Sample records for fluorescein diacetate staining

  1. Use of an adenosine triphosphate assay, and simultaneous staining with fluorescein diacetate and propidium iodide, to evaluate the effects of cryoprotectants on hard coral ( Echinopora spp.) oocytes

    Microsoft Academic Search

    S. Tsai; E. Spikings; F. W. Kuo; N. C. Lin; C. Lin

    2010-01-01

    The objective was to examine the effects of cryoprotectants on oocytes of hard corals (Echinopora spp.) to obtain basic knowledge for cryopreservation procedures. Oocytes were exposed to various concentrations of cryoprotectants (0.25 to 5.0M) for 20min at room temperature (25°C). Two tests were used to assess ovarian follicle viability: fluorescein diacetate (FDA)+propidium iodide (PI) staining, and adenosine triphosphate (ATP) assay.

  2. Use of an adenosine triphosphate assay, and simultaneous staining with fluorescein diacetate and propidium iodide, to evaluate the effects of cryoprotectants on hard coral (Echinopora spp.) oocytes.

    PubMed

    Tsai, S; Spikings, E; Kuo, F W; Lin, N C; Lin, C

    2010-03-15

    The objective was to examine the effects of cryoprotectants on oocytes of hard corals (Echinopora spp.) to obtain basic knowledge for cryopreservation procedures. Oocytes were exposed to various concentrations of cryoprotectants (0.25 to 5.0M) for 20 min at room temperature (25 degrees C). Two tests were used to assess ovarian follicle viability: fluorescein diacetate (FDA)+propidium iodide (PI) staining, and adenosine triphosphate (ATP) assay. Both FDA+PI staining and ATP assay indicated that cryoprotectant toxicity to oocytes increased in the order methanol, dimethyl sulfoxide (DMSO), propylene glycol (PG), and ethylene glycol (EG). The no observed effect concentrations for Echinopora spp. oocytes were 1.0, 0.5, 0.25, and 0.25 M for methanol, DMSO, PG, and EG, respectively, when assessed with FDA+PI. The ATP assay was more sensitive than FDA+PI staining (P<0.05). Oocyte viability after 1.0M methanol, DMSO, EG, or PG treatment for 20 min at room temperature assessed with FDA+PI tests and ATP assay were 88.9+/-3.1% and 72.2+/-4.4%, 66.2+/-5.0% and 23.2+/-4.9%, 58.9+/-5.4% and 1.1+/-0.7%, and 49.1+/-5.1% and 0.9+/-0.5%, respectively. We inferred that the ATP assay was a valuable measure of cellular injury after cryoprotectant incubation. The results of this study provided a basis for development of protocols to cryopreserve coral oocytes. PMID:20005561

  3. Aging of the erythrocyte. V. Hydrolysis of fluorescein diacetate in red cells.

    PubMed

    Bartosz, G

    1982-01-01

    In contrast to situation found in other cell types, no linear dependence of product fluorescence vs time is observed when fluorescein diacetate (FDA) is hydrolysed by erythrocytes and hemolysates. The rate of hydrolysis is increased by high concentrations of sucrose suggesting a positive effect of viscosity on the rate of the reaction. These peculiarities can be explained by assumption of a two-step hydrolysis of FDA. The FDA-hydrolytic activity decreases with increasing cell density (age). PMID:7093429

  4. Effects of Fluorescein Staining on Laser In Vivo Confocal Microscopy Images of the Cornea

    PubMed Central

    Sindt, Christine W.; Critser, D. Brice; Grout, Trudy K.; Kern, Jami R.

    2012-01-01

    This study was designed to identify whether topical fluorescein, a common ophthalmic tool, affects laser in vivo confocal microscopy of the cornea, a tool with growing applications. Twenty-five eye care specialists were asked to identify presence or absence of fluorescein in 99 confocal micrographs of healthy corneas. Responses were statistically similar to guessing for the epithelium (48%?±?14% of respondents correct per image) and the subbasal nerve plexus (49%?±?11% correct), but results were less clear for the stroma. Dendritic immune cells were quantified in bilateral images from subjects who had been unilaterally stained with fluorescein. Density of dendritic immune cells was statistically similar between the unstained and contralateral stained eyes of 24 contact lens wearers (P = .72) and of 10 nonwearers (P = .53). Overall, the results indicated that fluorescein staining did not interfere with laser confocal microscopy of corneal epithelium, subbasal nerves, or dendritic immune cells. PMID:22363837

  5. 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

  6. Dual appearance of fluorescein staining in vivo of diseased human corneal epithelium. A non-contact photomicrographic study.

    PubMed Central

    Tabery, H M

    1992-01-01

    Adherence of fluorescein sodium dye to diseased epithelial cells, a hitherto unreported phenomenon, was captured in photomicrographs in severe herpes zoster and keratoconjunctivitis sicca keratopathies. It is notable that this phenomenon differs completely from the well known fluorescent property of the dye penetrating into defective corneal epithelium, and that the staining pattern shown by adherent fluorescein correlates well with the staining pattern shown by rose bengal dye. Images PMID:1371224

  7. Fluorescein isothiocyanate staining intensity as a probe of hyperthermia-induced changes in chromatin conformation.

    PubMed

    Dyson, J E; Britten, R A; Battersby, I; Surrey, C R

    1989-03-01

    In a previous report we presented evidence for large increases in fluorescein isothiocyanate (FITC) fluorescent intensity caused by hyperthermia which were not associated with synthesis of heat-shock proteins. We have now refined and considerably extended the measurements of increases in FITC fluorescent intensity caused by hyperthermia within the range 41.0 degrees C to 50.0 degrees C, and associated these with the extent of cell death caused by the hyperthermia. It appears that cell death ensues when the FITC fluorescent intensity has not returned to its baseline value within the time of one cell cycle. If thermotolerance is induced, there is a concomitant reduction in the increase in FITC staining intensity and the extent of cell death. When hyperthermia is followed by acid extraction, an additional increase in FITC staining intensity (above that due to hyperthermia alone) is observed, indicating separate sites of action on basic nuclear proteins. Hyperthermia and acid extraction have related effects on the relationship between FITC and propidium iodide staining. Hyperthermia-induced increases in FITC staining intensity are almost completely reversed by 6.7 mM formaldehyde with a marginal effect on the control FITC staining at this formaldehyde concentration. We suggest that hyperthermia causes extensive dissociation of basic protein-protein binding within nuclear chromatin, and that this may be a contributory cause of hyperthermia-induced cell death. PMID:2469557

  8. Effects of hyperthermia, irradiation, and cytotoxic drugs on fluorescein isothiocyanate staining intensity for flow cytofluorometry

    SciTech Connect

    Dyson, J.E.; McLaughlin, J.B.; Surrey, C.R.; Simmons, D.M.; Daniel, J.

    1987-01-01

    Measurement of fluorescein isothiocyanate (FITC) staining intensity of cultured lymphoblastoid cells following hyperthermia showed large increases without concomitant increases in nuclear protein. Similar measurements of cells following incubation with cytotoxic drugs showed fluorescent intensity increases that exceeded the increases in nuclear protein that were due to the cell cycle blocking action of the drug. The reverse, however, was true for cells following irradiation. In contrast, FITC staining intensity and nuclear protein measurements of cells proceeding through the cell cycle after removal of the cycle blocking agent showed nearly parallel changes, although there were reproducible minor differences, especially following blocking with hydroxyurea. These results suggest that FITC staining intensity is a function not only of nuclear protein content but also of stain access to the reaction sites of the protein constituents of the chromatin. Thus, it is possible that FITC staining may be used as a probe of changes in chromatin structure following experimental manipulation of cells in vitro or treatment of tumors in vivo.

  9. 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.

  10. Rapid and simple method for double staining of bacteria with 4',6-diamidino-2-phenylindole and fluorescein isothiocyanate-labeled antibodies.

    PubMed Central

    Hoff, K A

    1988-01-01

    Bacteria were either heat fixed on microscope slides or filtered with 0.2 micron-pore-size Nuclepore filters. The samples were stained with 4',6-diamidino-2-phenylindole (DAPI) for total staining and with polyvalent rabbit antibodies and fluorescein isothiocyanate-conjugated swine anti-rabbit antibodies for specific staining. By switching between two different optical filter packages in the microscope, only one sample was needed for determining both total and specific counts of bacteria. False-positive counts and other artifacts that occur with antibody staining were easily distinguished when individual fluorescent particles were checked for DAPI fluorescence. The method for applying the general stain to membrane filters was performed quickly and simply by using a DAPI-soaked polypropylene filter that lay beneath the Nuclepore filter which collected the sample. Images PMID:2464967

  11. Fluorescein. Physiochemical factors affecting its fluorescence.

    PubMed

    Romanchuk, K G

    1982-01-01

    Fluorescein's property of fluorescence is reviewed. Of the many factors which affect its fluorescence, concentration is probably the most important and it best explains why leaking aqueous turns fluorescein bright green during Seidel's test. The intensity and pattern of fluorescein staining of corneal lesions is probably due to the concentration and distribution of fluorescein in the cornea. The concentration of fluorescein achieved in the retinal blood vessels during fluorescein angiography affects its fluorescence. PMID:7046118

  12. A preservable two color staining procedure to detect toxicant impacts on algae (Selenastrum capricornutum) using flow cytometry

    SciTech Connect

    Faber, M.; Smith, L.; Boermans, H.; Stephenson, G.; Thompson, D.G. [Univ. of Guelph, Ontario (Canada). Dept. of Environmental Biology

    1995-12-31

    Over the last several years, the use of flow cytometry to assess the impacts of toxicants on algae has increased. Previous studies have tested cell viability using chlorophyll autofluorescence or single stain flow cytometric analysis in fresh algal cultures. A rapid, two-color flow cytometric assay to evaluate viability and cytotoxicity of Selenastrum capricomutum in preserved samples is described. The staining procedure involved fluorescein diacetate, a fluorogenic esterase substrate cleaved in viable cells to fluorescein (green 525 nm) and ethidium homodimer-1 dye which passes through plasma membranes of compromised and dead cells staining DNA (red 620 nm). The auto fluorescence of chlorophyll-a (deep red 675 nm) was used to assess cell viability and examined for use as an internal comparison with the staining procedure. For storage, stained cells were fixed in glutaraldehyde, flash frozen in liquid nitrogen and stored frozen ({minus}20C) for assessment at a later date. Weekly analysis of stored samples showed that the fluorescence of fluorescein diacetate and ethidium homodimer-1 stained cells was stable under these conditions for the 2 months used in this study. A prepared culture of Selenastrum capricomutum containing a 50% (v/v) mixture of live and heat killed cells showed 36.1 % stained live, 13.2% as compromised, 12% unlabeled, and 38.7% dead algal cells. This two-color flow cytometric procedure has proven to be a reliable, sensitive assay to determine viability of Selenastrum capricomutum in preserved samples. The sensitivity of this dual-color assay and the ability to store samples for later analysis are significant improvements over current techniques. This method is being tested for detection of chemical induced algal cytotoxicity and for potential adaptations to field studies.

  13. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 2013-04-01 2013-04-01 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....

  14. 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....

  15. 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....

  16. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 2012-04-01 2012-04-01 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....

  17. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 2014-04-01 2014-04-01 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....

  18. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 2013-04-01 2013-04-01 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....

  19. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 2012-04-01 2012-04-01 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....

  20. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 2014-04-01 2014-04-01 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....

  1. 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....

  2. 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....

  3. 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

  4. Fluorescein Derivatives in Intravital Fluorescence Imaging

    PubMed Central

    Robertson, Thomas A.; Bunel, Florestan; Roberts, Michael S.

    2013-01-01

    Intravital fluorescence microscopy enables the direct imaging of fluorophores in vivo and advanced techniques such as fluorescence lifetime imaging (FLIM) enable the simultaneous detection of multiple fluorophores. Consequently, it is now possible to record distribution and metabolism of a chemical in vivo and to optimise the delivery of fluorophores in vivo. Recent clinical applications with fluorescein and other intravital fluorescent stains have occurred in neurosurgery, dermatology [including photodynamic therapy (PDT)] and endomicroscopy. Potential uses have been identified in periodontal disease, skin graft and cancer surgery. Animal studies have demonstrated that diseased tissue can be specifically stained with fluorophore conjugates. This review focuses on the fluorescein derived fluorophores in common clinical use and provides examples of novel applications from studies in tissue samples. PMID:24709799

  5. Seidel's test using 10% fluorescein.

    PubMed

    Romanchuk, K G

    1979-10-01

    A 10% solution of fluorescein applied topically shows a leak from the anterior chamber better than a 2% solution. Fluorescein changes color because it is diluted by the leaking aqueous and not because its pH is changed. PMID:550919

  6. Gram stain

    MedlinePLUS

    A Gram stain is a test used to identify bacteria. It is one of the most common ways to quickly diagnose ... Urethral discharge Gram stain; Feces Gram stain; Stool Gram stain; Joint fluid Gram stain; Pericardial fluid Gram stain; Gram stain of urethral discharge; ...

  7. Wood stains

    MedlinePLUS

    Wood stains are products used for wood finishing. Wood stain poisoning occurs when someone swallows these substances. This is ... Various wood stains Note: This list does not include all sources of wood stain.

  8. "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

  9. 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.

  10. Hypersensitivity to mercuric fluorescein compounds.

    PubMed

    Barranco Sanz, P; Martín Muñoz, F; López Serrano, C; Martín Esteban, M; Ojeda Casas, J A

    1989-01-01

    The mercurial compounds are known to be a common cause of allergic contact dermatitis. Immediate hypersensitivity is rarely induced by mercurial organic derivatives. However, none of the published cases showed both immediate and delayed hypersensitivity as was the case of one of our two patients. A boy and a woman experienced urticaria and anaphylaxis, respectively after topical application of Merchromine (Merbromine). We were able to demonstrate immediate hypersensitivity in both patients to mercuric fluorescein compounds by skin test and histamine liberation. In addition, the child but not the woman, showed delayed hypersensitivity to mercurial compounds. PMID:2479247

  11. Gram Stain

    MedlinePLUS

    ... may be present are categorized by color and shape during the microscopic evaluation: Color — typically bacteria may be either "Gram positive" (purple) ... cells (intracellular) are also noted. The Gram stain color and the bacterial shape give clues as to what bacteria might be ...

  12. 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

    ...Ethinyl Estradiol; Ethynodiol Diacetate) Tablet and Four Other Drug Products Were Not...ethynodiol diacetate) Pkwy., Skokie, Tablet, 0.05 mg; 1 mg. IL 60077. NDA 018160...ethinyl estradiol; ethynodiol diacetate) Tablet, 0.035 mg; 1 mg. [[Page...

  13. Studies on fluorescein-III The acid strengths of fluorescein as shown by potentiometric titration.

    PubMed

    Diehl, H; Horchak-Morris, N; Hefley, A J; Munson, L F; Markuszewski, R

    1986-11-01

    Potentiometric back-titration of yellow solid fluorescein (H(2)Fl) and of red solid fluorescein in alkali with acid yielded titration curves that were practically identical in shape and position. The end-points at pH 8.5, 5.40 and 3.3 corresponded, respectively, to titration of the excess of standard alkali, and the successive protonations Fl(2-) + H(+) = HFl(-) and HFl(-) + H(+) = H(2)Fl. The pH at the mid-point of the first protonation yielded a value of 6.36 for pK(HFl) (ionic strength 0.10). Because of precipitation of yellow fluorescein during the second protonation step, a value for pK(H(2)Fl) could not be obtained. The total concentration of fluorescein at the first appearance of the precipitate fell on the curve for the solubility of yellow fluorescein as a function of pH. The titrations and the pK values found for the three acid groups of protonated fluorescein (H(3)Fl(+)) have been interpreted on the basis that in water fluorescein exists in only one structural form the yellow zwitterion. Similar back-titrations of alkalinized solutions of yellow or red fluorescein in 50% aqueous ethanol showed that in this medium fluorescein is present in only one form, presumably the quinonoid structure, with much weaker apparent acid functions, pK'(1) = 6.38 and PK'(2) = 7.16 (ionic strength 0.10). PMID:18964223

  14. Advantage of microscope integrated for both indocyanine green and fluorescein videoangiography on aneurysmal surgery: case report.

    PubMed

    Yoshioka, Hideyuki; Kinouchi, Hiroyuki; Nishiyama, Yoshihisa; Kanemaru, Kazuya; Yagi, Takashi; Hanihara, Mitsuto; Horikoshi, Toru

    2014-01-01

    Neck clipping of a large middle cerebral artery aneurysm was performed using a newly developed surgical microscope integrated with modules for both indocyanine green (ICG) and fluorescein videoangiography. During surgery, ICG and fluorescein videoangiography by intra-arterial or intravenous injection were safely carried out without interrupting the surgical procedure. Based on the findings obtained from the case, we evaluated the differences between the dyes and the injection routes. With intra-arterial injection, fluorescein offered sharper contrast images and was better at depicting fine arteries than ICG. Patchy staining of vessel walls was observed in intravenous fluorescein videoangiography, while it was not evident in ICG. Intra-arterial injection method had a great advantage in the rapid clearance of the dyes, which allowed us to perform repeated videoangiography within a short period, and was useful in detecting incomplete clipping in this case; however, catheter insertion requires additional work and carries a potential risk. Use of a microscope integrated for both ICG and fluorescein videoangiography would be another method for repeated evaluation. Namely, alternate use of the dyes enables us to perform videoangiography in a short time even via intravenous injection. PMID:24097092

  15. Preparation of Fluorescein Isothiocyanate-Labeled ?-Globulin by Dialysis, Gel Filtration, and Ion-Exchange Chromatography in Combination

    PubMed Central

    Dedmon, Robert E.; Holmes, Albert W.; Deinhardt, Friedrich

    1965-01-01

    Dedmon, Robert E. (Presbyterian-St. Luke's Hospital, Chicago, Ill.), Albert W. Holmes, and Friedrich Deinhardt. Preparation of fluorescein isothiocyanate-labeled ?-globulin by dialysis, gel filtration, and ion-exchange chromatography in combination. J. Bacteriol. 89:734–739. 1965.—Antiviral immune ?-globulins isolated from rabbit and guinea pig sera were labeled through dialysis membranes with fluorescein isothiocyanate and purified in several ways to eliminate nonspecific staining. Gel filtration of the conjugate with Sephadex G-25 coarse beads followed by column fractionation with diethylaminoethyl-Sephadex yielded consistently highly specific staining materials. Fluorescein-protein ratios varied between 1.0 and 4.0. This technique has proved to be simple and reliable, and is less time-consuming than previous techniques. PMID:14273654

  16. Der Einfluß von Na-Fluorescein auf die Feinstruktur der Wurzelhaare von Lepidium sativum

    Microsoft Academic Search

    Udo Kristen

    1972-01-01

    Cytoplasmic disarrangements in the root hairs of Lepidium sativum caused by the vital stain Na-fluorescein (Uranine) after applications of different duration were analyzed by electron microscopy. After an application time of eight min there appear microbody-like structures and vacuoles in the cytoplasm. After a 16-min application severe disorganizations of membranes are brought about. There are distortions and dissolutions of the

  17. Clinical staining of the ocular surface: mechanisms and interpretations.

    PubMed

    Bron, A J; Argüeso, P; Irkec, M; Bright, F V

    2015-01-01

    In this article we review the mechanism of ocular surface staining. Water-soluble dyes are excluded from the normal epithelium by tight junctions, the plasma membranes and the surface glycocalyx. Shed cells can take up dye. A proportion of normal corneas show sparse, scattered time-dependent, punctate fluorescein uptake, which, we hypothesise, is due to a graded loss of the glycocalyx barrier, permitting transcellular entry into pre-shed cells. In pathological staining, there is little evidence of 'micropooling' at sites of shedding and the term 'punctate erosion' may be a misnomer. It is more likely that the initial event involves transcellular dye entry and, in addition, diffusion across defective tight junctions. Different dye-staining characteristics probably reflect differences in molecular size and other physical properties of each dye, coupled with differences in visibility under the conditions of illumination used. This is most relevant to the rapid epithelial spread of fluorescein from sites of punctate staining, compared to the apparent confinement of dyes to staining cells with dyes such as lissamine green and rose bengal. We assume that fluorescein, with its lower molecular weight, spreads initially by a paracellular route and then by transcellular diffusion. Solution-Induced Corneal Staining (SICS), related to the use of certain contact lens care solutions, may have a different basis, involving the non-pathological uptake of cationic preservatives, such as biguanides, into epithelial membranes and secondary binding of the fluorescein anion. It is transient and may not imply corneal toxicity. Understanding the mechanism of staining is relevant to the standardisation of grading, to monitoring disease and to the conduct of clinical trials. PMID:25461622

  18. 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 ...

  19. 40 CFR 180.1058 - Sodium diacetate; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...Sodium diacetate, when used postharvest as a fungicide, is exempt from the requirement of a tolerance for residues in or on alfalfa, hay; Bermudagrass, hay; bluegrass, hay; bromegrass, hay; clover,hay; corm, field, grain; corn, pop,...

  20. 40 CFR 180.1058 - Sodium diacetate; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...Sodium diacetate, when used postharvest as a fungicide, is exempt from the requirement of a tolerance for residues in or on alfalfa, hay; Bermudagrass, hay; bluegrass, hay; bromegrass, hay; clover,hay; corm, field, grain; corn, pop,...

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

    PubMed

    Marrero, María Teresa; Estévez, Sara; Negrín, Gledy; Quintana, José; López, Mariana; Pérez, Francisco J; Triana, Jorge; León, Francisco; Estévez, Francisco

    2012-11-01

    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(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(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. PMID:23063980

  2. Direct observation and validation of fluorescein tear film break-up patterns by using a dual thermal-fluorescent imaging system.

    PubMed

    Su, Tai-Yuan; Chang, Shu-Wen; Yang, Chiao-Ju; Chiang, Huihua Kenny

    2014-08-01

    The fluorescein tear film break-up test is a common tear film stability test for dry eye diagnosis. This test requires applying fluorescein sodium drops to a tear film to observe the tear film break-up. However, this test is limited by using the fluorescein sodium drops, which can induce reflex tearing and reduce the reliability of the diagnosis results. This paper proposes that tear film evaporation accelerates on the fluorescein tear film break-up area (FTBA), resulting in a lower temperature area (LTA) on the tear film. A dual modality system was established to capture the thermal and fluorescent image of fluorescein-stain tear films for 48 participants. Observations showed that the LTA and FTBA were highly correlated in their location (r = 0.82) and size (r = 0.91). This is first study to show that the FTBA and LTA are essentially the same region. This study demonstrated the feasibility of using the noncontact thermograph method to evaluate tear film stability without using a fluorescein sodium drop. PMID:25136489

  3. 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

  4. Ultra-widefield fluorescein angiography of white without pressure

    PubMed Central

    Orlin, Anton; Fatoo, Aalya; Ehrlich, Joshua; D’Amico, Donald J; Chan, RV Paul; Kiss, Szilárd

    2013-01-01

    Purpose To describe ultra-widefield fluorescein angiography (UWFA) findings in eyes with white without pressure (WWOP) and in eyes without any obvious peripheral chorioretinal disease, and to determine if a difference exists between these two groups. Methods A retrospective review of 379 eyes undergoing diagnostic UWFA using the Optos 200Tx imaging system. Eyes were excluded if the quality of the color photograph or UWFA prevented reliable evaluation. Eyes were also excluded if there was any evidence of peripheral retinal or choroidal disease, which was thought to have an effect on UWFA (eg, peripheral background diabetic or hypertensive retinopathy, vein occlusion, or any other peripheral vascular disorder). Eyes were determined to have WWOP, based on a dilated fundus examination and color fundus photography that contained areas of peripheral retinal whitening consistent with the diagnosis. UWFA was evaluated by trained masked graders, and determined to have or not have peripheral vascular leakage and/or staining. Results Of the 379 eyes evaluated, 45 eyes were included in the study. Twelve eyes were determined to have peripheral WWOP; 33 eyes did not have WWOP on examination or color fundus photography. Three common UWFA peripheral patterns were visualized. Eyes with and without WWOP were grouped into one of three patterns. The majority of eyes without WWOP demonstrated UWFA pattern one (69.7%), while those in the WWOP group demonstrated pattern three (50%). The distribution of UWFA patterns is statistically different between those with and without WWOP (P = 0.002). In eyes without WWOP, in patients with no documented systemic microvascular disease (diabetes, hypertension), 71.4% of eyes had UWFA pattern one while 14.3% had both patterns two and three. Conclusion This study is one of the first to specifically evaluate peripheral vascular leakage/staining in eyes with WWOP as well as in eyes without any obvious peripheral chorioretinal disease. We demonstrate that a significant portion of WWOP eyes exhibit peripheral findings on UWFA (pattern one) compared to eyes without WWOP. Importantly, even in eyes that are apparently unremarkable in the periphery on exam and color photography, UWFA can still show peripheral vascular abnormalities. These results warrant further investigation. PMID:23737658

  5. 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

  6. Nanoparticle Stained Glass

    NSDL National Science Digital Library

    2014-06-18

    In this activity/demo, learners are introduced to the connection between medieval stained glass artisans and nanotechnology. Learners discover that the red and yellow colors in stained glass windows come from nanoparticles of gold and silver embedded in the glass. This activity/demo consists of two hands-on activities: making a collaborative stained glass window with pre-made nanoparticle solutions containing silver or gold and making a take-away card that contains a small piece of nanoparticle stained “glass."

