of this in vitro study was to evaluate surface discoloration of three microhybrid composite resins (Esthet•X HD, Clearfil AP-X, Gradia Direct) and five nanohybrid composite resins (Ceram•X, GC Kalore, G-aenial, Grandio, GrandioSO), after staining and bleaching procedures. Materials and Methods: The composite resins were polymerized with a curing light (Celalux II, Voco, Cuxhaven, Germany) into 160 silicon molds (6,4 mm in diameter and 2 mm in thickness) to obtain identical specimens. Twenty samples for each composite resin were prepared. The specimens were polished using an automated polishing machine with the sequence of 600-, 800-, 1000-grit abrasive paper under water irrigation. The specimens were immersed in tea and distilled water: the specimens were dipped for 20 min, once a day (every 24 h), for 14 days into the drinks. The specimens were then bleached with carbamide peroxide at 17% (Perfect Bleach-Voco). The color of specimens was measured with a spectrophotometer according to the CIE L*a*b* system after light-polymerization of composite resin specimens, after 7 days, after 14 days, and after bleaching. The color difference h index (DEab*) between each measurement was calculated. Statistical analysis was made using analysis of variance (ANOVA). Results: All specimens showed a significant increase in staining with a similar trend and no significant differences between microhybrid and nanohybrid composite resins. After whitening procedures, materials tested showed both significant and unsignificant differences of the h index. Conclusions: Microhybrid and nanohybrid composite resins had similar in vitro surface discoloration in tea. After bleaching, discoloration was removed from some composite resins tested. PMID:23559921

  14. A Novel Contrast Stain for the Rapid Diagnosis of Pityriasis Versicolor: A Comparison of Chicago Sky Blue 6B Stain, Potassium Hydroxide Mount and Culture

    PubMed Central

    Lodha, Nikita; Poojary, Shital Amin

    2015-01-01

    Background: The mycological study of pityriasis versicolor is usually done by potassium hydroxide (KOH) mount and culture. However, KOH mount lacks a color contrast and requires a trained eye to interpret, while culture is difficult to perform, time consuming and has low sensitivity. Chicago Sky Blue 6B (CSB) is a new contrast stain that highlights the fungal hyphae and spores, blue against a purplish background. Aims and Objectives: This study was done to compare the utility of a novel contrast stain (CSB stain) with KOH mount and culture. Materials and Methods: Skin scrapings from the lesions of 100 clinically diagnosed cases of P. versicolor were subjected to (1) KOH mount and CSB stain for direct microscopic examination and (2) culture using Sabouraud's dextrose agar. The statistical analysis of CSB stain and culture was done using KOH mount as the reference method, as it is the most commonly performed and practical diagnostic test available for P. versicolor. An interrater reliability analysis using the Cohen's Kappa statistic was performed to determine consistency (agreement) among the different modalities. Observations and Results: Direct microscopy with CSB stain, KOH mount and mycological culture showed positive results in 98 (98%), 92 (92%) and 56 (56%) patients, respectively. Using KOH mount as the reference method, CSB stain had a sensitivity of 100% which was significantly higher than culture (60.9%). Statistically significant fair agreement was found between CSB stain and KOH mount (94% with ?=0.38, P < 0.001). Negligible agreement was found between CSB stain and culture (66%, ?=0.199, P = 0.001) as well as between KOH mount and culture (64%, ?=0.051, P = 0.107). Conclusion: CSB staining of skin scrapings is the most sensitive method for the diagnosis of pityriasis versicolor. Due to the distinct contrast provided by CSB, it is easy to perform, rapid and qualitatively superior to KOH mount. PMID:26288400

  15. Metal ion complexes of N,N'-bis(2-pyridylmethyl)-1,3-diaminopropane-N,N'-diacetic acid, H2bppd.

    PubMed

    Kissel, Daniel S; Florián, Jan; McLauchlan, Craig C; Herlinger, Albert W

    2014-04-01

    A higher yield synthesis of N,N'-bis(2-pyridylmethyl)-1,3-diaminopropane-N,N'-diacetic acid (H2bppd) and its complexation of trivalent metal ions (Al(III), Ga(III), In(III)) and selected lanthanides (Ln(III)) are reported. H2bppd and the metal-bppd(2-) complexes, isolated as hexafluorophosphate salts, were characterized by elemental analysis, mass spectrometry, IR, and (1)H and (13)C NMR spectroscopy. [Ga(bppd)]PF6, [Ga(C19H22N4O4)]PF6, was crystallized as colorless needles by slow evaporation from anhydrous methanol; its molecular structure was solved by direct X-ray crystallography methods. The compound crystallized in the monoclinic space group P21/c, with a = 9.6134(2) Å, b = 20.2505(4) Å, c = 11.6483(3) Å, ? = 97.520(1)(o), and Z = 4. Ga is coordinated in a distorted octahedral geometry provided by a N4O2 donor atom set with cis-monodentate acetate groups and cis-2-pyridylmethyl N atoms. Quantum mechanical calculations were performed for the three possible geometric isomers of a pseudo-octahedral metal-bppd(2-) complex with five different metal ions. The results indicate, that in aqueous solution, the stability of the trans-O,O isomer is similar to that of the cis-O,O; cis-Npy,Npy isomer but is greater than that of the trans-Npy,Npy isomer. Calculations for a six-coordinate La(III)-bppd(2-) complex converge to a structure with a very large Npy-La-Npy bond angle (146.4°), a high metal charge (2.28 au), and a high solvation free energy (-79.4 kcal/mol). The most stable geometric arrangement for bppd(2-) around the larger La(III) is best described as an open nestlike structure with space available for additional ligands. IR spectroscopy was used to investigate the nature of the H2bppd-metal complexes isolated in the solid state and the binding modes of the carboxylate functionalities. The spectra indicate that fully deprotonated [M(bppd)](+) complexes as well as partially protonated complexes [M(Hbppd)Cl](+) were isolated. The (1)H and (13)C assignments for H2bppd and metal-bppd(2-) complexes were made on the basis of 2D COSY, NOESY, and (1)H-(13)C HSQC experiments, which were used to differentiate among the cis (C1 symmetry) and the two trans (C2 symmetry) isomers. PMID:24649926

  16. Spectrofluorometric determination of intracellular levels of reactive oxygen species in drug-sensitive and drug-resistant cancer cells using the 2?,7?-dichlorofluorescein diacetate assay

    NASA Astrophysics Data System (ADS)

    Loetchutinat, Chatchanok; Kothan, Suchart; Dechsupa, Samarn; Meesungnoen, Jintana; Jay-Gerin, Jean-Paul; Mankhetkorn, Samlee

    2005-02-01

    This article examines a non-invasive spectrofluorometric method using the 2',7'-dichlorofluorescein diacetate (DCHF-DA) assay for quantifying the intracellular reactive oxygen species (ROS i) produced in four cultured cancer cell lines: drug-sensitive (K562) and drug-resistant (K562/ adr) human erythromyelogenous leukemia cell lines, and drug-sensitive (GLC4) and drug-resistant (GLC4/ adr) human small cell lung carcinoma cell lines. The oxidation of the probe to the fluorescent dichlorofluorescein (DCF) was continuously monitored by following the DCF fluorescence intensity as a function of time using a standard spectrofluorometer in the presence of an extracellular DCF fluorescence quencher (Co 2+). By fitting the spectrofluorometric data to a kinetic model based on the following two reactions: (i) deacetylation of DCHF-DA to the oxidant-sensitive compound 2',7'-dichlorofluorescein (DCHF) by cellular esterase enzymes (pseudo-first-order rate constant: ke) and (ii) oxidation of DCHF by ROS i (second-order rate constant: k2), the parameters intervening in DCF formation, ke and the product of k2 by the ROS i concentration, were quantitatively determined for the different cell lines studied. The results revealed that the intracellular esterase content or activity is similar in K562, K562/ adr, and GLC4 cells, but 5-fold higher in GLC4/ adr cells. The product k2[ROS i] was found to be similar in the four cell lines considered, with a mean value of (5.3±0.9)×10 -7 cell -1 s -1. Assuming that H 2O 2 (in combination with peroxidases) is the primary responsible species for DCHF oxidation in intact cells, and using the rate constant value k2=790±62 M s established in our laboratory for the reaction of DCHF with H 2O 2 in the presence of horseradish peroxidase, the mean value of the intracellular levels of ROS i in those cells was estimated to be 0.67±0.16 nM per cell. Such a value compares favorably to H 2O 2 intracellular steady-state concentrations that have been estimated in the literature for a few other cell types.

  17. Detection of infection or infectious agents by use of cytologic and histologic stains.

    PubMed Central

    Woods, G L; Walker, D H

    1996-01-01

    A wide variety of stains are useful for detection of different organisms or, for viruses, the cytopathologic changes they induce, in smears prepared directly from clinical specimens and in tissue sections. Other types of stains, such as hematoxylin and eosin, are used routinely to stain tissue sections and are most valuable for assessing the immunologic response of the host to the invading pathogen. In many cases, the pattern of inflammation provides important clues to diagnosis and helps to guide the selection of additional "special" stains used predominantly for diagnosis of infectious diseases. A stain may be nonspecific, allowing detection of a spectrum of organisms, as do the Papanicolaou stain and silver impregnation methods, or detection of only a limited group of organisms, as do the different acid-fast techniques. Some nonspecific stains, such as the Gram stain, are differential and provide valuable preliminary information concerning identification. Immunohistochemical stains, on the other hand, are specific for a particular organism, although in some cases cross-reactions with other organisms occur. Despite the wealth of information that can be gleaned from a stained smear or section of tissue, however, the specific etiology of an infection often cannot be determined on the basis of only the morphology of the organisms seen; culture data are essential and must be considered in the final diagnosis. PMID:8809467

  18. Analysis of the Microbiota of Black Stain in the Primary Dentition

    PubMed Central

    Li, Yue; Zhang, Qian; Zhang, Fangfei; Liu, Ruoxi; Liu, He; Chen, Feng

    2015-01-01

    Black tooth stain is a characteristic extrinsic discoloration commonly seen on the cervical enamel following the contour of the gingiva. To investigate the relationship between black tooth stain and the oral microbiota, we used 16S rRNA gene sequencing to compare the microbial composition of dental plaque and saliva among caries-free children with and without black stain. Dental plaque and saliva, as well as black stain, were sampled from 10 children with and 15 children without black stain. Data were analyzed using the pipeline tool MOTHUR. Student’s t-test was used to compare alpha diversities and the Mann-Whitney U test to compare the relative abundances of the microbial taxa. A total of 10 phyla, 19 classes, 32 orders, 61 families and 102 genera were detected in these samples. Shannon and Simpson diversity were found to be significantly lower in saliva samples of children with black stain. Microbial diversity was reduced in the black stain compared to the plaque samples. Actinomyces, Cardiobacterium, Haemophilus, Corynebacterium, Tannerella and Treponema were more abundant and Campylobacter less abundant in plaque samples of children with black stain. Principal component analysis demonstrated clustering among the dental plaque samples from the control group, while the plaque samples from the black stain group were not and appeared to cluster into two subgroups. Alterations in oral microbiota may be associated with the formation of black stain. PMID:26340752

  19. Staining of in vivo subsurface degradation in dental composites with silver nitrate

    SciTech Connect

    Mair, L.H. )

    1991-03-01

    A previously reported technique for staining areas of degradation in dental composite restorations was evaluated in 51 removed restorations. The staining reagent was silver nitrate, which penetrated the degraded subsurface as ionic silver and was subsequently developed into colored deposits of metallic silver. Several artefacts were recognized that resulted in an apparent image of subsurface stain. Most importantly, the presence of a layer of adsorbed silver on the edge of the specimen exaggerated the extent of staining. In order for the true depth of stain to be determined, thin sections of the materials should first be examined with a stereomicroscope to distinguish any contribution from adsorbed silver on the specimen edge. With this regimen, no stain was present in 41% of the restorations, and in a further 30%, the depth of stain was less than 50 microns. In two composites, the depth of stain was greater than 900 microns, and in a number of specimens, localized stain was found in association with attrition scars. Energy-dispersive x-ray analysis indicated that the amount of silver present in the degraded layers was very small. Overall, the results indicated that the staining technique is useful in the study of composite degradation.

  20. The propensity of different brands of tea and coffee to cause staining associated with chlorhexidine.

    PubMed

    Leard, A; Addy, M

    1997-02-01

    Dental staining is a well known and probably the most problematic side effect of using chlorhexidine oral products. Whatever mechanisms are involved, there is no doubt that cationic antiseptics, such as chlorhexidine, can precipitate or bind to surfaces anionic chromogens contained in foods and beverages. The aim of this study in vitro was to determine whether, under controlled conditions, different brands of tea or coffee varied in their propensity to cause staining associated with chlorhexidine. Optically clear acrylic specimens were cycled through saliva, chlorhexidine and different tea and coffee solutions. Staining was measured using a spectrophotometer. After 15 cycles, it was apparent that staining varied at the extreme, both within and between the tea and coffee groups. All coffee brands produced less staining than the tea brands. The least staining coffee and least staining tea brands were approximately 3x less chromogenic than the most staining equivalent beverage. Previous randomised controlled clinical trials have indicated that tea and coffee contribute to dental and tongue staining associated with chlorhexidine mouthrinses. Additionally, abstinence from tea and coffee significantly reduces staining. The results of this study in vitro suggest that when abstinence is difficult, tea and coffee brands of low chromogenicity may be recommended. Clearly these data in vitro require validation in vivo. PMID:9062858

  1. Use of environmental scanning electron microscopy to evaluate dental stain removal.

    PubMed

    Zammitti, S; Habib, C; Kugel, G

    1997-01-01

    The purpose of the study was to assess the usefulness of environmental scanning electron microscopy (ESEM) to evaluate stain removal from extracted teeth. The ESEM differs from conventional SEM in that no sample preparation is needed, eliminating artifactual changes. Furthermore, the same sample can be viewed on multiple occasions, allowing "before" and "after" pictures of the same tooth. As a model stain removal device, we tested the Sonicare sonic toothbrush, which has previously been shown to remove dental stain in vivo. Twelve freshly extracted teeth with extrinsic coffee, tea or tobacco stain were obtained for the study. Nine of these had heavy stain (stain covering more than one-third buccal or lingual surface) and were used without further modification. Three teeth were treated in vitro with chlorhexidine and a mixture of coffee and tea to enhance staining. All teeth were examined by ESEM at three times: prior to brushing, after 15-30 seconds of brushing, and after 60-80 seconds of brushing. Light microscopy and 35 mm photography was also done to correlate the ultrastructural changes with those visible at low magnification. Water, mouthwash and 30% slurry of toothpaste were used as fluid vehicles during brushing, but little difference in stain removal was noted among these three fluids. Approximately half the stain was removed within 15-30 seconds, and most visible stain was removed in 60-80 seconds of brushing. Pits and crevices of tooth enamel that were smaller than the bristle diameter, and thus would be inaccessible to abrasive cleaning by direct bristle contact, were generally found to be stain-free. These findings confirm previous reports of the stain removal effectiveness of the Sonicare, and demonstrate the usefulness of ESEM for stain removal studies. PMID:9487841

  2. Flow cytometric detection of human immunodeficiency virus type 1 proviral DNA by the polymerase chain reaction incorporating digoxigenin- or fluorescein-labeled dUTP

    SciTech Connect

    Yang, Gang; Olson, J.C.; Pu, R.; Vyas, G.N.

    1995-10-01

    Serological assays are routinely used in the laboratory diagnosis of human immunodeficiency virus type-1 (HrV-1) infection, but the polymerase chain reaction (PCR) is ultimately the most sensitive and direct method for establishing definitive diagnosis. As an alternative to the conventional radioactive PCR procedure we have developed and evaluated a pair of rapid nonradioisotopic flow cytometric detection methods. Using heminested PCR we directly incorporated fluorescein-12-dUTP (fluo-dUTP) or digoxigenin-11-dUTP (dig-dUTP) into the PCR-amplicons. The labeled amplicons were hybridized with biotinylated antisense and sense probes, followed by capture of the hybrid DNA using streptavidin-coated beads which were finally analyzed in a flow cytometer by (1) direct detection of the fluorescence intensity of the amplicons incorporating fluo-dUTP and (2) immunodetection of the amplicons incorporating dig-dUTP by anti-digoxigenin IgG labeled with fluorescein isothiocyanate (FITC). Although both assays were functionally comparable with radiolabeled probe in reliably detecting as low as five copies of HIV-1 proviral DNA sequences, the immunodetection of dig-dUTP consistently yielded higher mean channel fluorescence and gave a stable signal over an extended period of 12-14 weeks. In testing a panel of 20 pedigreed PBMC specimens from blood donors with or without HIV-1 infection, the results of both flow cytometric assays were identical with those of the conventional radioactive procedure. Therefore, we conclude that the dig-dUTP incorporation in amplicons, hybridization with a pair of sense-antisense biotinylated probes and immunodetection of hybrids by flow cytometric analyses is the nonisotopic method of choice for PCR-diagnosis of HIV-1 infection. 21 refs., 2 figs., 4 tabs.

  3. A level-set method for pathology segmentation in fluorescein angiograms and en face retinal images of patients with age-related macular degeneration

    NASA Astrophysics Data System (ADS)

    Mohammad, Fatimah; Ansari, Rashid; Shahidi, Mahnaz

    2013-03-01

    The visibility and continuity of the inner segment outer segment (ISOS) junction layer of the photoreceptors on spectral domain optical coherence tomography images is known to be related to visual acuity in patients with age-related macular degeneration (AMD). Automatic detection and segmentation of lesions and pathologies in retinal images is crucial for the screening, diagnosis, and follow-up of patients with retinal diseases. One of the challenges of using the classical level-set algorithms for segmentation involves the placement of the initial contour. Manually defining the contour or randomly placing it in the image may lead to segmentation of erroneous structures. It is important to be able to automatically define the contour by using information provided by image features. We explored a level-set method which is based on the classical Chan-Vese model and which utilizes image feature information for automatic contour placement for the segmentation of pathologies in fluorescein angiograms and en face retinal images of the ISOS layer. This was accomplished by exploiting a priori knowledge of the shape and intensity distribution allowing the use of projection profiles to detect the presence of pathologies that are characterized by intensity differences with surrounding areas in retinal images. We first tested our method by applying it to fluorescein angiograms. We then applied our method to en face retinal images of patients with AMD. The experimental results included demonstrate that the proposed method provided a quick and improved outcome as compared to the classical Chan-Vese method in which the initial contour is randomly placed, thus indicating the potential to provide a more accurate and detailed view of changes in pathologies due to disease progression and treatment.

  4. Specimen block counter-staining for localization of GUS expression in transgenic arabidopsis and tobacco

    NASA Technical Reports Server (NTRS)

    Kim, M. K.; Choi, J-W; Jeon, J-H; Franceschi, V. R.; Davin, L. B.; Lewis, N. G.

    2002-01-01

    A simple counter-staining procedure has been developed for comparative beta-glucuronidase (GUS) expression and anatomical localization in transgenic herbaceous arabidopsis and tobacco. This protocol provides good anatomical visualization for monitoring chimeric gene expression at both the organ and tissue levels. It can be used with different histochemical stains and can be extended to the study of woody species. The specimens are paraffin-embedded, the block is trimmed to reveal internal structure, safranin-O staining solution is briefly applied to the surface of the block, then washed off and, after drying, a drop of immersion oil is placed on the stained surface for subsequent photographic work. This gives tissue counter-staining with good structural preservation without loss of GUS staining product; moreover, sample observation is rapid and efficient compared to existing procedures.

  5. Sensitive silver-staining detection of bacterial lipopolysaccharides in polyacrylamide gels.

    PubMed

    Kittelberger, R; Hilbink, F

    1993-02-01

    For sensitive detection of bacterial lipopolysaccharides in the nanogram range, three almost identical silver-staining methods are often used, which are based on ammoniacal silver solutions and an acidic developer. We modified a method used for proteins, based on neutral silver nitrate solution and an alkaline developer, for the visualization of lipopolysaccharides in polyacrylamide gels, which yields better sensitivity and is much less prone to formation of non-specific background staining than the acidic developer-based silver stains. PMID:8387076

  6. An epidemiologic study of factors affecting extrinsic staining of teeth in an English population.

    PubMed

    Ness, L; Rosekrans, D de L; Welford, J F

    1977-01-01

    A survey is described in which extrinsic staining of teeth was assessed by practising dentists during routine clinical examinations. Habits information was also supplied by patients. The results show that extrinsic staining is markedly influenced by smoking habits and age, and that males tend to have more staining than females. The effect of tea and coffee consumption was not pronounced. No effect attributable to the use of a stannous fluoride toothpaste was observed. PMID:264419

  7. A comparative study of stain removal with two electric toothbrushes and a manual brush.

    PubMed

    Moran, J M; Addy, M; Newcombe, R G

    1995-01-01

    Recent studies have suggested that a sonic electric toothbrush is more effective than a manual brush at removing extrinsic dental stain. There have been few studies of the comparative stain removal properties of different electric brushes. The study reported here was conducted to compare the efficacy of the sonic toothbrush (Sonicare) with an oscillating/rotating brush (Braun Oral-B Plaque Remover) and a conventional manual brush (Crest Complete). The study was a single-blind, randomized, cross-over design, balanced for residual effects and employing 24 subjects. Stain was enhanced over a 21-day period by twice-daily rinses with chlorhexidine and frequent intakes of tea and/or coffee. At the end of each period, tooth stain intensity and area, tongue stain intensity and area, lower lingual calculus and subjective tooth sensitivity were recorded together with preference for the brushes determined at the study's completion. Similar levels of tongue staining were recorded for the three periods, with no significant differences between the three groups. Tooth stain intensity, for most sites, was not significantly different between the three groups. For mean total stain area and for lingual and lingual interproximal sites, a significant reduction in stain was seen following use of the oscillating/rotating brush compared to the manual brush. The reductions in stain with the sonic brush were not significantly different from the manual brush. With the exception of maximum stain intensity, there were no significant differences between the oscillating/rotating and sonic brushes. Significantly less tooth sensitivity was found following use of the oscillating/rotating brush compared to both the manual and sonic brushes. All three brushes were found to be safe, but volunteer preference significantly and predominantly favored the oscillating/rotating brush. The results suggest that the oscillating/rotating brush is superior to a manual brush for stain removal. PMID:8624230

  8. Effect of staining solutions on discoloration of resin nanocomposites

    PubMed Central

    Park, Jeong-Kil; Kim, Tae-Hyong; Ko, Ching-Chang; García-Godoy, Franklin; Kim, Hyung-Il; Kwon, Yong Hoon

    2011-01-01

    Purpose To examine the effect of staining solutions on the discoloration of resin nanocomposites. Methods Three resin nanocomposites (Ceram X, Grandio, and Filtek Z350) were light cured for 40 seconds at a light intensity of 1000 mW/cm2. The color of the specimens was measured in %R (reflectance) mode before and after immersing the specimens in four different test solutions [distilled water (DW), coffee (CF), 50% ethanol (50ET) and brewed green tea (GT)] for 7 hours/day over a 3-week period. The color difference (?E*) was obtained based on the CIEL*a*b* color coordinate values. Results The specimens immersed in DW, 50ET and GT showed a slight increase in L* value. However, the samples immersed in CF showed a decrease in the L* value and an increase in the b* value. CF induced a significant color change (?E*: 3.1~5.6) in most specimens but the other solutions induced only a slight color change. Overall, coffee caused unacceptable color changes to the resin nanocomposites. PMID:20437726

  9. The challenges of analysing blood stains with hyperspectral imaging

    NASA Astrophysics Data System (ADS)

    Kuula, J.; Puupponen, H.-H.; Rinta, H.; Pölönen, I.

    2014-06-01

    Hyperspectral imaging is a potential noninvasive technology for detecting, separating and identifying various substances. In the forensic and military medicine and other CBRNE related use it could be a potential method for analyzing blood and for scanning other human based fluids. For example, it would be valuable to easily detect whether some traces of blood are from one or more persons or if there are some irrelevant substances or anomalies in the blood. This article represents an experiment of separating four persons' blood stains on a white cotton fabric with a SWIR hyperspectral camera and FT-NIR spectrometer. Each tested sample includes standardized 75 _l of 100 % blood. The results suggest that on the basis of the amount of erythrocytes in the blood, different people's blood might be separable by hyperspectral analysis. And, referring to the indication given by erythrocytes, there might be a possibility to find some other traces in the blood as well. However, these assumptions need to be verified with wider tests, as the number of samples in the study was small. According to the study there also seems to be several biological, chemical and physical factors which affect alone and together on the hyperspectral analyzing results of blood on fabric textures, and these factors need to be considered before making any further conclusions on the analysis of blood on various materials.

  10. Development of a stained cell nuclei counting system

    NASA Astrophysics Data System (ADS)

    Timilsina, Niranjan; Moffatt, Christopher; Okada, Kazunori

    2011-03-01

    This paper presents a novel cell counting system which exploits the Fast Radial Symmetry Transformation (FRST) algorithm [1]. The driving force behind our system is a research on neurogenesis in the intact nervous system of Manduca Sexta or the Tobacco Hornworm, which was being studied to assess the impact of age, food and environment on neurogenesis. The varying thickness of the intact nervous system in this species often yields images with inhomogeneous background and inconsistencies such as varying illumination, variable contrast, and irregular cell size. For automated counting, such inhomogeneity and inconsistencies must be addressed, which no existing work has done successfully. Thus, our goal is to devise a new cell counting algorithm for the images with non-uniform background. Our solution adapts FRST: a computer vision algorithm which is designed to detect points of interest on circular regions such as human eyes. This algorithm enhances the occurrences of the stained-cell nuclei in 2D digital images and negates the problems caused by their inhomogeneity. Besides FRST, our algorithm employs standard image processing methods, such as mathematical morphology and connected component analysis. We have evaluated the developed cell counting system with fourteen digital images of Tobacco Hornworm's nervous system collected for this study with ground-truth cell counts by biology experts. Experimental results show that our system has a minimum error of 1.41% and mean error of 16.68% which is at least forty-four percent better than the algorithm without FRST.

  11. Diffuse reflectance FTIR of stains on grit blasted metals

    SciTech Connect

    Powell, G.L.; Hallman, R.L. Jr.; Cox, R.L.

    1997-08-09

    Diffuse reflectance mid-infrared Fourier transform (DRIFT) spectroscopy has been applied to the detection of oil contamination on grit-blasted metals. The object of this application is to detect and discriminate between silicone and hydrocarbon oil contamination at levels approaching 10 mg/m{sup 2}. A portable FTIR spectrometer with dedicated diffuse reflectance optics was developed for this purpose. Using translation devices positioned by instructions from the spectrometer operating system, images of macroscopic substrates were produced with millimeter spatial resolution. The pixels that comprise an image are each a full mid-infrared spectrum with excellent signal-to-noise, each determined as individual files and uniquely saved to disc. Reduced spectra amplitudes, based on peak height, area, or other chemometric techniques, mapped as a function of the spatial coordinates of the pixel are used to display the image. This paper demonstrates the application of the technique to the analysis of stains on grit-blasted metals, including the calibration of the method, the inspection of substrates, and the migration of oil contamination.

  12. Multispectral image enhancement for H&E stained pathological tissue specimens

    NASA Astrophysics Data System (ADS)

    Bautista, Pinky A.; Abe, Tokiya; Yamaguchi, Masahiro; Ohyama, Nagaaki; Yagi, Yukako

    2008-03-01

    The presence of a liver disease such as cirrhosis can be determined by examining the proliferation of collagen fiber from a tissue slide stained with special stain such as the Masson's trichrome(MT) stain. Collagen fiber and smooth muscle, which are both stained the same in an H&E stained slide, are stained blue and pink respectively in an MT-stained slide. In this paper we show that with multispectral imaging the difference between collagen fiber and smooth muscle can be visualized even from an H&E stained image. In the method M KL bases are derived using the spectral data of those H&E stained tissue components which can be easily differentiated from each other, i.e. nucleus, cytoplasm, red blood cells, etc. and based on the spectral residual error of fiber weighting factors are determined to enhance spectral features at certain wavelengths. Results of our experiment demonstrate the capability of multispectral imaging and its advantage compared to the conventional RGB imaging systems to delineate tissue structures with subtle colorimetric difference.

  13. PCR-based forensic testing of DNA from stained cytological smears.

    PubMed

    Dimo-Simonin, N; Grange, F; Brandt-Casadevall, C

    1997-05-01

    Experiments were performed to evaluate the efficiency of PCR-STR (Short Tandem Repeats) and PCR-sequence polymorphisms for the identification of stained pap smears and postcoital slides stained with cytological and forensic techniques. HLA-DQA1, PolyMarker, Amelogenin, HUMTH01, HUMVWFA31, HUMF13B, and HUMFES/FPS were determined. With the exception of the forensic Baecchi stain, all the PCR-systems gave consistent results in comparison with the reference blood from the donors. Cytological stained smears can be important evidence for identification in sexual assault cases and in missing person cases. PMID:9144941

  14. Karyotype analysis of four jewel-beetle species (Coleoptera, Buprestidae) detected by standard staining, C-banding, AgNOR-banding and CMA3/DAPI staining

    PubMed Central

    Karagyan, Gayane; Lachowska, Dorota; Kalashian, Mark

    2012-01-01

    Abstract The male karyotypes of Acmaeodera pilosellae persica Mannerheim, 1837 with 2n=20 (18+neoXY), Sphenoptera scovitzii Faldermann, 1835 (2n=38–46), Dicerca aenea validiuscula Semenov, 1895 – 2n=20 (18+Xyp) and Sphaerobothris aghababiani Volkovitsh et Kalashian, 1998 – 2n=16 (14+Xyp) were studied using conventional staining and different chromosome banding techniques: C-banding, AgNOR-banding, as well as fluorochrome Chromomycin A3 (CMA3) and DAPI. It is shown that C-positive segments are weakly visible in all four species which indicates a small amount of constitutive heterochromatin (CH). There were no signals after DAPI staining and some positive signals were discovered using CMA3 staining demonstrating absence of AT-rich DNA and presence of GC-rich clusters of CH. Nucleolus organizing regions (NORs) were revealed using Ag-NOR technique; argentophilic material mostly coincides with positive signals obtained using CMA3 staining. PMID:24260661

  15. Morphometry of cupromeronic blue-stained proteoglycan molecules in animal corneas, versus that of purified proteoglycans stained in vitro, implies that tertiary structures contribute to corneal ultrastructure.

    PubMed Central

    Scott, J E

    1992-01-01

    Isolated, purified small chondroitin (dermatan) sulphate proteoglycans from corneas of cow and rabbit and cow sclera were stained with Cupromeronic blue in 'model' experiments. The lengths and thicknesses of the images were compared with those of the same proteoglycans stained in the tissue, using the critical electrolyte concentration principle to give specificity for sulphated proteoglycans, and keratanase 1 or chondroitinase ABC digestion to distinguish between chondroitin and keratan sulphate. Corrections for orientation of the stained glycan filaments within the section plane were made to convert the observed lengths to true average lengths. Observed lengths of stained chondroitin (dermatan) sulphate were greater than those of keratan sulphate, both in models and tissues, in agreement with published data from biochemical and rotary-shadowing studies, in both species. Corrected (true) average lengths of stained isolated chondroitin (dermatan) sulphate proteoglycans were slightly, but not significantly, longer than expected from rotary shadowing or biochemical measurements. Keratan sulphate lengths were similarly somewhat longer. The data support the idea that Cupromeronic blue acts as a scaffold that helps maintain polyanion shape against distortion on staining. Stained filaments in tissues were sometimes over twice the length of isolated stained proteoglycans, suggesting that 2 glycan chains were aligned end-to-end. Thicknesses of proteoglycan filaments suggested that at least 2 glycan chains were aligned side-by-side, both in models and in tissues. A scheme for proteoglycan tertiary structure in cornea is proposed, in which glycan chains may bridge collagen fibrils in duplexed forms similar to those observed in rotary shadowed preparations. Images Fig. 1 Fig. 2 PMID:1452471

  16. An evaluation of Retaine™ ophthalmic emulsion in the management of tear film stability and ocular surface staining in patients diagnosed with dry eye.

    PubMed

    Ousler, George; Devries, Douglas K; Karpecki, Paul M; Ciolino, Joseph B

    2015-01-01

    A single-center, open-label study consisting of two visits over the course of approximately 2 weeks was conducted to evaluate the efficacy of Retaine™ ophthalmic emulsion in improving the signs and symptoms of dry eye. Forty-two subjects were enrolled and received 1-2 drops twice daily of Retaine™ beginning at the first visit (day 1) and ending at the second visit. Subjects were instructed to complete a symptomatology diary twice daily prior to drop instillation through the morning of the second visit. Ocular sign and symptom assessments, visual acuity procedures, and comfort assessments were conducted during both visits. A statistically significant reduction was observed in mean breakup area on the second visit between the predose time and the postdose time (P=0.026). On the second visit, subjects had significantly less corneal fluorescein staining in the superior (P=0.002), central (P=0.017), corneal sum (P=0.011), and all ocular regions combined (P=0.038) than on the first visit. On the second visit, statistically significant reductions in dryness (P<0.001), grittiness (P=0.0217), ocular discomfort (P=0.0017), and all symptoms (P<0.001) were also seen as measured by the Ora Calibra™ Ocular Discomfort and 4-Symptom Questionnaire (0-5 scale). Subjects reported a statistically significant improvement in their abilities to work with a computer at night (P=0.044). Mean drop comfort scores ranged from 1.29-1.81 on the Ora Calibra™ 0-10 Drop Comfort Scale, on which 0 is very comfortable and 10 is very uncomfortable. Retaine™ demonstrates promising results as a novel artificial tear option for individuals suffering from dry eye. The unique mechanism of action of Retaine™ provides enhanced comfort and improves the quality of life of dry eye subjects while reducing the ocular signs of dry eye. PMID:25709384

  17. Platinum blue as an alternative to uranyl acetate for staining in transmission electron microscopy.

    PubMed

    Inaga, Sumire; Katsumoto, Tetsuo; Tanaka, Keiichi; Kameie, Toshio; Nakane, Hironobu; Naguro, Tomonori

    2007-04-01

    This paper introduces an aqueous solution of platinum blue (Pt-blue) as an alternative to uranyl acetate (UA) for staining in transmission electron microscopy (TEM). Pt-blue was prepared from a reaction of cis-dichlorodiamine-platinum (II) (cis-platin) with thymidine. When Pt-blue was dried on a microgrid and observed by TEM it showed a uniform appearance with tiny particles less than 1 nm in diameter. The effect of Pt-blue as an electron stain was then examined not only for positive staining of conventional ultrathin resin sections and counterstaining of post-embedding immuno-electron microscopy but also for negative staining. In ultrathin sections of the rat liver and renal glomerulus, Pt-blue provided good contrast images, especially in double staining combined with a lead stain (Pb). Almost all cell organelles were clearly observed with high contrast in these sections. Glycogen granules in the hepatic parenchymal cells were particularly electron dense in Pt-blue stained sections compared with those treated with UA. In longitudinal and transverse sections of budding influenza A viruses, a specific arrangement of rod-like structures, which correspond to the ribonucleoprotein complexes, was clearly shown in each virion stained with Pt-blue and Pb. When post-embedding immunoelectron microscopy was performed in ultrathin sections of HeLa cells embedded in Lowicryl K4M, the localization of Ki-67 protein was sufficiently detected even after Pt-blue and Pb staining. The present study also revealed that Pt-blue could be used for the negative staining of E. coli, allowing the visualization of a flagellum. These findings indicate that Pt-blue is a useful, safe, and easily obtainable electron stain that is an alternative to UA for TEM preparations. PMID:17558143

  18. Clinical and computer-assisted evaluations of the stain removal ability of the Sonicare electronic toothbrush.

    PubMed

    McInnes, C; Johnson, B; Emling, R C; Yankell, S L

    1994-01-01

    Two single-blind clinical studies investigated the stain removal properties of Sonicare, a new electronic toothbrush that combines sonic vibrations and dynamic fluid activity with mechanical scrubbing to clean tooth surfaces. In one study, 30 subjects used a 0.12% chlorhexidine mouthrinse (Peridex) for two weeks to accumulate stain, and then were assigned to either Sonicare or a manual toothbrush (Oral-B P-35). The subjects brushed with their assigned device for 2 minutes twice a day. In a second study, 19 subjects with extrinsic stain due to coffee, tea, or tobacco (CTT) causes were randomly assigned to either Sonicare or a manual toothbrush (Crest Complete). These subjects also brushed for 2 minutes twice a day, with additional brushing on the stained areas. Stain on the labial surfaces of the subjects' anterior teeth was evaluated with the Lobene index at the pretrial, 2-week, and 4-week periods. Clinical analysis indicated that use of Sonicare resulted in Peridex stain reductions of 54% and 50% after 2 and 4 weeks, respectively, and reductions in CTT stain of 39% and 82% at similar time points. The manual toothbrush resulted in stain increases of 4% and 24% in the Peridex study and CTT stain decreases of 41% and 39% after 2- and 4-week brushing periods. Computer image analysis was performed on photographic records from the CTT stain study and showed a high correlation with the Lobene index (r = 0.82). The results of these two independent studies indicate that Sonicare is superior to the manual toothbrushes studied in removing both Peridex and CTT stains.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8031484

  19. Analysis of Staining Observed on Structures in the Georgetown, South Carolina Area

    SciTech Connect

    Cramer, Stephen D.; Covino, Bernard S. Jr.; Govier, R. Dale

    2002-05-01

    Beginning around 1970, the Georgetown, SC, community complained about black dust and red stains collecting on houses, cars, boats, and other structures. The community, through the South Carolina Department of Health and Environmental Control (SCDHEC), seeks to identify the source or cause of the staining and ways to reduce or eliminate it in the future.

  20. Staining and etching: a simple method to fabricate inorganic nanostructures from tissue slices.

    PubMed

    Dai, Xiaoshu; Schalek, Richard; Xu, Qiaobing

    2012-01-17

    Staining and etching is developed for the fabrication of two-dimensional arrays and three-dimensional foam-like metallic nanostructures from a piece of tissue using a combination of metal staining and oxygen plasma etching. This novel nanofabrication technique is repeatable and cost effective, which are very attractive features from the perspective of nanomanufacturing process. PMID:22174186

  1. Silver Staining SDS Gels Remove the stacking gel before the first fixative step.

    E-print Network

    Aris, John P.

    37 Silver Staining SDS Gels · Remove the stacking gel before the first fixative step. · Do, for frequent silver staining: make up solutions as 10X stocks ahead of time. 1. Fix in 50% methanol, 10% acetic. Replace silver nitrate solution in same tray and gently agitate for 15 minutes again. 6. Rinse the gel 2 X

  2. MP 33250 Coomassie FluorTM Orange Protein Gel Stain Product Information

    E-print Network

    Lebendiker, Mario

    MP 33250 Coomassie FluorTM Orange Protein Gel Stain Product Information Storage upon receipt Introduction Molecular Probes proprietary Coomassie FluorTM Orange protein gel stain provides fast, simple-to-protein variability. Because Coomassie Fluor Orange dye interacts with the SDS coat around proteins in the gel

  3. Tear staining in pigs: a potential tool for welfare assessment on commercial farms

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tear staining or chromodacryorrhea refers to a dark stain below the inner corner of the eye, caused by porphyrin-pigmented secretion from the Harderian gland. It been shown to be a consistent indicator of stress in rats, and recently it has been shown to correlate with social stress and a barren env...

  4. Numerical modeling of spray cooling-assisted dermatologic laser surgery for treatment of port wine stains

    E-print Network

    Aguilar, Guillermo

    Numerical modeling of spray cooling-assisted dermatologic laser surgery for treatment of port wine to the epidermis during dermatologic laser surgery (DLS) for removal of port wine stain (PWS) birthmarks) to treat vascu- lar superficial lesions and abnormalities, such as port wine stains (PWS) birthmarks

  5. Evaporation stains: suppressing the coffee-ring effect by contact angle hysteresis.

    PubMed

    Li, Yueh-Feng; Sheng, Yu-Jane; Tsao, Heng-Kwong

    2013-06-25

    A ring-shaped stain is frequently left on a substrate by a drying drop containing colloids as a result of contact line pinning and outward flow. In this work, however, different patterns are observed for drying drops containing small solutes or polymers on various hydrophilic substrates. Depending on the surface activity of solutes and the contact angle hysteresis (CAH) of substrates, the pattern of the evaporation stain varies, including a concentrated stain, a ringlike deposit, and a combined structure. For small surface-inactive solutes, the concentrated stain is formed on substrates with weak CAH, for example, copper sulfate solution on silica glass. On the contrary, a ringlike deposit is developed on substrates with strong CAH, for example, a copper sulfate solution on graphite. For surface-active solutes, however, the wetting property can be significantly altered and the ringlike stain is always visible, for example, Brij-35 solution on polycarbonate. For a mixture of surface-active and surface-inactive solutes, a combined pattern of a ringlike and concentrated stain can appear. For various polymer solutions on polycarbonate, similar results are observed. Concentrated stains are formed for weak CAH such as sodium polysulfonate, and ring-shaped patterns are developed for strong CAH such as poly(vinyl pyrrolidone). The stain pattern is actually determined by the competition between the time scales associated with contact line retreat and solute precipitation. The suppression of the coffee-ring effect can thus be acquired by the control of CAH. PMID:23721254

  6. A comparison of silver staining protocols for detecting DNA in polyester-backed polyacrylamide gel.

    PubMed

    Qiu, Shanlian; Chen, Jichen; Lin, Si; Lin, Xinjian

    2012-04-01

    Eight silver-staining protocols were applied to detect DNA in polyester-backed gels to select the optimal. Results showed important differences in staining quality and that four methods were well-suited for TGGE gels due to high sensitivity and low background, including the Bassam et al. methods, the manufacturer method and our improved method. PMID:24031876

  7. Use of supravital toluidine blue staining to improve the efficiency of fine-needle aspiration cytology reporting in comparison to papanicolaou stain

    PubMed Central

    Saba, Kanwal; Niazi, Shahida; Bukhari, Mulazim Hussain; Imam, Sardar Fakhar

    2015-01-01

    Objective: To see the efficiency, adequacy and accuracy of toluidine blue stained smears of FNAC of Breast thyroid and salivary glands swelling with comparison to conventional stained FNAC smears with Papanicolaou. Methods: A total of 114 aspirates from various sites were included in the study. The smears were stained with toluidine blue and conventional Papanicolaou stain and the cytomorphology of both the smears were compared. The values were tabulated and statistical tests of significance was applied. Results: Of the 114 aspirates included in our study the diagnostic accuracy by using papanicolaou was 78%, while it was upto 100% with supravital toluidine blue stained smears. The percentage of inadequacy was reduced to just 25%. The observations were statistically significant. Breast 37/48 (77%) and Salivary glands 11/48 (23%) respectively. The most commonly used categorization of a five-tier system was used for reporting of breast cytology, with categories ranging from insufficient materials (C1), benign (C2), atypical (C3), suspicious of malignancy (C4), or (C5) frankly malignant. Most of breast lesions were benign 25 (67.56%). There were only 9 (24.32%) malignant cases followed by 2 cases of C-4 and one case of C-3. Benign thyroid lesion were more frequent comprising of 51 (72.27%) cases. One case (1.5%) of papillary carcinoma was found while 13 case were follicular lesions. There were 4 (36.4%) cases of pleomorphic adenoma and 3 (27.3) cases of non-specific sialadenitis. There was one case (9%) of each lesion for mucoepidermoid carcinoma, adenoidcytic carcinoma and benign cyst. Conclusion: Toluidine blue stained study of FNAC improves the diagnostic accuracy by minimizing the smearing and drying artifact, loss of cell sample during fixation and staining which influences the diagnostic accuracy.

  8. Characterisation of medieval yellow silver stained glass from Convento de Cristo in Tomar, Portugal

    NASA Astrophysics Data System (ADS)

    Delgado, J.; Vilarigues, M.; Ruivo, A.; Corregidor, V.; Silva, R. C. da; Alves, L. C.