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

    SciTech Connect

    Marrero, Maria Teresa; Estevez, Sara; Negrin, Gledy; Quintana, Jose [Departamento de Bioquimica, Unidad Asociada al Consejo Superior de Investigaciones Cientificas, Universidad de Las Palmas de Gran Canaria, Plaza Dr. Pasteur s/n, 35016 Las Palmas de Gran Canaria (Spain)] [Departamento de Bioquimica, Unidad Asociada al Consejo Superior de Investigaciones Cientificas, Universidad de Las Palmas de Gran Canaria, Plaza Dr. Pasteur s/n, 35016 Las Palmas de Gran Canaria (Spain); Lopez, Mariana; Perez, Francisco J.; Triana, Jorge [Departamento de Quimica, Universidad de Las Palmas de Gran Canaria, Instituto Canario de Investigacion del Cancer, 35017 Las Palmas de Gran Canaria (Spain)] [Departamento de Quimica, Universidad de Las Palmas de Gran Canaria, Instituto Canario de Investigacion del Cancer, 35017 Las Palmas de Gran Canaria (Spain); Leon, Francisco [Instituto de Productos Naturales y Agrobiologia, Consejo Superior de Investigaciones Cientificas, Avda. Astrofisico F. Sanchez 3, 38206 La Laguna, Tenerife (Spain)] [Instituto de Productos Naturales y Agrobiologia, Consejo Superior de Investigaciones Cientificas, Avda. Astrofisico F. Sanchez 3, 38206 La Laguna, Tenerife (Spain); Estevez, Francisco, E-mail: festevez@dbbf.ulpgc.es [Departamento de Bioquimica, Unidad Asociada al Consejo Superior de Investigaciones Cientificas, Universidad de Las Palmas de Gran Canaria, Plaza Dr. Pasteur s/n, 35016 Las Palmas de Gran Canaria (Spain)] [Departamento de Bioquimica, Unidad Asociada al Consejo Superior de Investigaciones Cientificas, Universidad de Las Palmas de Gran Canaria, Plaza Dr. Pasteur s/n, 35016 Las Palmas de Gran Canaria (Spain)

    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.

  8. 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.

  9. Gram-Stained Fusobacterium

    NSDL National Science Digital Library

    American Society For Microbiology

    2006-06-30

    A gram-negative stained Fusobacterium spp. with spindle-shaped morphology. This bacterium may be spindle shaped or coccobacilli; it is often pleomorphic. Without spindle shapes, it is often difficult to differentiate from other nonmotile anaerobe

  10. Port-Wine Stain

    MedlinePLUS

    ... limb. The syndrome is most frequently diagnosed in infancy or early childhood. Who's At Risk Port-wine ... difference between a port-wine stain and other birthmarks, such as a salmon patch or a hemangioma, ...

  11. Candida, fluorescent stain (image)

    MedlinePLUS

    This microscopic film shows a fluorescent stain of Candida. Candida is a yeast (fungus) that causes mild disease, but in immunocompromised individuals it may cause life-threatening illness. (Image ...

  12. Port-Wine Stains

    MedlinePLUS

    ... their own, they can be treated. In fact, laser therapies can make many port-wine stains much ... mark might be. The good news is that lasers (highly concentrated light energy) can make many kids' ...

  13. Diflorasone diacetate: vasoconstrictor activity and clinical efficacy of a new topical corticosteroid.

    PubMed

    Bluefarb, S M; Howard, F M; Leibsohn, E; Schlagel, C A; Wexler, L

    1976-01-01

    Diflorasone diacetate, a new topical corticosteroid, was generally more potent than three high potency reference standards (fluocinonide, beta-methasone 17-valerate and fluocinolone acetonide) when the compounds were dissolved in 95% alcohol and applied in vasoconstrictor assays in healthy volunteers. On the basis of additional vasoconstrictor assay results, a 0-05% concentration of the steroid in a cream vehicle containing 15% propylene glycol was developed for therapeutic evaluation. In a double-blind comparison in 384 patients with dermatoses, 0-05% diflorasone diacetate cream was as effective as 0-05% fluocinonide cream in the therapy of lesions of psoriasis or atopic/neurodermatitis. PMID:800385

  14. Case Report of Bullous Pemphigoid following Fundus Fluorescein Angiography.

    PubMed

    Demirci, Goktug; Demirci, Gulsen Tukenmez; Gulkilik, Gokhan

    2010-01-01

    PURPOSE: To report a first case of bullous pemphigoid (BP) following intravenous fluorescein for fundus angiography. Clinical Features: A 70-year-old male patient was admitted to the intensive care unit with BP and sepsis. He reported a history of fundus fluorescein angiography with a pre-diagnosis of senile macular degeneration 2 months prior to presentation. At that time, fluorescein extravasated at the antecubital region. Following the procedure, pruritus and erythema began at the wrists bilaterally, and quickly spread to the entire body. The patient also reported a history of allergy to human albumin solution (Plamasteril(R); Abbott) 15 years before, during bypass surgery. On dermatologic examination, erythematous patches were present on the scalp, chest and anogenital region. Vesicles and bullous lesions were present on upper and lower extremities. On day 2 of hospitalization, tense bullae appeared on the upper and lower extremities. The patient was treated with oral methylprednisolone 48 mg (Prednol(R); Mustafa Nevzat), topical clobetasol dipropionate 0.05% cream (Dermovate(R); Glaxo SmithKline), and topical 4% urea lotion (Excipial Lipo(R); Orva) for presumptive bullous pemphigoid. Skin punch biopsy provided tissue for histopathology, direct immunofluorescence examination, and salt extraction, which were all consistent with BP. After 1 month, the patient was transferred to the intensive care unit with sepsis secondary to urinary tract infection; he died 2 weeks later from sepsis and cardiac failure. CONCLUSIONS: To our knowledge, this is the first reported case of BP following fundus fluorescein angiography in a patient with known human albumin solution allergy. Consideration should be made to avoid fluorescein angiography, change administration route, or premedicate with antihistamines in patients with known human albumin solution allergy. The association between fundus fluorescein angiography and BP should be further investigated. PMID:20737052

  15. Stain Removal Guide

    NSDL National Science Digital Library

    Recently retired from "the most successful contract manufacturing Detergent and Sanitiser company in New Zealand," Allan Campbell, PhC MPS, has decided to share his knowledge of detergent chemistry with the world. And what better source for fabric stain tips than a chemist? Visitors can browse the guide, which covers everything from acids to wood saps (including cod liver oil, soy sauce, and chutney), via a frame on the left-hand side of their browsers, Removal tips are listed on the right. A handy site, especially for users facing an ever-spiralling variety of stains in their youngsters's frocks.

  16. 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...

  17. 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…

  18. Stained Glass Glue

    NSDL National Science Digital Library

    American Chemical Society

    2001-01-01

    In this activity on page 6 of the PDF, learners use glue instead of glass to create artwork that can be hung in a window. Discover how the chemicals in various materials mix together to make a colorful, translucent "stained glass" creation.

  19. "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…

  20. A Photostable, pH-Invariant Fluorescein Derivative for Single-Molecule Microscopy

    Microsoft Academic Search

    Baoxu Liu; Steven Fletcher; Miriam Avadisian; Patrick T. Gunning; Claudiu C. Gradinaru

    2009-01-01

    We herein report the comprehensive characterization of the spectral and single-photon fluorescence properties of a recently\\u000a synthesized fluorescein derivative and its biotinylated analog. The fluorophore displays significant increases in photostability\\u000a compared to the known fluorescein label fluorescein isothiocyanate (FITC), as well as superb pH independence. This fluorescein\\u000a variant has two readily accessible functional groups (aniline NH2 and phenol OH) that

  1. Lineage labeling of zebrafish cells with laser uncagable fluorescein dextran.

    PubMed

    Clanton, Joshua A; Shestopalov, Ilya; Chen, James K; Gamse, Joshua T

    2011-01-01

    A central problem in developmental biology is to deduce the origin of the myriad cell types present in vertebrates as they arise from undifferentiated precursors. Researchers have employed various methods of lineage labeling, such as DiI labeling and pressure injection of traceable enzymes to ascertain cell fate at later stages of development in model systems. The first fate maps in zebrafish (Danio rerio) were assembled by iontophoretic injection of fluorescent dyes, such as rhodamine dextran, into single cells in discrete regions of the embryo and tracing the labeled cell's fate over time. While effective, these methods are technically demanding and require specialized equipment not commonly found in zebrafish labs. Recently, photoconvertable fluorescent proteins, such as Eos and Kaede, which irreversibly switch from green to red fluorescence when exposed to ultraviolet light, are seeing increased use in zebrafish. The optical clarity of the zebrafish embryo and the relative ease of transgenesis have made these particularity attractive tools for lineage labeling and to observe the migration of cells in vivo. Despite their utility, these proteins have some disadvantages compared to dye-mediated lineage labeling methods. The most crucial is the difficulty we have found in obtaining high 3-D resolution during photoconversion of these proteins. In this light, perhaps the best combination of resolution and ease of use for lineage labeling in zebrafish makes use of caged fluorescein dextran, a fluorescent dye that is bound to a quenching group that masks its fluorescence. The dye can then be "uncaged" (released from the quenching group) within a specific cell using UV light from a laser or mercury lamp, allowing visualization of its fluorescence or immunodetection. Unlike iontophoretic methods, caged fluorescein can be injected with standard injection apparatuses and uncaged with an epifluorescence microscope equipped with a pinhole. In addition, antibodies against fluorescein detect only the uncaged form, and the epitope survives fixation well. Finally, caged fluorescein can be activated with very high 3-D resolution, especially if two-photon microscopy is employed. This protocol describes a method of lineage labeling by caged fluorescein and laser uncaging. Subsequently, uncaged fluorescein is detected simultaneously with other epitopes such as GFP by labeling with antibodies. PMID:21559005

  2. Fullerene–fluorescein–anthracene hybrids: a model for artificial photosynthesis and solar energy conversion

    Microsoft Academic Search

    Bingwen Jing; Daoben Zhu

    2004-01-01

    Two novel C60–fluorescein–anthracene hybrids have been synthesized. Fluorescence quenching in the hybrids indicates an energy transfer from the excited state of anthracene to fluorescein and photoinduced intramolecular electron transfer from the excited state of fluorescein to the C60 moiety.

  3. Fabrication and characterization of vacuum deposited fluorescein thin films

    Microsoft Academic Search

    Pasi Jalkanen; Sampo Kulju; Konstantin Arutyunov; Liisa Antila; Pasi Myllyperkiö; Teemu Ihalainen; Tommi Kääriäinen; Marja-Leena Kääriäinen; Jouko Korppi-Tommola

    2011-01-01

    Simple vacuum evaporation technique for deposition of dyes on various solid surfaces has been developed. The method is compatible with conventional solvent-free nanofabrication processing enabling fabrication of nanoscale optoelectronic devices. Thin films of fluorescein were deposited on glass, fluorine–tin–oxide (FTO) coated glass with and without atomically layer deposited (ALD) nanocrystalline 20nm thick anatase TiO2 coating. Surface topology, absorption and emission

  4. 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

    Microbial safety and quality attributes were evaluated for frankfurters formulated with potassium lactate/sodium diacetate (0 or 3%) and inoculated with a four-strain Listeria monocytogenes cocktail before and after treatment with pasteurizing doses...

  5. 7.G Stained Glass

    NSDL National Science Digital Library

    This is a task from the Illustrative Mathematics website that is one part of a complete illustration of the standard to which it is aligned. Each task has at least one solution and some commentary that addresses important asects of the task and its potential use. Here are the first few lines of the commentary for this task: The students in Mr. Rivera's art class are designing a stained-glass window to hang in the school entryway. The window will be 2 feet tall and 5 feet w...

  6. 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

  7. Length of stain dosimeter

    NASA Technical Reports Server (NTRS)

    Lueck, Dale E. (inventor)

    1994-01-01

    Payload customers for the Space Shuttle have recently expressed concerns about the possibility of their payloads at an adjacent pad being contaminated by plume effluents from a shuttle at an active pad as they await launch on an inactive pad. As part of a study to satisfy such concerns a ring of inexpensive dosimeters was deployed around the active pad at the inter-pad distance. However, following a launch, dosimeters cannot be read for several hours after the exposure. As a consequence factors such as different substrates, solvent systems, and possible volatilization of HCl from the badges were studied. This observation led to the length of stain (LOS) dosimeters of this invention. Commercial passive LOS dosimeters are sensitive only to the extent of being capable of sensing 2 ppm to 20 ppm if the exposure is 8 hours. To map and quantitate the HCl generated by Shuttle launches, and in the atmosphere within a radius of 1.5 miles from the active pad, a sensitivity of 2 ppm HCl in the atmospheric gases on an exposure of 5 minutes is required. A passive length of stain dosimeter has been developed having a sensitivity rendering it capable of detecting a gas in a concentration as low as 2 ppm on an exposure of five minutes.

  8. Recognition of carotid stenosis with bilateral simultaneous retinal fluorescein angiography.

    PubMed

    Choromokos, E A; Raymond, L A; Sacks, J G

    1982-10-01

    In order to study the arm to retina circulation time (ARCT), we simultaneously photographed both fundi in patients with suspected or known carotid artery occlusive disease. Noninvasive carotid artery tests, selective carotid arteriography, or both, were performed on all of the patients. In the 11 patients with arteriographically proven significant carotid stenosis, or with noninvasive carotid tests demonstrating carotid occlusive disease, a triad of fluoroangiographic features (delayed ARCT, disparity of ARCT between eyes, and delayed ciliary circulation at the optic disc) was found. We believe that bilateral simultaneous retinal fluorescein angiography is a sensitive and accurate method for the recognition of patients with carotid artery occlusive disease. PMID:7155525

  9. Removing Stains from Washable Fabrics.

    E-print Network

    Beard, Ann Vanderpoorten

    1988-01-01

    I UUL. Z TA24S.7 8873 NO.1616 B.1616 / Texas Agricultural Extension Service LIBRARY FEB 0 1 1989 Texas A&M University Removing Stains from Washable Fabrics Ann Vanderpoorten 8eard* Most spots and stains can be removed by prompt... warded by extending the life of your clothing and other textile products. Guidelines for Removing Stains ? Know as much about the fabric as possible. Read the care label to determine safe procedures, and re member that the procedures described...

  10. Biotransformation of oral contraceptive ethynodiol diacetate with microbial and plant cell cultures

    PubMed Central

    2012-01-01

    Background Biotransformation by using microbial and plant cell cultures has been applied effectively for the production of fine chemicals on large scale. Inspired by the wealth of literature available on the biotransformation of steroids, we decided to investigate the biotransformation of ethynodiol diacetate (1) by using plant and microbial cultures. Results The biotransformation of ethynodiol diacetate (1) with Cunninghamella elegans and plant cell suspension cultures of Ocimum basilicum and Azadirachta indica is being reported here for the first time. Biotransformation of 1 with Cunninghamella elegans yielded three new hydroxylated compounds, characterized as 17?-ethynylestr-4-en-3?,17?-diacetoxy-6?-ol (2), 17?-ethynylestr-4-en-3?,17?-diacetoxy-6?-ol (3), and 17?-ethynylestr-4-en-3?,17?-diacetoxy-10?-ol (4) and a known metabolite, 17?-ethynyl-17?-acetoxyestr-4-en-3-one (5). The biotransformation of 1 with Ocimum basilicum included hydrolysis of the ester group, oxidation of alcohol into ketone, and rearrangement of the hydroxyl group. Thus four major known metabolites were characterized as 17?-ethynyl-17?-acetoxyestr-4-en-3-one (5), 17?-ethynyl-17?-hydroxyestr-4-en-3-one (6), 17?-ethynyl-3 ?-hydroxy-17?-acetoxyestr-4-ene (7) and 17?-ethynyl-5?,17?-dihydroxyestr-3-ene (8). Biotransformation of 1 with Azadirachta indica culture yielded compounds 5 and 6. Spectroscopic data of compound 8 is being reported for the first time. Structure of compound 6 was unambiguously deduced through single-crystal x-ray diffraction studies. Conclusion Biotransformation of an oral contraceptive, ethynodiol diacetate (1), by using microbial and plant cell cultures provides an efficient route to the synthesis of a library of new steroids with potential contraceptive properties. These methods can be employed in the production of such compounds with high stereoselectivity. PMID:23021311

  11. Can 1% chlorhexidine diacetate and ethanol stabilize resin-dentin bonds?

    PubMed Central

    Manso, Adriana Pigozzo; Grande, Rosa Helena Miranda; Bedran-Russo, Ana Karina; Reis, Alessandra; Loguercio, Alessandro D.; Pashley, David Henry; Carvalho, Ricardo Marins

    2014-01-01

    Objectives To examine the effects of the combined use of chlorhexidine and ethanol on the durability of resin-dentin bonds. Methods Forty-eight flat dentin surfaces were etched (32% phosphoric acid), rinsed (15 s) and kept wet until bonding procedures. Dentin surfaces were blot-dried with absorbent paper and re-wetted with water (Water, control), 1% chlorhexidine diacetate in water (CHD/Water), 100% ethanol (Ethanol), or 1% chlorhexidine diacetate in ethanol (CHD/Ethanol) solutions for 30 s. They were then bonded with All Bond 3 (AB3, Bisco) or Excite (EX, Ivoclar-Vivadent) using a smooth, continuous rubbing application (10 s), followed by 15 s gentle air stream to evaporate solvents. The adhesives were light-cured (20 s) and resin composite build-ups constructed for the microtensile method. Bonded beams were obtained and tested after 24-hours, 6-months and 15-months of water storage at 37°C. Storage water was changed every month. Effects of treatment and testing periods were analyzed (ANOVA, Holm-Sidak, p<0.05) for each adhesive. Results There were no interactions between factors for both etch-and-rinse adhesives. AB3 was significantly affected only by storage (p = 0.003). Excite was significantly affected only by treatments (p = 0.048). AB3 treated either with ethanol or CHD/ethanol resulted in reduced bond strengths after 15 months. The use of CHD/ethanol resulted in higher bond strengths values for Excite. Conclusions Combined use of ethanol/1% chlorhexidine diacetate did not stabilize bond strengths after 15 months. PMID:24815823

  12. 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.

  13. 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.

  14. Silver stain for electron microscopy

    NASA Technical Reports Server (NTRS)

    Corbett, R. L.

    1972-01-01

    Ammoniacal silver stain used for light microscopy was adapted advantageously for use with very thin biological sections required for electron microscopy. Silver stain can be performed in short time, has more contrast, and is especially useful for low power electron microscopy.

  15. 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...

  16. Diabetic Macular Edema: Passive and Active Transport of Fluorescein through the Blood-Retina Barrier

    Microsoft Academic Search

    Birgit Sander; Michael Larsen; Birgitte Moldow; Henrik Lund-Andersen

    PURPOSE. To investigate the passive bidirectional and active outward transport of fluorescein through the blood-retina bar- rier (BRB) in diabetic patients with clinically significant macu- lar edema and in healthy controls. METHODS. The passive and active transport of fluorescein through the BRB was quantitated by vitreous fluorometry. A previously developed method was used to model passive trans- port. A new

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

    E-print Network

    Castro, Juan C.

    2010-07-14

    Castro, B.S., California State University, San Bernardino Chair of Advisory Committee: Dr. Kevin Burgess A series of fluorescein and rosamine derivatives have been prepared and their spectroscopical properties analyzed to determine their usefulness... CHAPTER IV MICROWAVE-ASSISTED FUNCTIONALIZATION OF BROMO-FLUORESCEIN AND BROMO-RHODAMINE DERIVATIVES ..................................40 CHAPTER V SYNTHESIS OF REGIOISOMERICALLY PURE 5-FUNCTIONALIZED 2...

  18. Spore Stain of Bacillus cereus

    NSDL National Science Digital Library

    American Society For Microbiology

    2002-01-01

    This strain of Bacillus cereus was isolated from a sample of gasoline-contaminated soil and cultured on blood agar. This picture allows students to see spores utilizing a simple, reliable method of staining.

  19. SILVER STAINING OF RIBOSOMAL PROTEINS

    Microsoft Academic Search

    J. W. SMITH; R. J. STUART

    1971-01-01

    SUMMARY The effects of staining several tissues with silver nitrate were studied in the electron micro- scope. Tissues were fixed in 2% glutaraldehyde buffered to pH 7-3 and containing 02 2 M sucrose, some being post-osmicated. Staining was effected by immersion of Araldite sections in unbuffered 5 % silver nitrate for 30 min. Increase in electron density was restricted to

  20. Biological staining: mechanisms and theory.

    PubMed

    Horobin, R W

    2002-01-01

    New staining techniques continue to be introduced, and older ones continue to be used and improved. Several factors control specificity, selectivity and visibility of the end product in any procedure using dyes, fluorochromes, inorganic reagents or histochemical reactions applied to sections or similar preparations. Local concentration of the tissue target often determines the intensity of the observed color, as does the fine structure within the object being stained, which may facilitate or impede diffusion of dyes and other reagents. Several contributions to affinity control the specificity of staining. These include electrical forces, which result in accumulation of dye ions in regions of oppositely charged tissue polyions. Weaker short-range attractions (hydrogen bonding, van der Waals forces or hydrophobic bonding, depending on the solvent) hold dyes ions and histochemical end products in contact with their macromolecular substrates. Nonionic forces can also increase visibility of stained sites by causing aggregation of dye molecules. Covalent bonds between dye and tissue result in the strongest binding, such as in methods using Schiff's reagent and possibly also some mordant dyes. The rate at which a reagent gains access to or is removed from targets in a section or other specimen affect what is stained, especially when more then one dye is used, together or sequentially. Rate-controlled staining is greatly influenced by the presence and type of embedding medium, such as a resin, that infiltrates the tissue. The rates of chemical reactions are major determinants of outcome in many histochemical techniques. Selective staining of different organelles within living cells is accomplished mainly with fluorochromes and is controlled by mechanisms different from those that apply to fixed tissues. Quantitative structure-activity relations (QSAR) of such reagents can be derived from such molecular properties as hydrophilic-hydrophobic balance, extent of conjugated bond systems, acid-base properties and ionic charge. The QSAR correlates with staining of endoplasmic reticulum, lysosomes, mitochondria, DNA, or the plasma membranes of living cells. PMID:11991329

  1. Control of Listeria monocytogenes on commercially-produced frankfurters prepared with and without potassium lactate and sodium diacetate and surface .....using the Sprayed Lethality in Container (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 (low = 0.68% lactate and 0.097% diacetate and high = 1.36% lactate and 0.19% diacetate) and then treated with 22 or ...

  2. 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...

  3. 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...

  4. 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...

  5. 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 ...

  6. 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.

  7. 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.