    2011-10-01

    Yellow decoration effects in stained glasses using silver staining were first applied in the beginning of the 14th century. The glass piece being decorated was usually painted on its side intended to be facing the exterior environment, and then fired to temperatures between 500 and 650 °C, resulting in colours ranging from pale lemon to deep orange. Stained glass fragments painted by this process and belonging to the Convento de Cristo, in Tomar, Portugal, were characterised using micro-PIXE, and complemented with other analytical techniques, namely UV-Vis spectroscopy and XRF. Preliminary analysis showed that a mixture of Ag and Cu was used for the production of the yellow staining. In order to understand this staining process and the influence of the firing temperature on the resulting colours, several soda and potash glasses with compositions similar to those of medieval glasses were produced and characterised. The role played by the addition of Cu in the final colours was also investigated.

  9. [KI-67 PAP stain for histologic grading of brain tumors by squash preparations].

    PubMed

    Shibata, T; Ostertag, C B; Volk, B

    1989-01-01

    Using Ki-67 monoclonal antibody to the nuclear antigen of proliferating cells, the squash preparations of 141 brain tumors and the 36 frozen sections from corresponding tumor tissues were stained with peroxidase-antiperoxidase method. The staining indices in squash preparations correlated well to those in the corresponding frozen sections. There were good correlations between the percentage of stained cells and the histologic grade in agreement with known biologic behavior. In order of decreasing malignancy, the averages of Ki-67 staining indices were 26.7% for metastatic carcinomas, 12.6% for glioblastomas, and less than 2.8% for benign mesodermal tumor groups. Ki-67 staining with squash preparations could be applied to small tissues obtained by transsphenoidal surgery and stereotactic surgery as valuable adjuncts to intraoperative histologic diagnosis and the estimation of histologic malignancy. PMID:2470401

  10. Ki67/KIT double immunohistochemical staining in cutaneous mast cell tumors from Boxer dogs.

    PubMed

    Fonseca-Alves, Carlos Eduardo; Bento, Daniel Diola; Torres-Neto, Rafael; Werner, Juliana; Kitchell, Barbara; Laufer-Amorim, Renée

    2015-10-01

    Cutaneous mast cell tumors (MCTs) are among the most frequent malignant tumors in dogs and Boxer breed dogs have a higher incidence of this disease. Ki67 staining and KIT staining are widely used to predict natural behavior in canine MCT but no previous study has evaluated double staining of these proteins as a prognostic factor. Based on biological behavior predictors in canine MCT, the purpose of this study was to determine the Ki67 proliferative index in KIT positive cells using double stain immunohistochemistry technique. Sixty-nine MCTs from Boxer dogs were selected and a tissue microarray was constructed for the double stained immunohistochemistry. Double positivity (Ki67(+)/KIT(+)) was observed in 20/69 (29%) MCT, with a mean of 9.06 double positive cells per tissue core (range 0.48%-43.97%) and Ki67(-)/KIT(+) animals had a longer survival time than Ki67(+)/KIT(+) animals (p=0.03). PMID:26412531

  11. Influence of some aldehyde blocking agents on staining of depurinized DNA with cationic dyes.

    PubMed

    Erenpreisa, J; Freivalds, T

    1979-01-01

    Rat liver, spleen and Walker carcinosarcoma imprints were subjected to depurinizing Feulgen hydrolysis and then treated with blocking agents of aldehyde groups. Such blockators as sodium bisulfite and hydroxylamine which multiplay additionally anionic groups in DNA and intensify the reactions with cationic dyes, ensuring anisotropic staining. Hydrazine lowers the binding of carionic dyes to DNA, instead phenylhydrazine, completely blocks both aldehyde and phosphate groups. When the imprints were treated with 2.4-dinitrophenylhydrazine, aldehyde and phosphate groups of apurinic acid were blocked, and DNA staining by cationic dyes occurred only on account of nitrogroups of the blocking agents which have been used. The staining reaction of cationic dyes after the use of anionogenic blocking agents of aldehyde groups is prospective not only for revealing DNA but also for several other compounds with natural or potential aldo- and ketogroups. However the reaction with phenylhydrazine can serve as a staining without removal of DNA prior to staining as an optional procedure. PMID:86483

  12. Preservation of calcium pyrophosphate dihydrate crystals: effect of Mayer's haematoxylin staining period

    PubMed Central

    Ohira, T; Ishikawa, K

    2001-01-01

    OBJECTIVE—To clarify the deleterious effects of Mayer's haematoxylin staining procedure which result in a decrease in, or complete loss of, the number of calcium pyrophosphate dihydrate (CPPD) crystals, and to determine the proper staining period for preserving the crystals in a histological paraffin section of articular tissues.?METHODS—Paraffin sections of CPPD crystal-bearing articular tissues of six patients were stained with Mayer's haematoxylin for 3, 8, or 15 minutes, and subsequently with eosin for one minute. The specimens were examined with an Olympus BHS polarised light microscope. The pH of Mayer's haematoxylin solution was measured with a TOA pH meter.?RESULTS—Positive birefringent CPPD crystals were seen clearly in all specimens stained with Mayer's haematoxylin for three minutes. The specimens stained for eight minutes showed a reduced number of crystals. No crystals were seen in the specimens stained for 15 minutes. Ordinary light microscopy showed no notable differences in the stainability of nucleus, cell membrane, and their surrounding tissues among specimens when stained with Mayer's haematoxylin for either 3, 8, or 15 minutes. The pH of Mayer's haematoxylin solution was 2.31.?CONCLUSIONS—To find CPPD crystals in the paraffin sections of articular tissues, the staining period with Mayer's haematoxylin should be limited to three minutes. The longer the staining period, the greater the reduction in the number of crystals owing to the strong acidity of the haematoxylin solution. A staining period of 15 minutes causes a complete loss of CPPD crystals.?? PMID:11114290

  13. Surface and statistical analysis of CaCO 3 crystals synthesized in the presence of fluorescein-tagged starch grafted with polyacrylic acid

    NASA Astrophysics Data System (ADS)

    Matahwa, H.; Sanderson, R. D.

    2009-02-01

    Crystallization of CaCO 3 was carried out in the presence of fluorescein-tagged starch grafted with polyacrylic acid (PAA) as polymeric additive. The crystal morphology at high polymeric additive concentration was spherical, with vaterite and calcite polymorphs. Mixed crystal morphologies were obtained at lower concentration of the polymeric additive and calcite was obtained. The CaCO 3 crystals obtained fluoresced under UV irradiation showing the presence of grafted starch expected on the surface of CaCO 3. The zeta potentials of the synthesized crystals were negative and addition of cationic starch lead to inversion of the zeta potential. Rodamine B-tagged cationic starch was successfully deposited onto the surface of the anionic-grafted starch-coated CaCO 3 crystals. Statistical analysis of the crystals showed that the fluorescence intensities of the crystals increased with increasing granularity and size of the CaCO 3 crystals. The high granularity and large-size crystals were due to aggregation of secondary crystals as observed with fluorescence microscopy.

  14. Analysis of Fundus Fluorescein Angiogram Based on the Hessian Matrix of Directional Curvelet Sub-bands and Distance Regularized Level Set Evolution

    PubMed Central

    Soltanipour, Asieh; Sadri, Saeed; Rabbani, Hossein; Akhlaghi, Mohammad Reza

    2015-01-01

    This paper presents a new procedure for automatic extraction of the blood vessels and optic disk (OD) in fundus fluorescein angiogram (FFA). In order to extract blood vessel centerlines, the algorithm of vessel extraction starts with the analysis of directional images resulting from sub-bands of fast discrete curvelet transform (FDCT) in the similar directions and different scales. For this purpose, each directional image is processed by using information of the first order derivative and eigenvalues obtained from the Hessian matrix. The final vessel segmentation is obtained using a simple region growing algorithm iteratively, which merges centerline images with the contents of images resulting from modified top-hat transform followed by bit plane slicing. After extracting blood vessels from FFA image, candidates regions for OD are enhanced by removing blood vessels from the FFA image, using multi-structure elements morphology, and modification of FDCT coefficients. Then, canny edge detector and Hough transform are applied to the reconstructed image to extract the boundary of candidate regions. At the next step, the information of the main arc of the retinal vessels surrounding the OD region is used to extract the actual location of the OD. Finally, the OD boundary is detected by applying distance regularized level set evolution. The proposed method was tested on the FFA images from angiography unit of Isfahan Feiz Hospital, containing 70 FFA images from different diabetic retinopathy stages. The experimental results show the accuracy more than 93% for vessel segmentation and more than 87% for OD boundary extraction. PMID:26284170

  15. Analysis of Fundus Fluorescein Angiogram Based on the Hessian Matrix of Directional Curvelet Sub-bands and Distance Regularized Level Set Evolution.

    PubMed

    Soltanipour, Asieh; Sadri, Saeed; Rabbani, Hossein; Akhlaghi, Mohammad Reza

    2015-01-01

    This paper presents a new procedure for automatic extraction of the blood vessels and optic disk (OD) in fundus fluorescein angiogram (FFA). In order to extract blood vessel centerlines, the algorithm of vessel extraction starts with the analysis of directional images resulting from sub-bands of fast discrete curvelet transform (FDCT) in the similar directions and different scales. For this purpose, each directional image is processed by using information of the first order derivative and eigenvalues obtained from the Hessian matrix. The final vessel segmentation is obtained using a simple region growing algorithm iteratively, which merges centerline images with the contents of images resulting from modified top-hat transform followed by bit plane slicing. After extracting blood vessels from FFA image, candidates regions for OD are enhanced by removing blood vessels from the FFA image, using multi-structure elements morphology, and modification of FDCT coefficients. Then, canny edge detector and Hough transform are applied to the reconstructed image to extract the boundary of candidate regions. At the next step, the information of the main arc of the retinal vessels surrounding the OD region is used to extract the actual location of the OD. Finally, the OD boundary is detected by applying distance regularized level set evolution. The proposed method was tested on the FFA images from angiography unit of Isfahan Feiz Hospital, containing 70 FFA images from different diabetic retinopathy stages. The experimental results show the accuracy more than 93% for vessel segmentation and more than 87% for OD boundary extraction. PMID:26284170

  16. Exploitation of phosphorescent labelling reagent of fullerol-fluorescein isothiocyanate and new method for the determination of trace alkaline phosphatase as well as forecast of human diseases.

    PubMed

    Liu, Jia-Ming; Huang, Xiao-Mei; Liu, Zhen-Bo; Lin, Shao-Qin; Li, Fei-Ming; Gao, Fei; Li, Zhi-Ming; Zeng, Li-Qing; Li, Lian-Ying; Ouyang, Ying

    2009-08-26

    A new phosphorescent labelling reagent consisting of fullerol, fluorescein isothiocyanate and N,N-dimethylaniline (F-ol-(FITC)(n)-DMA) was developed. The mode of action is based on the reactivity of the active -OH group in F-ol with the -COOH group of FITC to form an F-ol-(FITC)(n)-DMA complex containing several FITC molecules. F-ol-(FITC)(n)-DMA increased the number of luminescent molecules in the biological target of WGA-AP-WGA-F-ol-(FITC)(n)-DMA (WGA and AP are wheat germ agglutinin and alkaline phosphatase, respectively) which improved the sensitivity using solid substrate room temperature phosphorimetry (SSRTP) detection. The proposed method provided high sensitivity and strong specificity for WGA-AP. The limit of detection (LD) was 0.15 ag AP spot(-1) for F-ol and 0.097 ag AP spot(-1) for FITC in F-ol-(FITC)(n)-DMA, which was lower than the method using single luminescent molecules of F-ol-DMA and FITC-DMA to label WGA (0.20 ag AP spot(-1) for F-ol-DMA and 0.22 ag AP spot(-1) for FITC-DMA). Results for the determination of AP in human serum were in good agreement with those obtained by enzyme-linked immunosorbent assay. The mechanism of F-ol-(FITC)(n)-DMA labelling of WGA was discussed. PMID:19646588

  17. Fullerol-fluorescein isothiocyanate-concanavalin agglutinin phosphorescent sensor for the detection of alpha-fetoprotein and forecast of human diseases.

    PubMed

    Liu, Jia-ming; Lin, Li-ping; Jiang, Shu-lian; Cui, Ma Lin; Jiao, Li; Zhang, Xiao Yang; Zhang, Li-hong; Zheng, Zhi Yong; Lin, Xuan; Lin, Shao-qin

    2013-11-01

    Based on the reaction of the active -OH group in fullerol (F) with the dissociated -COOH group in fluorescein isothiocyanate (FITC) to form an F-FITC and the enhanced effect of N, N-dimethylaniline (DMA) on phosphorescence signal of F-FITC, a new phosphorescent labeling reagent (DMA-F-FITC) was developed. What's more, a phosphorescent sensor for the determination of alpha-fetoprotein variant (AFP-V) has been designed via the coupling technique of the high sensitivity for affinity adsorption-solid substrate-room temperature phosphorimetry (AA-SS-RTP) with the strong specificity reaction between DMA-F-FITC-Con A and AFP-V. The DMA-F-FITC increased the number of luminescent molecules in the biological target which improved the sensitivity of phosphorescent sensor. The proposed sensor was responsive, simple, selective and sensitive, and it has been applied to the determination of trace AFP-V in human serum and the forecast of human diseases using phosphorescence emission wavelength of F or FITC, with the results agreed well with those obtained by enzyme-linked immunoassay (ELISA). Meanwhile, the mechanisms for the labeling reaction and the sensing detection of AFP-V were discussed. PMID:23832221

  18. Fullerol-fluorescein isothiocyanate-concanavalin agglutinin phosphorescent sensor for the detection of alpha-fetoprotein and forecast of human diseases

    NASA Astrophysics Data System (ADS)

    Liu, Jia-ming; Lin, Li-ping; Jiang, Shu-Lian; Cui, Ma Lin; Jiao, Li; Zhang, Xiao Yang; Zhang, Li-hong; Zheng, Zhi Yong; Lin, Xuan; Lin, Shao-qin

    2013-11-01

    Based on the reaction of the active -OH group in fullerol (F) with the dissociated -COOH group in fluorescein isothiocyanate (FITC) to form an F-FITC and the enhanced effect of N, N-dimethylaniline (DMA) on phosphorescence signal of F-FITC, a new phosphorescent labeling reagent (DMA-F-FITC) was developed. What's more, a phosphorescent sensor for the determination of alpha-fetoprotein variant (AFP-V) has been designed via the coupling technique of the high sensitivity for affinity adsorption-solid substrate-room temperature phosphorimetry (AA-SS-RTP) with the strong specificity reaction between DMA-F-FITC-Con A and AFP-V. The DMA-F-FITC increased the number of luminescent molecules in the biological target which improved the sensitivity of phosphorescent sensor. The proposed sensor was responsive, simple, selective and sensitive, and it has been applied to the determination of trace AFP-V in human serum and the forecast of human diseases using phosphorescence emission wavelength of F or FITC, with the results agreed well with those obtained by enzyme-linked immunoassay (ELISA). Meanwhile, the mechanisms for the labeling reaction and the sensing detection of AFP-V were discussed.

  19. A highly sensitive solid substrate room temperature phosphorimetry for carbaryl detection based on its activating effect on NaIO4 oxidizing fluorescein.

    PubMed

    Liu, Jiaming; Huang, Qitong; Liu, Zhen-bo; Lin, Xiaofeng; Zhang, Li-Hong; Lin, Chang-Qing; Zheng, Zhi-Yong

    2014-11-01

    Fluorescein (HFin) could emit strong and stable room temperature phosphorescence (RTP) signal on polyamide membrane (PAM) using Pb(2+) as the ion perturber. Carbaryl could activate effect on NaIO4 oxidating HFin, which caused the RTP signal of the system to quench sharply. The phosphorescence intensity (?I p) of activating system higher 3.3 times (119.4/36.0) than that of non-activating system, and is directly proportional to the content of carbaryl. Thus, an activating solid substrate room temperature phosphorimetry (SSRTP) for carbaryl detection has been established. This sensitive (the limit of quantification (LOQ) was 2.0?×?10(-13) g mL(-1)), selective, simple and rapid method has been applied to determine trace carbaryl in water samples with the results consisting with those obtained by fluorimetry, showing its high accuracy. The apparent activation energy (E) and rate constant (k) of this activating reaction were 20.77 kJ mol(-1) and 1.85?×?10(-4) s(-1), respectively. Meanwhile, the mechanism of activating SSRTP for carbaryl detection was also discussed using infrared spectra (IR). PMID:25294182

  20. Mechanistic study of decreased skin penetration using a combination of sonophoresis with sodium fluorescein-loaded PEGylated liposomes with d-limonene

    PubMed Central

    Rangsimawong, Worranan; Opanasopit, Praneet; Rojanarata, Theerasak; Ngawhirunpat, Tanasait

    2015-01-01

    The effect of low frequency sonophoresis (SN, 20 kHz) on the skin transport of sodium fluorescein (NaFI)-loaded liposomes was investigated. An in vitro skin penetration study in open and blocked hair follicles was performed, and confocal laser scanning microscopy and scanning electron microscopy were used to visualize the penetration pathways. The results showed that SN significantly increased the flux of NaFI solution, whereas it significantly decreased the flux of NaFI-loaded polyethylene glycol-coated (PEGylated) liposomes with D-limonene (PL-LI). SN did not significantly affect the flux of NaFI-loaded conventional liposomes and PEGylated liposomes. In the blocked follicles, the flux of NaFI-loaded PL-LI both with and without SN decreased, indicating that NaFI-loaded PL-LI penetrated the skin via the transfollicular pathway. A confocal laser scanning microscopy image showed that in the skin without SN, the fluorescence intensity of NaFI-loaded PL-LI was observed in the skin and along the length of hair inside the skin, whereas in the skin with applied SN, the fluorescence intensity was detected only on the top of hair outside the skin. From scanning electron microscopy images, SN dislocated the corneocytes and reduced the deposition of PL-LI around hair follicles. These results revealed that SN may partially plug hair follicle orifices and reduce percutaneous absorption through the follicular pathway. PMID:26719685

  1. Effect of chloroquine and methylamine on endocytosis of fluorescein-labelled controlled IgG and of anti-(plasma membrane) IgG by cultured fibroblasts.

    PubMed

    Schneider, Y J; Trouet, A

    1981-08-01

    We report here the effect of chloroquine and methylamine two lysosomotropic drugs, on the binding, uptake and subcellular localization of fluorescein-labelled control immunoglobulin G (control IgG) a marker for non-specific adsorptive endocytosis and of anti-(plasma membrane) IgG (specific IgG), a specific ligand of cell-surface antigens. At 4 degrees C, methylamine and chloroquine inhibit the binding of control IgG to the cell surface, probably by a reversible competition. These two drugs, methylamine more than chloroquine, considerably slow down the rate at which control IgG is transferred from its binding sites on the phagosomal membrane to the lysosomal compartment; both drugs block almost completely the intralysosomal digestion of this IgG as well as the release of degradation products into the culture medium. They do not affect the binding and uptake of the specific IgG. In addition, methylamine seems to inhibit partially the return of the cell surface of membrane antigens and of membrane fragments bearing 5'-nucleotidase or binding sites for control IgG. We conclude that important steps (binding to cell surface, delivery to lysosomes, digestion and recycling of plasma membrane) involved in the uptake and the processing of IgG by fibroblasts are inhibited by these two substances. The effects of lysosomotropic agents on the regulation and function of the endocytic pathway and of lysosomes could have many pharmacological and therapeutic implications. PMID:6793366

  2. Effects of pretreatment of needle puncture and sandpaper abrasion on the in vitro skin permeation of fluorescein isothiocyanate (FITC)-dextran.

    PubMed

    Wu, Xue-Ming; Todo, Hiroaki; Sugibayashi, Kenji

    2006-06-19

    Microneedle systems have gained attention as having many advantages over transdermal patches and hypodermic needles. The procedure provides adequate skin permeation rates without pain or severe infection. To obtain information for designing a microneedle system, macroneedles were used instead of microneedles to investigate the effects of pretreatment of needle puncture in the skin barrier stratum corneum on in vitro skin permeation of fluorescein isothiocyanate (FITC)-dextrans (4.3, 9.6 and 42.0 kDa) (FD-4, FD-10 and FD-40). The effect of sandpaper abrasion was also investigated for comparison. Both pretreatments on the skin barrier significantly increased the skin permeation of FDs. Lactate dehydrogenase (LDH) leaching was measured after pretreatment of macroneedle and sandpaper abrasion on the skin to evaluate the skin damage by these pretreatment methods. Lower leaching of LDH was observed after macroneedle puncture than after sandpaper abrasion. Next, a parallel permeation-resistance model of the skin barrier was established. Skin permeation of FD-10 was predicted by the model as a function of the number of pores in the skin barrier. Our results suggest that needle puncture may provide a safe, efficient and controllable alternative for increasing transdermal drug delivery. PMID:16597490

  3. Permeability of blood-retinal barriers in urethane-induced rat retinopathy: a fluorescein angiographic, vitreous fluorophotometric, and fluorescence microscopic study.

    PubMed Central

    Kritzinger, E E; Bellhorn, R W

    1982-01-01

    Urethane-induced rat retinopathy, characterised by permeability abnormalities of the blood-retinal barriers (BRB), was studied during the developmental phases by fluorescein fundus angiography (FFA), vitreous fluorophotometry (VF), and fluorescence microscopy (FM). A distinction based on VF values could be made at the p less than 0.02 confidence level between the retinopathic rats as a group and the control rats. Fluorescence microscopy provided a basis, however, for subdividing the test group into those rats with evidence of intraretinal leakage of NaFl and those without. Statistical analysis of the VF values of the control (A), nonleaky retinopathic (B), and leaky retinopathic (C) rats showed no significance between groups A and B, but highly significant differences between groups A and C (p less than 0.001) and between groups B and C (p less than 0.01). Fluorescence microscopy also showed that leakage of NaFl from retinal vessels occurred only after the retinopathy has progressed to the point where retinal vessels had become incorporated into the pigment epithelium. We conclude from this fluorescent marker clinicopathological study that breakdown of the blood-retinal barriers is a result of an interaction between the retinal pigment epithelium (RPE) and the retinal vessels after the vessels become incorporated into the RPE. Images PMID:7115647

  4. Species-specific interaction of HIV protease inhibitors with accumulation of cholyl-glycylamido-fluorescein (CGamF) in sandwich-cultured hepatocytes.