  8. 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

  9. Drop Coating Deposition Raman Spectroscopy of Fluorescein Isothiocyanate Labeled Protein

    PubMed Central

    Vangala, Karthikeshwar; Jiang, Dongping; Zou, Sige; Pechan, Tibor

    2011-01-01

    Using bovine serum albumin (BSA) as the model protein normal Raman spectra of Fluorescein isothiocyanate (FITC) -conjugated protein was systematically studied for the first time using both solution and the drop coating deposition Raman (DCDR) sampling techniques. The FITC-BSA Raman spectra are dominated by the FITC Raman features that are strongly pH dependent. Current DCDR detection sensitivity obtained with a 10:1 FITC-BSA conjugate is 45 fmol in terms of total protein consumption and ~15 attomol at laser probed volume. Unlike the FITC-BSA solution Raman spectra where the FITC Raman features are photostable, concurrent FITC fluorescence and Raman photobleaching is observed in the DCDR spectra of FITC-BSA. While the FITC Raman photobleaching follows a single exponential decay function with a time constant independent of the FITC labeling ratio, the fluorescence background photobleaching is much more complicated and it depends strongly on the FITC labeling ratio and sample conditions. Mechanistically, the FITC Raman photobleaching is believed to be due to photochemical reaction of the FITC molecules in the electronically excited state. The FITC fluorescence photobleaching involves both concentration quenching and photochemical quenching, and the latter may involve a photochemical intermediate that is fluorescence inactive but Raman active. PMID:20925976

  10. Extraction of Capillary Non-perfusion from Fundus Fluorescein Angiogram

    NASA Astrophysics Data System (ADS)

    Sivaswamy, Jayanthi; Agarwal, Amit; Chawla, Mayank; Rani, Alka; Das, Taraprasad

    Capillary Non-Perfusion (CNP) is a condition in diabetic retinopathy where blood ceases to flow to certain parts of the retina, potentially leading to blindness. This paper presents a solution for automatically detecting and segmenting CNP regions from fundus fluorescein angiograms (FFAs). CNPs are modelled as valleys, and a novel technique based on extrema pyramid is presented for trough-based valley detection. The obtained valley points are used to segment the desired CNP regions by employing a variance-based region growing scheme. The proposed algorithm has been tested on 40 images and validated against expert-marked ground truth. In this paper, we present results of testing and validation of our algorithm against ground truth and compare the segmentation performance against two others methods.The performance of the proposed algorithm is presented as a receiver operating characteristic (ROC) curve. The area under this curve is 0.842 and the distance of ROC from the ideal point (0,1) is 0.31. The proposed method for CNP segmentation was found to outperform the watershed [1] and heat-flow [2] based methods.

  11. Automated single-slide staining system

    NASA Technical Reports Server (NTRS)

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

    1974-01-01

    Apparatus developed to Gram-stain single slides automatically is flexible enough to accommodate other types of staining procedures. Method frees operator and eliminates necessity for subjective evaluations as to length of staining or decolorizing time.

  12. Tear Film, Contact Lens, and Patient Factors Associated with Corneal Staining

    PubMed Central

    Sinnott, Loraine T.

    2011-01-01

    Purpose. The purpose of this study was to examine ocular surface and tear film, contact lens, care solution, medical, and patient-related factors that are associated with corneal staining in contact lens wearers. Methods. In this cross-sectional/nested case–control study, in addition to the assessment of corneal staining with fluorescein, a variety of tear film and ocular surface, contact lens, and patient-related factors were examined. Poisson regression models were used to examine the relation between corneal staining and these factors. Results. Data from 413 patients were eligible for the analyses described. The average age was 30.6 ± 11.1 years, and 277 (67.1%) of the patients were women. Several factors were shown to be related to increased corneal staining in multivariate modeling, including increased daily wearing times (P = 0.0006), lower income (P = 0.0008), lissamine green conjunctival staining (P = 0.002), contact lens deposition (P = 0.007), increased tear meniscus height (P = 0.007), and decreased hydrogel nominal water content (P = 0.02). The wearing of silicone hydrogels (as opposed to hydrogels) was protective against corneal staining (P = 0.0004). Notably, neither contact lens care solutions nor disinfectants were associated with corneal staining. Conclusions. Corneal staining in contact lens wearers continues to be a frequent, but not well understood, outcome. These data suggest that contact lens factors (water content, material, wearing time, and deposition) are more generally associated with corneal staining than are contact lens care solutions or other ocular surface and tear film, demographic, or medical factors. PMID:21087960

  13. Templating Water Stains for Nanolithography

    E-print Network

    -2 electronics,3 material science,4 and biotechnology.5-10 A variety of methods now exist to create nanoscale for creating twin features in polymers and metals. The process works by combining evaporative staining with a templating process. Well-ordered hexagonally arrayed double rings were fabricated using hydrophobic spherical

  14. 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.

  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. CAPILLARY ELECTROPHORESIS/LASER-INDUCED FLUORESCENCE DETECTION OF FLUORESCEIN AS A GROUNDWATER MIGRATION TRACER

    EPA Science Inventory

    Capillary electrophoresis (CE) has been applied to the determination of the groundwater migration tracer dye fluorescein based on laser-induced fluorescence (LIF) detection and compared to determinations obtained with traditional spectrofluorimetry. Detection limits of injected d...

  17. Effects of Acetazolamide on Passive and Active Transport of Fluorescein across the Normal BRB

    Microsoft Academic Search

    Birgitte Moldow; Birgit Sander; Michael Larsen; Henrik Lund-Andersen

    1999-01-01

    PURPOSE. To investigate the effect of the carbonic anhydrase inhibitor acetazolamide (AZM) on passive permeability and active transport of fluorescein across the blood-retina barrier in healthy subjects. The study may have implications for the understanding of the edema-reducing effect of AZM. METHODS. The effect of AZM on the blood-retina barrier function was assessed by differential vitreous spectrofluorometry using fluorescein as

  18. 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.

  19. Method of staining target interphase chromosomal DNA

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel

    2005-03-29

    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 methods for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes.

  20. 68Ga-N,N'-bis[2-Hydroxy-5-(carboxyethyl)benzyl] ethylenediamine-N,N'-diacetic acid-polyethylene glycol-single-

    E-print Network

    Levin, Judith G.

    68Ga-N,N'-bis[2-Hydroxy-5-(carboxyethyl)benzyl] ethylenediamine-N,N'-diacetic acid-polyethylene-N,N'-bis[2-Hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N'- diacetic-polyethylene glycol-single-chain Cys,4,7,10- tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)-polyethylene glycol (PEG)-scVEGF (64Cu-DOTA-PEG-scVEGF), 99m

  1. A combined spectroscopic and theoretical study of dibutyltin diacetate and dilaurate in supercritical CO2.

    PubMed

    Renault, Benjamin; Cloutet, Eric; Cramail, Henri; Hannachi, Yacine; Tassaing, Thierry

    2008-09-11

    Two organotin catalysts, namely, dibutyltin dilaurate (DBTDL) and dibutyltin diacetate (DBTDA), commonly used in the synthesis of polyurethanes, have been investigated combining vibrational spectroscopic measurements with molecular modeling. The structure and vibrational spectra of the DBTDA molecule have been simulated using density functional theory. Thus, because of the Sn...O interactions, the lowest energy conformer reveals an asymmetrically chelated structure of the acetate groups with a C2v symmetry. The experimental IR spectra of DBTDA and DBTDL diluted in carbon tetrachloride and in supercritical CO2 show unambiguously that these molecules adopt the asymmetrically chelated conformation in the solvent. A new attribution of the main peaks constituting the respective IR spectra of the catalysts could be carried out. Finally, from the IR spectra of the two catalysts diluted in supercritical CO2 reported as a function of time, it was found that both molecules react slightly with CO2. However, their spectrum remains unchanged at the earliest stage of the polymerization, indicating that these molecules preserve a catalytic activity similar to that noted in conventional organic solvent. PMID:18707063

  2. 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.

  3. Templating water stains for nanolithography.

    PubMed

    Liao, Wei-Ssu; Chen, Xin; Chen, Jixin; Cremer, Paul S

    2007-08-01

    Herein, a nanoscale patterning technique is demonstrated for creating twin features in polymers and metals. The process works by combining evaporative staining with a templating process. Well-ordered hexagonally arrayed double rings were fabricated using hydrophobic spherical templates. The diameter of the rings, the width of individual rings, and the spacing between concentric and adjacent rings could be tuned by varying the solution conditions. Arrays could be made without the outer ring by employing hydrophilic templates. PMID:17637019

  4. 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

  5. Modification and further evaluation of a fluorescein-labeled peanut agglutinin test for identification of Haemonchus contortus eggs.

    PubMed

    Jurasek, Megan E; Bishop-Stewart, Janell K; Storey, Bobby E; Kaplan, Ray M; Kent, Michael L

    2010-04-19

    Gastrointestinal nematodes of the family Trichostrongylidae are the most important parasites of sheep, goats and other ruminants worldwide. Of this group, Haemonchus contortus is usually the most damaging species, particularly in warmer climates or during the summer. It is therefore useful to be able to rapidly differentiate infections with this nematode from other, less pathogenic, species. However, aside from Nematodirus spp., there are only subtle differences between the egg morphology within the trichostrongyles, making it very difficult to identify eggs to the species level. It has been shown previously that peanut agglutinin specifically binds to Haemonchus eggs and not those of other trichostrongyle species. By using this lectin conjugated to fluorescein isothiocyanate, binding to Haemonchus eggs can be visualized under ultraviolet illumination. We adapted this test using eggs purified by routine sugar centrifugation methods and evaluated 26 diagnostic samples from ruminants submitted to our laboratories in Oregon and Georgia. Very good correlations were seen between this test and larval culture (adjusted R(2)=0.72132; F(1,25)=65.7; p<0.001). There was little variability between two different diagnosticians reading the same sample, suggesting that the test is robust and not subject to reader bias. An additional benefit is that eggs can be examined following preservation; fixation of H. contortus eggs in formalin at 0.5-5% for to up to 4wk did not affect their staining. PMID:20060646

  6. Fluorescein: a rapid, sensitive, nonlethal method for detecting skin ulceration in fish.

    PubMed

    Noga, E J; Udomkusonsri, P

    2002-11-01

    There is a need to develop simple, rapid, and accurate methods for assessing health in fish populations. In this study we demonstrate that use of fluorescein, a nontoxic fluorescent dye, can rapidly and easily detect the presence of skin ulcers in all fish tested, including rainbow trout (Oncorhynchus mykiss), channel catfish (Ictalurus punctatus), goldfish (Carassius auratus), and hybrid striped bass (Morone saxatilis male X M. chrysops female). Exposure of fish to as little as 0.10 mg fluorescein per milliliter of water for 3 minutes was sufficient to identify experimentally induced lesions, even pinpoint ulcerations. Such lesions were not visible to the naked eye but were clearly demarcated with fluorescein treatment. Examination of fish that appeared clinically normal often revealed the presence of focal ulcerations, which might have been a consequence of damage during capture, but it also might suggest that skin ulceration may be common even in "clinically normal" fish. Exposure of either nonulcerated or experimentally ulcerated hybrid striped bass to an excessively high concentration of fluorescein had no apparent effect on health or survival. Our studies suggest that fluorescein may be a highly useful tool for rapid health screening in fish populations. PMID:12450204

  7. 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

  8. Inactivation of Listeria innocua on frankfurters that contain potassium lactate and sodium diacetate by flash pasteurization.

    PubMed

    Sommers, C H; Geveke, D J; Fan, X

    2008-03-01

    Listeria monocytogenes, a psychrotrophic foodborne pathogen, is a recurring postprocess 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 incorporated into the formulation of fine emulsion sausage. Flash (steam) pasteurization (FP) has been shown to reduce levels of L. monocytogenes, and its surrogate L. innocua, on frankfurter surfaces. The ability of FP to inactivate and prevent the growth of the L. monocytogenes surrogate L. innocua in a pilot plant setting was investigated. FP treatment (1.5 s, 121 degrees C) of single layers of frankfurters that were surface-inoculated with either 5, 4, or 3 log CFU/g of L. innocua immediately before FP (1.5 s, 121 degrees C) resulted in log reductions of 1.97 (+/- 0.11), 2.03 (+/- 0.10), or 2.07 (+/- 0.14), respectively. Inoculum level had no effect on the inactivation of L. innocua. Following 8 wk of refrigerated storage (4 degrees C), L. innocua levels decreased by 0.5 log in non-FP-treated frankfurter packs, while the 2 log reduction of L. innocua was maintained for FP-treated frankfurters. FP (1.5 s, 121 degrees C) had no effect on frankfurter color or texture. Because the numbers of L. monocytogenes associated with contaminations of ready-to-eat meats are typically very low, the use of FP in combination with PL and SDA has the potential to reduce the number of frankfurter recalls and foodborne illness outbreaks. PMID:18298739

  9. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2013-01-01

    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.

  11. 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

  12. Pro-Q Emerald Glycoprotein Stain Kits

    E-print Network

    Lebendiker, Mario

    Pro-Q Emerald Glycoprotein Stain Kits The most advanced technology for staining glycoproteins steps -- fixation, oxidation and staining More sensitive than any other nonradioactive glycoprotein Kits provide the most advanced technology available for detection of glycoproteins in gels and on blots

  13. Scoring of dual fluorescein and ICG inflammatory angiographic signs for the grading of posterior segment inflammation (dual fluorescein and ICG angiographic scoring system for uveitis)

    Microsoft Academic Search

    Ilknur Tugal-Tutkun; Carl P. Herbort; Moncef Khairallah

    2010-01-01

    Purpose To propose a semiquantitative dual fluorescein angiography (FA) and indocyanine green angiography (ICGA) scoring system for\\u000a uveitis that would assist in the follow-up of disease progression and monitoring response to treatment. Methods The scoring system was based on the FA scoring systems, the standardized ICGA protocol, and schematic interpretation of ICGA\\u000a findings in posterior uveitis that have been previously

  14. The Fluorescein-derived Dye Aminophenyl Fluorescein Is a Suitable Tool to Detect Hypobromous Acid (HOBr)-producing Activity in Eosinophils*

    PubMed Central

    Flemmig, Jörg; Zschaler, Josefin; Remmler, Johannes; Arnhold, Jürgen

    2012-01-01

    The specific detection of peroxidase activity in human granulocytes is essential to elucidate their role in innate immune responses, immune regulation, and inflammatory diseases. The halogenating activity of myeloperoxidase in neutrophils can be determined by the novel fluorescent probe aminophenyl fluorescein (APF). Thereby non-fluorescent APF is oxidized by HOCl to form fluorescein. We successfully verified that APF equally detects the hypobromous acid (HOBr)-producing activity of eosinophil granulocytes. This was revealed by three different approaches. First, we investigated the conversion of non-fluorescent APF into fluorescein by HOCl and HOBr by means of fluorescence and mass spectrometry approaches. Thereby comparable chemical mechanisms were observed for both acids. Furthermore in vitro kinetic studies were used to detect the halogenating activity of myeloperoxidase and eosinophil peroxidase by using APF. Here the dye well reflected the different substrate specificities of myeloperoxidase and eosinophil peroxidase regarding chloride and bromide. Finally, peroxidase activities were successfully detected in phorbol ester-stimulated neutrophils and eosinophils using flow cytometry. Thereby inhibitory studies confirmed the peroxidase-dependent oxidation of APF. To sum up, APF is a promising tool for further evaluation of the halogenating activity of peroxidases in both neutrophils and eosinophils. PMID:22718769

  15. 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.

  16. Chromatin fluorescence after carmine staining.

    PubMed

    Stockert, J C; Llorente, A R; Del Castillo, P; Gómez, A

    1990-01-01

    After staining with dilute solutions (0.1 mg/ml in distilled water) of commercial carmine, a strong reddish orange fluorescence was observed in nuclei from cell smears and frozen and paraffin tissue sections. Optimal exciting light was 436 nm (violet-blue) or 450-490 nm (blue). Compact chromatin from interphase nuclei, mitotic and meiotic chromosomes and the kinetoplast of Trypanosoma cruzi showed the highest fluorescence, while the basophilic cytoplasm appeared weakly fluorescent. No emission was observed in cartilage matrix, mast cell granules or goblet cell mucin. This selective method could be valuable in microscopic and cytochemical studies on chromatin because the carmine fluorescence is stable and preparations can be dehydrated and mounted permanently without changes in the fluorescence pattern. PMID:2080525

  17. Phenolic acridine orange fluorescent stain for mycobacteria.

    PubMed Central

    Smithwick, R W; Bigbie, M R; Ferguson, R B; Karlix, M A; Wallis, C K

    1995-01-01

    A new fluorescence acid-fast staining method with acridine orange as the specific stain is presented. Only two reagents are required: the acridine orange-specific stain and a destaining-counterstaining reagent. Compared with auramine fluorescence acid-fast staining, there was less nonspecific staining of non-acid-fast debris which fluoresced a pale green contrasting color to provide a background in which to search for the red-to-orange fluorescing acid-fast bacilli. The results of the study indicate that the acridine orange method is superior to the auramine method in detecting acid-fast bacilli in specimen smears. PMID:8567921

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

    E-print Network

    Lebendiker, Mario

    expression comparisons Staining is compatible with mass spectrometry or microsequencing Simple staining protein can be recovered from the gel and accurately identified using MALDI-TOF mass spectrometry.3SYPRO Ruby Protein Gel Stain Advanced staining technology for 2-D gels and proteomics ® Detection

  19. IN VIVO STAINING OF ASTROCYTES Specific in vivo staining of astrocytes in the whole brain

    E-print Network

    Boyer, Edmond

    become a reference for in vivo staining of the whole astrocytes population in animal modelsIN VIVO STAINING OF ASTROCYTES 1 Specific in vivo staining of astrocytes in the whole brain after or electroencephalographic recordings in vivo. The high contrast of the staining facilitates the image processing and allows

  20. Fluoresceinated FKBP12 ligands for a high-throughput fluorescence polarization assay

    Microsoft Academic Search

    Gene M Dubowchik; Jonathan L Ditta; John J Herbst; Sagarika Bollini; Alexander Vinitsky

    2000-01-01

    Several fluoresceinated FKBP12 ligands have been prepared for a high-throughput fluorescence polarization assay. Kis for FKBP12 rotamase inhibition by these ligands range from 1.3 ?M to 32 nM, and their design is based on X-ray crystal structures of FKBP12 complexed with known immunophilin ligands.

  1. Patterns of blood flow in episcleral vessels studied by low-dose fluorescein videoangiography

    Microsoft Academic Search

    Paul A R Meyer

    1988-01-01

    The blood supply of the ocular anterior segment arises from a saggittal arterial ring composed of the long posterior ciliary arteries, the muscular and anterior ciliary arteries and perforating scleral arteries. This ring supplies coronal arterial circles within and outside the globe.Low dose anterior segment fluorescein videoangiography demonstrates arterial and venous flow, recording its characteristics and direction.Videoangiograms were performed at

  2. Experimental occlusion of the central artery of the retina. I. Ophthalmoscopic and fluorescein fundus angiographic studies

    Microsoft Academic Search

    S S Hayreh; T A Weingeist

    1980-01-01

    Transient experimental occlusion of the central artery of the retina (OCAR), lasting from 15 to 270 minutes, was produced by clamping the artery in the orbit in 63 eyes of rhesus monkeys. Ophthalmoscopic and fluorescein angiographic studies were performed before and during clamping of the artery, as well as periodically after unclamping, for periods of up to 22 weeks. The

  3. Mapping Retinal Fluorescein Leakage With Confocal Scanning Laser Fluorometry of the Human Vitreous

    Microsoft Academic Search

    Conceicao L. Lobo; Rui C. Bernardes; Fernando J. Santos; Jose G. Cunha-Vaz

    Objective: To demonstrate an objective, quantitative, and sensitive method of mapping retinal fluorescein leakage into the vitreous while simultaneously imaging the retina. Methods: A prototype Zeiss confocal scanning laser oph- thalmoscope was modified to obtain fluorometric measure- ments from 18 optical planes across the retina and cor- tical vitreous, separated from each other by 150 µm, and parallel to the

  4. Fluorescein mercuric acetate—A novel sensor for oral malodour detection

    Microsoft Academic Search

    Sujatha Jayaraman; Ritu Walia; Nethaji Alagirisamy

    2010-01-01

    Volatile sulphur compounds (VSC) like hydrogen sulphide, methyl mercaptan and dimethyl sulphide are the primary constituents of oral malodour. We have developed a fluorimetric assay, using fluorescein mercuric(II) acetate (FMA), for the quantification of VSC in mouth air. The assay is based on the quenching of fluorescence of FMA on reaction with VSC. The detection limit of the sensor is

  5. Reese/Doering/Pierini 4/2004 CELL WALL TAGGING WITH FLUORESCEIN

    E-print Network

    Doering, Tamara

    )fluorescein, hydrochloride (Molecular Probes A-1351), store at 4 °C - EDAC = 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide particles. Remove the top 100 µl to a fresh tube for use. 3) Measure EDAC to have ready for use). The final-weighed EDAC, mix, and add to cells. Incubate 5 min at RT. 6) Wash samples 2x with PBS, where each wash

  6. Exploring the optimal fluorescein dose in probe-based confocal laser endomicroscopy for colonic imaging

    PubMed Central

    Shahid, Muhammad W; Crook, Julia E; Meining, Alexander; Perchant, Aymeric; Buchner, Anna; Gomez, Victoria

    2011-01-01

    Background Probe-based confocal laser endomicroscopy (pCLE) is an emerging method for in-vivo imaging of the gastrointestinal tract and requires a contrast agent. Fluorescein is the most commonly used agent. The optimal dose of fluorescein for pCLE in colon is unknown. Objective Exploration of optimal dose of fluorescein for pCLE in colon. Design Comparative, prospective pilot trail. Setting Tertiary-care center. Patients 18 participants underwent colonoscopy without complications. Interventions pCLE videos were recorded in normal cecum, using 10% fluorescein intravenously. Main Outcome Measurements For subjective analysis, pCLE videos were scored for quality, by 2 observers, independently and blinded to fluorescein dose. For objective analysis, signal-to-noise ratios (SNR) were calculated for each video by an expert. Results 6 fluorescein doses were used, including 0.5 mL, 1 mL, 2.5 mL, 5 mL, 7.5 mL and 10 mL and each dose was used in three patients. For each dose, median image quality score was 2.5, 2.0, 3.25, 4.0, 4.0 and 3.5 by first observer and 2.0, 3.0, 4.0, 5.0, 4.0 and 4.0 by second observer, respectively. The subjective quality scores increased from 0.5 mL to 5.0 mL, with no evidence of further improved quality at 7.5 mL and 10 mL doses. SNR were not significantly different between doses but trended higher for higher doses. Limitations Small sample size. The results can not be applied to other parts of gastrointestinal tract i.e. duodenum, esophagus with different blood supply. Conclusion This preliminary study suggests that the optimal dose of fluorescein for high quality pCLE imaging in colon is approximately 5.0 mL. PMID:22586530

  7. The combined effects of temperature, pH and NaCl on growth of Debaryomyces hansenii analyzed by flow cytometry and predictive microbiology

    Microsoft Academic Search

    Birgitte Bjørn Sørensen; Mogens Jakobsen

    1997-01-01

    Flow cytometry was applied to determine growth of Debaryomyces hansenii in a laboratory medium. Viable yeasts were enumerated after staining with the fluorogenic ester fluorescein diacetate (FDA). Initial studies showed that the flow cytometric determinations correlated well with viable yeast populations determined as colony forming units (CFU) whereas the relationship between CFU and optical density was only linear over a

  8. Bacterial Succession in Glacial Forefield Soils Characterized by Community Structure, Activity and Opportunistic Growth Dynamics

    Microsoft Academic Search

    W. V. Sigler; S. Crivii; J. Zeyer

    2002-01-01

    The succession of bacterial communities inhabiting the forefield of the Dammaglacier (Switzerland) was investigated in soils ranging in successional age from 0 to 100 years since deglaciation. Overall activity per bacterial cell was estimated by the amount of fluorescein diacetate (FDA) hydrolyzed per DAPI-stained cell, and an index of \\

  9. Quantitative Spatial and Temporal Analysis of Fluorescein Angiography Dynamics in the Eye

    PubMed Central

    Hui, Flora; Nguyen, Christine T. O.; Bedggood, Phillip A.; He, Zheng; Fish, Rebecca L.; Gurrell, Rachel; Vingrys, Algis J.; Bui, Bang V.

    2014-01-01

    Purpose We describe a novel approach to analyze fluorescein angiography to investigate fluorescein flow dynamics in the rat posterior retina as well as identify abnormal areas following laser photocoagulation. Methods Experiments were undertaken in adult Long Evans rats. Using a rodent retinal camera, videos were acquired at 30 frames per second for 30 seconds following intravenous introduction of sodium fluorescein in a group of control animals (n?=?14). Videos were image registered and analyzed using principle components analysis across all pixels in the field. This returns fluorescence intensity profiles from which, the half-rise (time to 50% brightness), half-fall (time for 50% decay) back to an offset (plateau level of fluorescence). We applied this analysis to video fluorescein angiography data collected 30 minutes following laser photocoagulation in a separate group of rats (n?=?7). Results Pixel-by-pixel analysis of video angiography clearly delineates differences in the temporal profiles of arteries, veins and capillaries in the posterior retina. We find no difference in half-rise, half-fall or offset amongst the four quadrants (inferior, nasal, superior, temporal). We also found little difference with eccentricity. By expressing the parameters at each pixel as a function of the number of standard deviation from the average of the entire field, we could clearly identify the spatial extent of the laser injury. Conclusions This simple registration and analysis provides a way to monitor the size of vascular injury, to highlight areas of subtle vascular leakage and to quantify vascular dynamics not possible using current fluorescein angiography approaches. This can be applied in both laboratory and clinical settings for in vivo dynamic fluorescent imaging of vasculature. PMID:25365578

  10. [Thyroid papillary cancer using TPO staining].

    PubMed

    Nygaard, Birte; Frisch, Thomas; Kiss, Katalin

    2008-04-28

    Immunostaining for TPO (MoAb47) has been used to predict the risk of thyroid cancer in thyroid adenomas without uptake in thyroid 99m pertechnetate scintigraphy. This case describes a 16-year-old girl with thyroid papillary cancer staining 95% positive using TPO staining. The case indicates that TPO staining can not be used as the only parameter in the evaluation of the risk of cancer in the thyroid gland. PMID:18454932

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. 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...

  16. Systemic lipopolysaccharide compromises the blood-labyrinth barrier and increases entry of serum fluorescein into the perilymph.

    PubMed

    Hirose, Keiko; Hartsock, Jared J; Johnson, Shane; Santi, Peter; Salt, Alec N

    2014-10-01

    The blood vessels that supply the inner ear form a barrier between the blood and the inner ear fluids to control the exchange of solutes, protein, and water. This barrier, called the blood-labyrinth barrier (BLB) is analogous to the blood-brain barrier (BBB), which plays a critical role in limiting the entry of inflammatory and infectious agents into the central nervous system. We have developed an in vivo method to assess the functional integrity of the BLB by injecting sodium fluorescein into the systemic circulation of mice and measuring the amount of fluorescein that enters perilymph in live animals. In these experiments, perilymph was collected from control and experimental mice in sequential samples taken from the posterior semicircular canal approximately 30 min after systemic fluorescein administration. Perilymph fluorescein concentrations in control mice were compared with perilymph fluorescein concentrations after lipopolysaccharide (LPS) treatment (1 mg/kg IP daily for 2 days). The concentration of perilymphatic fluorescein, normalized to serum fluorescein, was significantly higher in LPS-treated mice compared to controls. In order to assess the contributions of perilymph and endolymph in our inner ear fluid samples, sodium ion concentration of the inner ear fluid was measured using ion-selective electrodes. The sampled fluid from the posterior semicircular canal demonstrated an average sodium concentration of 145 mM, consistent with perilymph. These experiments establish a novel technique to assess the functional integrity of the BLB using quantitative methods and to provide a comparison of the BLB to the BBB. PMID:24952083

  17. A NEGATIVE STAIN FOR ELECTRON MICROSCOPIC TOMOGRAPHY

    PubMed Central

    Fera, Andrea; Farrington, Jane E.; Zimmerberg, Joshua; Reese, Thomas S.

    2013-01-01

    Negative staining can provide detailed, two-dimensional images of biological structures, but combining tomography with negative staining can provide three-dimensional images. Basic requirements for a negative stain for tomography are that the density and atomic number of the stain are optimal, and that the stain is not degraded with the intensive electron dose needed to collect a full set of tomographic images. A commercially available, tungsten-based stain, methylamine tungstate, appears to satisfy these prerequisites. Tomograms derived from multiple projections of EM images of the same structure yielded detailed images of single proteins on the surface of influenza A virus. Comparison of these images with published results from other methods served to evaluate this negative stain tomography. Images of surface renderings of the virus are a good fit to images derived from cryomicroscopy, as well as to the shapes of crystallized surface proteins. Thus, negative stain tomography provides realistic and detailed images of individual molecules in their normal setting on the surface of influenza A virus. PMID:22364718

  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. 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

  20. 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...

  1. Viability of Listeria Monocytogenes on commercially-prepared uncured turkey breast, formulated with and without potassium levulinate, potassium diacetate and potassium propionate, during extended refrigerated storage

    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...

  2. Separating the isomers—Efficient synthesis of the N-hydroxysuccinimide esters of 5 and 6-carboxyfluorescein diacetate and 5 and 6-carboxyrhodamine B

    PubMed Central

    Brunet, Aurélie; Aslam, Tashfeen; Bradley, Mark

    2014-01-01

    Diacetate protection of 5 and 6-carboxyfluorescein followed by synthesis of the N-hydroxysuccinimide esters allowed ready separation of the two isomers on a multi-gram scale. The 5 and 6-carboxyrhodamine B N-hydroxysuccinimide esters were also readily synthesised and separated. PMID:24856065

  3. 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...

  4. 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...

  5. 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, ...

  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. 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...

  8. Efficacy of a food grade blend of lactate-diacetate-propionate as ingredients to control Listeria monocytogenes on commericially produced frankfurters

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Further research is warranted to evaluate different levels/types of food grade antimicrobials to control Listeria monocytogenes (Lm) on RTE meats. Purpose: Determine viability of Lm on frankfurters formulated with a blend of lactate-diacetate-propionate (0, 0.5, 0.75, or 1.0%) and then...

  9. Fluorescein-Trp 25-Exendin-4, a Biologically Active Fluorescent Probe for the Human GLP-1 Receptor

    Microsoft Academic Search

    G. G. Chicchi; M. A. Cascieri; M. P. Graziano; T. Calahan; M. R. Tota

    1997-01-01

    Chicchi, G. G., M. A. Cascieri, M. P. Graziano, T. Calahan and M. R. Tota. Fluorescein-Trp25-exendin-4, a biologically active fluorescent probe for the human Glp-1 receptor. Peptides 18(2) 319–321, 1997.—Exendin-4, a reptilian GLP-1 analogue, has been fluorescently labeled by covalently linking a fluorescein moiety onto the Trp residue yielding fluorescein-Trp25-exendin-4 (FLEX). FLEX is equipotent to GLP-1(7–36)-amide and exendin-4 as an

  10. Influence of ionization states of antigen on anti-fluorescein antibodies

    NASA Astrophysics Data System (ADS)

    Fukunishi, Hiroaki

    2012-10-01

    Ratios of anion and di-anion states of fluorescein (FLU(-1) and FLU(-2)) are 21.2% and 78.8%, respectively, in the neutral pH. We investigated the influence of ionization states of antigen on anti-fluorescein antibodies. For this purpose, steered molecular dynamics (SMD) simulations were performed. Potential of mean forces (PMF) based on Jarzynski equality showed that wild-type (4-4-20) more strongly binds to FLU(-1) than FLU(-2), whereas its femtomolar-affinity mutant (4M5.3) more strongly binds to FLU(-2) than FLU(-1). It was speculated that the environment or the process of in vivo antibody production had been different from those of the protein engineering.

  11. Inhibition of housefly oxidative detoxication by phthaleins, fluoresceins and related compounds.

    PubMed

    Jordan, T W; Smith, J N

    1981-01-01

    1. Phenolphthalein, halogenated fluoresceins, and other triphenylmethane and diphenylmethane derivatives inhibited biphenyl hydroxylation, aldrin epoxidation and several O-dealkylations in insect abdomen homogenates. Phenolphthalein and eosin (50 muM) were 2-3 times more effective than SKF 525-A and piperonyl butoxide (50 muM) as inhibitors of biphenyl hydroxylation in vitro. 2. The phthaleins, Aurin and Aluminon, inhibited both epoxidation and hydroxylation to similar extents, but fluoresceins, Rhodamine B, Malachite Green, and basic diphenylmethane derivatives preferentially inhibited hydroxylation. 3. Tetrabromophenolphthalein ethyl ester and bis-(N-dimethyl-4-aminophenyl-methane inhibited biphenyl hydroxylation in vivo. Bis-(N-dimethyl-4-aminophenyl) methane synergized the toxic effects of 1-naphthyl N-methylcarbamate in live houseflies. PMID:7222726

  12. Studies on Dental Stains Induced by Antibacterial Agents and Rational Approaches for Bleaching Dental Stains

    Microsoft Academic Search

    S. A. Nathoo; A. Gaffar

    1995-01-01

    Extrinsic stain resides in the dental pellicle and can be caused by introduction of chromogenic materials or therapeutic agents into the oral cavity. In contrast, intrinsic tooth stain is found within the tooth structure and can be caused by a variety of agents, including hematological and developmental abnormalities and drugs such as tetracycline. The mechanisms of extrinsic stain formation differ

  13. Preparation and Characterization of Self-Assembled Aminated Agarose Loaded with Fluorescein Isothiocyanate Nanoparticles

    Microsoft Academic Search

    Lingmin Zhang; Xiaohui Peng; Shunqing Tang

    2011-01-01

    A number of organic fluorescent molecules self-assembling into nanoparticles can significantly promote their fluorescent effects. In this paper, a novel fluorescein isothiocyanate labeling aminated agarose (FITC-AA) is prepared and tested as an effective fluorescent labeling agent. FITC-AA self-assembled into nanoparticles with a diameter between 104.54 ? 164.94 nm at below 37°C by the gelling effect of agarose. FITC-AA nanoparticles could

  14. Flow microfluorometric system for screening gynecologic cytology specimens using propidium iodide-fluorescein isothiocyanate.

    PubMed

    Fowlkes, B J; Herman, C J; Cassidy, M

    1976-01-01

    Seventy cervical cytology specimens have been screened by a xero resolution flow analyzer-sorter using propidium iodide and fluorescein isothiocyanate as fluorochromes for nucleus and cytoplasm, respectively. This system shows a 1% sensitivity for detection of abnormal cells using only crude visual data analysis. Screening of clinical specimens was performed on the instrument with a 5.8% false negative rate and a 11.8% false positive rate by comparison with routine visual cytologic evaluation of the same samples. PMID:1254927

  15. Multiple and sensitive fluorescence in situ hybridization with rhodamine-, fluorescein-, and coumarin-labeled DNAs

    Microsoft Academic Search

    J. Wiegant; C. C. Wiesmeijer; J. M. N. Hoovers; E. Schuuring; A. d’Azzo; J. Vrolijk; H. J. Tanke; A. K. Raap

    1993-01-01

    We have tested the use of several newly developed red, green, and blue fluorescent dUTPs in direct, multiple, and sensitive fluorescence in situ hybridization procedures. Among the ones tested, the tetramethylrhodamine-dUTP proved to give the best sensitivity; using conventional epifiuorescence microscopy, cosmids could be visualized with a hybridization efficiency of 90%. Fluorescein-dUTP permitted visual cosmid detection with 50% efficiency, and,

  16. Protolytic equilibria of fluorescein halo derivatives in aqueous-organic systems

    Microsoft Academic Search

    N. O. Mchedlov-Petrosyan; V. I. Kukhtik; S. I. Egorova

    2006-01-01

    The dissociation constants of fluorescein halo derivatives containing substituents in the xanthene ring and\\/or in the phthalic\\u000a acid moiety were determined in 91.4 wt % aqueous ethanol. The tautomerism of the dyes was inferred from the electronic absorption\\u000a spectra of ions and molecules in this solvent and of the dichloromethane and chloroform extracts of associates of the anions\\u000a with tetra-n-butylammonium

  17. Spectral Optical Coherence Tomography vs. fluorescein pattern for rigid gas-permeable lens fit

    PubMed Central

    Piotrowiak, Ilona; Ka?u?ny, Bart?omiej J.; Danek, Beata; Chwi?dacz, Adam; Sikorski, Bartosz ?.; Malukiewicz, Gra?yna

    2014-01-01

    Background This study aimed to evaluate anterior segment spectral optical coherence tomography (AS SOCT) for assessing the lens-to-cornea fit of rigid gas-permeable (RGP) lenses. The results were verified with the fluorescein pattern method, considered the criterion standard for RGP lens alignment evaluations. Material/Methods Twenty-six eyes of 14 patients were enrolled in the study. Initial base curve radius (BCR) of each RGP lens was determined on the basis of keratometry readings. The fluorescein pattern and AS SOCT tomograms were evaluated, starting with an alignment fit, and subsequently, with BCR reductions in increments of 0.1 mm, up to 3 consecutive changes. AS SOCT examination was performed with the use of RTVue (Optovue, California, USA). Results The average BCR for alignment fits, defined according to the fluorescein pattern, was 7.8 mm (SD=0.26). Repeatability of the measurements was 18.2%. BCR reductions of 0.1, 0.2, and 0.3 mm resulted in average apical clearances detected with AS SOCT of 12.38 (SD=9.91, p<0.05), 28.79 (SD=15.39, p<0.05), and 33.25 (SD=10.60, p>0.05), respectively. Conclusions BCR steepening of 0.1 mm or more led to measurable changes in lens-to-cornea fits. Although AS SOCT represents a new method of assessing lens-to-cornea fit, apical clearance detection with current commercial technology showed lower sensitivity than the fluorescein pattern assessment. PMID:24995686

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

    PubMed

    Zhang, Li; Zhang, Xianhong

    2014-12-10

    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 Cu(2+). In the presence of Cu(2+) the BMSFH exhibits a rapid color change from colorless to yellow together with an obvious new band appeared at 502nm 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 Cu(2+) in a 1:1 stoichiometry and this binding to Cu(2+) is reversible, as indicated by the bleaching of the color when the Cu(2+) is extracted with EDTA. Experimental results indicate that the BMSFH can provide a rapid, selective and sensitive response to Cu(2+) with a linear dynamic range 3.0-330?mol/L and can be used as a potential Cu(2+) colorimetric probe in aqueous solution. PMID:24929315

  19. 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.

  20. Thiol reactive probe based on fluorescence resonance energy transfer between fluorescein and Au nanoparticles.

    PubMed

    Qi, Li; Song, Juan; Wu, Fang-Ying; Wan, Yi-Qun

    2014-01-01

    Sensitive and selective fluorescent probe of thiols with lower limit of detection based on fluorescence resonance energy transfer (FRET) between fluorescein and Au nanoparticles (AuNPs) is presented. The fluorescein-AuNPs complex emits weak fluorescence. Upon chemically binding to organosulfur compound that contains a carbon-bonded sulfhydryl (-C-SH or R-SH) thiols, a stable enhancement of fluorescence is observed due to the competitive binding on AuNPs between thiols and fluorescein. The magnitude of fluorescence enhancement is linearly proportional to the logarithm of the thiols concentration. We use cysteine as an example to show how this useful analytical assay works selectively, which is closely nonresponsive to 20 other amino acids even though they are in solution at a concentration 10 times greater than the thiols. The detection limit for cysteine is 7.27 × 10-9 mol L-1. The possible mechanism of this assay is discussed in details. The proposed method was successfully applied for the determination of Cys in urine. PMID:24664329

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

    PubMed

    Sommers, C H; Cooke, P H; Fan, X; Sites, J E

    2009-04-01

    Listeria monocytogenes, a psychrotrophic foodborne pathogen, is an occasional postprocess contaminant on ready-to-eat meat (RTE) products including frankfurters. Ultraviolet C light (UVC) is an FDA-approved technology for the decontamination of food surfaces. In this study, the ability of UVC to inactivate L. monocytogenes on frankfurters that contained potassium lactate (PL) and sodium diacetate (SDA), either before or after packaging, was investigated. UVC irradiation of frankfurters that were surface-inoculated with L. monocytogenes resulted in a 1.31, 1.49, and 1.93 log reduction at doses of 1, 2, and 4 J/cm(2), respectively. UVC treatment had no effect on frankfurter color or texture at UVC doses up to 4 J/cm(2). Frankfurter meat treated with UVC doses up to 16 J/cm(2) did not increase mutagenesis in bacterial or human cells, either with or without exogenous metabolic activation. UVC treatment of single-layer frankfurter packs at a dose of 2 J/cm(2) resulted in a 0.97 (+/- 0.14) log reduction of L. monocytogenes. Following 8 wk of refrigerated storage L. monocytogenes levels decreased by only 0.65 log in non-UVC-treated frankfurter packs compared with 2.5 log in the UVC-treated packs. Because the numbers of L. monocytogenes associated with contaminations of ready-to-eat meats are typically very low, the use of UVC in combination with potassium lactate and sodium diacetate has the potential to reduce the number of frankfurter recalls and foodborne illness outbreaks. PMID:19397726

  2. 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

  3. 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...

  4. Gram staining apparatus for space station applications

    NASA Technical Reports Server (NTRS)

    Molina, T. C.; Brown, H. D.; Irbe, R. M.; Pierson, D. L.

    1990-01-01

    A self-contained, portable Gram staining apparatus (GSA) has been developed for use in the microgravity environment on board the Space Station Freedom. Accuracy and reproducibility of this apparatus compared with the conventional Gram staining method were evaluated by using gram-negative and gram-positive controls and different species of bacteria grown in pure cultures. A subsequent study was designed to assess the performance of the GSA with actual specimens. A set of 60 human and environmental specimens was evaluated with the GSA and the conventional Gram staining procedure. Data obtained from these studies indicated that the GSA will provide the Gram staining capability needed for the microgravity environment of space.

  5. Gram staining apparatus for space station applications.

    PubMed Central

    Molina, T C; Brown, H D; Irbe, R M; Pierson, D L

    1990-01-01

    A self-contained, portable Gram staining apparatus (GSA) has been developed for use in the microgravity environment on board the Space Station Freedom. Accuracy and reproducibility of this apparatus compared with the conventional Gram staining method were evaluated by using gram-negative and gram-positive controls and different species of bacteria grown in pure cultures. A subsequent study was designed to assess the performance of the GSA with actual specimens. A set of 60 human and environmental specimens was evaluated with the GSA and the conventional Gram staining procedure. Data obtained from these studies indicated that the GSA will provide the Gram staining capability needed for the microgravity environment of space. Images PMID:1690529

  6. A ‘Magnetic’ Gram Stain for Bacterial Detection

    PubMed Central

    Budin, Ghyslain; Chung, Hyun Jung; Lee, Hakho

    2012-01-01

    Magnetic stain. Bacteria are often classified into Gram-positive and Gram-negative strains by their visual staining properties using crystal violet (CV), a triarylmethane dye. Here we show, that bioorthogonal modification of crystal violet with transcyclooctene (TCO) can be used to render Gram-positive bacteria magnetic with magneto-nanoparticles-Tetrazine (MNP-Tz). This allows for class specific automated magnetic detection, magnetic separation or other magnetic manipulations. PMID:22744868

  7. New Grocott Stain without Using Chromic Acid

    PubMed Central

    Shiogama, Kazuya; Kitazawa, Kayo; Mizutani, Yasuyoshi; Onouchi, Takanori; Inada, Ken-ichi; Tsutsumi, Yutaka

    2015-01-01

    We established a new “ecological” Grocott stain for demonstrating fungi, based upon a 4R principle of refusal, reduction, reuse, and recycle of waste management. Conventional Grocott stain employs environmentally harsh 5% chromic acid for oxidization. Initially, we succeeded in reducing the concentration of chromic acid from 5% to 1% by incubating the solution at 60°C and using five-fold diluted chromic acid solution at which point it was reusable. Eventually, we reached the refusal level where 1% periodic acid oxidization was efficient enough, when combined with preheating of sections in the electric jar, microwave oven, or pressure pan. For convenience sake, we recommend pressure pan heating in tap water for 10 min. Stainability of fungi in candidiasis and aspergillosis was comparable with conventional Grocott stain, while Mucor hyphae showed enhanced staining. The modified sequence was further applicable to detecting a variety of mycotic pathogens in paraffin sections. Our environmentally-friendly Grocott stain also has the advantage of avoiding risk of human exposure to hexavalent chromium solution in the histopathology laboratory. The simple stain sequence is can be easily applied worldwide. PMID:25861133

  8. The Luna stain, an improved selective stain for detection of microsporidian spores in histologic sections

    PubMed Central

    Peterson, Tracy S.; Spitsbergen, Jan M.; Feist, Stephen W.; Kent, Michael L.

    2014-01-01

    Microsporidia in histologic sections are most often diagnosed by observing spores in host tissues. Spores are easy to identify if they occur in large aggregates or xenomas when sections are stained with hematoxylin and eosin (H&E). However, individual spores are not frequently detected in host tissues with conventional H&E staining, particularly if spores are scattered within the tissues, areas of inflammation or small spores in nuclei (i.e., Nucleospora salmonis). Hence, a variety of selective stains that enhance visualization of spores are recommended. We discovered that the Luna stain, used to highlight eosinophils, red blood cells and chitin in arthropods and other invertebrates, also stains spores of Pseudoloma neurophilia. We compared this stain to the Gram, Fite’s acid fast, Giemsa, and H&E stains on eight aquatic microsporidian organisms that were readily available in our two laboratories: Loma salmonae, Glugea anomala, Pseudoloma neurophilia, Pleistophora hyphessobryconis, Pleistophora vermiformis, Glugea sp., Steinhausia mytilovum and an unidentified microsporidian from E. sinensis, UK. Based on tinctorial properties and background staining, the Luna stain performed better for detection of 6 of the 8 microsporidia. Gram stain was superior for the two microsporidia from invertebrates, Steinhausia mytilovum and the unidentified microsporidian from E. sinensis. PMID:21848126

  9. Luna stain, an improved selective stain for detection of microsporidian spores in histologic sections.

    PubMed

    Peterson, Tracy S; Spitsbergen, Jan M; Feist, Stephen W; Kent, Michael L

    2011-06-16

    Microsporidia in histologic sections are most often diagnosed by observing spores in host tissues. Spores are easy to identify if they occur in large aggregates or xenomas when sections are stained with hematoxylin and eosin (H&E). However, individual spores are not frequently detected in host tissues with conventional H&E staining, particularly if spores are scattered within the tissues, areas of inflammation, or small spores in nuclei (i.e. Nucleospora salmonis). Hence, a variety of selective stains that enhance visualization of spores is recommended. We discovered that the Luna stain, used to highlight eosinophils, red blood cells, and chitin in arthropods and other invertebrates, also stains spores of Pseudoloma neurophilia. We compared this stain to the Gram, Fite's acid fast, Giemsa, and H&E stains on 8 aquatic microsporidian organisms that were readily available in our 2 laboratories: Loma salmonae, Glugea anomala, Pseudoloma neurophilia, Pleistophora hyphessobryconis, Pleistophora vermiformis, Glugea sp., Steinhausia mytilovum, and an unidentified microsporidian from UK mitten crabs Eriocheir sinensis. Based on tinctorial properties and background staining, the Luna stain performed better for detection of 6 of the 8 microsporidia. Gram stain was superior for the 2 microsporidia from invertebrates: S. mytilovum and the unidentified microsporidian from E. sinensis. PMID:21848126

  10. Classification of Human Retinal Microaneurysms Using Adaptive Optics Scanning Light Ophthalmoscope Fluorescein Angiography

    PubMed Central

    Dubow, Michael; Pinhas, Alexander; Shah, Nishit; Cooper, Robert F.; Gan, Alexander; Gentile, Ronald C.; Hendrix, Vernon; Sulai, Yusufu N.; Carroll, Joseph; Chui, Toco Y. P.; Walsh, Joseph B.; Weitz, Rishard; Dubra, Alfredo; Rosen, Richard B.