    PubMed

    Ye, Zhi-wei; Van Pelt, Jos; Camus, Sandrine; Snoeys, Jan; Augustijns, Patrick; Annaert, Pieter

    2010-06-01

    Using sandwich-cultured hepatocytes from rat, dog, pig, and human, we investigated the species-specificity of interaction of HIV protease inhibitors (PI) with in vitro hepatic accumulation of the bile salt analogue cholyl-glycylamido-fluorescein (CGamF). Extracellular sodium depletion or coincubation with the OATP/Oatp inhibitors rifampicin and digoxin revealed that about 35% of active CGamF accumulation was mediated by Ntcp/NTCP in rat and human hepatocytes, while the contribution of this sodium-dependent transporter reached 50-60% in dog and pig hepatocytes. One or more sodium-independent transporters, likely belonging to the Oatp/OATP family, constitute a major transport mechanism for CGamF accumulation. Various HIV PI (0.5, 5, 25 microM) exhibited pronounced species differences in their interaction with active CGamF accumulation (1 microM), although some similarity was observed between the dog and human interaction profiles when HIV PI were tested at 0.5 microM. Atazanavir, indinavir, and darunavir were the most potent inhibitors of CGamF accumulation in human hepatocytes. Potent inhibition of CGamF accumulation by ritonavir in rat hepatocytes contrasted with a weak effect in human hepatocytes. Thorough characterization of in vitro disposition of probe substrates in preclinical species compared to human hepatocytes will ultimately support a better insight in species-specific mechanisms underlying drug interactions and drug-mediated toxicity. PMID:20014428

  5. Staining potential of acidulated phosphate fluoride (APF) foam on dental restorations in vitro

    PubMed Central

    Lin, David; Huang, Boyen

    2015-01-01

    Objectives: To identify the staining potential of acidulated phosphate fluoride (APF) foam on restorations in vitro. Materials and Methods: Two hundred ovine molars were used. Except 40 teeth remained unrestored as the controls, each was randomly selected to receive one of four restorative materials including preparation without restoration, glass ionomer cement (GIC), resin modified glass ionomer cement (RMGIC), or composite resin (CR). Following the procedure, topical APF was applied with a predetermined frequency. Staining formation was then evaluated. Results: APF-treated teeth and restorations appeared with a darker shade, an orange-colored surface and/or a brown margin. The staining rates on GIC, RMGIC, and CR were 50%, 27.5%, and 17.5%, respectively. GIC had a higher staining potential than RMGIC (?2 = 4.266, df = 1, P = 0.039) and CR (?2 = 9.448, df = 1, P = 0.002), whereas the difference between RMGIC and CR was indiscernible (?2 = 1.147, df = 1, P = 0.284). Repeated applications of topical APF increased the risk of staining on RMGIC (?2 = 8.436 df = 1, P = 0.004) and CR (?2 = 6.873, df = 1, P = 0.009) but not on GIC (?2 = 0, df = 1, P = 1) and the controls (?2 = 4.051, df = 3, P = 0.256). Conclusions: APF-foam-related staining was confirmed in vitro. GIC was more susceptible to fluoride staining. This study suggested aesthetic implications when applying fluorides to restored teeth. PMID:25657523

  6. Cell-death assessment by fluorescent and nonfluorescent cytosolic and nuclear staining techniques.

    PubMed

    Atale, N; Gupta, S; Yadav, U C S; Rani, V

    2014-07-01

    Apoptosis, a genetically programmed cellular event leads to biochemical and morphological changes in cells. Alterations in DNA caused by several factors affect nucleus and ultimately the entire cell leading to compromised function of the organ and organism. DNA, a master regulator of the cellular events, is an important biomolecule with regards to cell growth, cell death, cell migration and cell differentiation. It is therefore imperative to develop the staining techniques that may lead to visualize the changes in nucleus where DNA is housed, to comprehend the cellular pathophysiology. Over the years a number of nuclear staining techniques such as propidium iodide, Hoechst-33342, 4', 6-diamidino-2-phenylindole (DAPI), Acridine orange-Ethidium bromide staining, among others have been developed to assess the changes in DNA. Some nonnuclear staining techniques such as Annexin-V staining, which although does not stain DNA, but helps to identify the events that result from DNA alteration and leads to initiation of apoptotic cell death. In this review, we have briefly discussed some of the most commonly used fluorescent and nonfluorescent staining techniques that identify apoptotic changes in cell, DNA and the nucleus. These techniques help in differentiating several cellular and nuclear phenotypes that result from DNA damage and have been identified as specific to necrosis or early and late apoptosis as well as scores of other nuclear deformities occurring inside the cells. PMID:24831993

  7. Acridine orange staining reaction as an index of physiological activity in Escherichia coli

    NASA Technical Reports Server (NTRS)

    McFeters, G. A.; Singh, A.; Byun, S.; Callis, P. R.; Williams, S.

    1991-01-01

    The assumption that the acridine orange (AO) color reaction may be used as an index of physiological activity was investigated in laboratory grown Escherichia coli. Spectrofluorometric observations of purified nucleic acids, ribosomes and the microscopic color of bacteriophage-infected cells stained with AO confirmed the theory that single-stranded nucleic acids emit orange to red fluorescence while those that are double-stranded fluoresce green in vivo. Bacteria growing actively in a rich medium could be distinguished from cells in stationary phase by the AO reaction. Cells from log phase appeared red, whereas those in stationary phase were green. However, this differentiation was not seen when the bacteria were grown in a minimal medium or when a variation of the staining method was used. Also, shifting bacteria in stationary phase to starvation conditions rapidly changed their AO staining reaction. Boiling and exposure to lethal concentrations of azide and formalin resulted in stationary-phase cells that appeared red after staining but bacteria killed with chlorine remained green. These findings indicate that the AO staining reaction may be suggestive of physiological activity under defined conditions. However, variables in staining and fixation procedures as well as uncertainties associated with mixed bacterial populations in environmental samples may produce results that are not consistent with the classical interpretation of this reaction. The importance of validating the putative physiological implications of this staining reaction is stressed.

  8. A full color system for quantitative assessment of histochemical and immunohistochemical staining patterns.

    PubMed

    Ma, W; Lozanoff, S

    1999-01-01

    Quantitative measures of staining distributions are important to compare the presence and patterns of cells or macromolecules. Typically, achromatic thresholding systems are used to compare staining distributions. Achromatic video signals, however, lack sufficient resolution to identify and compare chromatic changes. The purpose of this study is to describe a full color system for analysis of chromatic staining distributions. The hardware system includes a Leitz Diaplan microscope, video camera, GVP videoboard and Amiga 3000 computer. Software was developed in "C" to partition the video signal into hue (H), saturation (S) and value (V). Also, percentage of stained area was determined. Kodak color filters were used to assess the accuracy and precision of the system. Craniofacial tissues were stained with varying concentrations of toluidine blue and primary anti-BrdU antibodies. HSV and the percentage of stained areas were determined and displayed low coefficients of error. HSV values also performed as expected for standard filters as well as cellular staining concentrations. This system is easily implemented and should be useful for comparing chromatic changes with any color resulting from histochemical or immunohistochemical procedures. PMID:10190254

  9. The application of visible wavelength reflectance hyperspectral imaging for the detection and identification of blood stains.

    PubMed

    Li, Bo; Beveridge, Peter; O'Hare, William T; Islam, Meez

    2014-12-01

    Current methods of detection and identification of blood stains rely largely on visual examination followed by presumptive tests such as Kastle-Meyer, Leuco-malachite green or luminol. Although these tests are useful, they can produce false positives and can also have a negative impact on subsequent DNA tests. A novel application of visible wavelength reflectance hyperspectral imaging has been used for the detection and positive identification of blood stains in a non contact and non destructive manner on a range of coloured substrates. The identification of blood staining was based on the unique visible absorption spectrum of haemoglobin between 400 and 500 nm. Images illustrating successful discrimination of blood stains from nine red substances are included. It has also been possible to distinguish between blood and approximately 40 other reddish stains. The technique was also successfully used to detect latent blood stains deposited on white filter paper at dilutions of up to 1 in 512 folds and on red tissue at dilutions of up to 1 in 32 folds. Finally, in a blind trial, the method successfully detected and identified a total of 9 blood stains on a red T-shirt. PMID:25498930

  10. Differentiating Taenia eggs found in human stools - Does Ziehl Neelsen staining help?

    PubMed Central

    Jimenez, Juan A.; Rodriguez, Silvia; Moyano, Luz M.; Castillo, Yesenia; García, Héctor H.

    2012-01-01

    SUMMARY Introduction Unlike other tapeworms, T. solium infections carry risk for neurocysticercosis. Differential diagnosis of human tapeworm infections relies on morphology of the scolex or proglottids, frequently unavailable. DNA-based assays are poorly available in endemic areas. Ziehl Neelsen staining has been suggested but not tested in controlled designs. We validated whether Ziehl Neelsen staining could differentiate T. solium and T. saginata eggs. Methods Tapeworm proglottids (33 specimens, 23 T. solium and 10 T. saginata) and eggs (31 specimens, 13 T. solium and 10 T. saginata) were stained. Four eggs from each sample were measured and average diameters were recorded. Results T. saginata eggs stained entirely magenta in seven of 13 cases. T. solium eggs stained entirely blue/purple in 4/18 cases and entirely magenta in one. Eggs of T. saginata were slightly larger and always ovoid, while T. solium eggs were smaller and were mostly spheric. Conclusions Ziehl Neelsen staining can occasionally distinguish fully mature T. solium from T. saginata eggs. This distinction is poorly sensitive and not completely specific. Differential staining suggest differences in embryophore components between species, evident along egg maturation. In this small series, egg morphology (shape, maximal diameter) provided appropriate differentiation between T. solium and T. saginata eggs. PMID:20579318

  11. Staining of hybrid composites with coffee, oolong tea, or red wine.

    PubMed

    Omata, Yo; Uno, Shigeru; Nakaoki, Yasuko; Tanaka, Toru; Sano, Hidehiko; Yoshida, Shigemitsu; Sidhu, Sharanbir K

    2006-03-01

    This study examined the surface staining mechanism of a photopolymerized composite by coffee, oolong tea, and red wine. Dental composite was subjected to an experimental 24-hour staining cycle: 17-hour immersion in artificial saliva solution containing 0.3% mucin followed by 7-hour immersion in coffee, tea, or wine. After one, two, and four weeks, digital images of the composite surface were analyzed in grayscale mode with an imaging analyzer. Specimens polished but not immersed were used as a baseline measurement for color change. Additionally, the effects of mechanical brushing and chlorhexidine on drink-induced staining were examined. Wine caused the most severe staining, followed by tea and coffee. After four weeks of immersion, brushing reduced surface staining by wine. On the contrary, chlorhexidine increased the staining effect of tea and coffee (p<0.05) when compared to the control specimens. In conclusion, we showed that common drinks stained the dental composite, but each by a specific mechanism that depended on external conditions such as the presence of chlorhexidine. PMID:16706307

  12. Lower azure B methylene blue ratios in Giemsa type blood and malaria stains.

    PubMed

    Russo, A; Donaldson, P T; Lillie, R D

    1978-01-01

    Starting from ancient reports that rare samples of methylene blue were apparently sufficiently contaminated with azures to give red plasmodial and red purple nuclear chromatin in Chenzinsky type methylene blue eosin stains, it was decided to determine how little azure B would suffice for such staining in methylene blue eosin stains. The traditional 1902 Giemsa had an azure : methylene blue : eosin ratio of about 6 : 3 : 6.3 : 10; Lillie's 1943 formula had a 5 : 7 : 10 ratio. In the current series of tests 5 : 7 : 10 (I), 4 : 8 : 10 (II), 3 : 9 : 10 (III), 2 : 10 : 10 (IV), 1 : 11 : 10 (V), and 0 : 12 : 10 (VI) were used. Malaria and blood stains were better than the standard 5 : 7 : 10 (I) in III, IV and II in that order. Normal and leukemic human blood, mouse blood with Plasmodium berghei, and monkey blood with the CDC strain of Pl. falciparum were used as test materials. The staining mixtures were made from highly purified samples of azure B and methylene blue. Staining mixtures contained 12 ml 0.1% thiazin dye, 10 ml 0.1% eosin, 2 ml each of glycerol, methanol and 0.1 M phosphate buffer pH 6.5, 3 ml acetone as accelerator, and distilled water to make 40 ml; staining times of 10--30 min were used. PMID:663946

  13. [Statistical analyses of different staining methods for viability assessment in cystic echinococcosis].

    PubMed

    Yildirim, Alparslan; Iça, Anil; Düzlü, Onder; Inci, Abdullah

    2007-01-01

    The viability assessment in cystic echinococcosis is important for in vitro and in vivo studies and also drug experiments. Several vital and avital stains like neutral red, methylene blue, eosin, Papanicolao, Giemsa, Ziehl-Neelsen, toluidine blue and trypan blue have been used to assess viability of protoscolexes in fertile cysts. In this study, the viability of 10 different fertile cysts were analyzed by trypan blue, eosin and methylene blue staining techniques which have been used frequently in recent years. A total of 2000 protoscolexes per hydatid cyst were examined by all three staining techniques. The obtained results were analyzed statistically by the one-way ANOVA test. The mean numbers of viable protoscolexes in examined cysts according to trypan blue, methylene blue and eosin staining techniques were 1430+/-172.9 (1150-1709), 1376.4+/-101.1 (1169-1521) and 1342+/-147.9 (1119-1608), respectively. The highest viability rate was determined with the trypan blue staining technique (71.5%), followed by 68.8% with methylene blue and 67.1% with eosin. No statistically significant differences (p > 0.05) were observed among the staining techniques. As a result, it was found that all three staining methods have similar results in viability assessment of hydatid cysts. PMID:17594648

  14. Immunohistochemistry staining for mismatch repair proteins: the endoscopic biopsy material provides useful and coherent results.

    PubMed

    Vilkin, Alex; Leibovici-Weissman, Ya'ara; Halpern, Marisa; Morgenstern, Sara; Brazovski, Eli; Gingold-Belfer, Rachel; Wasserberg, Nir; Brenner, Baruch; Niv, Yaron; Sneh-Arbib, Orly; Levi, Zohar

    2015-11-01

    Immunohistochemistry (IHC) testing for mismatch repair proteins (MMRP) in patients with colorectal cancer can be performed on endoscopic biopsy material or the surgical resection material. Data are continuing to accumulate regarding the deleterious effect of neoadjuvant chemoradiation on MMRP expression. However, despite continuing rise in the use of endoscopic biopsies for IHC, most pathology departments still use mainly the surgical materials for IHC testing. In this study we compared the quality of stains among 96 colon cancer subjects with paired endoscopic and surgical material available for MLH1, MSH2, MSH6, and PMS2 stains (96 × 4, yielding 384 paired stains). Each slide received both a quantitative score (immunoreactivity [0-3] × percent positivity [0-4]) and a qualitative score (absent; weak and focal; strong). The quantitative scores of all MMRP were significantly higher among the endoscopic material (P<.001 for all). In 358 pairs (93.2%), both the endoscopic and operative material stained either strong (322, 83.9%) or absent (36, 9.4%). In 26 pairs (6.8%), the endoscopic material stained strong, whereas the operative material stained focal and weak. No endoscopic biopsy materials stained focal and weak. Our findings indicate that the biopsy material may provide more coherent results. Although these results may indicate that biopsy material provides coherent and useful results, it is yet to be determined if the demonstrated differences pose a real clinical problem in interpreting final results of IHC staining of such kind. Hence, we suggest that when available, the endoscopic material rather than the operative one should serve as the primary substrate for IHC staining. PMID:26359539

  15. Tobacco Stained Fingers and Its Association with Death and Hospital Admission: A Retrospective Cohort Study

    PubMed Central

    John, Gregor; Genné, Daniel

    2015-01-01

    Background Among smokers, the presence of tobacco stains on fingers has recently been associated with a high prevalence of tobacco related conditions and alcohol abuse. Objective we aimed to explore tobacco stains as a marker of death and hospital readmission. Method Seventy-three smokers presenting tobacco-tar staining on their fingers and 70 control smokers were followed during a median of 5.5 years in a retrospective cohort study. We used the Kaplan-Meier survival analysis and the log-rank test to compare mortality and hospital readmission rates among smokers with and smokers without tobacco stains. Multivariable Cox models were used to adjust for confounding factors: age, gender, pack-year unit smoked, cancer, harmful alcohol use and diabetes. The number of hospital admissions was compared through a negative binomial regression and adjusted for the follow-up time, diabetes, and alcohol use. Results Forty-three patients with tobacco-stained fingers died compared to 26 control smokers (HR 1.6; 95%CI: 1.0 to 2.7; p 0.048). The association was not statistically significant after adjustment. Patients with tobacco-stained fingers needed a readmission earlier than smokers without stains (HR 2.1; 95%CI: 1.4 to 3.1; p<0.001), and more often (incidence rate ratio (IRR) 1.6; 95%CI: 1.1 to 2.1). Associations between stains and the first hospital readmission (HR 1.6; 95%CI: 1.0 to 2.5), and number of readmissions (IRR 1.5; 95%CI: 1.1 to 2.1) persisted after adjustment for confounding factors. Conclusions Compared to other smokers, those presenting tobacco-stained fingers have a high unadjusted mortality rate and need early and frequent hospital readmission even when controlling for confounders. PMID:26375287

  16. Effect of Staining Solutions on Color Stability of Silorane & Methacrylate Restorative Material

    PubMed Central

    S. Madhyastha, Prashanthi; G. Naik, Dilip; Kotian, Ravindra; Srikant, N.; M. R. Bhat, Kumar

    2015-01-01

    Color stability throughout the functional lifetime of restorations is important for the durability of treatment and of cosmetic importance. The purpose of this study was to evaluate the discoloration properties of a silorane-based (Filtek P90) and methacrylate-based (Z100) composites upon exposure to different staining solutions that are used on day to day basis (turmeric, tea, coffee, cocoa, lime, yoghurt and distilled water) for different immersion periods (1, 7, 14 and 28 days). The colors of all specimens before and after storage in the solutions were measured by a reflectance spectrophotometer based on CIE Lab system and the color differences were calculated. Data were statistically analyzed by repeated measures of ANOVA and sidak post hoc test (for immersion period);‘t’ test (for each material) and one way ANOVA (for staining agents). All the staining agents showed significant difference in staining over time in both the materials. However, Z100 showed higher quantum of discoloration at all time periods at each staining agents (p<0.005). In conclusion, the silorane-based resin (Filtek P90) composites exhibited better color stability (less change in ?E) after exposure to the staining solutions. Among the staining agents cocoa was found to be least staining followed by lime, yoghurt, coffee, tea whereas turmeric discolored the composites to the maximum. Highest discoloration was seen at day 28 in all staining agents. Cocoa and lime discolored to maximum at early stages but remained stable thereafter whereas tea, coffee and turmeric progressively discolored the composite over time.

  17. Comparison of the bisbiguanide antiseptics alexidine and chlorhexidine. II. Clinical and in vitro staining properties.

    PubMed

    Addy, M; Roberts, W R

    1981-06-01

    A blind cross-over trial was carried out to compare the tooth and tongue staining associated with the use of a 0.035% alexidine and a 0.2% chlorhexidine mouthrinse. Twenty-two volunteers were divided into two groups termed "tea drinkers" and "non-tea drinkers". All volunteers were requested to refrain from oral hygiene measures throughout two 10-day periods when they rinsed twice a day with the preparation randomly allocated for the respective period. During both periods the members of the groups excluded coffee, red wine and port from their diet. The tea drinking group consumed seven cups of tea per day. Tooth and tongue staining was recorded for extent and severity at the end of each period. The amount of stain accumulating in the two groups was similar following the use of chlorhexidine and alexidine. However, for both chlorhexidine and alexidine the extent and severity of tooth and tongue staining were significantly increased in the tea drinking group. An in vitro study of tea staining of perspex blocks exposed twice a day to 0.035% solutions of alexidine or chlorhexidine throughout a 5-day period demonstrated significantly more staining with alexidine when measured spectrophotometrically. Visually however, the differences in the specimens were minimal. Saliva treatment of the perspex did not significantly alter the staining by alexidine or chlorhexidine. The results provide further evidence for a dietary aetiology to the staining associated with cationic antiseptics. However, alexidine at the concentration used offered no advantage in reducing the side effect of staining when compared with chlorhexidine. PMID:6947988

  18. Laboratory and clinical stain removal evaluations of two tartar control dentifrices.

    PubMed

    Yankell, S L; Emling, R C; Prencipe, M; Rustogi, K; Volpe, A R

    1995-01-01

    The purpose of this research was to evaluate Colgate original Tartar Control dentifrice (TC) and new Colgate Micro Cleansing Tartar Control (MCTC) dentifrice in a laboratory and clinical study for their ability to remove induced stain. In the laboratory study, polished and etched bovine enamel specimens were stained for 4 days with a coffee, tea, and mucin, Sarcina lutea tartox culture in trypticase soy broth. Sixteen specimens were then brushed for 200 strokes with a sodium carboxymethyl cellulose solution to remove loose stain, and then brushed for 300 strokes with a 1:1 toothpaste slurry. Tooth color was measured by reflectance with the CLE L*a*b* scale. The calculated stain removal, 37% for MCTC and 24% for TC, was significantly different (p<0.01) favoring the MCTC dentifrice. In the clinical study, eight-six subjects were given Peridex (0.12% chlorhexidine gluconate) mouthrinse, a low abrasive dentifrice and a soft toothbrush to use twice a day for 8 weeks. Stain was monitored on the buccal and lingual surfaces of the anterior teeth using the Lobene index. The seventy-seven subjects with sufficient total stain were stratified into two balanced groups, given a new soft toothbrush and were randomly assigned to use one of the dentifrices tested, twice a day for 8 weeks. At 4, 6 and 8 weeks in the study, the MCTC dentifrice product had consistent, significantly (p<0.05-0.001) lower total stain scores than the TC dentifrice. In the laboratory and clinical studies conducted, even though stain induction and evaluation procedures differed, the new Colgate Micro Cleansing Tartar Control dentifrice was significantly more effective at removing stain than the Colgate original Tartar Control dentifrice formula. PMID:8624234

  19. Topical dual-stain difference imaging for rapid intra-operative tumor identification in fresh specimens

    PubMed Central

    Davis, Scott C.; Gibbs, Summer L.; Gunn, Jason R.; Pogue, Brian W.

    2014-01-01

    Assessing tumor margin status during surgery is critical to ensure complete resection of cancer tissue; however, current approaches are ineffective and often result in repeat surgery. We present a novel optical imaging approach for margin assessment using topical application of two fluorescent stains, one targeted to a tumor biomarker and the other a non-targeted reference, to freshly excised specimens. Computing a normalized difference image from fluorescence images of the targeted and untargeted stains suppresses the confounding effects of non-specific uptake. Applying this approach in excised breast tumor models produced promising tumor-to-normal tissue contrasts that were significantly higher than single-targeted-stain imaging. PMID:24281541

  20. SiR–Hoechst is a far-red DNA stain for live-cell nanoscopy

    PubMed Central

    Lukinavi?ius, Gražvydas; Blaukopf, Claudia; Pershagen, Elias; Schena, Alberto; Reymond, Luc; Derivery, Emmanuel; Gonzalez-Gaitan, Marcos; D'Este, Elisa; Hell, Stefan W.; Wolfram Gerlich, Daniel; Johnsson, Kai

    2015-01-01

    Cell-permeable DNA stains are popular markers in live-cell imaging. Currently used DNA stains for live-cell imaging are either toxic, require illumination with blue light or are not compatible with super-resolution microscopy, thereby limiting their utility. Here we describe a far-red DNA stain, SiR–Hoechst, which displays minimal toxicity, is applicable in different cell types and tissues, and is compatible with super-resolution microscopy. The combination of these properties makes this probe a powerful tool for live-cell imaging. PMID:26423723

  1. The effect of osmium staining on lamellar spacing in thin polystyrene-polyisoprene diblock copolymer films

    NASA Astrophysics Data System (ADS)

    Staniewicz, L.; Donald, A. M.; Stokes, D. J.

    2010-07-01

    Thin films of lamellar-structured polystyrene-block-polyisoprene were vapour stained with osmium tetroxide and compared using transmission imaging in an environmental scanning electron microscope to unstained films of the same polymer. Staining was found to swell the affected phase by a factor of 1.15 in-plane, compressing the unstained phase to a factor of 0.71 of its original size as it did so. Additionally, images exhibiting contrast between phases in the unstained sample were successfully taken with conventional transmission electron microscopy, showing that staining is not necessary for imaging such samples.

  2. SiR-Hoechst is a far-red DNA stain for live-cell nanoscopy.

    PubMed

    Lukinavi?ius, Gražvydas; Blaukopf, Claudia; Pershagen, Elias; Schena, Alberto; Reymond, Luc; Derivery, Emmanuel; Gonzalez-Gaitan, Marcos; D'Este, Elisa; Hell, Stefan W; Gerlich, Daniel Wolfram; Johnsson, Kai

    2015-01-01

    Cell-permeable DNA stains are popular markers in live-cell imaging. Currently used DNA stains for live-cell imaging are either toxic, require illumination with blue light or are not compatible with super-resolution microscopy, thereby limiting their utility. Here we describe a far-red DNA stain, SiR-Hoechst, which displays minimal toxicity, is applicable in different cell types and tissues, and is compatible with super-resolution microscopy. The combination of these properties makes this probe a powerful tool for live-cell imaging. PMID:26423723

  3. Influence of chromatin condensation on the absorption spectra of nuclei stained with toluidine blue.

    PubMed

    Erenpreisa, J; Freivalds, T; Selivanova, G

    1992-01-01

    To study the influence of chromatin condensation on the absorption spectra of nuclei stained with toluidine blue, DNA staining methods--which favour or prevent dye polymerization--were applied to the imprints of rat tissues that differed greatly in the density of chromatin packing. It is stated that all factors promoting dye polymerization cause a left shift of the spectra while the factors preventing it, a right one. It was found that condensation of the chromatin can raise prerequisites that both enhance and hinder polymerization, and that the final result depends on the staining method, the manner of chromatin folding, and the density of its packing. PMID:1365771

  4. Large-volume en-bloc staining for electron microscopy-based connectomics

    PubMed Central

    Hua, Yunfeng; Laserstein, Philip; Helmstaedter, Moritz

    2015-01-01

    Large-scale connectomics requires dense staining of neuronal tissue blocks for electron microscopy (EM). Here we report a large-volume dense en-bloc EM staining protocol that overcomes the staining gradients, which so far substantially limited the reconstructable volumes in three-dimensional (3D) EM. Our protocol provides densely reconstructable tissue blocks from mouse neocortex sized at least 1?mm in diameter. By relaxing the constraints on precise topographic sample targeting, it makes the correlated functional and structural analysis of neuronal circuits realistic. PMID:26235643

  5. Phosphoprotein staining for sodium dodecyl sulfate-polyacrylamide gel electrophoresis using fluorescent reagent morin hydrate.

    PubMed

    Wang, Xu; Hwang, Sun-Young; Cong, Wei-Tao; Jin, Li-Tai; Choi, Jung-Kap

    2013-04-01

    A fluorescence-based stain with 3,5,7,2',4'-pentahydroxyflavone (morin hydrate, MH) was designed to stain phosphoproteins in one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Al(3+) was applied as a "fixed bridge," providing an efficient energy transfer channel between phosphoprotein and MH, to produce a strong fluorescent complex for the determination of phosphoprotein. As little as 62.5ng of ?-casein (7 or 8 phosphates) and ?-casein (5 phosphates), 125ng of ovalbumin (2 phosphates), and ?-casein (1 phosphate) could be visualized with a wide linear dynamic range. In comparison with conventional methods, MH stain is a time-saving method that takes just 90min. It also has good compatibility with routine protein stainings such as Coomassie Brilliant Blue R (CBBR) and SYPRO Ruby for total protein analysis. PMID:23274386

  6. Limonite-stained Siltite near the Blackbird Cobalt-Copper Mine

    USGS Multimedia Gallery

    Limonite-stained outcrop of the banded siltite unit of the Apple Creek Formation, near Blackbird Creek, south of the Blackbird cobalt-copper mine area, in the Salmon River Mountains of east-central Idaho....

  7. Identification of malaria parasites by fluorescence microscopy and acridine orange staining

    PubMed Central

    Shute, G. T.; Sodeman, T. M.

    1973-01-01

    The need for a technique that is more sensitive than the use of Romanowsky-stained thick blood films for detecting malaria parasites at low concentration in the blood is well recognized. One of the more promising methods appeared to be fluorochrome staining with acridine orange. However, reports on the efficacy of the technique were contradictory and it was not clear to what extent blood films taken under survey conditions would contain fluorescing artefacts that might confuse diagnosis. An investigation indicated that, provided reasonable care was taken, blood films made under survey conditions contained few confusing artefacts. However, it was found that, while acridine orange staining might have a slight advantage when large malaria parasites were present, it was inferior to routine Romanowsky staining for the detection of young trophozoites, the inferiority becoming more pronounced as the parasite concentration decreased. PMID:4130021

  8. High contrast en bloc staining of neuronal tissue for field emission scanning electron microscopy

    PubMed Central

    Tapia, Juan C.; Kasthuri, Narayanan; Hayworth, Kenneth; Schalek, Richard; Lichtman, Jeff W.; Smith, Stephen J; Buchanan, JoAnn

    2013-01-01

    Conventional heavy metal post staining methods on thin sections lend contrast but often cause contamination. To avoid this problem, we tested several en bloc staining techniques to contrast tissue in serial sections mounted on solid substrates for examination by Field Emission Scanning Electron Microscope (FESEM). Because FESEM section imaging requires that specimens have higher contrast and greater electrical conductivity than transmission electron microscope (TEM) samples, our technique utilizes osmium impregnation (OTO) to make the samples conductive while heavily staining membranes for segmentation studies. Combining this step with other classic heavy metal en bloc stains including uranyl acetate, lead aspartate, copper sulfate and lead citrate produced clean, highly contrasted TEM and SEM samples of insect, fish, and mammalian nervous system. This protocol takes 7–15 days to prepare resin embedded tissue, cut sections and produce serial section images. PMID:22240582

  9. Targeted alteration of real and imaginary refractive index of biological cells by histological staining

    E-print Network

    Taflove, Allen

    Targeted alteration of real and imaginary refractive index of biological cells by histological of epithe- lial cells caused by histological stains such as hematox- ylin and eosin-containing cytostain. We

  10. Electron tomography of negatively stained complex viruses: application in their diagnosis

    PubMed Central

    Mast, Jan; Demeestere, Lien

    2009-01-01

    Background Electron tomographic analysis can be combined with the simple and rapid negative staining technique used in electron microscopy based virus diagnosis. Methods Standard negative staining of representative examples of parapoxviruses and paramyxoviruses was combined with electron tomographic analysis. Results Digital sectioning of reconstructions of these viruses at a selected height demonstrated the viral ultrastructure in detail, including the characteristic diagnostic features like the surface threads on C-particles of a parapoxvirus and individual glycoproteins and the internal nucleoprotein strand of Newcastle disease virus. For both viruses, deformation and flattening were observed. Conclusion The combination of negative staining of complex viruses with electron tomographic analysis, allows visualizing and measuring artifacts typical for negative staining. This approach allows sharp visualisation of structures in a subnanometer-thick plane, avoiding blurring due to superposition which is inherent to TEM. In selected examples, such analyses can improve diagnosis of viral agents. PMID:19208223

  11. What Poisoned the Apple Juice? A Gram Staining and Selective Media Lab.

    ERIC Educational Resources Information Center

    Hammond, Paul; Brown, Nikole; Hauser, Doug; Pomart, Katrina; Karcher, Sue; Balschweid, Mark

    2002-01-01

    Introduces an inquiry-based laboratory experiment in which students identify an unknown bacterial species by using techniques such as Gram staining. Uses an authentic problem solving approach in a scenario entitled, "What poisoned the apple juice?" (YDS)

  12. Identification of senescence and death in Emiliania huxleyi and Thalassiosira pseudonana: Cell staining, chlorophyll alterations, and

    E-print Network

    Berges, John A.

    staining, chlorophyll alterations, and dimethylsulfoniopropionate (DMSP) metabolism Daniel J. Franklin pigment degradation and transformation occurred in both species after growth; chlorophyll a (Chl a, total DMSP and DMS increased in E. huxleyi. Our data show a novel chlorophyll alteration product

  13. Under-air staining of the anterior capsule using Trypan blue with a 30 G needle.

    PubMed

    Giammaria, Daniele; Giannotti, Michele; Scopelliti, Angelo; Pellegrini, Giacomo; Giannotti, Bruno

    2013-01-01

    The original technique of staining the anterior capsule of the lens with Trypan blue involves the injection of an air bubble in the anterior chamber. A drawback of this technique is the possible instability of the anterior chamber caused by the sudden exit of air when the dye is injected with the cannula through the side-port incision. Other staining techniques that use viscoelastic substances to increase the stability of the anterior chamber and to dose the injected dye have been described. The authors present an under-air staining technique of the anterior capsule using one drop of Trypan blue injected with a 30 G needle through the peripheral cornea. This procedure prevents the air bubble from escaping the anterior chamber and allows fast and selective staining of the capsule. PMID:23386783

  14. Assessing Nezara viridula (Hemiptera: Pentatomidae) feeding damage in macadamia nuts by using a biological stain.

    PubMed

    Golden, Mary; Follett, Peter A; Wright, Mark G

    2006-06-01

    Damage caused by southern green stink bug, Nezara viridula (L.), to macadamia nuts, Macadamia integrifolia Maiden & Betche, is normally determined after nuts are harvested and processed, which may be many months after damage occurred in the field. We developed a method using ruthenium red dye to stain stink bug feeding probes and indirectly assess feeding activity in macadamia nuts. By using the staining method, feeding probes were easily detected on the husk, shell, and kernel. Husk probing was highly correlated (0.80-0.90) with feeding and damage to the kernel. Failure rate to detect kernel damage from stained husk probes was generally <6%. The staining method was equally effective for immature and mature nuts; therefore, N. viridula feeding activity can be monitored throughout the season to evaluate pest management tactics and forecast outbreak populations. PMID:16813317

  15. A staining method for assessing the viability of Esteya vermicola conidia.

    PubMed

    Wang, Yunbo; Thang, NguyenTrong; Li, Zheng; Zhang, Yongan; Li, Jingjie; Xue, Jianjie; Gu, Lijuan; Hong, VuThuy; Mira, Lee; Sung, Changkeun

    2014-07-01

    The viability of conidia of Esteya vermicola, a potentially important biocontrol agent against the pinewood nematode Bursaphelenchus xylophilus, is usually determined by cultivation for 18-48 h in culture medium. As an alternative to this labor-intensive method, we have developed a rapid, simple, and low-cost staining method for assessing E vermicola conidia survival rates. A mixture of neutral red and methylene blue was found to be the most optimal among several stains that also included safranin O and Janus green B. This mixture stained nonviable conidia blue, in contrast to viable conidia, which were stained red in the cytoplasm and blue in the cell wall. This method may be particularly useful for traditional research laboratories, as it provides rapid results using common, relatively inexpensive laboratory equipment. PMID:24585076

  16. At-home vital bleaching: effects on stained enamel and dentin.

    PubMed

    Scherer, W; Penugonda, B; Styner, D; Georgescu, M

    1992-03-01

    In a scanning electron microscope study evaluating the effects of a 10% carbamide peroxide gel on stained enamel and dentin, no surface morphologic changes were observed after 72 hours of contact with the bleaching gel. PMID:1525369

  17. Screening for cervical neoplasia: a community-based trial comparing Pap staining, human papilloma virus testing, and the new bi-functional Celldetect® stain.

    PubMed

    Idelevich, Pavel; Kristt, Don; Schechter, Eduardo; Lew, Sylvia; Elkeles, Adi; Terkieltaub, Dov; Rivkin, Ilia; Bruchim, Ilan; Fishman, Ami

    2012-12-01

    Although cytological screening for cervical neoplasia has lowered mortality rates, current screening methods are plagued by sub-optimal sensitivity and/or specificity. The purpose of this study was to compare the performance of the new CellDetect® staining technology as a potential screening tool. This initial, non-blinded study, utilized samples are taken at a community-based clinic. The diagnostic results using CellDetect® were compared with the performance of Pap staining and human papilloma virus (HPV) testing on the same material, as well as the follow-up biopsies. These data were statistically analyzed in terms of sensitivity, specificity, predictive value (N.P.V and P.P.V), and inter-observer agreement. Bi-functional CellDetect® staining revealed morphological details and tinctorial properties that permitted recognition of neoplasia even at low magnification. Performance-wise, CellDetect® demonstrated non-inferiority for all statistical parameters to both Pap and HPV tests. Importantly, superior sensitivity compared with Pap staining was observed, as well as higher specificity than HPV testing with near equivalent sensitivity. We conclude that CellDetect® is a promising approach to early detection of cervical cancer because of its bi-functional capabilities that afford high sensitivity and specificity. The data suggest that this new methodology warrants further and more extensive clinical evaluation. PMID:21630482

  18. Transient receptor potential ankyrin 1 activation enhances hapten sensitization in a T-helper type 2-driven fluorescein isothiocyanate-induced contact hypersensitivity mouse model

    SciTech Connect

    Shiba, Takahiro; Tamai, Takuma; Sahara, Yurina; Kurohane, Kohta; Watanabe, Tatsuo; Imai, Yasuyuki

    2012-11-01

    Some chemicals contribute to the development of allergies by increasing the immunogenicity of other allergens. We have demonstrated that several phthalate esters, including dibutyl phthalate (DBP), enhance skin sensitization to fluorescein isothiocyanate (FITC) in a mouse contact hypersensitivity model, in which the T-helper type 2 (Th2) response is essential. On the other hand, some phthalate esters were found to activate transient receptor potential ankyrin 1 (TRPA1) cation channels on sensory neurons. We then found a positive correlation between the enhancing effects of several types of phthalate esters on skin sensitization to FITC and their ability to activate TRPA1. Here we examined the involvement of TRPA1 in sensitization to FITC by using TRPA1 agonists other than phthalate esters. During skin sensitization to FITC, the TRPA1 agonists (menthol, carvacrol, cinnamaldehyde and DBP) augmented the ear-swelling response as well as trafficking of FITC-presenting dendritic cells to draining lymph nodes. We confirmed that these TRPA1 agonists induced calcium influx into TRPA1-expressing Chinese hamster ovary (CHO) cells. We also found that TRPA1 antagonist HC-030031 inhibited DBP-induced calcium influx into TRPA1-expressing CHO cells. After pretreatment with this antagonist upon skin sensitization to FITC, the enhancing effect of DBP on sensitization was suppressed. These results suggest that TRPA1 activation will become a useful marker to find chemicals that facilitate sensitization in combination with other immunogenic haptens. -- Highlights: ? Role of TRPA1 activation was revealed in a mouse model of skin sensitization to FITC. ? TRPA1 agonists enhanced skin sensitization as well as dendritic cell trafficking. ? Dibutyl phthalate (DBP) has been shown to enhance skin sensitization to FITC. ? TRPA1 activation by DBP was inhibited by a selective antagonist, HC-030031. ? HC-030031 inhibited the enhancing effect of DBP on skin sensitization to FITC.

  19. Evaluating the use of 3'-(p-Aminophenyl) fluorescein for determining the formation of highly reactive oxygen species in particle suspensions

    PubMed Central

    2009-01-01

    Background Given the importance of highly reactive oxygen species (hROS) as reactants in a wide range of biological, photochemical, and environmental systems there is an interest in detection and quantification of these species. The extreme reactivity of the hROS, which includes hydroxyl radicals, presents an analytical challenge. 3'-(p-Aminophenyl) fluorescein (APF) is a relatively new probe used for measuring hROS. Here, we further evaluate the use of APF as a method for the detection of hydroxyl radicals in particle suspensions. Results Particle-generated hROS can be quantified with an estimated detection limit of 50 nM. Measurements of hROS in two National Institute of Standards and Technology (NIST 2709 and 2710) soil suspensions and a pyrite suspension show non-linear particle dose-response curves for hROS generation. APF can also be used in solutions containing no dissolved molecular oxygen (O2) to determine the role of O2 in the formation of hROS. Results confirm that O2 is mechanistically important in the formation of hROS by dissolved ferrous iron and in pyrite suspensions. Conclusion Given the non-linear dose-response curves for particle generation of hROS, we recommend using several particle loadings in experiments aimed to compare particles for their hROS generation potential. The method presented here is specific to hROS and simple to perform. The analysis can be conducted in mobile labs as only basic laboratory equipment is required. PMID:19671165

  20. Cellular accumulation of cholyl-glycylamido-fluorescein in sandwich-cultured rat hepatocytes: kinetic characterization, transport mechanisms, and effect of human immunodeficiency virus protease inhibitors.

    PubMed

    Ye, Zhi-Wei; Augustijns, Patrick; Annaert, Pieter

    2008-07-01

    The present study was aimed at characterizing the in vitro cellular uptake mechanism and kinetics of the bile salt analog cholylglycylamido-fluorescein (CGamF) in sandwich-cultured rat hepatocytes (SCRHs). Concentration-dependent inhibition of active CGamF accumulation by seven human immunodeficiency virus (HIV) protease inhibitors (PIs) was also determined and compared with inhibition data obtained with taurocholate (TC) as a substrate. A K(m) value of 9.3 +/- 2.6 microM was obtained for saturable CGamF accumulation in SCRHs. The organic anion-transporting polypeptide (Oatp) inhibitor rifampicin (100 microM) inhibited CGamF (1 microM) accumulation in SCRHs by 72%; sodium depletion did not further reduce CGamF accumulation. In contrast, TC accumulation was reduced by only 25% in the presence of rifampicin, whereas additional sodium depletion resulted in a complete loss of TC accumulation. These data imply that Oatp(s) and sodium taurocholate-cotransporting polypeptide preferentially mediate hepatic uptake of CGamF and TC, respectively. Coincubation of CGamF with HIV PIs (amprenavir, atazanavir, darunavir, indinavir, nelfinavir, ritonavir, saquinavir) revealed that five of them had a concentration-dependent inhibitory effect on CGamF accumulation in SCRHs, with IC(50) values between 0.25 +/- 0.07 and 43 +/- 12 microM. The rank order for inhibition of CGamF accumulation in SCRHs was: ritonavir > saquinavir > atazanavir > darunavir > amprenavir. Indinavir (up to 100 microM) did not alter CGamF accumulation, whereas nelfinavir solubility was limited to 10 microM. Taken together, these findings illustrate the utility of CGamF as a suitable probe (complementary to TC) for rapid in vitro determination of interaction potential with sodium-independent uptake mechanisms (likely Oatps) in rat liver. PMID:18420783

  1. Fluorescein-5-isothiocyanate-conjugated protein-directed synthesis of gold nanoclusters for fluorescent ratiometric sensing of an enzyme-substrate system.