    2014-01-01

    Purpose. Microaneurysms (MAs) are considered a hallmark of retinal vascular disease, yet what little is known about them is mostly based upon histology, not clinical observation. Here, we use the recently developed adaptive optics scanning light ophthalmoscope (AOSLO) fluorescein angiography (FA) to image human MAs in vivo and to expand on previously described MA morphologic classification schemes. Methods. Patients with vascular retinopathies (diabetic, hypertensive, and branch and central retinal vein occlusion) were imaged with reflectance AOSLO and AOSLO FA. Ninety-three MAs, from 14 eyes, were imaged and classified according to appearance into six morphologic groups: focal bulge, saccular, fusiform, mixed, pedunculated, and irregular. The MA perimeter, area, and feret maximum and minimum were correlated to morphology and retinal pathology. Select MAs were imaged longitudinally in two eyes. Results. Adaptive optics scanning light ophthalmoscope fluorescein angiography imaging revealed microscopic features of MAs not appreciated on conventional images. Saccular MAs were most prevalent (47%). No association was found between the type of retinal pathology and MA morphology (P = 0.44). Pedunculated and irregular MAs were among the largest MAs with average areas of 4188 and 4116 ?m2, respectively. Focal hypofluorescent regions were noted in 30% of MAs and were more likely to be associated with larger MAs (3086 vs. 1448 ?m2, P = 0.0001). Conclusions. Retinal MAs can be classified in vivo into six different morphologic types, according to the geometry of their two-dimensional (2D) en face view. Adaptive optics scanning light ophthalmoscope fluorescein angiography imaging of MAs offers the possibility of studying microvascular change on a histologic scale, which may help our understanding of disease progression and treatment response. PMID:24425852

  11. Fluorescence microplate-based assay for tumor necrosis factor activity using SYTOX Green stain.

    PubMed

    Jones, L J; Singer, V L

    2001-06-01

    We have developed a simple, sensitive, fluorescence microplate-based assay for tumor necrosis factor (TNF) biological activity. The assay employs SYTOX Green nucleic acid stain to detect TNF-induced cell necrosis in actinomycin D sensitized cultured cell lines. SYTOX Green stain is a cationic unsymmetrical cyanine dye that is excluded from live cells but can readily penetrate cells with compromised cell membranes. Upon binding to cellular nucleic acids, the dye exhibits a large enhancement in fluorescence, which is monitored at fluorescein wavelengths. We detected 2.5 pg/mL and quantitated 25-500 pg/mL recombinant murine (rm) and recombinant human (rh) TNF-alpha, using mouse fibroblast-derived WEHI 164, WEHI 13var, and L929 cell lines. The procedure can also be used to detect agents that modulate TNF activity. We demonstrated complete inhibition of rhTNF-alpha using monoclonal anti-human TNF-alpha antibody and determined that approximately 20 ng/mL antibody was sufficient to neutralize 50% of the biological activity of 250 pg/mL rhTNF-alpha in these cell lines. Reagents are added in a single step, followed by a 6- to 8-h incubation period, during which the cytokine exhibits its effects. There are no wash steps, and the assay is readily amenable to automation and high-throughput screening procedures. PMID:11373072

  12. 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.

  13. 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.

  14. 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

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

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

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...2012-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food...HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

  1. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...2013-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food...HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

  2. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...2014-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food...HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

  3. Original article Quantitation of histochemical staining of salivary

    E-print Network

    Paris-Sud XI, Université de

    alone. Quantitative assessment of normal salivary gland histochemistry allows com- parison with stainingOriginal article Quantitation of histochemical staining of salivary gland mucin using image were employed to complement subjective assessment of patterns of mucin staining (by periodic acid

  4. 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...

  5. Fluorescein isothiocynate-dextran uptake by chinese hamster ovary cells in a 1.5 MHz ultrasonic standing wave in the presence of contrast agent.

    PubMed

    Khanna, Sanjay; Hudson, Benjamin; Pepper, Christopher J; Amso, Nazar N; Coakley, W Terence

    2006-02-01

    Uptake of fluorescein isothiocynate-dextran (FITC-dextran) by Chinese hamster ovary cells was studied after exposure to ultrasonic standing wave (USW) in presence of Optison, an ultrasound contrast agent. Confluent Chinese hamster ovary cells were harvested and suspended in phosphate-buffered saline + 0.1% bovine serum albumin containing FITC-dextran (10, 40, and 500 kDa) at 10 microM final concentration. The suspension was seeded with contrast agent (75 microL/mL) and exposed to a 1.5 MHz USW system at acoustic pressures ranging from 0.98 to 4.2 MPa. Macromolecular uptake was assessed by fluorescent microscopy and quantified by flow cytometry 10 min after exposure. FITC-dextran positive cells, as assessed by flow cytometry, were 1 +/- 0.05% and 2.58 +/- 0.27% for acoustic pressures of 1.96 and 4.2 MPa, respectively (p = 0.006). Fluorescent microscopy indicated a degree of macromolecular loading at 0.98 MPa with 46% of peripherally FITC-dextran- and/or propidium iodide-stained cells coincident with the appearance of significant frequency (f0/2 and 2 f0) emission signals. At higher pressures, high macromolecular loading with 6% peripherally stained cells at 1.96 MPa was associated with lower order emission signals and white noise. The study conclusively demonstrates macromolecular loading in an USW, a significantly higher macromolecular loading at higher pressures and indicates potential of emission signals for a feedback loop to control the acoustic power outputs and fine-tune the biologic effects associated with sonoporation. PMID:16464674

  6. Laser treatment of port-wine stains

    PubMed Central

    Brightman, Lori A; Geronemus, Roy G; Reddy, Kavitha K

    2015-01-01

    Port-wine stains are a type of capillary malformation affecting 0.3% to 0.5% of the population. Port-wine stains present at birth as pink to erythematous patches on the skin and/or mucosa. Without treatment, the patches typically darken with age and may eventually develop nodular thickening or associated pyogenic granuloma. Laser and light treatments provide improvement through selective destruction of vasculature. A variety of vascular-selective lasers may be employed, with the pulsed dye laser being the most common and well studied. Early treatment produces more optimal results. Advances in imaging and laser treatment technologies demonstrate potential to further improve clinical outcomes. PMID:25624768

  7. 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.

  8. 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.

  9. Wound dressings based on chitosans and hyaluronic acid for the release of chlorhexidine diacetate in skin ulcer therapy.

    PubMed

    Rossi, Silvia; Marciello, Marzia; Sandri, Giuseppina; Ferrari, Franca; Bonferoni, Maria Cristina; Papetti, Adele; Caramella, Carla; Dacarro, Casare; Grisoli, Pietro

    2007-01-01

    In the present work wound dressings, based on chitosan hydrochloride (HCS), 5-methyl-pyrrolidinone chitosan (MPC), and their mixtures with an anionic polymer, hyaluronic acid (HA), were prepared by freeze-drying. Chlorhexidine diacetate (CX) was used as an antimicrobic drug. The mechanical properties of the wound dressings were investigated. In particular, the wound dressings were subjected to dynamic hydration measurements to evaluate their capability to absorb wound exudate and to rheological analysis to investigate their resistance to mechanical stresses on hydration. The wound dressings were also characterized for drug release properties. The antioxidant and antimicrobial activities of medicated and non-medicated wound dressings were also investigated. All the wound dressings are characterized by mechanical resistance suitable for skin application. The addition of hyaluronic acid to chitosans leads to a reduction in wound dressing hydration properties and a modulation of drug release. The wound dressing based on MPC is characterized by the highest elastic properties and by the best scavenger activity. Antimicrobial activity against bacteria and C. albicans is shown by the dressing based on chitosan also in absence of chlorhexidine. PMID:17763146

  10. Synthesis and Characterization of Pt(IV) Fluorescein Conjugates to Investigate Pt(IV) Intracellular Transformations

    PubMed Central

    Song, Ying; Suntharalingam, Kogularamanan; Yeung, Jessica S.; Royzen, Maksim; Lippard, Stephen J.

    2013-01-01

    Pt(IV) anticancer compounds typically operate as prodrugs that are reduced in the hypoxic environment of cancer cells, losing two axial ligands in the process to generate active Pt(II) species. Here we report the synthesis of two fluorescent Pt(IV) prodrugs of cisplatin in order to image and evaluate the Pt(IV) reduction process in simulated and real biological environments. Treatment of the complexes dissolved in PBS buffer with reducing agents typically encountered in cells, glutathione or ascorbate, afforded a 3- to 5-fold fluorescence turn-on owing to reduction and loss of their fluorescein-based axial ligands, which are quenched when bound to platinum. Both Pt(IV) conjugates displayed moderate cytotoxicity against human cancer cell lines, with IC50 values higher than that of cisplatin. Immunoblotting and DNA flow cytometry analyses of one of the complexes, Pt(IV)FL2, revealed that it damages DNA, causing cell cycle arrest in S or G2/M depending on exposure time, with ultimately triggering of apoptotic cell death. Fluorescence microscopic studies prove that Pt(IV)FL2 enters cells intact and undergoes reduction intracellularly. The results are best interpreted in terms of a model in which the axial fluorescein ligands are expelled through lysosomes, with the platinum(II) moiety generated in the process binding to genomic DNA, which results in cell death. PMID:23957697

  11. Original article Blue-stain fungi associated

    E-print Network

    Paris-Sud XI, Université de

    , Hormonema dematioides, Leptographium wingfieldii and Ophiostoma minus. The latter 2 species were most active dematioides, Leptographium wingfieldii, et Ophiostoma minus. Les 2 der- nières nommées se sont avérées les (L) and the blue-stain fungus Ophiostoma polonicum Siem (Horntvedt et al, 1983; Christiansen

  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. 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.

  14. The Science and Application of Hematoxylin and Eosin Staining

    E-print Network

    Chisholm, Rex L.

    The Science and Application of Hematoxylin and Eosin Staining Skip Brown, M.Div, HT (ASCP) Robert H and Staining Systems Living up to Life #12;Routine Staining (Hematoxylin & Eosin) Nuclear detail/definition - Hematoxylin Contrasting counterstain - Eosin #12;#12;The Hematoxylin and Eosin Stain · Long history of use

  15. A Chemical Stain for Identifying Arsenic-Treated Wood

    E-print Network

    Florida, University of

    A Chemical Stain for Identifying Arsenic-Treated Wood (FINAL) Submitted June 23, 2006 Amy Omae.2 Motivation 4 I.3 Objectives 5 CHAPTER II, DEVELOPMENT OF A CHEMICAL STAIN FOR IDENTIFYING ARSENIC-TREATED WOOD II.1 Applying Phosphate Stains to Arsenate Stains 7 II.2 A Potential Arsenic-Test Kit 14 II.3

  16. Comparison of corneal sensitivity, tear function and corneal staining following laser in situ keratomileusis with two femtosecond laser platforms

    PubMed Central

    Petznick, Andrea; Chew, Annabel; Hall, Reece C; Chan, Cordelia ML; Rosman, Mohamad; Tan, Donald; Tong, Louis; Mehta, Jodhbir S

    2013-01-01

    Purpose To evaluate longitudinal changes in corneal sensitivity, tear function, and corneal staining in patients who underwent laser in situ keratomileusis (LASIK) using two different femtosecond lasers. Methods In a prospective, randomized clinical trial, contralateral eyes of 45 patients underwent flap creation by either VisuMax or IntraLase™ femtosecond laser. Corneal sensitivity, tear break up time (TBUT), Schirmer’s test, and corneal fluorescein staining were assessed preoperatively and at 1 week, 1 month, and 3 months postoperatively. Results There were no statistical differences in any clinical outcome measure between the two femtosecond lasers (P > 0.05), although there was a trend towards slightly lower reductions for corneal sensitivity and TBUT in VisuMax-operated eyes. Overall, corneal sensitivity was significantly reduced at 1 week (P < 0.05), 1 month (P < 0 .001), and 3 months (P < 0.001) postoperatively. A significantly greater reduction of corneal sensitivity was noted in eyes with a myopic spherical equivalent of ?6.00 diopters (D) to ?11.25 D as compared with eyes that had a relatively lower level of myopia of less than ?6.00 D (P < 0.001). TBUT and Schirmer’s test values were significantly diminished at 1 week postoperatively (P < 0.04). Overall, corneal staining was significantly increased at 1 week postoperatively (P < 0.001). The level of myopia did not significantly affect postoperative changes in TBUT, Schirmer’s test values, or corneal staining (P > 0.05). Conclusion This study showed that changes in corneal sensitivity, tear function, and corneal staining were statistically similar in LASIK using VisuMax and IntraLase femtosecond lasers for flap creation. However, the trend towards faster recovery of corneal sensitivity and TBUT observed in VisuMax-operated eyes may be attributable to improved technical specifications. PMID:23576858

  17. 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.

  18. Structure of complexes formed by dissolution of palladium diacetate in methanol and chloroform. In situ NMR study.

    PubMed

    Nosova, Valentina M; Ustynyuk, Yuri A; Bruk, Lev G; Temkin, Oleg N; Kisin, Alexander V; Storozhenko, Pavel A

    2011-10-01

    The behavior of palladium diacetate cyclic trimer [Pd(OAc)(2)](3) (1) upon its dissolution in methanol and wet chloroform was studied by (1)H and (13)C NMR including 2D-HSQC and 2D-DOSY techniques. Upon dissolution, trimer 1 reacts with methanol and is completely transformed first into the methoxo complex Pd(3)(?-OMe)(OAc)(5) (2), which already at -18 °C undergoes a slow exchange of second bridging acetate ligand between the same palladium atoms to form the symmetric dimethoxo complex Pd(3)(?-OMe)(2)(OAc)(4), the maximum relative concentration of which reaches 20-30 mol % of initial loading trimer 1. Along with the dimethoxo complex, both soluble and insoluble polynuclear palladium clusters are gradually formed at -18 °C, and their total amount reaches up to 60% of the starting Pd(2+) loading. The increase of temperature to 27 °C results in the reduction of palladium(II) to Pd metal by methanol, which is oxidized and transformed into formaldehyde hemiacetal and methyl formate. Upon dissolution in wet chloroform, trimer 1 is reversibly hydrolyzed to the hydroxo complex Pd(3)(?-OH)(OAc)(5) (10) in ratio 1/10 ? 3/1. The temperature decrease and addition of acetic acid shift the equilibrium in this system toward trimer 1, and addition of water shifts it in the opposite direction. Addition of methanol to the equilibrium mixture of 1 and 10 results in the fast exchange of bridging acetate in trimer 1 by the ?-OMe group. Substitution of the ?-OH ligand by ?-OMe in 10 occurs in parallel but more slowly. Complex 2 formed in both cases is more stable in chloroform than in methanol. PMID:21894919

  19. Comparison of different staining methods for polyvinylidene difluoride membranes.

    PubMed

    Christiansen, J; Houen, G

    1992-03-01

    Several new staining methods for polyvinylidene difluoride membranes, including mercurochrome, silver and dimethylaminoazobenzene isothiocyanate staining were compared with Coomassie Brilliant Blue and gold staining. Of these, Coomassie was most versatile and completely compatible with ensuing microsequencing, immunostaining or other visualization methods, while gold and silver staining were more sensitive. Mercurochrome allows selective detection of sulfhydryl-containing proteins while dimethylaminoazobenzene isothiocyanate staining may allow quantitation of sequenceable protein. PMID:1375557

  20. 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.

  1. Improved diagnostics by automated matching and enhancement in fluorescein angiography of the ocular fundus

    NASA Astrophysics Data System (ADS)

    Noordmans, Herke Jan; van den Biesen, Pieter; de Roode, Rowland; Verdaasdonk, Rudolf

    2008-02-01

    An interactive image matching program has been developed to help ophthalmologists in perceiving subtle differences between sequential images obtained during fluorescein angiography. In a pilot experiment, it appeared that the image matching program could effectively correct camera alignment errors. By offering simple tools like image overlay, blinking and image subtraction, differences between angiograms can be greatly enhanced and interpreted. It appeared that newly formed, leaking blood vessels could be detected at an earlier stage of the disease process using these tools. Treatment can be initiated right away, thereby preventing the patient from having additional visual loss. The matching program seems to improve the quality of fundus diagnostics but needs to be validated in future studies.

  2. Development of an alcohol optode membrane based on fluorescence enhancement of fluorescein derivatives.

    PubMed

    Zeng, H H; Wang, K M; Li, D; Yu, R Q

    1994-06-01

    A new type of optode membrane for the determination of alcohols, such as methanol, ethanol, propan-1-ol and propan-2-ol etc, is presented that can be exploited in optical and fibre-optic sensors. It is based on the use of a new lipophilic fluorescent reagent, namely fluorescein octadecyl ester (FODE), which is immobilized in a plasticized PVC membrane. The response mechanism relies on the fact that FODE can reversibly recognize alcohol molecules due to the hydrogen bonding formation between FODE and alcohol molecules. The analytical information in this membrane is the enhancement of the relative fluorescence intensity measured at 527 nm (463 nm, excitation). Under the optimum condition, the membrane has a wide measuring range for alcohol samples. The membrane has been applied to determine the concentration of ethanol in alcoholic drinks with satisfactory results. PMID:18966024

  3. Novel Fluorescein Angiography-Based Computer-Aided Algorithm for Assessment of Retinal Vessel Permeability

    PubMed Central

    Chassidim, Yoash; Parmet, Yisrael; Tomkins, Oren; Knyazer, Boris; Friedman, Alon; Levy, Jaime

    2013-01-01

    Purpose To present a novel method for quantitative assessment of retinal vessel permeability using a fluorescein angiography-based computer algorithm. Methods Twenty-one subjects (13 with diabetic retinopathy, 8 healthy volunteers) underwent fluorescein angiography (FA). Image pre-processing included removal of non-retinal and noisy images and registration to achieve spatial and temporal pixel-based analysis. Permeability was assessed for each pixel by computing intensity kinetics normalized to arterial values. A linear curve was fitted and the slope value was assigned, color-coded and displayed. The initial FA studies and the computed permeability maps were interpreted in a masked and randomized manner by three experienced ophthalmologists for statistical validation of diagnosis accuracy and efficacy. Results Permeability maps were successfully generated for all subjects. For healthy volunteers permeability values showed a normal distribution with a comparable range between subjects. Based on the mean cumulative histogram for the healthy population a threshold (99.5%) for pathological permeability was determined. Clear differences were found between patients and healthy subjects in the number and spatial distribution of pixels with pathological vascular leakage. The computed maps improved the discrimination between patients and healthy subjects, achieved sensitivity and specificity of 0.974 and 0.833 respectively, and significantly improved the consensus among raters for the localization of pathological regions. Conclusion The new algorithm allows quantification of retinal vessel permeability and provides objective, more sensitive and accurate evaluation than the present subjective clinical diagnosis. Future studies with a larger patients’ cohort and different retinal pathologies are awaited to further validate this new approach and its role in diagnosis and treatment follow-up. Successful evaluation of vasculature permeability may be used for the early diagnosis of brain microvascular pathology and potentially predict associated neurological sequelae. Finally, the algorithm could be implemented for intraoperative evaluation of micovascular integrity in other organs or during animal experiments. PMID:23626701

  4. The spots and stains of plate tectonics

    NASA Astrophysics Data System (ADS)

    Oliver, Jack

    1992-01-01

    This paper describes a synthesis characterized by broad scope, substantial support, and some speculation. The framework for the synthesis is the speculative concept that the process of convergence and collision of large landmasses disrupts the fluid regime of the collision zone and adjoining areas. The disturbed fluids leave a record of their disruption and transport in great spots and stains. Some of these spots and stains persist in the modern geologic record where they are known as mineral deposits, mineral occurrences, oil fields, gas fields, tar sands, diagenesis, authigenesis, metamorphism, dolomitization, fluid inclusions, and paleoremagnetization. Various observed characteristics of these phenomena provide supporting evidence of such diversity and consistency that the concept seems firmly rooted in observation. Nevertheless, many opportunities for further testing remain. If the synthesis is more or less correct, then a major link between plate tectonics, or global-scale geodynamics, and a wide variety of terrestrial geological observations of lesser scale is in hand.

  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. Hematoxyliin & Eosin Staining 1. Xylene 3 min

    E-print Network

    Oliver, Douglas L.

    Hematoxyliin & Eosin Staining 1. Xylene 3 min 2. Xylene up & down 3. 100% EtOH up & down 4. 100% Et to dark 14. Ammonia water (bluing)c 5-6 dips 15. Running tap water 2 min 16. Eosin Yd quick dip 17. 70% Et concentrated HCl in 500 ml 70% EtOH c Ammonia water: 0.25% NH4OH d Eosin Y (Richard-Allan Scientific, #7111

  7. Hematoxylin & Eosin (HE) staining -Hiroki Kuroda (Sectioning)

    E-print Network

    De Robertis, Eddy M.

    Hematoxylin & Eosin (HE) staining - Hiroki Kuroda (Sectioning) Time courses are different in sample for 15 min · 50-60ºC tap water for 15 min · Eosin solution for 1 min · Tap water for a while · 70% Et% Ethylene Glycol, with water. Eosin solution (1L) Eosin Y 2.5g, 10% Acetic Acid 2.5 ml in 1L of 60% Et

  8. Stain etching of silicon pillars and macropores

    Microsoft Academic Search

    David Mills; Mona Nahidi; Kurt W. Kolasinski

    2005-01-01

    We used a XeCl excimer laser (308 nm, 3 J cm-2) and chemically enhanced laser ablation in the presence of SF6 to create arrays of silicon pillars. Etching of these pillars with KOH leads to arrays of macropores whose morphology depends on surface crystallography. Several new stain etchants - containing some combination of HF, NH4HF2, HCl, HNO3, Fe(III), Mn(VII) and

  9. Correlation between the Uptake of Sodium Fluorescein in the Tissue and Xenon133 Clearance and Laser Doppler Fluxmetry in Measuring Changes in Skin Circulation

    Microsoft Academic Search

    E. Proano; L. Svensson; L. Perbeck

    1997-01-01

    We have measured the plantar forefoot skin circulation by the uptake of sodium fluorescein (fluorescein flowmetry), 133Xe clearance and laser Doppler fluxmetry in 24 healthy subjects and correlated measurements under basal conditions and after provocation by alcohol intake and application of external heat. To assess the change in skin circulation between the initial measurement at rest and the second measurement

  10. A bifunctional converter: fluorescein quenching scFv/fluorogen activating protein for photostability and improved signal to noise in fluorescence experiments.

    PubMed

    Saunders, Matthew J; Block, Ethan; Sorkin, Alexander; Waggoner, Alan S; Bruchez, Marcel P

    2014-08-20

    Monoclonal antibodies are one of the most useful and ubiquitous affinity reagents used in the biological sciences. Immunostaining of fixed and live cells for microscopy or cytometry measurements frequently employs fluorescently labeled antibodies, in particular fluorescein-labeled antibodies. This dye emits light at a wavelength overlapping with cellular autofluorescence, making it difficult to measure antibody binding to proteins of relatively low copy number or in cells of high green autofluorescence. A number of high affinity fluorescein binding antibodies and antibody domains have been developed that quench the dye's fluorescence. Using a fluorescein-binding recombinant antibody domain genetically fused to a fluorogen activating protein (FAP), we demonstrate a molecular converter capable of binding and quenching fluorescein, while binding and activating a fluorogenic triarylmethane dye. This reagent converts fluorescein conjugates to far-red fluorescent probes, where cellular autofluorescence is low, improving signal-to-background of cell-based antibody binding measurements by ?7-fold. Microscopy experiments show colocalization of both fluorescein and MG fluorescence. This dual affinity fluorescein-quenching-FAP can also be used to convert fluorescein to the red fluorescing MG fluorogen on biological molecules other than antibodies. PMID:25072845

  11. Investigation of binding of nanomarkers of fluorescein family to bovine serum albumin at various values of pH: Spectroscopic study

    NASA Astrophysics Data System (ADS)

    Vlasova, Irina M.; Kuleshova, Anna A.; Vlasov, Alexander A.; Saletsky, Alexander M.

    2013-11-01

    This work is dedicated to investigation of influence of different values of pH on binding of nanomarkers of fluorescein family (fluorescein, erythrosin, eosin and Bengal rose) to bovine serum albumin (BSA). For this purpose dependences of nanomarkers fluorescence, of nanomarkers molecular association, of types of chemical bonds between BSA and nanomarkers on pH are detected. The red shift of fluorescence spectra and the quenching of fluorescence of nanomarkers of fluorescein family in BSA solutions are observed. The decrease of degree of molecular association of nanomarkers in BSA solutions is found out. The dependences of fluorescence intensity and dependences of degree of molecular association on pH of halogen-derivatives of fluorescein differ dramatically from that of fluorescein.

  12. Supporting Information Tuning the pKa of Fluorescein to Optimize Binding Assays

    E-print Network

    Raines, Ronald T.

    , Thomas J. Rutkoski, and Ronald T. Raines* Departments of Chemistry and Biochemistry, University illumination or staining with I2, ceric ammonium molybdate, or phosphomolybdic acid. Flash chromatography maintaining the water-bath temperature below 40 °C. The term "concentrated under high vacuum" refers

  13. Strategic Tool Use: Pencil Box Staining

    NSDL National Science Digital Library

    WGHB Boston

    2013-01-01

    This professional development video clip of students engaged in Common Core Practice Standard #5—Use appropriate tools strategically, shows students making estimates of the amount of stain needed for 26 pencil boxes. Mr. Levy presents the class with the problem and a set of tools with which to choose how to solve the problem, the video clips shows the students engaged in problem solving through their interactions with one another and their teacher. Additional resources include a video transcript, teaching tips, and a link to a professional development reflection activity based upon the video.