    PubMed

    Ke, Chen-Yi; Wu, Yun-Tse; Tseng, Wei-Lung

    2015-07-15

    This study describes the synthesis of a dual emission probe for the fluorescent ratiometric sensing of hydrogen peroxide (H2O2), enzyme activity, and environmental pH change. Green-emitting fluorescein-5-isothiocyanate (FITC) was conjugated to the amino groups of bovine serum albumin (BSA). This FITC-conjugated BSA acted as a template for the synthesis of red-emitting gold nanoclusters (AuNCs) under alkaline conditions. Under single wavelength excitation, FITC/BSA-stabilized AuNCs (FITC/BSA-AuNCs) emitted fluorescence at 525 and 670nm, which are sensitive to changes in solution pH and H2O2 concentration, respectively. The effective fluorescence quenching of AuNCs by H2O2 enabled FITC/BSA-AuNCs to ratiometrically detect the H2O2 product-related enzyme system and its inhibition, including glucose oxidase-catalyzed oxidation of glucose, acetylcholinesterase/choline oxidase-mediated hydrolysis and oxidation of acetylcholine, and paraoxon-induced inhibition of acetylcholinesterase activity. When pH-insensitive AuNCs were used as an internal standard, FITC/BSA-AuNCs offered a sensitive and reversible ratiometric sensing of a 0.1-pH unit change in the pH range 5.0-8.5. The pH-induced change in FITC fluorescence enabled FITC/BSA-AuNCs to detect an ammonia product-related enzyme system. This was exemplified with the determination of urea in plasma by urease-mediated hydrolysis of urea. PMID:25703728

  2. Systematic investigation of drip stains on apparel fabrics: The effects of prior-laundering, fibre content and fabric structure on final stain appearance.

    PubMed

    de Castro, Therese C; Taylor, Michael C; Kieser, Jules A; Carr, Debra J; Duncan, W

    2015-05-01

    Bloodstain pattern analysis is the investigation of blood deposited at crime scenes and the interpretation of that pattern. The surface that the blood gets deposited onto could distort the appearance of the bloodstain. The interaction of blood and apparel fabrics is in its infancy, but the interaction of liquids and apparel fabrics has been well documented and investigated in the field of textile science (e.g. the processes of wetting and wicking of fluids on fibres, yarns and fabrics). A systematic study on the final appearance of drip stains on torso apparel fabrics (100% cotton plain woven, 100% polyester plain woven, blend of polyester and cotton plain woven and 100% cotton single jersey knit) that had been laundered for six, 26 and 52 cycles prior to testing was investigated in the paper. The relationship between drop velocity (1.66±0.50m/s, 4.07±0.03m/s, 5.34±0.18m/s) and the stain characteristics (parent stain area, axes 1 and 2 and number of satellite stains) for each fabric was examined using analysis of variance. The experimental design and effect of storing blood were investigated on a reference sample, which indicated that the day (up to five days) at which the drops were generated did not affect the bloodstain. The effect of prior-laundering (six, 26 and 52 laundering cycles), fibre content (cotton vs. polyester vs. blend) and fabric structure (plain woven vs. single jersey knit) on the final appearance of the bloodstain were investigated. Distortion in the bloodstains produced on non-laundered fabrics indicated the importance of laundering fabrics to remove finishing treatments before conducting bloodstain experiments. For laundered fabrics, both the cotton fabrics and the blend had a circular to oval stain appearance, while the polyester fabric had a circular appearance with evidence of spread along the warp and weft yarns, which resulted in square-like stains at the lowest drop velocity. A significant (p<0.001) increase in the stain size on laundered blend fabric was identified. Bloodstain characteristics varied due to fibre content (p<0.001) and fabric structure (p<0.001). Blood on polyester fabric, after impact, primarily moved due to capillary force and wicking of the blood along the fibres/yarns, while for the cotton fabrics wicking was accompanied by movement of blood into the fibres/yarns. This study highlights the importance for forensic analysts of apparel evidence to consider the age, the fibre type and the fabric structure before interpreting bloodstain patterns. PMID:25828382

  3. Natural fruit extracts as non-toxic fluorescent dyes for staining fungal chlamydospores.

    PubMed

    Vujanovic, Silva; Goh, Yit Kheng; Vujanovic, Vladimir

    2012-01-01

    Currently, most synthetic dyes utilized for fungal fluorescent staining are toxic, carcinogenic, or harmful to animals, humans, and the environment. This study proposes non-toxic extracts of fruits from the genera Rhamnus, Ribes, Sambucus, Viburnum, Sorbus and Beta as simple, safe, and ecological alternatives to chemical fluorescent dye for efficient staining of Fusarium chlamydospore cells using, as test strains, five different pathogenic Fusarium species. PMID:22806815

  4. Silver Staining Protocol Fixer: 50% MeOH, 12% HAc, 0.05% formalin

    E-print Network

    Lamond, Angus I.

    Silver Staining Protocol Buffers Fixer: 50% MeOH, 12% HAc, 0.05% formalin (35% Formaldehyde) Volume Milli-Q 200 ml Silver Nitrate: 0.2% AgNO3 , 0.076% formalin (35% Formaldehyde) Volume 200 ml AgNO3 0.4 g. Repeat 3 times. 5. Stain gel with Silver nitrate for 20mins. 6. Wash the gel with Milli-Q for 1 minute

  5. Detection of alkali-silica reaction swelling in concrete by staining

    DOEpatents

    Guthrie, Jr., George D. (Santa Fe, NM); Carey, J. William (Santa Fe, NM)

    1998-01-01

    A method using concentrated aqueous solutions of sodium cobaltinitrite and rhodamine B is described which can be used to identify concrete that contains gels formed by the alkali-silica reaction (ASR). These solutions present little health or environmental risk, are readily applied, and rapidly discriminate between two chemically distinct gels; K-rich, Na--K--Ca--Si gels are identified by yellow staining, and alkali-poor, Ca--Si gels are identified by pink staining.

  6. Detection of alkali-silica reaction swelling in concrete by staining

    DOEpatents

    Guthrie, G.D. Jr.; Carey, J.W.

    1998-04-14

    A method using concentrated aqueous solutions of sodium cobalt nitrite and rhodamine B is described which can be used to identify concrete that contains gels formed by the alkali-silica reaction (ASR). These solutions present little health or environmental risk, are readily applied, and rapidly discriminate between two chemically distinct gels; K-rich, Na-K-Ca-Si gels are identified by yellow staining, and alkali-poor, Ca-Si gels are identified by pink staining.

  7. Comparison of modified Chicago sky blue stain and potassium hydroxide mount for the diagnosis of dermatomycoses and onychomycoses.

    PubMed

    Liu, Zhong; Sheng, Ping; Yang, Yan-Ping; Li, Wen; Huang, Wen-Ming; Wang, Jie-Di; Fan, Yi-Ming

    2015-05-01

    The diagnostic value of modified Chicago sky blue (CSB) stain and potassium hydroxide (KOH) mount for superficial mycoses was compared using fungal culture as gold standard. The sensitivity and screening time of the CSB stain were superior to the KOH mount. The CBS stain is simple, quick and reliable for diagnosing superficial mycoses. PMID:25765148

  8. Lasers in Surgery and Medicine 39:118127 (2007) Laser Surgery of Port Wine Stains Using Local Vaccum

    E-print Network

    Aguilar, Guillermo

    2007-01-01

    Lasers in Surgery and Medicine 39:118­127 (2007) Laser Surgery of Port Wine Stains Using Local port wine stain (PWS) laser treatment. Study Design/ Materials and Methods: Mathematical models INTRODUCTION The goal of cutaneous laser surgery during treatment of port wine stain (PWS) birthmarks

  9. Experimental study of multiple-intermittent cryogen spurts and laser pulses for the treatment of port wine stain birthmarks

    E-print Network

    Aguilar, Guillermo

    of port wine stain birthmarks Guillermo Aguilar1,2 , Bernard Choi1 , John A. Viator1 , Dan Andersen3 of epidermal damage during pulsed laser treatment of port wine stain (PWS) birthmarks. Unfortunately such as port wine stain (PWS) birthmarks (150 to 500 µm deep) [1-2]. Clinical studies [2-4] have demonstrated

  10. Fluorescent viability stains to probe the metabolic status of aflatoxigenic fungus in dual culture of Aspergillus flavus and Pichia anomala

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The metabolic activity of aflatoxigenic fungus, Aspergillus flavus co-cultured with a biocontrol yeast, Pichia anomala was examined using several vital stains. Both the FUN-1 stain and the combined use of DiBAC4(5) with CDFA-AM stains demonstrated that P. anomala inactivated the ATP generating syst...

  11. A six-week clinical study to compare the stain removal efficacy of three dentifrices.

    PubMed

    Nathoo, Salim; Petrone, Margaret E; DeVizio, William; Chaknis, Patricia; Volpe, Anthony R

    2002-01-01

    The objective of this double-blind clinical study was to compare the tooth whitening efficacy (stain removal) of a new commercially available tooth whitening dentifrice (Colgate Total Plus Whitening Toothpaste) containing 0.2% triclosan and 3.0% PVM/MA copolymer in a 0.243% sodium fluoride/high cleaning silica base, with that of two commercially available dentifrices, Crest Multi-Care Advanced Cleaning Toothpaste and Colgate Winterfresh Gel Fluoride Toothpaste. Following a baseline examination to assess extrinsic tooth stain, qualifying adult male and female subjects were randomized into three treatment groups which were balanced for gender, age and level of extrinsic tooth stain. Subjects were asked to brush their teeth twice (morning and evening) for one minute with their assigned dentifrice using a soft-bristled toothbrush. Examinations for extrinsic tooth stain were repeated after six weeks' use of the study dentifrices. One-hundred and twenty-three (123) subjects complied with the protocol and completed the study. At the six-week examination, subjects assigned to the Colgate Total Plus Whitening Toothpaste treatment group exhibited statistically significant reductions in extrinsic tooth stain area and extrinsic tooth stain intensity relative to those subjects assigned to the Crest Multi-Care Advanced Cleaning Toothpaste and the Colgate Winterfresh Gel Fluoride Toothpaste. PMID:11695214

  12. Balancing Image Quality and Compression Factor for Special Stains Whole Slide Images

    PubMed Central

    Sharma, Anurag; Bautista, Pinky; Yagi, Yukako

    2012-01-01

    The objective is to find a practical balance between quality and performance for daily high volume whole slide imaging. We evaluated whole slide images created by various scanners at different compression factors to determine the best suitable quality factor (QF) needed for pathological images of special stains. Method: We scanned two sets of eight special stains slides each at 0.50 ?m/pixel resolution in Hamamatsu scanner at six and five QF levels respectively to generate 72 images which were observed at a calibrated monitor by imaging specialists, a histo-technician, and a pathologist to find the most suitable QF level for special stains in digital slides. Results: Most special stains images were acceptable at QF 30 except for the stain Reticulin where the lowest acceptable QF was 50. The compression of images from QF 90 to QF 50 reduced the size of the images by 62.73%. Conclusion: 0.50 ?m/pixel images at QF 50 or above were found suitable 12 special stain. PMID:21987586

  13. X-gal staining of canine skin tissues: A technique with multiple possible applications

    PubMed Central

    Pati, Soumyaranjan; Jain, Sumeet; Behera, Monalisa; Acharya, Aditya Prasad; Panda, Susen K.; Senapati, Shantibhusan

    2014-01-01

    Background: Estimation of ?-galactosidase (?gal) activity in human cells and tissues indicate its possible use as a marker of senescence. Objectives: This study was done to detect senescence-associated ?gal (SA-?gal) activity in canine skin tissue by using its substrate 5-bromo-4-chloro-3-indolyl ?-D-galactosidase (X-gal). Materials and Methods: Skin samples were collected through rapid necropsy process. The X-gal staining was done by altering different factors of the staining procedure like pH of the reagents and incubation time. Further, effect of tissue preservation procedure was also evaluated. Results: Typical X-gal staining was detected in old dogs’ skin samples and it was detectable both at pH 6 and pH 7.3. The cells present in the inner lining of the hair follicles and sebaceous glands are the major cells that have high SA-?gal activity. The X-gal staining intensity was more prominent in tissues preserved in liquid nitrogen at -196°C than in -80°C freezer. Prolonged incubation period increased the intensity of staining. Conclusions: This study indicates possibility of X-gal staining in canine tissues and opens an avenue for further in-depth studies that might be useful for different research and clinical studies like determination of dog's approximate age. PMID:25097391

  14. Efficacy of LED versus KTP laser activation of photodynamic bleaching of tetracycline-stained dentine.

    PubMed

    Bennett, Zackary Y; Walsh, Laurence J

    2015-09-01

    In some well-established laser applications where large spot sizes are used, an array of high-intensity light emitting diodes (LED) emitting at similar wavelength could potentially replace the laser. This situation applies for the photodynamic bleaching of stains in teeth. This study compared the relative efficacy of an array of visible green LED (535 nm?±?15 nm) with a KTP laser in photodynamic bleaching of tetracycline-stained dentine in human tooth roots. After establishing consistent staining in 96 roots using a validated method, the roots were sectioned into 2-3-mm thick horizontal slices that were treated with gels containing rhodamine B (Smartbleach® or Smartbleach® 3LT). Colour changes were tracked up to 1 month after treatment. While both systems were effective in bleaching the tetracycline-stained dentine, KTP laser activation gave greater bleaching efficacy than LED activation, enhancing the action of the gel. Use of the KTP laser would be preferable over an LED system when confronted with tetracycline staining. Use of this photodynamic bleaching method offers valuable means to reduce the severity of tetracycline staining. PMID:25288264

  15. Quest for An Ideal, Simple and Cost-Effective Stain for Morphological Assessment of Sperms

    PubMed Central

    Govindashetty, Abhishek Mandya; Krishnamurthy, Anoosha; Puttaveerachary, Ashok Kagathur; Manchaiah, Sanjay; Shimoga, Indira Channagangappa; Mallaradhya, Sushma Hulikere; Gowda, Sarvesh Ballekoppa Mukunda

    2015-01-01

    Background Recent alarming trends of a substantial rise in the number of cases of infertility with as many as 30-40% being attributed to male-factor associated causes have created a need for further studies and advancements in semen analysis. Despite the focus on semen analysis over the years, assessment of sperm morphology has not been given due importance although it is a simple, standard and baseline diagnostic modality. It can be used to predict the need and outcome of Artificial Reproductive Techniques such as Invitro Fertilization, Gamete Intra Fallopian Tube Transfer and Intra Cytoplasmic Sperm Injection. Aim To find the ideal, simple and cost-effective basic stain for assessment of sperm morphology in a rural tertiary care set- up where advanced equipment for assessment of sperm morphometry are inaccessible. Materials and Methods An updated way of determining sperm shape is called the Kruger’s strict morphology method. Keeping this as the standard criterion, we studied semen samples of 62 healthy male subjects using four basic staining techniques and the consensus of four independent observers was tabulated. Results We found that Haematoxylin and Eosin stain was the best stain for assessment of sperm head morphology. Rapid Papanicolau stain was the most ideal, simple and cost-effective stain for overall assessment of sperm morphology. Conclusion Sperm morphology assessment remains the baseline necessity for the diagnosis and management of male factor associated infertility when advanced techniques are unavailable, inaccessible or unaffordable. PMID:26557524

  16. Cell wall staining with Trypan blue enables quantitative analysis of morphological changes in yeast cells

    PubMed Central

    Liesche, Johannes; Marek, Magdalena; Günther-Pomorski, Thomas

    2015-01-01

    Yeast cells are protected by a cell wall that plays an important role in the exchange of substances with the environment. The cell wall structure is dynamic and can adapt to different physiological states or environmental conditions. For the investigation of morphological changes, selective staining with fluorescent dyes is a valuable tool. Furthermore, cell wall staining is used to facilitate sub-cellular localization experiments with fluorescently-labeled proteins and the detection of yeast cells in non-fungal host tissues. Here, we report staining of Saccharomyces cerevisiae cell wall with Trypan Blue, which emits strong red fluorescence upon binding to chitin and yeast glucan; thereby, it facilitates cell wall analysis by confocal and super-resolution microscopy. The staining pattern of Trypan Blue was similar to that of the widely used UV-excitable, blue fluorescent cell wall stain Calcofluor White. Trypan Blue staining facilitated quantification of cell size and cell wall volume when utilizing the optical sectioning capacity of a confocal microscope. This enabled the quantification of morphological changes during growth under anaerobic conditions and in the presence of chemicals, demonstrating the potential of this approach for morphological investigations or screening assays. PMID:25717323

  17. Persistence of DNA from laundered semen stains: Implications for child sex trafficking cases.

    PubMed

    Brayley-Morris, Helen; Sorrell, Amber; Revoir, Andrew P; Meakin, Georgina E; Court, Denise Syndercombe; Morgan, Ruth M

    2015-11-01

    In sexual assault cases, particularly those involving internal child sex trafficking (ICST), victims often hide their semen-stained clothing. This can result in a lag time of several months before the items are laundered and subsequently seized during a criminal investigation. Although it has been demonstrated previously that DNA can be recovered from clothing washed immediately after semen deposition, laundered items of clothing are not routinely examined in ICST cases, due to the assumption that the time delay and washing would result in no detectable DNA. The aim of this study was to examine whether viable DNA profiles could be recovered from laundered semen stains where there has been a significant lag time between semen deposition from one or more individuals and one or more washes of the stained clothing. Items of UK school uniform (T-shirts, trousers, tights) were stained with fresh semen (either from a single donor or a 1:1 mixture from two donors) and stored in a wardrobe for eight months. Stained and unstained items (socks) were then washed at 30°C or 60°C and with non-biological or biological detergent. DNA samples extracted from the semen-stained sites and from the unstained socks were quantified and profiled. High quantities of DNA, (6-18?g) matching the DNA profiles of the semen donors, were recovered from all semen-stained clothing that had been laundered once, irrespective of wash conditions. This quantity,and profile quality,did not decline significantly with multiple washes. The two donor semen samples yielded ?10-fold more DNA from the T-shirts than from the trousers. This disparity resulted in the T-shirts yielding a ?1:1 mixture of DNA from the two donors, whereas the trousers yielded a major DNA profile matching only that of the second donor. The quantities of DNA recovered from the unstained socks were an order of magnitude lower, with most of the DNA being attributable to the donor of the semen on the stained clothing within the same wash, demonstrating the transfer of semen-derived DNA among items of clothing in the washing machine. This study demonstrates that complete DNA profiles can be obtained from laundered semen stains on school uniform-type clothing, with an eight-month lag time between semen deposition and laundering, despite multiple washes and stains from two semen donors. These data emphasise the need to recover and examine the clothing of victims for semen and DNA evidence, even if the clothing has been stored for several months or washed multiple times since the sexual offence took place. PMID:26232275

  18. Metal Ion Complexes of N,N'-Bis(2-Pyridylmethyl)-trans-1,2-Diaminocyclohexane-N,N'-Diacetic Acid, H2bpcd: Cis/Trans Isomerization Equilibria.

    PubMed

    Florián, Jan; McLauchlan, Craig C; Kissel, Daniel S; Eichman, Chad C; Herlinger, Albert W

    2015-11-01

    The synthesis of N,N'-bis(2-pyridylmethyl)-trans-1,2-diaminocyclohexane-N,N'-diacetic acid (H2bpcd) and its complexation of Ga(III) and Co(III) are reported. H2bpcd and the metal-bpcd(2-) complexes, isolated as hexafluorophosphate salts, were characterized by elemental analysis, X-ray crystallography, IR spectroscopy, and (1)H and (13)C NMR spectroscopy. [Ga(bpcd)]PF6, [Ga(C22H26N4O4)]PF6, crystallized in the orthorhombic space group Ibca, with a = 13.8975(7) Å, b = 15.0872(7) Å, c = 22.2418(10) Å, and Z = 8. Ga is coordinated in a distorted octahedral geometry provided by a N4O2 donor atom set with trans-monodentate acetate groups and cis-2-pyridylmethyl N atoms, i.e., the trans-O,O isomer. The diamagnetic [Co(bpcd)]PF6, [Co(C22H26N4O4)]PF6, also crystallized from solution in the Ibca space group as the trans-O,O isomer. The (1)H and (13)C assignments for H2bpcd and metal-bpcd(2-) complexes were made on the basis of 2D COSY and HSQC experiments, which were used to differentiate among three possible isomers, i.e., one cis (C1 symmetry) and two trans (C2 symmetry). NMR results indicate that the [Ga(bpcd)](+), [Co(bpcd)](+), and cis-O,O, cis-Npy,Npy-[Ga(bppd)](+) cations, where bppd(2-) stands for bis(2-pyridylmethyl)-1,3-diaminopropane diacetate, are present in solution as isomers with the same symmetry as observed in the solid state. The crystallographic data and the dramatic shift that occurs in the position of the cis/trans isomerization equilibria for the [Ga(bpad)](+) cations simply by increasing the number of bridging CH2 groups in the ligand's diamine backbone represent a unique opportunity to assess the accuracy of modern computational methods. The performance of several local density functionals using a pseudopotential-based SDD basis set was compared with the more rigorous HF and MP2 ab initio calculations. The SVWN5 and SV5LYP functionals provide significantly better Ga-O and Ga-N distances than the HF method or the nonlocal BLYP functional. However, to provide proper isomerization energies the pseudopotential-DFT calculations must be augmented by MP2 single-point energies and calculations of solvation free energies. PMID:26478942

  19. Iodine vapor staining for atomic number contrast in backscattered electron and X-ray imaging.

    PubMed

    Boyde, Alan; Mccorkell, Fergus A; Taylor, Graham K; Bomphrey, Richard J; Doube, Michael

    2014-12-01

    Iodine imparts strong contrast to objects imaged with electrons and X-rays due to its high atomic number (53), and is widely used in liquid form as a microscopic stain and clinical contrast agent. We have developed a simple technique which exploits elemental iodine's sublimation-deposition state-change equilibrium to vapor stain specimens with iodine gas. Specimens are enclosed in a gas-tight container along with a small mass of solid I2 . The bottle is left at ambient laboratory conditions while staining proceeds until empirically determined completion (typically days to weeks). We demonstrate the utility of iodine vapor staining by applying it to resin-embedded tissue blocks and whole locusts and imaging them with backscattered electron scanning electron microscopy (BSE SEM) or X-ray microtomography (XMT). Contrast is comparable to that achieved with liquid staining but without the consequent tissue shrinkage, stain pooling, or uneven coverage artefacts associated with immersing the specimen in iodine solutions. Unmineralized tissue histology can be read in BSE SEM images with good discrimination between tissue components. Organs within the locust head are readily distinguished in XMT images with particularly useful contrast in the chitin exoskeleton, muscle and nerves. Here, we have used iodine vapor staining for two imaging modalities in frequent use in our laboratories and on the specimen types with which we work. It is likely to be equally convenient for a wide range of specimens, and for other modalities which generate contrast from electron- and photon-sample interactions, such as transmission electron microscopy and light microscopy. PMID:25219801

  20. Al adjuvants can be tracked in viable cells by lumogallion staining.

    PubMed

    Mile, Irene; Svensson, Andreas; Darabi, Anna; Mold, Matthew; Siesjö, Peter; Eriksson, Håkan

    2015-07-01

    The mechanism behind the adjuvant effect of aluminum salts is poorly understood notwithstanding that aluminum salts have been used for decades in clinical vaccines. In an aqueous environment and at a nearly neutral pH, the aluminum salts form particulate aggregates, and one plausible explanation of the lack of information regarding the mechanisms could be the absence of an efficient method of tracking phagocytosed aluminum adjuvants and thereby the intracellular location of the adjuvant. In this paper, we want to report upon the use of lumogallion staining enabling the detection of phagocytosed aluminum adjuvants inside viable cells. Including micromolar concentrations of lumogallion in the culture medium resulted in a strong fluorescence signal from cells that had phagocytosed the aluminum adjuvant. The fluorescence appeared as spots in the cytoplasm and by confocal microscopy and co-staining with probes presenting fluorescence in the far-red region of the spectrum, aluminum adjuvants could to a certain extent be identified as localized in acidic vesicles, i.e., lysosomes. Staining and detection of intracellular aluminum adjuvants was achieved not only by diffusion of lumogallion into the cytoplasm, thereby highlighting the presence of the adjuvant, but also by pre-staining the aluminum adjuvant prior to incubation with cells. Pre-staining of aluminum adjuvants resulted in bright fluorescent particulate aggregates that remained fluorescent for weeks and with only a minor reduction of fluorescence upon extensive washing or incubation with cells. Both aluminum oxyhydroxide and aluminum hydroxyphosphate, two of the most commonly used aluminum adjuvants in clinical vaccines, could be pre-stained with lumogallion and were easily tracked intracellularly after incubation with phagocytosing cells. Staining of viable cells using lumogallion will be a useful method in investigations of the mechanisms behind aluminum adjuvants' differentiation of antigen-presenting cells into inflammatory cells. Information will be gained regarding the phagosomal pathways and the events inside the phagosomes, and thereby the ultimate fate of phagocytosed aluminum adjuvants could be resolved. PMID:25896212

  1. Iodine Vapor Staining for Atomic Number Contrast in Backscattered Electron and X-ray Imaging

    PubMed Central

    Boyde, Alan; Mccorkell, Fergus A; Taylor, Graham K; Bomphrey, Richard J; Doube, Michael

    2014-01-01

    Iodine imparts strong contrast to objects imaged with electrons and X-rays due to its high atomic number (53), and is widely used in liquid form as a microscopic stain and clinical contrast agent. We have developed a simple technique which exploits elemental iodine's sublimation-deposition state-change equilibrium to vapor stain specimens with iodine gas. Specimens are enclosed in a gas-tight container along with a small mass of solid I2. The bottle is left at ambient laboratory conditions while staining proceeds until empirically determined completion (typically days to weeks). We demonstrate the utility of iodine vapor staining by applying it to resin-embedded tissue blocks and whole locusts and imaging them with backscattered electron scanning electron microscopy (BSE SEM) or X-ray microtomography (XMT). Contrast is comparable to that achieved with liquid staining but without the consequent tissue shrinkage, stain pooling, or uneven coverage artefacts associated with immersing the specimen in iodine solutions. Unmineralized tissue histology can be read in BSE SEM images with good discrimination between tissue components. Organs within the locust head are readily distinguished in XMT images with particularly useful contrast in the chitin exoskeleton, muscle and nerves. Here, we have used iodine vapor staining for two imaging modalities in frequent use in our laboratories and on the specimen types with which we work. It is likely to be equally convenient for a wide range of specimens, and for other modalities which generate contrast from electron- and photon-sample interactions, such as transmission electron microscopy and light microscopy. Microsc. Res. Tech. 77:1044–1051, 2014. © 2014 The Authors. Microscopy Research Technique published by Wiley Periodocals, Inc. PMID:25219801

  2. Critical tonicity determination of sperm using fluorescent staining and flow cytometry

    SciTech Connect

    Noiles, E.E.; Ruffing, N.A.; Kleinhans, F.W.; Mark, L.A.; Watson, P.F.; Critser, J.K. ); Horstman, L. . School of Veterinary Medicine); Mazur, P. )

    1990-01-01

    The use of cryopreserved, rather than fresh, mammalian semen for artificial insemination confers several important medical and/or economic advantages. However, current methods for cryopreservation of both human and bovine spermatozoa result in approximately only a 50% survival rate with thawing, obviously reducing the fertilizing capacity of the semen. A primary consideration during the cooling process is to avoid intracellular ice crystal formation with its lethal consequences to the cell. Current techniques achieve this by controlling the cooling rate. Computation of the time necessary for this dehydration, and hence, the cooling rate, is dependent upon knowledge of the water permeability coefficient (L{sub {rho}}) and its activation energy. The fluorophore, 6-carboxyfluoroscein diacetate (CFDA), which is nonfluorescent, readily crosses the intact plasma membrane. Intracellular esterases hydrolyze CFDA to 6-carboxyfluoroscein, a fluorescent, membrane-impermeable fluorophore. Consequently, spermatozoa with intact plasma membranes fluoresce bright green (Garner et. al., 1986), but those with disrupted membranes do not. Therefore, the purpose of this study was to use loss of CFDA fluorescence to determine the osmolality at which 50% of the spermatozoa will swell and lyse (critical tonicity, CT). These data will then be used to determine the L{sub {rho}} and its activation energy for sperm, thus increasing the knowledge available in cellular cryopreservation. 15 refs., 3 figs.

  3. Neuroenteric Staining as a Tool in the Evaluation of Pediatric Motility Disorders.

    PubMed

    Waseem, Shamaila H; Idrees, Muhammed T; Croffie, Joseph M

    2015-08-01

    The diagnosis of enteric neuromuscular disorders has come a long way since the first description of an enteric neuropathic disorder by the Danish physician Harald Hirschsprung in 1886. Advances in specialized enteric histopathological staining techniques have made it possible to identify subtle neuropathies and myopathies that cause intestinal motility disorders, from the common and now better understood and relatively easily diagnosed Hirschsprung's disease to the less common and more severe and not well-characterized chronic idiopathic intestinal pseudoobstruction, which continues to present a diagnostic challenge to the gastroenterologist and histopathologist alike. This article will discuss the common gastrointestinal motility disorders and some of the specialized histological stains, such as the relatively common enzyme stain, acetylcholinesterase, used to diagnose Hirschsprung's disease; advanced tinctorial stains, such as Masson trichrome, which may aid in diagnosis of enteric myopathies causing pseudoobstruction; and immunohistochemical stains such as C-Kit or PG 9.5, which may aid in the diagnosis of enteric neuropathies causing pseudoobstruction. PMID:26143629

  4. Comparison of Gram stain and Pap smear procedures in the diagnosis of bacterial vaginosis.

    PubMed Central

    Vardar, Enver; Maral, Izzet; Inal, Murat; Ozgüder, Ozgül; Tasli, Funda; Postaci, Hakan

    2002-01-01

    OBJECTIVE: The purpose of this study was to examine the characteristics of Gram stain versus Pap smear in diagnosis of bacterial vaginosis (BV). METHODS: One-thousand and sixty women were enrolled in this study. All cases with symptoms of BV were determined by Amsel's criteria, which were accepted as the gold standard for diagnosis of BV. Pap smear and Gram stain evaluations were compared according to Amsel's criteria, without viewing the clinical results of the patients. Gram stain and Pap smear results were determined as negative or positive according to Amsel's criteria. Sensitivity, specifity and positive predictive values were calculated. RESULTS: After accepting the cases that were diagnosed as BV according to Amsel's criteria as reference cases, the sensitivity of the Gram stain method was calculated as 97% and the sensitivity of the Pap smear method as 93%. Similar specificity rates were obtained with both methods in diagnosis of BV related to the clinical results. There were no statistically significant differences in diagnosis of BV between these two groups. CONCLUSION: If Amsel's criteria are accepted as the gold standard for diagnosis of BV, Gram stain and Pap smear methods will give similar results in diagnosis. PMID:12648314

  5. Evaluation of sorption, solubility and staining of universal and silorane resin-based composites.

    PubMed

    Anfe, T E de Almeida; Agra, C M; Vieira, G F

    2011-12-01

    Resin-based composite staining is a multifactoral phenomenon and can be caused by intrinsic and extrinsic factors. The purpose of this study was to compare staining, sorption and solubility of silorane resin-based and universal resin-based composites. Five different resin-based composites (4 Seasons, Charisma, Filtek Silorane, Filtek Supreme and Grandio) were tested. Twenty five specimens were prepared (10 mm diameter and 1.5 mm thick). To staining test, the specimens were divided into 3 groups (n = 5): distilled water (control), coffee and red wine. The specimens were immersed in one of the solutions at 37 degrees C for 7 days. Using the values of L*, a*, b*, color variation (CIEDE2000) was determined. For sorption and solubility test, the specimens were divided into 2 groups (n = 5): with previous desiccation (Group 1) and with no previous desiccation (Group 2). The methodology used for sorption and solubility test was based on ISO 4049:2000. The results presented no significant difference in staining between composites. In sorption and solubility test, Filtek Silorane presented the smallest values, followed by Grandio. Under tested experimental conditions, it is not possible to assert the dependence of staining in sorption that composites are undergone. There was no significant correlation between colour change and sorption values. PMID:22645799

  6. Lipopolysaccharide aggregates in native agarose gels detected by reversible negative staining with imidazole and zinc salts.

    PubMed

    Rodríguez, Caridad; Hardy, Eugenio

    2015-09-15

    We investigated the use of imidazole and zinc salts for the detection of lipopolysaccharide (LPS) aggregates separated by native agarose gel electrophoresis (NAGE). As a result, a new staining procedure was established by which as little as 1.5 ?g of Escherichia coli O55:B5 LPS aggregates was detected by means of inducing a clear, transparent pattern, contrasted against an opaque background. E. coli O55:B5 LPS preparations treated with nucleases and proteinase K proved that the reverse-stained LPS pattern is not related to any potential artifacts caused by unrelated biomolecules (e.g., nucleic acids, proteins). After this, we showed that the procedure is applicable to two-dimensional LPS separation using NAGE/SDS-PAGE, while at the same time confirming that real polydisperse LPS aggregates are represented by the stained profile. Also, we demonstrated the general applicability of this stain to the detection of different NAGE-separated LPS aggregates (e.g., from E. coli 026:B6, E. coli 0111:B4, Salmonella minnesota Re595). Finally, using lysozyme as a model protein, we found that imidazole-zinc may be combined with Coomassie brilliant blue R-250 into a double-staining process to enable the use of NAGE for investigating the interaction of cationic proteins and LPS aggregates and protein or LPS concentration effects on protein-LPS binding. PMID:26095395

  7. Hereditary Cystinosis Detected by CD68 Staining of the Bone Marrow Biopsies of 2 Siblings.

    PubMed

    Erçin, Cengiz; Paksoy, Nadir; Gök, Nazli D

    2015-01-01

    Cystinosis is an autosomal recessive lysosomal storage disease. We present 2 siblings in whom cystinosis was detected by CD68 immunohistochemical analysis of bone marrow biopsies. The older patient was a 6-year-old boy who had been receiving erythrocyte suspension therapy for 5.5 years because of low hemoglobin levels. The patient was admitted to our hospital because of hepatomegaly, anemia, and thrombocytopenia and underwent a trephine bone marrow biopsy based on a preliminary diagnosis of lipid storage disease. Macrophage-like cells were observed in the hematoxylin/eosin-stained sections. These cells were stained for CD68 to confirm that they were macrophages. Some crystalline structures were seen in the cytoplasm of the macrophages after CD68 staining. These structures were thought to be cystine crystals. The diagnosis of cystinosis was confirmed by a clinical assessment. The 1-year-old sibling of the patient was also examined; this sibling exhibited renal disease and had a family history of consanguineous marriage. Cystinosis was also detected in this sibling by clinical assessment and staining of the patient's trephine bone marrow biopsy for CD68. The staining of the bone marrow biopsies for CD68 enabled the patient and his sibling to be diagnosed with cystinosis; these patients were not correctly diagnosed over the previous 6-year period. No similar report was found in the literature regarding this topic. PMID:25153500

  8. Diagnostic value of immunohistochemical staining of GP73, GPC3, DCP, CD34, CD31, and reticulin staining in hepatocellular carcinoma.

    PubMed

    Yao, Shuzhe; Zhang, Jianping; Chen, Haiyan; Sheng, Yan; Zhang, Xiaoying; Liu, Zhiyan; Zhang, Cuijuan

    2013-09-01

    It has been reported that Golgi protein-73 (GP73), glypican-3 (GPC3), and des-?-carboxy prothrombin (DCP) could serve as serum markers for the early detection of hepatocellular carcinoma (HCC). This study aimed to evaluate a panel of immunostaining markers (including GP73, GPC3, DCP, CD34, and CD31) as well as reticulin staining to distinguish HCC from the mimickers. Our results revealed that CD34 immunostaining and reticulin staining were highly sensitive for the diagnosis of HCC. A special immunoreaction pattern of GP73--a diffuse coarse-block pattern in a perinuclear region or a concentrated cluster-like or cord-like pattern in a certain part of the cytoplasm--was observed in HCC cells, in contrast to the cytoplasmic fine-granular pattern in surrounding non-tumor cells and non-malignant nodules. This coarse-block pattern correlated significantly with less differentiated HCC. In comparison, GPC3 displayed a good advantage in diagnosing well-differentiated HCC. In our study, DCP and CD31 showed little diagnostic value for HCC as an immunostaining marker. When GP73, GPC3, and CD34 were combined, the specificity improved to 96.6%. Our findings demonstrate for the first time that the immunohistochemical panel of GP73, GPC3, and CD34 as well as reticulin staining is highly specific for the pathological diagnosis of HCC. PMID:23686365

  9. New staining methods for yeast like fungi under special consideration of human pathogenic fungi

    NASA Astrophysics Data System (ADS)

    Paulitsch-Fuchs, Astrid; Treiber, Fritz; Grasser, Erik; Buzina, Walter; Rosker, Christian

    2010-11-01

    A new method for in-cellular staining of yeast like fungi with Oregon Green and SYTOX Green is presented enabling their detection as well as the observation of cellular details via confocal laser scanning microscopy. Fluorochromes play an important role in many scientific disciplines including medicine, cell biology and botany. For the visualisation of fungal cell walls Calcofluor White is the flourochrome of choice. The necessity of an UV laser for its excitation makes it unpracticable for daily use. Safranin O, DAPI, 2NBDG, Ethidium Bromide and Acridin-orange are commonly used stains for nuclei in fugal microscopy. The attention was given to the possibility of using the differences in staining patterns to distinguish certain pathogenic yeast species e.g. Candida albicans and Candida krusei. Our results show that high quality microscopy of yeast like organisms can readily be achieved by the use of two suitable fluorochromes.

  10. Physical markers for landmarking fluorescently stained gels that facilitate automated spot-picking.

    PubMed

    Fehring, V; Wandschneider, S; Löhr, M

    2001-08-01

    The quantitative comparison of spot patterns relies heavily on protein stains that do provide an appropriate dynamic range. Unfortunately most spot picking robot devices are still limited to nonfluorescent protein stains and the appropriate equipment is still quite expensive. These problems are solved by the application of a newly developed "GelMarker" that combines a spot picking robot device and a UV scanner. The "GelMarkers" are detectable in both the visible and UV range of light and permit the comparison of gel pictures taken by such different devices. The application of these "GelMarkers" together with the transformation of spot coordinates by using a "spot matching" procedure allows the automated excision of selected protein spots. The obtained picking accuracies are as good as those obtainable from visible stained gels due to the shape stability of the gels even over a longer time period. PMID:11565786

  11. The feasibility of digitally stained multimodal confocal mosaics to simulate histopathology

    PubMed Central

    Gareau, Daniel S.

    2010-01-01

    Fluorescence confocal mosaicing microscopy of tissue biopsies stained with acridine orange has been shown to accurately identify tumors and with an overall sensitivity of 96.6% and specificity of 89.2%. However, fluorescence shows only nuclear detail similar to hematoxylin in histopathology and does not show collagen or cytoplasm, which may provide necessary negative contrast information similar to eosin used in histopathology. Reflectance mode contrast is sensitive to collagen and cytoplasm without staining. To further improve sensitivity and specificity, digitally stained confocal mosaics combine confocal fluorescence and reflectance images in a multimodal pseudo-color image to mimic the appearance of histopathology with hematoxylin and eosin and facilitate the introduction of confocal microscopy into the clinical realm. PMID:19566342

  12. Application of the DNA-specific stain methyl green in the fluorescent labeling of embryos.

    PubMed

    Prieto, Daniel; Aparicio, Gonzalo; Machado, Matías; Zolessi, Flavio R

    2015-01-01

    Methyl green has long been known as a histological stain with a specific affinity for DNA, although its fluorescent properties have remained unexplored until recently. In this article, we illustrate the method for preparing a methyl green aqueous stock solution, that when diluted can be used as a very convenient fluorescent nuclear label for fixed cells and tissues. Easy procedures to label whole zebrafish and chick embryos are detailed, and examples of images obtained shown. Methyl green is maximally excited by red light, at 633 nm, and emits with a relatively sharp spectrum that peaks at 677 nm. It is very inexpensive, non-toxic, highly stable in solution and very resistant to photobleaching when bound to DNA. Its red emission allows for unaltered high resolution scanning confocal imaging of nuclei in thick specimens. Finally, this methyl green staining protocol is compatible with other cell staining procedures, such as antibody labeling, or actin filaments labeling with fluorophore-conjugated phalloidin. PMID:25993383

  13. Methods of staining target chromosomal DNA employing high complexity nucleic acid probes

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel; Kallioniemi, Ol'li-Pekka; Kallioniemi, Anne; Sakamoto, Masaru

    2006-10-03

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML), retinoblastoma, ovarian and uterine cancers, and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar but genetically different diseases, and for many prognostic and diagnostic applications.

  14. Western blotting using in-gel protein labeling as a normalization control: stain-free technology.

    PubMed

    Gilda, Jennifer E; Gomes, Aldrin V

    2015-01-01

    Western blotting is a commonly used laboratory technique for semi-quantifying protein amounts. It is important when quantifying protein expression to account for differences in the amount of total protein loaded onto the gel using a loading control. Common loading controls include housekeeping proteins, such as ?-actin or GAPDH, quantified by Western blot, or total protein, quantified using a stain such as Coomassie Brilliant Blue or Ponceau S. A more recently developed method for total protein quantification utilizes stain-free technology, which has a linear dynamic detection range and allows for protein detection on both gels and membranes. Here, we describe the theory and use of stain-free gels for total protein quantification and normalization of Western blots. PMID:25820735

  15. Immunohistochemical staining for endotoxin using horseshoe crab factor C in fecal peritonitis.

    PubMed

    Nakao, A; Yasui, M; Shen, S; Takagi, H

    1995-01-01

    The object of the present study was to apply a new immunohistochemical staining method to the in vivo determination of endotoxin localization. The immunohistochemical staining method requires factor C (an initiation factor in the Limulus clotting system which is mediated by endotoxin) as a specific ligand of endotoxin, and a newly developed murine monoclonal antibody to factor C. The blood endotoxin level and endotoxin localization in the rat were determined before and at 6, 12 and 24 h after intraperitoneal injection of 0.25 g/kg of fresh rat feces. The greatest blood endotoxin level was achieved at 12 h after the injection, and uptake of endotoxin was evident in Kupffer cells in the liver at 24 h after the injection. There has been no report on determining endotoxin localization in cases of endotoxemia attributed to fecal peritonitis. This new immunohistochemical staining method for determining endotoxin localization will contribute to the histopathological diagnosis of endotoxemia in humans. PMID:7544288

  16. Facile method to stain the bacterial cell surface for super-resolution fluorescence microscopy

    SciTech Connect

    Gunsolus, Ian L.; Hu, Dehong; Mihai, Cosmin; Lohse, Samuel E.; Lee, Chang-Soo; Torelli, Marco; Hamers, Robert J.; Murphy, Catherine; Orr, Galya; Haynes, Christy L.

    2014-01-01

    A method to fluorescently stain the surfaces of both Gram-negative and Gram-positive bacterial cells compatible with super-resolution fluorescence microscopy is presented. This method utilizes a commercially-available fluorescent probe to label primary amines at the surface of the cell. We demonstrate efficient staining of two bacterial strains, the Gram-negative Shewanella oneidensis MR-1 and the Gram-positive Bacillus subtilis 168. Using structured illumination microscopy and stochastic optical reconstruction microscopy, which require high quantum yield or specialized dyes, we show that this staining method may be used to resolve the bacterial cell surface with sub-diffraction-limited resolution. We further use this method to identify localization patterns of nanomaterials, specifically cadmium selenide quantum dots, following interaction with bacterial cells.

  17. Digital staining for histopathology multispectral images by the combined application of spectral enhancement and spectral transformation.