  14. Digital staining of pathological tissue specimens using spectral transmittance

    NASA Astrophysics Data System (ADS)

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

    2005-04-01

    Staining of tissue specimens is a classical procedure in pathological diagnosis to enhance the contrast between tissue components such that identification and classification of these components can be easily performed. In this paper, a framework for digital staining of pathological specimens using the information derived from the L-band spectral transmittance of various pathological tissue components is introduced, particularly the transformation of a Hematoxylin and Eosin (HE) stained specimen to its Masson-Trichrome (MT) stained counterpart. The digital staining framework involves the classification of tissue components, which are highlighted when the specimen is actually stained with MT stain, e.g. fibrosis, from the HE-stained image; and the linear mapping between specific sets of HE and MT stained transmittance spectra through pseudo-inverse procedure to produce the LxL transformation matrices that will be used to transform the HE stained transmittance to its equivalent MT stained transmittance configuration. To generate the digitally stained image, the decisions of multiple quadratic classifiers are pooled to form the weighting factors for the transformation matrices. Initial results of our experiments on liver specimens show the viability of multispectral imaging (MSI) for the implementation of digital staining in the pathological context.

  15. In Vivo Ocular Fluorophotometry: Delivery of Fluoresceinated Dextrans via Transscleral Diffusion in Rabbits

    PubMed Central

    Berezovsky, Damian E.; Patel, Samirkumar R.; McCarey, Bernard E.

    2011-01-01

    Purpose. To evaluate the transscleral delivery of fluoresceinated dextrans (FITC-D) with molecular mass up to 70 kDa to the rabbit posterior segment using sub-Tenon injections. Methods. Eighteen NZW rabbits received a unilateral 200-?L injection of 2 mg/mL sodium fluorescein (NaF), 25 mg/mL 40-kDa FITC-D, or 25 mg/mL 70-kDa FITC-D, with (n = 9) or without (n = 9) immediate euthanatization. In live animals, fluorescence was measured in the retina/choroid and mid-vitreous by fluorophotometry, immediately after injection and after 4, 24, 48, and 72 hours. Euthanatized animals were examined hourly through 5 or 6 hours. Results. In live animals, the average peak NaF concentration in the retina/choroid was 310.2 ng/mL, measured 3 hours after injection. Average 40- and 70-kDa FITC-D concentrations in the retina/choroid peaked at 5409.6 and 2375.6 ng/mL, respectively, 24 hours after injection. Fluorescence returned to baseline levels 6 hours after NaF injection, and 48 and 72 hours after 40- and 70-kDa FITC-D injections, respectively. Rabbits that received NaF followed by euthanatization exhibited a continuous increase in retina/choroid and mid-vitreous fluorescence, beginning 1 hour after injection, whereas FITC-D-injected eyes did not show elevated retina/choroid or mid-vitreous fluorescence through 6 hours. Conclusions. FITC-D weighing up to 70-kDa, as well as NaF, reached the posterior retina/choroid after sub-Tenon injections in live rabbits. NaF and 40-kDa FITC-D reached higher peak concentrations and were cleared from the eye more rapidly than was 70-kDa FITC-D. There was minimal penetration of NaF and FITC-D into the mid-vitreous in the in vivo experiments. PMID:21791594

  16. 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 include the ... be present anywhere on the body. Port wine stains may appear in association with other syndromes.

  17. 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.

  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. Polychromatic staining of plant cell walls by toluidine blue O

    Microsoft Academic Search

    T. P. O'Brien; N. Feder; M. E. McCully

    1964-01-01

    Summary 1.The polychromatic staining of plant cell walls by toluidine blue O is described and illustrated.2.The effects of various common fixatives and the effects of the pH of the staining solution are evaluated.3.Simple and rapid procedures are described for preparing stained temporary mounts of fresh material, or permanent mounts of embedded and sectioned material.4.The relationship between the polychromatic staining observed

  20. 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.

  1. 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

  2. Modified Genta triple stain for identifying Helicobacter pylori

    Microsoft Academic Search

    H. M. el-Zimaity; J. Wu; D. Y. Graham

    1999-01-01

    AIM: To evaluate whether lead nitrate could replace uranyl nitrate in the Genta stain for H pylori without sacrificing the advantages of the triple stain (Steiner silver impregnation combined with Alcian blue and haematoxylin\\/eosin (H&E)). METHODS: A comparison was made in 16 specimens between the original triple stain and the revised version. One pathologist evaluated all sections. RESULTS: Direct substitution

  3. Three color cDNA microarrays: quantitative assessment through the use of fluorescein-labeled probes

    PubMed Central

    Hessner, Martin J.; Wang, Xujing; Hulse, Katie; Meyer, Lisa; Wu, Yan; Nye, Steven; Guo, Sun-Wei; Ghosh, Soumitra

    2003-01-01

    Gene expression studies using microarrays have great potential to generate new insights into human disease pathogenesis, but data quality remains a major obstacle. In particular, there does not exist a method to determine prior to hybridization whether an array will yield high quality data, given good study design and target preparation. We have solved this problem through development of a three-color cDNA microarray platform where printed probes are fluorescein labeled, but are spectrally compatible with Cy3 and Cy5 dye-labeled targets when using confocal laser scanners possessing narrow bandwidths. This approach enables prehybridization evaluation of array/spot morphology, DNA deposition and retention and background levels. By using these measurements and the intra-slide coefficient of variation for fluorescence intensity we show that slides in the same batch are not equivalent and measurable prehybridization parameters can be predictive of hybridization performance as determined by replicate consistency. When hybridizing target derived from two cell lines to high and low quality replicate pairs (n = 50 pairs), a direct and significant relationship between prehybridization signal-to-background noise and post-hybridization reproducibility (R2 = 0.80, P < 0.001) was observed. We therefore conclude that slide selection based upon prehybridization quality scores will greatly benefit the ability to generate reliable gene expression data. PMID:12582259

  4. New Parametric Imaging Method with Fluorescein Angiograms for Detecting Areas of Capillary Nonperfusion

    PubMed Central

    Kim, Young Jae; Jeong, Chang Bu; Hwang, Jeong-Min; Yang, Hee Kyung; Lee, Seung Hyun

    2014-01-01

    Objectives Fluorescein angiography (FAG) is currently the most useful diagnostic modality for examining retinal circulation, and it is frequently used for the evaluation of patients with diabetic retinopathy, occlusive diseases, such as retinal venous and arterial occlusions, and wet macular degeneration. This paper presents a method for objectively evaluating retinal circulation by quantifying circulation-related parameters. Methods This method allows the semiautomatic preprocessing and registering of FAG images. The arterial input function is estimated from the registered set of FAG images using gamma-variate fitting. Then, the parameters can be computed by deconvolution on the basis of truncated singular value decomposition, and they can finally be presented as parametric color images in a combination of three colors, red, green, and blue. Results After the estimation of arterial input function, the parameters of relative blood flow and mean transit time were computed using deconvolution analysis based on truncated singular value decomposition. Conclusions The parametric color image is helpful to interpret the status of retinal blood circulation and provides quantitative data on retina ischemia without interobserver variability. This system easily provides the status of retinal blood circulation both qualitatively and quantitatively. It also helps to standardize FAG interpretation and may contribute to network-based telemedicine systems in the future. PMID:25152832

  5. Acute retinal pigment epitheliitis: spectral domain optical coherence tomography, fluorescein angiography, and autofluorescence findings.

    PubMed

    Aydo?an, Tu?ba; Güney, Esra; Akçay, Betül ?lkay Sezgin; Bozkurt, Tahir Kansu; Ünlü, Cihan; Ergin, Ahmet

    2015-01-01

    A 17-year-old presented with central and paracentral scotomas in his right eye for one week. There was no remarkable medical or ocular history. Blood analyses were within normal range. At presentation both eyes' best-corrected visual acuities were 20/20. Slit-lamp examination result was normal. Fundus examination revealed yellow-white hypopigmented areas in the macula. Fluorescein angiography (FA) showed hypofluorescence surrounded by ring of hyperfluorescence. Fundus autofluorescence (FAF) was slightly increased. Spectral domain optical coherence tomography (SD-OCT) showed disruption of IS/OS junction with expansion of abnormal hyperreflectivity from retinal pigment epithelium to the outer nuclear layer (ONL). One month later fundus examination showed disappearance of the lesions. FA revealed transmission hyperfluorescence. FAF showed increased autofluorescence and pigment clumping. Hyperreflective band in SD-OCT disappeared. Loss of photoreceptor segment layers was observed in some of the macular lesions. The diagnosis of acute retinal pigment epitheliitis can be challenging after disappearance of fundus findings. FA, FAF, and SD-OCT are important tests for diagnosis after resolution of the disease. PMID:25767511

  6. Acute Retinal Pigment Epitheliitis: Spectral Domain Optical Coherence Tomography, Fluorescein Angiography, and Autofluorescence Findings

    PubMed Central

    Aydo?an, Tu?ba; Güney, Esra; Akçay, Betül ?lkay Sezgin; Bozkurt, Tahir Kansu; Ünlü, Cihan; Ergin, Ahmet

    2015-01-01

    A 17-year-old presented with central and paracentral scotomas in his right eye for one week. There was no remarkable medical or ocular history. Blood analyses were within normal range. At presentation both eyes' best-corrected visual acuities were 20/20. Slit-lamp examination result was normal. Fundus examination revealed yellow-white hypopigmented areas in the macula. Fluorescein angiography (FA) showed hypofluorescence surrounded by ring of hyperfluorescence. Fundus autofluorescence (FAF) was slightly increased. Spectral domain optical coherence tomography (SD-OCT) showed disruption of IS/OS junction with expansion of abnormal hyperreflectivity from retinal pigment epithelium to the outer nuclear layer (ONL). One month later fundus examination showed disappearance of the lesions. FA revealed transmission hyperfluorescence. FAF showed increased autofluorescence and pigment clumping. Hyperreflective band in SD-OCT disappeared. Loss of photoreceptor segment layers was observed in some of the macular lesions. The diagnosis of acute retinal pigment epitheliitis can be challenging after disappearance of fundus findings. FA, FAF, and SD-OCT are important tests for diagnosis after resolution of the disease. PMID:25767511

  7. Major histocompatibility complex independent T cell receptor-antigen interaction: functional analysis using fluorescein derivatives

    PubMed Central

    1991-01-01

    We have isolated T cell receptor (TCR) cDNAs from fluorescein (FL)- specific human T cell clones (alpha FL beta FL), and transferred them to TCR beta- Jurkat cells in order to study direct FL-binding to the TCR. Using either FL-conjugated polymers (FL-polymer) or FL-substituted Sepharose beads, we are able to demonstrate the direct binding of antigen to the T cell surface, and the functional activation of the T cell transfectants. We present evidence against the involvement of major histocompatibility complex (MHC) molecules or antigen presentation in the interaction of FL with the alpha FL beta FL transfectants. Additionally, we have examined the effect of ring substitutions on the FL molecule as well as specific alterations of substituents attached to the 5' position, and we have found that all of them interfere with the functional recognition of the alpha FL beta FL TCR. These experiments demonstrate that TCRs like antibodies have intrinsic affinities for antigen, even without the involvement of MHC molecules. PMID:2056277

  8. Comparison of adaptive optics scanning light ophthalmoscopic fluorescein angiography and offset pinhole imaging

    PubMed Central

    Chui, Toco Y. P.; Dubow, Michael; Pinhas, Alexander; Shah, Nishit; Gan, Alexander; Weitz, Rishard; Sulai, Yusufu N.; Dubra, Alfredo; Rosen, Richard B.

    2014-01-01

    Recent advances to the adaptive optics scanning light ophthalmoscope (AOSLO) have enabled finer in vivo assessment of the human retinal microvasculature. AOSLO confocal reflectance imaging has been coupled with oral fluorescein angiography (FA), enabling simultaneous acquisition of structural and perfusion images. AOSLO offset pinhole (OP) imaging combined with motion contrast post-processing techniques, are able to create a similar set of structural and perfusion images without the use of exogenous contrast agent. In this study, we evaluate the similarities and differences of the structural and perfusion images obtained by either method, in healthy control subjects and in patients with retinal vasculopathy including hypertensive retinopathy, diabetic retinopathy, and retinal vein occlusion. Our results show that AOSLO OP motion contrast provides perfusion maps comparable to those obtained with AOSLO FA, while AOSLO OP reflectance images provide additional information such as vessel wall fine structure not as readily visible in AOSLO confocal reflectance images. AOSLO OP offers a non-invasive alternative to AOSLO FA without the need for any exogenous contrast agent. PMID:24761299

  9. Thermally activated delayed fluorescence of fluorescein derivative for time-resolved and confocal fluorescence imaging.

    PubMed

    Xiong, Xiaoqing; Song, Fengling; Wang, Jingyun; Zhang, Yukang; Xue, Yingying; Sun, Liangliang; Jiang, Na; Gao, Pan; Tian, Lu; Peng, Xiaojun

    2014-07-01

    Compared with fluorescence imaging utilizing fluorophores whose lifetimes are in the order of nanoseconds, time-resolved fluorescence microscopy has more advantages in monitoring target fluorescence. In this work, compound DCF-MPYM, which is based on a fluorescein derivative, showed long-lived luminescence (22.11 ?s in deaerated ethanol) and was used in time-resolved fluorescence imaging in living cells. Both nanosecond time-resolved transient difference absorption spectra and time-correlated single-photon counting (TCSPC) were employed to explain the long lifetime of the compound, which is rare in pure organic fluorophores without rare earth metals and heavy atoms. A mechanism of thermally activated delayed fluorescence (TADF) that considers the long wavelength fluorescence, large Stokes shift, and long-lived triplet state of DCF-MPYM was proposed. The energy gap (?EST) of DCF-MPYM between the singlet and triplet state was determined to be 28.36 meV by the decay rate of DF as a function of temperature. The ?E(ST) was small enough to allow efficient intersystem crossing (ISC) and reverse ISC, leading to efficient TADF at room temperature. The straightforward synthesis of DCF-MPYM and wide availability of its starting materials contribute to the excellent potential of the compound to replace luminescent lanthanide complexes in future time-resolved imaging technologies. PMID:24936960

  10. Assembly properties of fluorescein-labeled tubulin in vitro before and after fluorescence bleaching

    PubMed Central

    1984-01-01

    Brain tubulin has been conjugated with dichlorotriazinyl- aminofluorescein (DTAF) to form a visualizable complex for the study of tubulin dynamics in living cells. By using several assays we confirm the finding of Keith et al. (Keith, C. H., J. R. Feramisco, and M. Shelanski, 1981, J. Cell Biol., 88:234-240) that DTAF-tubulin polymerizes like control tubulin in vitro. The fluorescein moiety of the complex is readily bleached by the 488-nm line from an argon ion laser. When irradiations are performed over short times (less than 1 s) and in the presence of 2 mM glutathione, a mixture of DTAF-tubulin and control protein (as occurs after microinjection of the fluorescent conjugate into living cells) will retain full polymerization activity. Slow bleaching (approximately 5 min) or bleaching without glutathione promotes formation of covalent cross-links between neighboring polypeptides and kills the polymerization activity of DTAF-tubulin, including some molecules that are neither cross-linked nor bleached. Even under conditions that damage DTAF-tubulin, however, DTAF- microtubules are not destroyed by bleaching. They will continue to elongate by addition of DTAF-tubulin subunits to their free ends, and they neither bind nor exchange subunits along their lateral surfaces. These results suggest that DTAF-tubulin is a suitable analog for tubulin, both in studies of protein incorporation and for investigations of fluorescence redistribution after photobleaching. PMID:6438113

  11. Effect of lens implants on iris fluorescein angiography and the iris pigment layer.

    PubMed

    Krause, U

    1985-08-01

    Fourteen eyes underwent cataract operation and implantation of an iris fixated (Medallion) pseudophakos. Eleven had an uncomplicated per- and post-operative course (group 1), 3 had a subluxation of the lens implant (group 2). The control group (group 3) consisted of the 14 contralateral eyes from group 1 and 2. Two cadaver eyes had a similar lens implanted (group 4). Groups 1-3 were investigated by bilateral simultaneous iris fluorescein angiography and retroiridal stereo transillumination photography. In group 1, one case with pupillary and diffuse semiperipheral leakage areas was observed, in a clinically healthy eye 15 months after the operation. An other one had some leakage at the pupillary border, but not in the area of contact with the loops. Rubbing between iris surface and some parts of the lens did not provoke local leakage. No new vessel formation was found. Pigment layer defects were observed in every operated eye, but no leakage was related to them except in the two pupillary areas mentioned. Mayor pigment defects did not progress in a quiet eye. The iris seems to be to some degree resistant to contact with the parts of the intraocular lens implant. PMID:4050356

  12. Three color cDNA microarrays: quantitative assessment through the use of fluorescein-labeled probes.

    PubMed

    Hessner, Martin J; Wang, Xujing; Hulse, Katie; Meyer, Lisa; Wu, Yan; Nye, Steven; Guo, Sun-Wei; Ghosh, Soumitra

    2003-02-15

    Gene expression studies using microarrays have great potential to generate new insights into human disease pathogenesis, but data quality remains a major obstacle. In particular, there does not exist a method to determine prior to hybridization whether an array will yield high quality data, given good study design and target preparation. We have solved this problem through development of a three-color cDNA microarray platform where printed probes are fluorescein labeled, but are spectrally compatible with Cy3 and Cy5 dye-labeled targets when using confocal laser scanners possessing narrow bandwidths. This approach enables prehybridization evaluation of array/spot morphology, DNA deposition and retention and background levels. By using these measurements and the intra-slide coefficient of variation for fluorescence intensity we show that slides in the same batch are not equivalent and measurable prehybridization parameters can be predictive of hybridization performance as determined by replicate consistency. When hybridizing target derived from two cell lines to high and low quality replicate pairs (n = 50 pairs), a direct and significant relationship between prehybridization signal-to-background noise and post-hybridization reproducibility (R2 = 0.80, P < 0.001) was observed. We therefore conclude that slide selection based upon prehybridization quality scores will greatly benefit the ability to generate reliable gene expression data. PMID:12582259

  13. Extending applicability of the oxygen radical absorbance capacity (ORAC-fluorescein) assay.

    PubMed

    Dávalos, Alberto; Gómez-Cordovés, Carmen; Bartolomé, Begoña

    2004-01-14

    The ORAC-fluorescein (ORAC-FL) method recently validated using automatic liquid handling systems has now been adapted to manual handling and using a conventional fluorescence microplate reader. As calculated for Trolox, the precision of the method was <3.0, expressed as percent coefficient of variation. The accuracy of the method was <2.3, expressed as percent variation of the mean. The detection and quantification limits were those corresponding to 0.5- and 1-microM Trolox standard solutions, respectively. The method has been applied to 10 pure compounds (benzoic and cinnamic acids and aldehydes, flavonoids, and butylated hydroxyanisole), to 30 white, rose, and bottled- and oak-aged red wines, and to 7 commercial dietary antioxidant supplements. All samples exhibited a good linear response with concentration. As seen by other methodologies, the chemical structure of a compound determines its antioxidant activity (ORAC-FL value). Of particular interest were the results with oak-aged red wines from different vintages (1989-2002) that confirm influence of vintage, but not origin of the oak, in the antioxidant activity of wines from the same variety. Dietary antioxidant supplements presented a great variability (170-fold difference) in their antioxidant potency. This work proves applicability of the ORAC-FL assay in evaluating the antioxidant activity of diverse food samples. PMID:14709012

  14. A novel method of affinity-purifying proteins using a bis-arsenical fluorescein.

    PubMed Central

    Thorn, K. S.; Naber, N.; Matuska, M.; Vale, R. D.; Cooke, R.

    2000-01-01

    Genetically-encoded affinity tags constitute an important strategy for purifying proteins. Here, we have designed a novel affinity matrix based on the his-arsenical fluorescein dye FlAsH, which specifically recognizes short alpha-helical peptides containing the sequence CCXXCC (Griffin BA, Adams SR, Tsien RY, 1998, Science 281:269-272). We find that kinesin tagged with this cysteine-containing helix binds specifically to FlAsH resin and can be eluted in a fully active form. This affinity tag has several advantages over polyhistidine, the only small affinity tag in common use. The protein obtained with this single chromatographic step from crude Escherichia coli lysates is purer than that obtained with nickel affinity chromatography of 6xHis tagged kinesin. Moreover, unlike nickel affinity chromatography, which requires high concentrations of imidazole or pH changes for elution, protein bound to the FlAsH column can be completely eluted by dithiothreitol. Because of these mild elution conditions, FlAsH affinity chromatography is ideal for recovering fully active protein and for the purification of intact protein complexes. PMID:10716173

  15. Distinct patterns of cerebral extravasation by Evans blue and sodium fluorescein in rats.

    PubMed

    Yen, Lola Fenghuei; Wei, Vivi Chiali; Kuo, Eva Yuhua; Lai, Ted Weita

    2013-01-01

    The Evans blue dye (EBD; 961 Da) and the sodium fluorescein dye (NaF; 376 Da) are commonly used inert tracers in blood-brain barrier (BBB) research. They are both highly charged low molecular weight (LMW) tracers with similar lipophobic profiles. Nevertheless, the EBD binds to serum albumin (69,000 Da) to become a high molecular weight (HMW) protein tracer when injected into the circulation, whereas the NaF remains an unbound small molecule in the circulation. In this study, rats were injected with equal doses of either EBD or NaF to monitor their blood and tissue distribution. The EBD was largely confined to the circulation with little accumulation in the peripheral organ and even less accumulation in the central tissue, whereas the NaF distributed more evenly between the blood and the peripheral organ but was also largely excluded from the central tissue. Importantly, the EBD crossed the BBB most effectively at the prefrontal cortex and the cerebellum, and most poorly at the striatum. In marked contrast, the NaF was evenly distributed throughout the brain. Finally, the EBD exhibited this same peculiar tissue distribution profile when administered by either bolus injection or slow infusion. Our study suggests that different regions of the brain are equally permeable to LMW inert dyes like the NaF, but are markedly different in permeability to HMW proteins such as EBD-labelled serum albumin. PMID:23861924

  16. Effects of perfusion rate on permeability of frog and rat mesenteric microvessels to sodium fluorescein

    PubMed Central

    Montermini, D; Winlove, C P; Michel, C C

    2002-01-01

    The permeability, PS, to sodium fluorescein (Stokes-Einstein radius = 0.45 nm) has been measured in single mesenteric capillaries of pithed frogs and anaesthetised rats as perfusion velocity, U, was varied over a range from 400 up to 2000–10 000 ?m s?1. PS increased linearly with U. In 20 frog capillaries, mean (± S.E.M.) PS (in ?m s?1) = 9.35 (± 1.55)U × 10?5 + 0.244 (± 0.0291). Similarly, in nine rat venules, mean PS = 1.62 (± 0.385)U × 10?4 + 0.375 (± 0.025). The flow-dependent component of permeability could be reversibly abolished in frog capillaries by superfusing with 100 ?M noradrenaline and by superfusing rat venules with the nitric oxide synthase inhibitor, NG-nitro-L-arginine (20 ?M). It was shown that changes in microvascular pressure accompanying changes in U during free perfusion could account for only 15 % of the changes in PS, i.e. 85 % of the changes in PS were changes in the permeability coefficient itself. A comparison between the changes in PS with U and the previously described changes in microvascular permeability to K+ with U, suggest that if the flow-dependent component of permeability is modelled as a population of pores of constant size, these have radii of 0.8 nm. Such a pathway would limit flow-dependent permeability to small hydrophilic molecules and have minimal effect on net fluid exchange. PMID:12231651

  17. Differences in bull spermiograms using eosin-nigrosin stain, feulgen stain, and phase contrast microscopy methods

    Microsoft Academic Search

    D. J. Sprecher; P. H. Coe

    1996-01-01

    The Society for Theriogenology recently adopted a minimum standard of 70% normal spermatozoa for the bull breeding soundness examination (BSE). We conducted this study to determine if spermiograms derived by brightfield microscopy of eosin-nigrosin stained semen smears (Society method) overestimated the proportion of normal spermatozoa. Comparison of the above method was made with that of phase contrast microscopy (Phase method).

  18. Ultrafast tissue staining with chemical tags.

    PubMed

    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

    2014-09-01

    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

  19. Digistain: a digital staining instrument for histopathology.

    PubMed

    Amrania, Hemmel; Antonacci, Giuseppe; Chan, Che-Hung; Drummond, Laurence; Otto, William R; Wright, Nicholas A; Phillips, Chris

    2012-03-26

    We describe a new mid-infrared (mid-IR) imaging method specifically designed to augment the H + E tissue staining protocol. Images are taken with bespoke IR filters at wavelengths that enable chemical maps to be generated, corresponding to the cytoplasmic (amide) and nuclear (phosphodiester) components of unstained oesophageal tissue sections. A suitably calibrated combination of these generates false colour computer images that reproduce not only the tissue morphology, but also accurate and quantitative distributions of the nuclear-to-cytoplasmic ratio throughout the tissue section. This parameter is a well documented marker of malignancy, and because the images can be taken and interpreted by clinically trained personnel in a few seconds, we believe this new "digistain" approach makes spectroscopic mid-IR imaging techniques available for the first time as a practical, specific and sensitive augmentation to standard clinical cancer diagnosis methods. PMID:22453410

  20. 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.

  1. Tear Fluid Gelatinase B Activity Correlates with IL1a Concentration and Fluorescein Clearance in Ocular Rosacea

    Microsoft Academic Search

    Adolfo A. Afonso; Lucia Sobrin; Dagoberto C. Monroy; Marie Selzer; Balakrishna Lokeshwar; Stephen C. Pflugfelder

    1999-01-01

    RESULTS. Compared with normal control subjects, patients with ocular rosacea had a greater delay of tear fluorescein clearance (P , 0.001), a higher tear IL-1a concentration (P , 0.001), and a greater pro- gelatinase B (92 kDa) activity (P , 0.001) in their tear fluid. The 84-kDa active form of gelatinase B was observed in 46% of the rosacea tear

  2. Correlation of lipid layer thickness measurements with fluorescein tear film break-up time and Schirmer's test

    Microsoft Academic Search

    M A Isreb; J V Greiner; D R Korb; T Glonek; S S Mody; V M Finnemore; C V Reddy; Korb

    2003-01-01

    Purpose This study correlates measurement of lipid layer thickness (LLT) with two frequently used dry eye tests, fluorescein break-up time (FBUT) and Schirmer's test with anaesthesia (STA).Methods Subjects (n=44 eyes) with symptoms of dry eye and positive results for dry eye with either FBUT or STA or both were selected. Quantification of LLT was performed by the observation of colour

  3. Cloacal reflexes and uptake of fluorescein-labeled polystyrene beads in broiler chickens.

    PubMed

    van der Sluis, H J; Dwars, R M; Vernooij, J C M; Landman, W J M

    2009-06-01

    Experiments were performed (1) to quantify reflex movements and volume uptake of physiological salt solution by the cloaca and (2) to evaluate the conditioned cloacal uptake of fluorescein-labeled polystyrene. In experiment 1, measurements were done on birds (n = 12) once a day at 3, 4, and 5 d of age and during 5 consecutive days at wk 3, 5, 7, and 9 of age. The reflexes and volume uptake after applying saline droplets were studied simultaneously during 30 s. The median number and range of reflexes per 30 s during the first week of age were 45 (28 to 54), at 3 wk 35 (18 to 52), at 5 wk 44 (27 to 60), at 7 wk 47 (32 to 61), and at 9 wk 44 (23 to 56). The median volume uptake and range in wk 1, 3, 5, 7, and 9 were 0.10 (0.05 to 0.30), 0.25 (0.05 to 0.60), 0.58 (0.25 to 1.15), 1.05 (0.50 to 2.25), and 1.15 (0.30 to 3.05) mL per 30 s, increasing significantly with time. In experiment 2, a solution containing 10(7) polystyrene beads/mL was applied to the cloaca of broilers (3 aged 2 wk and 3 aged 9 wk) during 30 s. Most beads were found in the bursa of Fabricius. In the bursal lumen, a median of 10(6.43) beads/mL was found; the median number found in the follicular tissue was 5 (range 3 to 38) beads per tissue section. In the lumen content of ileum, cecum, and rectum of all birds together, it was 10(5.87), 0, and 10(6.32) beads/ ml, respectively. Polystyrene beads were never found intramuraly. PMID:19439636

  4. Specificity of the fluorescein transport process in Malpighian tubules of the cricket Acheta domesticus.

    PubMed

    Neufeld, Douglas S G; Kauffman, Ross; Kurtz, Zachary

    2005-06-01

    We demonstrate the presence of an efficient, multispecific transport system for excretion of organic anions in the Malpighian tubules of the cricket Acheta domesticus using fluorescein (FL) as a model substrate. Malpighian tubules rapidly accumulated FL via a high affinity process (Km = 7.75 micromol l(-1)); uptake was completely eliminated by the prototypical organic anion transport inhibitor probenecid (1 mmol l(-1)), but not by p-aminohippuric acid (3 mmol l(-1)). FL uptake was inhibited by monocarboxylic acids at a high concentration (3 mmol l(-1)), and inhibition was more effective with an increase in the carbon chain of the monocarboxylic acid (37% inhibition by 5-carbon valeric acid, and 89% inhibition by 7-carbon caprylic acid). Likewise, tests using a series of aliphatic glutathione conjugates indicated that only the compound with the longest side-chain (decyl-glutathione) significantly inhibited FL uptake (81% inhibition). FL uptake was inhibited by a number of xenobiotics, including a plant alkaloid (quinine), herbicides (2,4-dichlorophenoxyacetic acid and 4-(2,4-dichlorophenoxy)-butyric acid), and the insecticide metabolites malathion monocarboxylic acid (MMA) and 3-phenoxybenzoic acid (PBA), suggesting that this transport system plays an active role in excretion of xenobiotics from Acheta by Malpighian tubules. HPLC quantification of MMA and PBA accumulation into Malpighian tubules verified that MMA accumulation was via a mediated transport process, but suggested that PBA accumulation was by nonspecific binding. The presence of a transport system in Malpighian tubules that handles at least one pesticide metabolite (MMA) suggests that transport processes could be a mechanism conferring resistance to xenobiotic exposure in insects. PMID:15939766

  5. Spectroscopic characterization of fluorescein- and tetramethylrhodamine-labeled oligonucleotides and their complexes with a DNA template

    NASA Astrophysics Data System (ADS)

    Wang, L.; Gaigalas, A. K.; Blasic, J.; Holden, M. J.

    2004-10-01

    We measured absorption and emission spectra, fluorescence quantum yield, anisotropy, fluorescence resonance energy transfer (FRET), and melting temperature to characterize fluorescein- and tetramethylrhodamine (TMR)-labeled oligonucleotides in solution and when hybridized to a common DNA template. Upon hybridization to the template, both the absorption and emission spectra of TMR-labeled duplexes exhibited a shift with respect to those of labeled oligonucleotides, depending on the location of the TMR on the oligonucleotide. Measurements of quantum yield, anisotropy, and melting temperature indicated that TMR interacted with nucleotides within the duplexes in the order (T1>T5>T11, T16) that the oligonucleotide with TMR labeled at the 5' end (T1) is stronger than that labeled at position 5 from the 5' end (T5), which is also stronger than those labeled at the positions, 11 and 16, from the 5' end (T11, T16). In the case of the duplex formed between T1 and the template, fluorescence quenching was observed, which is attributed to the interaction between the dye molecule and guanosines located at the single-stranded portion of the template. A two-state model was suggested to describe the conformational states of TMR in the duplex. The melting temperatures of the four FRET complexes show the same pattern as those of TMR-labeled duplexes. We infer that the interactions between TMR and guanosine persist in the FRET complexes. This interaction may bring the donor and the acceptor molecules closely together, which could cause interaction between the two dye molecules shown in absorbance measurements of the FRET complexes.

  6. Fluorescein angiography-based diagnosis for retinopathy of prematurity: expert-non expert comparison.

    PubMed

    Guagliano, Rosanna; Barillà, Donatella; Bertone, Chiara; Maffia, Anna; Periti, Francesca; Spallone, Laura; Anselmetti, Giovanni; Giacosa, Elisabetta; Stronati, Mauro; Tinelli, Carmine; Bianchi, Paolo Emilio

    2013-05-27

    Purpose: To evaluate accuracy and inter-rater reliability of RetCam fundus images and digital camera fluorangioscopic images in acute retinopathy of prematurity (ROP) by comparing diagnoses given by trainee ophthalmologists with those provided by expert ophthalmologists.?Methods: This is a multicenter retrospective observational study of diagnostic data from 48 eyes of 24 premature infants with classical ROP, stage II, as evaluated by RetCam 3 and fluorescein angiography (FA). Average gestational age was 25.4 weeks, average weight 804.7 g. A staging grid (with ocular fundus divided into 3 concentric zones) and 24 15° sectors centered around the optic papilla were superimposed on 360° retina photomontages (Photoshop) made from RetCam and FA images. Non expert vs expert diagnosis agreement was measured for each sector by means of Cohen kappa (Fleiss, 1981).?Results: A high degree of concordance was found. Inter-rater agreement between expert and non expert interpretations of retinal photomontages was greater for fluorangiographic images than for RetCam images, with ? = 0.61-1 for 120/152 (78.9%) sectors examined on the RetCam images and ?? = 0.61-1 for 168/198 (84.8%) sectors examined on the FA images.?Conclusions: The FA images appear to be easier to interpret than RetCam images, both by expert and non expert ophthalmologists. The results confirm that FA is a good examination technique with a high degree of reliability, even where trainee practitioners are involved. This suggests that retinopathy management can be improved by entrusting diagnostic responsibilities to trainee ophthalmologists, in order to extend access to correct diagnosis, recognition of threshold lesions, and prompt treatment. PMID:23709330

  7. Rotational diffusion of markers of the fluorescein family in solutions of bovine serum albumin, according to the data from polarized fluorescence

    NASA Astrophysics Data System (ADS)

    Vlasova, I. M.; Kuleshova, A. A.; Vlasov, A. A.; Saletskii, A. M.

    2015-02-01

    The polarized fluorescence and rotational diffusion of markers of the fluorescein family (initial fluorescein and its halogenated derivatives erythrosine, eosin, and Rose Bengal) in solutions of bovine serum albumin (BSA) are studied. Elevated degrees of the polarization of fluorescence markers, increased times of rotational relaxation, and reduced coefficients of the rotational diffusion of markers are observed in solutions of BSA. It is shown that all four markers of the fluorescein family can be used to register binding with BSA, but fluorescein is better for studies of BSA with varied pH values and weakly electronegative hydrogen atoms in the structural formula. It is found that increasing the electronegativity of atoms in the structural formulas of markers raises the polarization of their fluorescence, reducing the coefficient of their rotational diffusion and lengthening their periods of rotational relaxation.

  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. Manual hematoxylin and eosin staining of mouse tissue sections.

    PubMed

    Cardiff, Robert D; Miller, Claramae H; Munn, Robert J

    2014-06-01

    The hematoxylin and eosin (H&E) stain is the standard used for microscopic examination of tissues that have been fixed, processed, embedded, and sectioned. It can be performed manually or by automation. For economic reasons, the manual technique is generally the method of choice for facilities with a low sample volume. This protocol describes manual H&E staining of fixed, processed, paraffin-embedded, and sectioned mouse tissues. In H&E-stained tissues, the nucleic acids stain dark blue and the proteins stain red to pink or orange. For accurate phenotyping and delineation of tissue detail, the protocol must be adhered to rigorously. This includes frequent reagent changes as well as the use of "in-date" reagents. Appropriate color in a good H&E stain allows for identification of many tissue subtleties that are necessary for accurate diagnosis. PMID:24890205

  11. New rapid staining methods of Cryptosporidium oocysts in stools.

    PubMed

    Chichino, G; Bruno, A; Cevini, C; Atzori, C; Gatti, S; Scaglia, M

    1991-01-01

    Two new extemporaneous negative-staining methods are proposed to detect Cryptosporidium oocysts in stools, using light-green and merbromine in 1% and 2% aqueous solution, respectively. A Ziehl-Neelsen stain as modified by Henriksen and Polhenz was used as a reference technique. A comparison between these two new stains and the reference, a modified Ziehl-Neelsen mod. method gave almost identical sensitivity and specificity. We propose their use in routine diagnosis for enteric cryptosporidiosis. PMID:1726326

  12. Factors involved in differential giemsa-staining of sister chromatids

    Microsoft Academic Search

    Keiko Goto; S. Maeda; Y. Kano; T. Sugiyama

    1978-01-01

    Microspectrophotometric evaluation of differentially stained sister chromatids made it possible to analyse precisely the factors involved in the Giemsa methods. The concentration of Hoechst 33258, pH of the mounting medium, temperature during UV-exposure and the quality (wavelength) of UV-light influenced the differential staining. Exposure of blacklight of 10-5 M Hoechst 33528-stained BrdU-labeled chromosome specimens mounted in McIlvaine buffer (pH 8.0)

  13. Enamel hypoplasia and essential staining of teeth from erythroblastosis fetalis.

    PubMed

    Atasu, M; Genc, A; Ercalik, S

    1998-01-01

    The dental, clinical, radiological, pedigree and dermatoglyphic findings of a patient showing hypoplasia of enamel and intrinsic staining of the teeth from erythroblastosis fetalis are presented. PMID:9641102

  14. Stain defect detection for mobile phone camera modules

    NASA Astrophysics Data System (ADS)

    Hong, Sehee; Lee, Chulhee

    2014-03-01

    In this paper, we present a stain defect detection algorithm based on the difference of window mean brightness. In particular, we use the maximum square value of the difference brightness in divided windows (MAXWDMS). Window shapes generally affect WDMS values and make stain images clearly distinguishable. The proposed method consists of three steps: window design, stain localization using MAXWDMS and setting the WDMS level. The proposed methodology has been successfully used in stain defect detection, achieving good detection rates in both quantitative evaluation and sensibility estimation. Experimental results show improved detection accuracy and a satisfactory processing time.

  15. Extrinsic stain removal with a toothpowder: A randomized controlled trial

    PubMed Central

    Khan, Muhammad Khalil; Bokhari, Syed Akhtar Hussain; Haleem, Abdul; Kareem, Abdul; Khan, Ayyaz Ali; Hosein, Tasleem; Khan, Muhammad Usama

    2014-01-01

    Objectives The efficacy of a commercially available toothpowder was compared with toothpaste in removing extrinsic dental stains. Methods In this single-blind, randomized controlled trial, 77 volunteers were included from a residential professional college. All study subjects (control toothpaste users and test toothpowder users) plaque control measures. All study subjects were instructed to rinse with 5 ml 0.12% chlorhexidine mouthwash for 1 minute, twice and one cup of double tea bag solution three times daily for three weeks. Subjects were randomized into test (n=36) and control (n=36) groups. Toothpaste (control) and toothpowder (test) was used for two weeks to see the effects on removing stains on the labial surfaces of 12 anterior teeth. For measuring dental extrinsic stains Lobene Stain Index (SI) was used. Results The amount of stain following the use of toothpaste and toothpowder was more controlled with the experimental toothpowder. For all sites combined, there was evidence that the experimental toothpowder was significantly superior to toothpaste in reducing stain area (p<.001), stain intensity (p<.001) and composite/product (area × intensity) (p<.001). Conclusion Stain removing efficacy of toothpowder was significantly higher as compared with toothpaste. A toothpowder may be expected to be of benefit in controlling and removing extrinsic dental staining. PMID:25505862

  16. Transmission electron microscopy staining methods for the cortex of human hair: a modified osmium method and comparison with other stains.

    PubMed

    Harland, D P; Vernon, J A; Walls, R J; Woods, J L

    2011-08-01

    For wool, superior staining of a wide range of ultrastructural components is achieved by en bloc treatment of fibres with a chemical reductant followed by osmium tetroxide. For human scalp hair, although staining quality is similar, the penetration of reagents is poor, resulting in large parts of the fibre cortex remaining unstained. Here we describe a modification to the reduction-osmication method in which reagents penetrate through a cut fibre end, allowing visualization of a wide range of features across the cortex. We compare the staining quality, artefacts and range of structure rendered visible using transmission electron microscopy for en bloc reduction-osmication to other staining alternatives including en bloc silver nitrate and section stains based on uranyl acetate and lead citrate, phosphotungstic acid, potassium permanganate, ammoniacal silver nitrate and some combinations of these stains. The effects of hair-care treatments are briefly examined. PMID:21477263

  17. Comparison of tetrachromic VOF stain to other histochemical staining techniques for characterizing stromal soft and hard tissue components.

    PubMed

    Belaldavar, C; Hallikerimath, S; Angadi, P V; Kale, A D

    2014-11-01

    The components of hard tissues including dentin, enamel, cementum, bone and other calcified deposits, and mature and immature collagen pose problems for identification in routine hematoxylin and eosin (H & E) stained sections. Use of combinations of stains can demonstrate the components of hard tissues and soft tissues distinctly. We assessed the efficacy of the Verde Luz-orange G-acid fuchsin (VOF) stain for differentiating hard and soft connective tissues and compared results with other histochemical staining techniques. Eighty tissue sections comprising developing tooth (30), ossifying fibroma (30) and miscellaneous pathologies (20) expected to contain varying types of calcified tissues were stained with H & E, VOF, and Masson's trichrome (MT). In developing tooth, VOF demonstrated better differentiation of hard tissues, while it was comparable to MT for ossifying fibroma and miscellaneous pathologies. The intensity of staining was greater with VOF than with the other stains studied. VOF stains hard tissue components distinctly and gives good contrast with the surrounding connective tissue. VOF is comparable to MT, but has added advantages including single step staining, rapid and easy procedures, and it distinguishes the maturity of the tissues. PMID:24830362

  18. Intra-arterial fluorescence angiography with injection of fluorescein sodium from the superficial temporal artery during aneurysm surgery: technical notes.

    PubMed

    Ichikawa, Tsuyoshi; Suzuki, Kyouichi; Watanabe, Yoichi

    2014-06-17

    Intra-arterial fluorescence angiography from a catheter inserted into the external carotid artery (ECA) via the superficial temporal artery (STA) allowed us to satisfactorily evaluate cerebral arterial and venous blood flow. We report this novel method that allowed for repeated angiography within minutes with a low risk of complications due to catheter placement from the STA. The STA was secured at the edge of the standard skin incision during cerebral aneurysm surgery. A 3 Fr catheter was inserted approximately 5 cm to 10 cm into the STA. After manual injection of 5 ml of 20 times diluted 10% fluorescein sodium (fluorescein), fluorescein reached the intracranial internal carotid artery (ICA) through the common carotid artery or anastomoses between the ECA and ICA. Fluorescence emission from the cerebral arteries, capillaries, and veins was clearly observed through the microscope and results were recorded. Quick dye clearance makes it possible to reexamine within 1 minute. In addition, we made a graph of the fluorescence emission intensity in the arteries, capillaries, and veins using fluorescence analysis software. With intravenous fluorescence angiography, dye remains in the vessels for a long time. When repeated examinations are necessary, intervals of approximately 10 minutes are required. There were some cases we could not correctly evaluate with intravenous injection due to weak fluorescence emission. Fluorescence angiography with intra-arterial injection from a catheter inserted into the carotid artery or another major vessel, like conventional angiography, has a risk of procedure-related complications. We report our new method since it solved these problems and is useful. PMID:24477067

  19. Solute concentration-dependent contact angle hysteresis and evaporation stains.

    PubMed

    Li, Yueh-Feng; Sheng, Yu-Jane; Tsao, Heng-Kwong

    2014-07-01

    The presence of nonvolatile solutes in a liquid drop on a solid surface can affect the wetting properties. Depending on the surface-activity of the solutes, the extent of contact angle hysteresis (CAH) can vary with their concentration and the pattern of the evaporation stain is altered accordingly. In this work, four types of concentration-dependent CAH and evaporation stains are identified for a water drop containing polymeric additives on polycarbonate. For polymers without surface-activity such as dextran, advancing and receding contact angles (?a and ?r) are independent of solute concentrations, and a concentrated stain is observed in the vicinity of the drop center after complete evaporation. For polymers with weak surface-activity such as poly(ethylene glycol) (PEG), both ?a and ?r are decreased by solute addition, and the stain pattern varies with increasing PEG concentration, including a concentrated stain and a mountain-like island. For polymers with intermediate surface-activity such as sodium polystyrenesulfonate (NaPSS), ?a descends slightly, but ?r decreases significantly after the addition of a substantial amount of NaPSS, and a ring-like stain pattern is observed. Moreover, the size of the ring stain can be controlled by NaPSS concentration. For polymers with strong surface-activity such as poly(vinylpyrrolidone) (PVP), ?a remains essentially a constant, but ?r is significantly lowered after the addition of a small amount of PVP, and the typical ring-like stain is seen. PMID:24933206

  20. Methylene blue selectively stains intestinal metaplasia in Barrett's esophagus

    Microsoft Academic Search

    Marcia Irene F. Canto; Sebouh Setrakian; Robert E. Petras; Edmond Blades; Amitabh Chak; Michael V. Sivak

    1996-01-01

    Background: Specialized columnar epithelium in Barrett's esophagus resembles gastric intestinal metaplasia, which selectively stains with methylene blue. Methods: We prospectively evaluated the safety, accuracy, reproducibility, cost, and diagnostic yield of methylene blue–directed biopsy in detecting specialized columnar epithelium and dysplasia in Barrett's esophagus. We performed upper endoscopy with methylene blue–directed biopsy and obtained 236 large cup biopsy specimens (145 stained,

  1. Silver Staining of 2D Electrophoresis Gels Thierry Rabilloud

    E-print Network

    Paris-Sud XI, Université de

    Silver Staining of 2D Electrophoresis Gels Thierry Rabilloud CEA-DSV-iRTSV/LCBM and UMR CNRS separation on polyacrylamide gels. It combines excellent sensitivity (in the low nanogram range) with the use spectrometry, quantification, polyacrylamide gels, protein visualisation, silver staining #12;1. Introduction

  2. 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...

  3. 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...

  4. 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....

  5. Dynamic staining of bacteria at a single-cell level

    NASA Astrophysics Data System (ADS)

    Nuñez, Vicente; Upadhyayula, Srigokul; Lin, Adam; Chau, Kenny; Vullev, Valentine I.

    2011-05-01

    Bacterial infectious diseases remain one of the major health hazards nation- and worldwide. The expedience of detection and identification of bacterial pathogens determines how early the diagnosis is, and hence, what the treatment and the outcome of the illness would be. As we have previously reported, the dynamics of fluorescence staining provides venues for the development of expedient assays for detection and identification of bacterial species[1]. We measured the kinetics of bacterial staining with cyanine and thioflavin dyes and investigated their photophysical properties. We demonstrated that the pseudo first-order kinetic constants of the fluorescence staining processes have species specificity without contrition dependence. Combining the dynamics of staining with real-time fluorescence microscopy we characterized the fluorescence staining process at the single-cell level with improved sensitivity and contrast.

  6. Mapping stain distribution in pathology slides using whole slide imaging

    PubMed Central

    Yeh, Fang-Cheng; Ye, Qing; Hitchens, T. Kevin; Wu, Yijen L.; Parwani, Anil V.; Ho, Chien

    2014-01-01

    Background: Whole slide imaging (WSI) offers a novel approach to digitize and review pathology slides, but the voluminous data generated by this technology demand new computational methods for image analysis. Materials and Methods: In this study, we report a method that recognizes stains in WSI data and uses kernel density estimator to calculate the stain density across the digitized pathology slides. The validation study was conducted using a rat model of acute cardiac allograft rejection and another rat model of heart ischemia/reperfusion injury. Immunohistochemistry (IHC) was conducted to label ED1+ macrophages in the tissue sections and the stained slides were digitized by a whole slide scanner. The whole slide images were tessellated to enable parallel processing. Pixel-wise stain classification was conducted to classify the IHC stains from those of the background and the density distribution of the identified IHC stains was then calculated by the kernel density estimator. Results: The regression analysis showed a correlation coefficient of 0.8961 between the number of IHC stains counted by our stain recognition algorithm and that by the manual counting, suggesting that our stain recognition algorithm was in good agreement with the manual counting. The density distribution of the IHC stains showed a consistent pattern with those of the cellular magnetic resonance (MR) images that detected macrophages labeled by ultrasmall superparamagnetic iron-oxide or micron-sized iron-oxide particles. Conclusions: Our method provides a new imaging modality to facilitate clinical diagnosis. It also provides a way to validate/correlate cellular MRI data used for tracking immune-cell infiltration in cardiac transplant rejection and cardiac ischemic injury. PMID:24672736

  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. Effects of chronic kidney disease on liver transport: quantitative intravital microscopy of fluorescein transport in the rat liver.