    PubMed

    Bautista, Pinky A; Yagi, Yukako

    2011-01-01

    In this paper we introduced a digital staining method for histopathology images captured with an n-band multispectral camera. The method consisted of two major processes: enhancement of the original spectral transmittance and the transformation of the enhanced transmittance to its target spectral configuration. Enhancement is accomplished by shifting the original transmittance with the scaled difference between the original transmittance and the transmittance estimated with m dominant principal component (PC) vectors;the m-PC vectors were determined from the transmittance samples of the background image. Transformation of the enhanced transmittance to the target spectral configuration was done using an nxn transformation matrix, which was derived by applying a least square method to the enhanced and target spectral training data samples of the different tissue components. Experimental results on the digital conversion of a hematoxylin and eosin (H&E) stained multispectral image to its Masson's trichrome stained (MT) equivalent shows the viability of the method. PMID:22256200

  18. Preliminary report: laboratory-induced stain removal as assessed by environmental scanning electron microscopy.

    PubMed

    Habib, C M; Kugel, G; Marcus, A

    1998-01-01

    Environmental scanning electron microscopy (ESEM) was employed to observe stain removal during brushing with Arm & Hammer Dental Care and Crest Regular Toothpaste. ESEM allows serial examinations of the same sample, and does not require a destructive preparative process. Three extracted molars were cleaned, placed into a 96-hour broth culture of Streptococcus mutans, and stain was produced with undiluted chlorhexidine rinse, concentrated coffee and tea for a period of 23 days. After staining, the teeth were examined by ESEM, then brushed using a toothbrushing machine. Imaging was repeated after 5, 10, 15 and 30 seconds of brushing. As seen with ESEM, the Arm & Hammer product had different effects than those from the distilled water control, suggesting something other than that expected from abrasive and mechanical forces alone. There were also differences from the Crest dentifrice removal on this single sample, suggesting a possible difference between the two products. Further studies are needed to confirm and explain these effects. PMID:10518864

  19. Light microscopic and color television image analysis of the development of staining on chlorhexidine-treated surfaces.

    PubMed

    Addy, M; Prayitno, S W

    1980-01-01

    Tooth staining with the use of chlorhexidine preparations is the major problem of long term application. Evidence suggests that the staining arises from a cationic/anionic interacation of chlorhexidine with components of certain dietary materials. The purpose of this in vitro study was to compare visually the development of tea and coffee staining on acrylic and tooth specimens treated with chlorhexidine and to follow the development of tea staining on perspex by light microscopy and color television image analysis. All specimens were maintained in their respective beverage for 5 days with test specimens removed three times a day and placed for 2 minutes in an 0.2% chlorhexidine solution. Both test tooth and acrylic specimens showed comparably and markedly increased staining by the beverages compared with control specimens. Color television image analysis of test specimens demonstrated more marked and rapid development of tea staining when studied on a daily basis. Microscopic examination revealed the staining to be made up of small particles of material which increased in size and coalesced with time. Again, marked differences were apparent in the stain on test and control specimens. The results of this in vitro method provided further evidence for a dietary aetiology to chlorhexidine staining and were consistent with clinical findings. Such a method may be useful to assess staining arising from the use of other anti-plaque agents. PMID:6928469

  20. Use of chloroacetate esterase staining for the histological diagnosis of prosthetic joint infection.

    PubMed

    Kashima, T G; Inagaki, Y; Grammatopoulos, G; Athanasou, N A

    2015-05-01

    A heavy neutrophil polymorph infiltrate [>5 per high-power field (HPF) after examination of at least 5 HPF by Musculoskeletal Infection Society (MSIS) criteria] is characteristically seen in peri-implant tissues of infected prosthetic hip and knee joints. We determined whether chloroacetate esterase (CAE) staining facilitated the identification of neutrophil polymorphs in peri-implant tissues in cases of hip and knee arthroplasty infection and reassessed MSIS criteria in the light of our findings. Frozen and paraffin sections of peri-prosthetic tissues of 76 cases of failed hip and knee arthroplasties classified as septic or aseptic loosening microbiologically were analysed histologically by both haematoxylin-eosin and CAE staining. The extent of the neutrophil polymorph infiltrate was determined semiquantitatively and correlated with the microbiological and clinical diagnosis. CAE staining facilitated identification of neutrophil polymorphs in arthroplasty tissues. All cases of aseptic loosening contained fewer than two neutrophil polymorphs per HPF. CAE staining showed that in some cases of septic loosening, fewer than five neutrophil polymorphs per HPF (on average) are present in peri-prosthetic tissues. The histological criterion of more than two neutrophil polymorphs per HPF showed increased sensitivity and accuracy for the diagnosis of septic loosening. CAE is a useful stain that facilitates the identification of neutrophil polymorphs in both frozen and paraffin sections of peri-implant tissues. CAE staining shows that some microbiologically confirmed cases of septic loosening contain relatively few neutrophil polymorphs, indicating that the MSIS histological criterion of more than five neutrophil polymorphs per HPF is too high an index figure for the diagnosis of all cases of hip and knee arthroplasty infection. PMID:25687172

  1. Solute accessibility to N epsilon-fluorescein isothiocyanate-lysine-23 cobra alpha-toxin bound to the acetylcholine receptor. A consideration of the effect of rotational diffusion and orientation constraints on fluorescence quenching.

    PubMed Central

    Johnson, D. A.; Yguerabide, J.

    1985-01-01

    To obtain information on the disposition of alpha-toxin when bound to the acetylcholine receptor (AChR), we evaluated the accessibility of solutes to fluorescein isothiocyanate (FITC) conjugated to alpha-toxin (siamensis 3) at lysine 23 (FITC-toxin) by measuring the rate constants for iodide quenching of the fluorescence of fluorescein free in solution and FITC-toxin free in solution and bound to AChR. Relative to the free fluorescein, we observed a 55% reduction in the quenching rate constant for the unbound FITC-toxin and 80% reduction for the AChR-bound FITC-toxin. It is tempting to interpret a decrease in the quenching rate constant as due to an increase in the masking of the labeling fluorophore, which in our case would then be indicative of masking of fluorescein conjugated to the free toxin and masking of FITC-toxin, in the region of lysine 23, when bound to AChR. However, elementary considerations indicate that the quenching rate depends not only on geometrical masking factors but also on the translational and rotational mobilities of the labeled molecules as well as orientational constraints. To evaluate these effects we have established quantitative relations between the rate of fluorescence quenching, the degree of masking of fluorophore, translational and rotational rates, and orientational constraints of the labeled macromolecules, using recent formulations for the rate of reaction between asymmetric molecules (Shoup et al., 1981, Biophys. J., 36:619-714). These relations predict that the decrease in quenching constant observed for the labeled FITC-toxin as well as the AChR-bound FITC-toxin is largely due to differences in translational and rotational rates and orientational constraints and not to significant increases in geometrical masking. Our theoretical formulation shows that the quenching rate can be decreased by a factor of 2-5 merely by immobilizing a fluorophore on the surface of a large protein without any significant increase in geometrical masking. PMID:3937557

  2. Greenish-blue staining of underclothing due to Pseudomonas aeruginosa infection of intertriginous dermatitis.

    PubMed

    Kalkan, Goknur; Duygu, Fazilet; Bas, Yalcin

    2013-09-01

    Intertrigo (intertriginous dermatitis) is an inflammatory condition of the skin folds, induced or aggravated by heat, moisture, maceration, friction, and lack of air circulation. Intertrigo can become secondarily infected. Three cases of intertrigo are described herein which presented with greenish-blue staining of underclothing. Cultures revealed Pseudomonas aeruginosa which is a gram-negative aerobic bacillus that produces the pigments pyocyanin and pyoverdin that responsible for this characteristic hue. All three patients responded to a course of oral ciprofloxacin with resolution of the intertriginous dermatitis. Therefore, greenish-blue staining of clothing is an important indicator for pseudomonal intertrigo. PMID:24601205

  3. [Morphology of low-velocity impact stains produced from single drops of blood].

    PubMed

    Benecke, Mark; Reibe, Saskia; Baumjohann, Kristina; Gulinski, Sarah; Wetzel, Waltraud; Schmidt, Kira; Pressler, Katharina; Lebküchner, Isabell; Streckenbach, Markus

    2012-01-01

    Systematic variation of blood droplet volume, the distance fallen and the surface (paper, wood, plastics, tiles) led to the conclusion that the size and the shape of the stains ("fingers", satellites) allowed to deduce the distance fallen but only if the actual surface structure was known. We found that detailed photography at the crime scene was necessary, yet experiments have to be performed due to the extreme influence of the actual surface texture on all characteristics (size, spines, peripheral spatter) of the blood stains. PMID:22924278

  4. Fast and automatic imaging of immunoenzyme-stained neuronal circuits in the whole brain of Drosophila

    NASA Astrophysics Data System (ADS)

    Tian, Qingping; Yuan, Jing; Li, Yuxin; Jiang, Tao; Gong, Hui; Zhou, Wei

    2014-09-01

    Knowledge of neuronal wiring and morphogenesis in Drosophila is essential to understand brain function and dysfunction. The immunoenzyme method based on horseradish peroxidase/diaminobenzidine (HRP/DAB) provides high-contrast images to resolve details underlying neuronal architecture. However, the poor staining penetration and a lack of corresponding three-dimensional imaging methodology limit its application. Herein, we modified the HRP/DAB method to stain neuronal circuits in the whole brain of Drosophila. Furthermore, we found that imaging with the micro-optical sectioning tomography system provided a fast and automatic method that could dissect cell-specific neuroanatomical architecture at a submicron voxel resolution.

  5. A method for determining supernatant-free spectra generated from fluorescently stained cells in suspension.

    PubMed

    Plášek, Jaromír; Gášková, Dana

    2014-12-01

    Here we present a fluorometric method for direct determination of supernatant-free fluorescence spectra generated from fluorescently stained cells in suspension. The key element in the new technique is the design of an adapter to a standard cuvette holder that makes it possible to measure front-face fluorescence spectra from thin layers of cells spun down to the bottom of a spectrofluorometric cuvette. We have demonstrated the applicability of this approach and its analytical potential using the suspensions of yeast cells stained with the potentiometric dye of 3,3'-dipropylthiadicarbocyanine, diS-C3(3), and with the specific cell-wall marker calcofluor. PMID:25463661

  6. Preference of Formosan subterranean termites for blue-stained southern yellow pine sapwood.

    PubMed

    Little, N S; Blount, N A; Londo, A J; Kitchens, S C; Schultz, T P; McConnell, T E; Riggins, J J

    2012-10-01

    Little research has been conducted to investigate interactions between the invasive Formosan subterranean termite, Coptotermes formosanus Shiraki, and pine bark beetles native to the southeastern United States. Facilitative interactions between these organisms could alter stand dynamics and impact wood utilization strategies. American Wood Protection Association Standard E1-09 choice tests were carried out to determine the feeding preference of Formosan subterranean termites for blue-stained versus unstained southern yellow pine sapwood. Three separate colonies of Formosan subterranean termites consumed on average twice as much air-dried blue-stained southern yellow pine sapwood over unstained air-dried controls. Additionally, Formosan subterranean termites consumed over five-times more kiln-dried blue-stained sapwood than unstained kiln-dried control wafers. The implications of these results are particularly relevant to pine forest ecology, nutrient cycling, and the utilization of blue-stained southern pine building products in the southeastern United States, where Formosan subterranean termites have become established. PMID:23156160

  7. Evaluation of Staining-Dependent Colour Changes in Resin Composites Using Principal Component Analysis

    PubMed Central

    Manojlovic, D.; Lenhardt, L.; Mili?evi?, B.; Antonov, M.; Miletic, V.; Drami?anin, M. D.

    2015-01-01

    Colour changes in Gradia Direct™ composite after immersion in tea, coffee, red wine, Coca-Cola, Colgate mouthwash, and distilled water were evaluated using principal component analysis (PCA) and the CIELAB colour coordinates. The reflection spectra of the composites were used as input data for the PCA. The output data (scores and loadings) provided information about the magnitude and origin of the surface reflection changes after exposure to the staining solutions. The reflection spectra of the stained samples generally exhibited lower reflection in the blue spectral range, which was manifested in the lower content of the blue shade for the samples. Both analyses demonstrated the high staining abilities of tea, coffee, and red wine, which produced total colour changes of 4.31, 6.61, and 6.22, respectively, according to the CIELAB analysis. PCA revealed subtle changes in the reflection spectra of composites immersed in Coca-Cola, demonstrating Coca-Cola’s ability to stain the composite to a small degree. PMID:26450008

  8. Iodamoeba butschlii in an anal pap test confirmed by iodine stain.

    PubMed

    Schuetz, Audrey N; Pritt, Bobbi S; Schreiner, Andrew M

    2014-09-01

    We report the finding of Iodamoeba butschlii amebic cysts on a liquid-based anal Pap smear from an HIV-positive male. Iodine staining of the smear confirmed the diagnosis. It is important to distinguish I. butschlii from pathogenic ameobae and other organisms seen on anal Pap smears. PMID:24167099

  9. Successful mRNA profiling of 23 years old blood stains.

    PubMed

    Kohlmeier, Fanni; Schneider, Peter M

    2012-03-01

    In this study, 23 years old human blood stains on different and problematic carrier materials, such as jeans, leather, wood, wallpaper, carpet, wool, and nylon fabric, were investigated by mRNA analysis of the blood markers HBB and SPTB. HBB turned out to be a very stable marker that could be detected in all blood samples from all carrier materials whereas SPTB showed no positive result in any of these old samples. Typing of co-extracted genomic DNA provided full STR profiles with the exception of one stain resulting in a partial profile. No differences were observed regarding the quality of results due to possible inhibitorial substances from carrier materials such as jeans or leather, whereas some difficulties to extract nucleic acids were caused by the physical properties of materials such as carpet fibres. HBB can be highly recommended for the identification of old blood stains in forensic casework as it was possible to detect HBB even in stains that failed to provide a full DNA profile. PMID:21612994

  10. A Simple and Efficient Method for Preparing Cell Slides and Staining without Using Cytocentrifuge and Cytoclips

    PubMed Central

    Hu, Xiaotang; Laguerre, Verronika; Packert, Daniel; Nakasone, Alice; Moscinski, Lynn

    2015-01-01

    Cell staining is a necessary and useful technique for visualizing cell morphology and structure under a microscope. This technique has been used in many areas such as cytology, hematology, oncology, histology, virology, serology, microbiology, cell biology, and immunochemistry. One of the key pieces of equipment for preparing a slide for cell staining is cytology centrifuge (cytocentrifuge) such as cytospin. However, many small labs do not have this expensive equipment and its accessory, cytoclips (also expensive relatively), which makes them difficult to study cell cytology. Here we present an alternative method for preparing a slide and cell staining in the absence of a cytocentrifuge (and cytoclips). This method is based on the principle that a regular cell centrifuge can be used to concentrate cells harvested from cell culture and then deposit the concentrated cell suspension to a slide evenly by using a cell spreader, followed by cell staining. The method presented is simple, rapid, economic, and efficient. This method may also avoid a possible change in cell morphology induced by cytocentrifuge.

  11. Periodic acid-Schiff staining demonstrates fungi in chronic anterior blepharitis.

    PubMed

    Dadaci, Z; K?l?nç, F; Ozer, T T; Sahin, G O; Acir, N O; Borazan, M

    2015-12-01

    PurposeTo evaluate the presence of fungi in patients with chronic anterior blepharitis with periodic acid-Schiff (PAS) staining of the eyelashes in addition to the conventional methods of fungal cultures and direct microscopy.MethodsNineteen patients with chronic anterior blepharitis of seborrheic or mixed seborrheic/staphylococcal type and 11 healthy age- and sex-matched controls were included in this prospective, nonrandomized, cross-sectional study. Blepharitis was diagnosed based on clinical evidence of greasy scales between the cilia, lid margin erythema, conjunctival hyperemia, telangiectasia, thickening, or irregularity of the eyelid margins by slit-lamp biomicroscopy. Eyelash samples were obtained by epilation with a sterile forceps and evaluated with PAS staining, fungal cultures, and direct microscopy.ResultsWe demonstrated fungal elements with PAS staining in 79% of the blepharitis group (hyphae and/or spores) and 18% of the control group. The difference was statistically significant (P=0.002). Four patients in the blepharitis group (21%) had positive cultures for fungi. The isolated fungi were Penicillium species (2 cases), Candida species (1 case), and Trichophyton verrucosum (1 case). Direct microscopic examination revealed Demodex mites in 42.1% of the blepharitis group. No culture growth or Demodex mites were observed in the control group.ConclusionsWe have shown fungi with PAS staining in the majority of patients with chronic anterior blepharitis. Further controlled studies are necessary to clarify the role of fungi in the etiopathogenesis of blepharitis. PMID:26293142

  12. Reaction of maturity group V soybean lines to purple seed stains in Mississippi 2010

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In 2009, soybean purple seed stain (PSS) caused 6.4 million bushels of yield losses in 16 southern states. This disease severely reduces seed market grade and affects seed germination and vigor. PSS is caused by Cercospora kikuchii and is an economy important disease. To identify new sources of resi...

  13. Alpha-amylase kinetic test in bodily single and mixed stains.

    PubMed

    Barni, Filippo; Berti, Andrea; Rapone, Cesare; Lago, Giampietro

    2006-11-01

    Recently, in Italy, a murder and a putative sexual violence was accomplished on a child. A bodily fluids mixture on the child's underwear between the victim (female) and the suspect (male) was ascertained by short tandem repeat (STR) DNA typing and, due to the absence of seminal fluid, saliva from the suspect and urine from the child was hypothesized. In order to investigate the possibility of specifically and rapidly detecting saliva stains both alone and mixed with other bodily fluids, we used a quantitative spectrophotometric technique, named Amylase test, for the detection of alpha-amylases. We determined alpha-amylase activity and reaction kinetic curves in several samples collected from the child's underwear. In order to confirm our intuition, we first tested saliva, perspiration, and urine, singularly and in mixtures; second, several forensic stains including saliva, perspiration, urine stains, saliva/perspiration, and saliva/urine mixture stains were tested. Evaluating alpha-amylase activity values and time-course curves' behavior of alpha-amylase reactions we were able to recognize successfully, in all cases, the presence of saliva and to distinguish it specifically from other bodily fluids containing alpha-amylase. A further confirmation of our result was provided by STR DNA typing on several areas of the underwear: a clear correlation between alpha-amylases activity and male DNA was detected on all the samples evaluated. PMID:17199626

  14. Design Implications from a Usability Study of GramStain-Tutor.

    ERIC Educational Resources Information Center

    Kim, Sara; Brock, Douglas; Orkand, Adam; Astion, Michael

    2001-01-01

    Describes a usability study conducted with health sciences students at the University of Washington that explored interface issues in the GramStain Tutor, an educational software program on CD-ROM, particularly the navigation of the program and the use of embedded design features. (LRW)

  15. 9 CFR 147.3 - The stained-antigen, rapid, whole-blood test. 3

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...-blood test. 3 147.3 Section 147.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE... Blood Testing Procedures § 147.3 The stained-antigen, rapid, whole-blood test. 3 3 The procedure... Secretary of Agriculture. (b) A loop for measuring the correct quantity of blood can usually be...

  16. Evaluation of maturity group III soybean lines for resistance to purple seed stain in Mississippi, 2010

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Purple seed stain (PSS) of soybean is an important disease caused by Cercospora kikuchii. PSS reduces seed quality and market grade, affects seed germination and vigor, and has been reported wherever soybeans are grown worldwide. In 2009, PSS caused 6.4 million bushels of yield losses in 16 southern...

  17. 9 CFR 147.3 - The stained-antigen, rapid, whole-blood test. 3

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false The stained-antigen, rapid, whole-blood test. 3 147.3 Section 147.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE LIVESTOCK IMPROVEMENT AUXILIARY PROVISIONS ON NATIONAL POULTRY IMPROVEMENT PLAN Blood Testing Procedures § 147.3...

  18. Quantitative analysis of stain variability in histology slides and an algorithm for standardization

    NASA Astrophysics Data System (ADS)

    Ehteshami Bejnordi, Babak; Timofeeva, Nadya; Otte-Höller, Irene; Karssemeijer, Nico; van der Laak, Jeroen A. W. M.

    2014-03-01

    This paper presents data on the sources of variation of the widely used hematoxylin and eosin (H&E) histological staining, as well as a new algorithm to reduce these variations in digitally scanned tissue sections. Experimental results demonstrate that staining protocols in different laboratories and staining on different days of the week are the major factors causing color variations in histopathological images. The proposed algorithm for standardizing histology slides is based on an initial clustering of the image into two tissue components having different absorption characteristics for different dyes. The color distribution for each tissue component is standardized by aligning the 2D histogram of color distribution in the hue-saturation-density (HSD) model. Qualitative evaluation of the proposed standardization algorithm shows that color constancy of the standardized images is improved. Quantitative evaluation demonstrates that the algorithm outperforms competing methods. In conclusion, the paper demonstrates that staining variations, which may potentially hamper usefulness of computer assisted analysis of histopathological images, can be reduced considerably by applying the proposed algorithm.

  19. Ruthenium Red Colorimetric and Birefringent Staining of Amyloid-? Aggregates in Vitro and in Tg2576 Mice

    PubMed Central

    2012-01-01

    Alzheimer’s disease (AD) is a devastating neurodegenerative disease most notably characterized by the misfolding of amyloid-? (A?) into fibrils and its accumulation into plaques. In this Article, we utilize the affinity of A? fibrils to bind metal cations and subsequently imprint their chirality to bound molecules to develop novel imaging compounds for staining A? aggregates. Here, we investigate the cationic dye ruthenium red (ammoniated ruthenium oxychloride) that binds calcium-binding proteins, as a labeling agent for A? deposits. Ruthenium red stained amyloid plaques red under light microscopy, and exhibited birefringence under crossed polarizers when bound to A? plaques in brain tissue sections from the Tg2576 mouse model of AD. Staining of A? plaques was confirmed via staining of the same sections with the fluorescent amyloid binding dye Thioflavin S. In addition, it was confirmed that divalent cations such as calcium displace ruthenium red, consistent with a mechanism of binding by electrostatic interaction. We further characterized the interaction of ruthenium red with synthetic A? fibrils using independent biophysical techniques. Ruthenium red exhibited birefringence and induced circular dichroic bands at 540 nm upon binding to A? fibrils due to induced chirality. Thus, the chirality and cation binding properties of A? aggregates could be capitalized for the development of novel amyloid labeling methods, adding to the arsenal of AD imaging techniques and diagnostic tools. PMID:23509974

  20. Computational model to evaluate port wine stain depth profiling using pulsed photothermal radiometry

    E-print Network

    Choi, Bernard

    laser therapy of port wine stain PWS lesions, which are congeni- tal vascular malformations present of pulsed dye laser light 585­595 nm , which is absorbed by hemoglo- bin constituents in blood and epidermal% of patients.5­8 A major limita- tion of current laser therapy is that treatment plans are based primarily