    PubMed

    Ryan, Jennifer C; Dunn, Kenneth W; Decker, Brian S

    2014-12-15

    Clinical studies indicate that hepatic drug transport may be altered in chronic kidney disease (CKD). Uremic solutes associated with CKD have been found to alter the expression and/or activity of hepatocyte transporters in experimental animals and in cultured cells. However, given the complexity and adaptability of hepatic transport, it is not clear whether these changes translate into significant alterations in hepatic transport in vivo. To directly measure the effect of CKD on hepatocyte transport in vivo, we conducted quantitative intravital microscopy of transport of the fluorescent organic anion fluorescein in the livers of rats following 5/6th nephrectomy, an established model of CKD. Our quantitative analysis of fluorescein transport showed that the rate of hepatocyte uptake was reduced by ?20% in 5/6th nephrectomized rats, consistent with previous observations of Oatp downregulation. However, the overall rate of transport into bile canaliculi was unaffected, suggesting compensatory changes in Mrp2-mediated secretion. Our study suggests that uremia resulting from 5/6th nephrectomy does not significantly impact the overall hepatic clearance of an Oatp substrate. PMID:25339682

  9. 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.

  10. Staining with methylthioninium chloride for the diagnosis of fungal keratitis

    PubMed Central

    LAN, LAN; WANG, FENG-YUN; ZENG, GUANGWEI

    2013-01-01

    The aim of this study was to investigate the potential efficacy of staining with methylthioninium chloride (MC) for the diagnosis of fungal keratitis. A total of 70 cases of fungal keratitis were included in the study from January 2009 to December 2010. The corneal scraping specimens of the patients were collected and stained with MC or a 10% potassium hydroxide (KOH)-based smear prior to microscopic examination. The staining results were confirmed with fungal culture and strain identification, which are recognized as ‘gold standards’ for the diagnosis of fungal keratitis. Among the 70 cases of fungal keratitis, 58 cases were positive and the positive rate was 82.86%. MC staining showed a positive rate of 62.86%, with higher levels of sensitivity (70.69%) and specificity (34.61%) compared with staining with the 10% KOH-based smear. The KOH-based smear showed a positive rate of 44.29% (P<0.05), a level of sensitivity of 44.83% (P<0.05) and a specificity of 17.95% (P<0.05). In addition, the MC staining showed false positive and negative rates of 25.00 and 29.31%, respectively, an accuracy index of 5.30% and positive and negative predictive values of 93.18 and 34.61%, respectively. The results indicate that MC staining is a fast and effective method for the early diagnosis of fungal keratitis. PMID:24223649

  11. DNA comet Giemsa staining for conventional bright-field microscopy.

    PubMed

    Osipov, Andreyan; Arkhangelskaya, Ekaterina; Vinokurov, Alexei; Smetanin?, Nadezhda; Zhavoronkov, Alex; Klokov, Dmitry

    2014-01-01

    This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R2>0.977). The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity) compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine. PMID:24727376

  12. DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy

    PubMed Central

    Osipov, Andreyan; Arkhangelskaya, Ekaterina; Vinokurov, Alexei; Smetanina, Nadezhda; Zhavoronkov, Alex; Klokov, Dmitry

    2014-01-01

    This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R2 > 0.977). The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity) compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine. PMID:24727376

  13. Scalable system for classification of white blood cells from Leishman stained blood stain images

    PubMed Central

    Mathur, Atin; Tripathi, Ardhendu S.; Kuse, Manohar

    2013-01-01

    Introduction: The White Blood Cell (WBC) differential count yields clinically relevant information about health and disease. Currently, pathologists manually annotate the WBCs, which is time consuming and susceptible to error, due to the tedious nature of the process. This study aims at automation of the Differential Blood Count (DBC) process, so as to increase productivity and eliminate human errors. Materials and Methods: The proposed system takes the peripheral Leishman blood stain images as the input and generates a count for each of the WBC subtypes. The digitized microscopic images are stain normalized for the segmentation, to be consistent over a diverse set of slide images. Active contours are employed for robust segmentation of the WBC nucleus and cytoplasm. The seed points are generated by processing the images in Hue-Saturation-Value (HSV) color space. An efficient method for computing a new feature, ‘number of lobes,’ for discrimination of WBC subtypes, is introduced in this article. This method is based on the concept of minimization of the compactness of each lobe. The Naive Bayes classifier, with Laplacian correction, provides a fast, efficient, and robust solution to multiclass categorization problems. This classifier is characterized by incremental learning and can also be embedded within the database systems. Results: An overall accuracy of 92.45% and 92.72% over the training and testing sets has been obtained, respectively. Conclusion: Thus, incremental learning is inducted into the Naive Bayes Classifier, to facilitate fast, robust, and efficient classification, which is evident from the high sensitivity achieved for all the subtypes of WBCs. PMID:23766937

  14. An overview of tooth discoloration: extrinsic, intrinsic and internalized stains.

    PubMed

    Sulieman, M

    2005-10-01

    The causes of tooth discoloration are varied and complex but are usually classified as being either intrinsic, extrinsic or internalized in nature. Dietary chromogens and other external elements deposit on the tooth surface or within the pellicle layer either directly or indirectly to form extrinsic discoloration. Stains within the dentine or intrinsic discoloration often results from systemic or pulpal origin, while internalized stains are the result of extrinsic stains entering the dentine via tooth defects such as cracks on the tooth surface. PMID:16262034

  15. 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

  16. 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

  17. 18. INTERIOR OF KITCHEN NO. 1 SHOWING STAINED CABINETRY ON ...

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

    18. INTERIOR OF KITCHEN NO. 1 SHOWING STAINED CABINETRY ON OPPOSITE WALL FROM PAINTED CABINETS. VIEW TO NORTHEAST. - Bishop Creek Hydroelectric System, Plant 6, Cashbaugh-Kilpatrick House, Bishop Creek, Bishop, Inyo County, CA

  18. VIEW OF THREE SOUTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ...

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

    VIEW OF THREE SOUTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ARE LOCATED ADJACENT TO THE ALTER. - U.S. Naval Base, Pearl Harbor, Chapel, Corner of Oakley & Nimitz Street, Pearl City, Honolulu County, HI

  19. INTERIOR VIEW OF ENTRY. SHOWING THE STAINED CONCRETE FLOOR AND ...

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

    INTERIOR VIEW OF ENTRY. SHOWING THE STAINED CONCRETE FLOOR AND WINDOW WITH DIAMOND PATTERN MUNTINS. VIEW FACING NORTHWEST. - Hickam Field, Officers' Housing Type F, 602 Beard Avenue, Honolulu, Honolulu County, HI

  20. VIEW OF THREE NORTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ...

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

    VIEW OF THREE NORTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ARE LOCATED ADJACENT TO THE ALTAR. - U.S. Naval Base, Pearl Harbor, Chapel, Corner of Oakley & Nimitz Street, Pearl City, Honolulu County, HI

  1. 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

  2. 4. September 1969 DETAIL OF STAINED GLASS WINDOWS IN EAST ...

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

    4. September 1969 DETAIL OF STAINED GLASS WINDOWS IN EAST WALL, INTERIOR VIEW FROM BALCONY - Mount Zion United Methodist Church, 1334 Twenty-ninth Street Northwest, Washington, District of Columbia, DC

  3. VIEW OF THREE NORTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ...

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

    VIEW OF THREE NORTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ARE LOCATED JUST BELOW THE CHOIR LOFT. - U.S. Naval Base, Pearl Harbor, Chapel, Corner of Oakley & Nimitz Street, Pearl City, Honolulu County, HI

  4. INTERIOR VIEW OF THE LANAI. SHOWING THE ORIGINAL STAINED CONCRETE ...

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

    INTERIOR VIEW OF THE LANAI. SHOWING THE ORIGINAL STAINED CONCRETE FLOOR WITH INCISED LINES, AND HINGED DOOR TO GARAGE WITH VERTICAL BOARD PANELING (BACKGROUND). VIEW FACING NORTHWEST. - Hickam Field, Officers' Housing Type J, 701 Beard Street, Honolulu, Honolulu County, HI

  5. Basal-cell carcinoma complicating a port-wine stain.

    PubMed

    Duhra, P; Foulds, I S

    1991-01-01

    A rare radio-resistant basal-cell carcinoma is described which presented as a recently enlarged non-ulcerated nodule on a port-wine stain. The literature on basal-cell carcinoma occurring on a port-wine stain is reviewed, and the aetiological significance of thorium-X, sun-exposure and vascular changes in the development of malignancy on this type of haemangioma is discussed. PMID:2025941

  6. Dyes and stains: from molecular structure to histological application.

    PubMed

    Veuthey, Tania; Herrera, Georgina; Dodero, Veronica I

    2014-01-01

    In the present review, the chemistry of dyes as well as the interaction mechanisms between tissue and dye has been detailed, and also some of the key factors affecting the selectivity of dyes by certain cellular structures have been mentioned. Moreover, due to the relevance that histological stains have acquired in biomedical research, some of the most common stains have been described, pointing out previous and current applications in basic and applied research. PMID:24389174

  7. Interior detail view, surviving stained glass panel in an east ...

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

    Interior detail view, surviving stained glass panel in an east aisle window. Most of the stained glass has been removed from the building and relocated to other area churches. (Similar to HABS No. PA-6694-25). - Acts of the Apostles Church in Jesus Christ, 1400-28 North Twenty-eighth Street, northwest corner of North Twenty-eighth & Master Streets, Philadelphia, Philadelphia County, PA

  8. Fate of pGFP-Bearing Escherichia coli O157:H7 in Ground Beef at 2 and 10°C and Effects of Lactate, Diacetate, and Citrate

    PubMed Central

    Ajjarapu, Srilatha; Shelef, Leora A.

    1999-01-01

    Although beef has been implicated in the largest outbreaks of Escherichia coli O157:H7 infection in the United States, studies on the fate of this pathogen have been limited. Problems in such studies are associated with detection of the pathogen at levels considerably lower than the levels of the competing microorganisms. In the present study, a green fluorescent protein-expressing E. coli O157:H7 strain was used, and the stable marker allowed us to monitor the behavior of the pathogen in ground beef stored aerobically from freshness to spoilage at 2 and 10°C. In addition, the effects of sodium salts of lactate (SL) (0.9 and 1.8%), diacetate (SDA) (0.1 and 0.2%), and buffered citrate (SC) (1 and 2%) and combinations of SL and SDA were evaluated. SC had negligible antimicrobial activity, and SL delayed microbial growth, while SDA and SL plus SDA were most inhibitory to the total-aerobe population in the meat. At 2°C, the initial numbers of E. coli O157:H7 (3 and 5 log10 CFU/g) decreased by ?1 log10 CFU/g when spoilage was manifest (>7 log10 CFU of total aerobes/g), irrespective of the treatment. There was no decline in the numbers of the pathogen during storage at 10°C. Our results showed that the pathogen was resistant to the salts tested and confirmed that refrigerated meat contaminated with the pathogen remains hazardous. PMID:10583994

  9. The cross-metathesis of methyl oleate with cis-2-butene-1,4-diyl diacetate and the influence of protecting groups

    PubMed Central

    Pérez Gomes, Jessica

    2011-01-01

    Summary Background: ?,?-Difunctional substrates are useful intermediates for polymer synthesis. An attractive, sustainable and selective (but as yet unused) method in the chemical industry is the oleochemical cross-metathesis with preferably symmetric functionalised substrates. The current study explores the cross-metathesis of methyl oleate (1) with cis-2-butene-1,4-diyl diacetate (2) starting from renewable resources and quite inexpensive base chemicals. Results: This cross-metathesis reaction was carried out with several phosphine and N-heterocyclic carbene ruthenium catalysts. The reaction conditions were optimised for high conversions in combination with high cross-metathesis selectivity. The influence of protecting groups present in the substrates on the necessary catalyst loading was also investigated. Conclusions: The value-added methyl 11-acetoxyundec-9-enoate (3) and undec-2-enyl acetate (4) are accessed with nearly quantitative oleochemical conversions and high cross-metathesis selectivity under mild reaction conditions. These two cross-metathesis products can be potentially used as functional monomers for diverse sustainable polymers. PMID:21286387

  10. Characterization of 12-molybdophosphoric acid supported on mesoporous silica MCM-41 and its catalytic performance in the synthesis of hydroquinone diacetate

    NASA Astrophysics Data System (ADS)

    Ahmed, Awad I.; Samra, S. E.; El-Hakam, S. A.; Khder, A. S.; El-Shenawy, H. Z.; El-Yazeed, W. S. Abo

    2013-10-01

    12-molybdophosphoric acid (PMA) was supported on mesoporous molecular sieves MCM-41 by impregnation of 12-molybdophosphoric acid followed by calcination. The nanochannels of MCM-41 provide a large surface area for the solid state dispersion of 12-molybdophosphoric acid. The samples have been characterized by N2 adsorption-desorption at -196 °C, transmission electron microscopy (TEM), powder X-ray diffraction (PXRD), and FT-IR measurements. The acidity and catalytic activity have been, respectively, examined by nonaqueous titration of n-butylamine in acetonitrile and synthesis of hydroquinone diacetate. The results showed that ordered hexagonal pore structure was observed in the synthesized MCM-41. Also the results indicate that PMA are highly dispersed on mesoporous silica MCM-41 spherical nanoparticles while PMA retains its Keggin structure. On the other hand, with increasing the introduced PMA amount, the specific surface area decreases, and the mesoporous ordering of the samples become poor. Both the surface acidity and the catalytic activity sharply increase with the modification of MCM-41 by PMA but decrease by increasing the calcination temperature. The sample with 55 wt% PMA/MCM-41 calcined at 350 °C shows the highest acidity and catalytic activity.

  11. Spreading Depression Increases Immunohistochemical Staining of Glial Fibrillary Acidic Protein

    PubMed Central

    Kraig, Richard P.; Dong, Liming; Thisted, Ronald; Jaeger, Christine B.

    2009-01-01

    Reactive astrocytosis is a process by which astrocytes respond to brain injury by showing an increase in glial fibrillary acidic protein (GFAP) staining that is associated with hypertrophy and/or hyperplasia of these cells. Because spreading depression (SD) is a perturbation uncomplicated by neuronal necrosis and is seen in both in vivo and in vitro neural structures, we sought to determine whether SD was a sufficient stimulus to induce enhanced GFAP staining. SD was elicited in anesthetized rats by application of KCI to parietal cortex for 3 hr; equimolar NaCl was applied to contralateral cortex. SD was confirmed by monitoring DC potentials in frontal neocortices. Animals were allowed to recover for 48 hr, and their brains were processed for semiquantitative and computer-based analyses of GFAP staining intensity. Experimental GFAP staining was referenced to contralateral control levels. Neocortical SD (13–37 SDs) was associated with a significant (p < 10?4), 43% increase in GFAP staining intensity, which remained statistically greater than normal for more than 2 weeks. If SD was inhibited by combined hyperoxia and hypercarbia, only a nonsignificant (p > 0.20), 7% increase in GFAP staining was seen. Thus, SD may be a useful physiologic process with which to begin to explore the cellular mechanisms that induce the transformation of normal astrocytes into reactive species. PMID:1906091

  12. Gram Staining for the Treatment of Peritonsillar Abscess

    PubMed Central

    Takenaka, Yukinori; Takeda, Kazuya; Yoshii, Tadashi; Hashimoto, Michiko; Inohara, Hidenori

    2012-01-01

    Objective. To examine whether Gram staining can influence the choice of antibiotic for the treatment of peritonsillar abscess. Methods. Between 2005 and 2009, a total of 57 cases of peritonsillar abscess were analyzed with regard to cultured bacteria and Gram staining. Results. Only aerobes were cultured in 16% of cases, and only anaerobes were cultured in 51% of cases. Mixed growth of aerobes and anaerobes was observed in 21% of cases. The cultured bacteria were mainly aerobic Streptococcus, anaerobic Gram-positive cocci, and anaerobic Gram-negative rods. Phagocytosis of bacteria on Gram staining was observed in 9 cases. The bacteria cultured from these cases were aerobic Streptococcus, anaerobic Gram-positive cocci, and anaerobic Gram-negative rods. The sensitivity of Gram staining for the Gram-positive cocci and Gram-negative rods was 90% and 64%, respectively. The specificity of Gram staining for the Gram-positive cocci and Gram-negative rods was 62% and 76%, respectively. Most of the Gram-positive cocci were sensitive to penicillin, but some of anaerobic Gram-negative rods were resistant to penicillin. Conclusion. When Gram staining shows only Gram-positive cocci, penicillin is the treatment of choice. In other cases, antibiotics effective for the penicillin-resistant organisms should be used. PMID:22518156

  13. An improved method for staining cell colonies in clonogenic assays

    PubMed Central

    Natale, Leanna; Markowitz, Sanford D.

    2007-01-01

    Clonogenic assay is a widely used experimental approach to test for the effects of drugs/genes on the growth and proliferative characteristics of cells in vitro. Accurate quantitation of treatment effects in clonogeneic assays depends on the ability to visualize and count cell colonies precisely. We report a novel method (referred as ETeB) for staining cell colonies grown on plastic and specially coated substrates like collagen. Using colon cancer cell lines grown on plastic and collagen, we compared the colony staining efficiencies of the widely used methylene blue, and Ethidium bromide (ETeB) stains. Results show that the ETeB protocol works well on plastic and is extremely effective for staining colonies on collagen when compared to methylene blue. The key features and advantages of ETeB technique are; (a) reduction in background for colonies grown on collagen and possibly other substrates, (b) the whole procedure takes less than a minute, (c) no post-stain washing step is required which eliminates colony losses for cell lines that are loosely adherent, (d) colony visualization and counting can be done immediately following the staining procedure using a standard UV illuminator and software, and (e) the method works across a wide variety of cell lines. The simplicity and robustness of this procedure should warrant its usage in both small and large-scale clonogenic experiments. PMID:19003022

  14. Spreading depression increases immunohistochemical staining of glial fibrillary acidic protein.

    PubMed

    Kraig, R P; Dong, L M; Thisted, R; Jaeger, C B

    1991-07-01

    Reactive astrocytosis is a process by which astrocytes respond to brain injury by showing an increase in glial fibrillary acidic protein (GFAP) staining that is associated with hypertrophy and/or hyperplasia of these cells. Because spreading depression (SD) is a perturbation uncomplicated by neuronal necrosis and is seen in both in vivo and in vitro neural structures, we sought to determine whether SD was a sufficient stimulus to induce enhanced GFAP staining. SD was elicited in anesthetized rats by application of KCI to parietal cortex for 3 hr; equimolar NaCI was applied to contralateral cortex. SD was confirmed by monitoring DC potentials in frontal neocortices. Animals were allowed to recover for 48 hr, and their brains were processed for semiquantitative and computer-based analyses of GFAP staining intensity. Experimental GFAP staining was referenced to contralateral control levels. Neocortical SD (13-37 SDs) was associated with a significant (p less than 10(-4)), 43% increase in GFAP staining intensity, which remained statistically greater than normal for more than 2 weeks. If SD was inhibited by combined hyperoxia and hypercarbia, only a nonsignificant (p greater than 0.20), 7% increase in GFAP staining was seen. Thus, SD may be a useful physiologic process with which to begin to explore the cellular mechanisms that induce the transformation of normal astrocytes into reactive species. PMID:1906091

  15. 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

  16. The Incidence of Staining of Permanent Teeth by the Tetracyclines

    PubMed Central

    Conchie, J. M.; Munroe, J. D.; Anderson, D. O.

    1970-01-01

    In this investigation an attempt has been made to determine the relationship between the staining of permanent teeth by tetracycline administered during the period of tooth formation with the dosage of the drug and the duration of therapy. Of 238 subjects whose hospital records indicated ingestion of stated doses of tetracycline, some 49 were seen to have staining which was confirmed by fluorescence, and a further six had staining which did not fluoresce and hence could not be confirmed. A definite relationship between total dosage and staining and duration of administration and staining was established; the condition occurred with greater frequency (in more than one-third of the children) when the total dosage exceeded 3 g. or the duration of treatment was longer than 10 days. However, as staining was seen at all dosage levels, whatever the duration, physicians should continue to follow previous advice and prescribe other antibiotics where possible for children under 8 years of age or for women in the last trimester of pregnancy. PMID:5447715

  17. Integrating photoacoustic ophthalmoscopy with scanning laser ophthalmoscopy, optical coherence tomography, and fluorescein angiography for a multimodal retinal imaging platform

    NASA Astrophysics Data System (ADS)

    Song, Wei; Wei, Qing; Liu, Tan; Kuai, David; Burke, Janice M.; Jiao, Shuliang; Zhang, Hao F.

    2012-06-01

    Photoacoustic ophthalmoscopy (PAOM) is a newly developed retinal imaging technology that holds promise for both fundamental investigation and clinical diagnosis of several blinding diseases. Hence, integrating PAOM with other existing ophthalmic imaging modalities is important to identify and verify the strengths of PAOM compared with the established technologies and to provide the foundation for more comprehensive multimodal imaging. To this end, we developed a retinal imaging platform integrating PAOM with scanning laser ophthalmoscopy (SLO), spectral-domain optical coherence tomography (SD-OCT), and fluorescein angiography (FA). In the system, all the imaging modalities shared the same optical scanning and delivery mechanisms, which enabled registered retinal imaging from all the modalities. High-resolution PAOM, SD-OCT, SLO, and FA images were acquired in both albino and pigmented rat eyes. The reported in vivo results demonstrate the capability of the integrated system to provide comprehensive anatomic imaging based on multiple optical contrasts.

  18. 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

  19. 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.

  20. A new in vivo staining method, cresyl violet staining, for fiberoptic magnified observation of carcinoma of the gastric mucosa

    Microsoft Academic Search

    Yuichi Furuta; Oichiro Kobori; Hisaaki Shimazu; Yasuhiko Morioka; Yamagi Okuyama

    1985-01-01

    Summary  A polypoid gastric mucosal lesion with a flat portion was observed by a fiberoptic magnifying endoscope with a new in vivo\\u000a staining method, cresyl violet staining. The fine surface structure of this lesion showed a characteristic “ruined sulciform\\u000a pattern” suggesting papillary carcinoma histologically. Successful application of the findings of dissecting microscopy to\\u000a clinical magnifying endoscopy by means of this new

  1. Unexpected sensitization effeciency of the near-infrared Nd3+, Er3+ and Yb3+ emission by fluorescein compared to eosin and erythrosin

    Microsoft Academic Search

    Gerald A. Hebbink; Lennart Grave; Leon A. Woldering; David N. Reinhoudt; Veggel van Frank C. J. M

    2003-01-01

    Near-infrared emissive lanthanide complexes were synthesized with covalently attached sensitizers that absorb in the visible. This functionalization was designed such that the sensitizer is in close proximity to the lanthanide ion, which is a prerequisite for efficient energy transfer from the excited sensitizer to the lanthanide ion. The sensitizers used were fluorescein, eosin, and erythrosin, which were linked via a

  2. The effects of ruthenium red, lanthanum, fluorescein isothiocyanate and trifluoperazine on vesicle transport, vesicle fusion and tip extension in pollen tubes

    Microsoft Academic Search

    Jill M. Picton; Martin W. Steer

    1985-01-01

    The effects of ruthenium red, lanthanum, fluorescein isothiocyanate and trifluoperazine, all antagonists of Ca2+ function in cells, have been studied in growing pollen tubes of Tradescantia virginiana. All four drugs inhibit pollen-tube growth but bring about different ultrastructural changes at the growing tips and within the cytoplasm. The results strongly support the hypothesis that Ca2+ plays a vital role in

  3. [Usefulness and limit of Gram staining smear examination].

    PubMed

    Nagata, Kuniaki; Mino, Hirotoshi; Yoshida, Shunsuke

    2010-05-01

    Gram staining is one of the most simple and inexpensive methods for the rapid diagnosis of bacterial and fungal infections. It yields results much faster than culture, and provides important data for the patient's treatment and prognosis. However, a difference exists in the quality and quantity of information yielded by Gram staining smears based on the experience and knowledge of those conducting the tests. Therefore, a risk of misdiagnosis based on the information obtained from Gram staining smears is also present. The Gram staining conditions and morphology of bacteria sometimes change due to antimicrobial therapy. Species of Gram-negative rods sometimes become filamentous and pleomorphic. Gram-positive bacteria may become gram variable (change in staining condition) after antimicrobial therapy. Even bacteria that are easy to mis-identify exist, because the morphology of bacteria may be similar. Enterococcus faecalis is a Gram-positive diplococcus, forming Gram-positive clustered cocci in specimens from blood culture bottles, resembling Streptococcus pneumoniae. Acinetobacter baumannii is a Gram-negative diplococcus in sputum, resembling Moraxella (Branhamella) catarrhalis. Pasteurella multocida is a small-sized, Gram-negative short rod in the sputum, resembling Haemophilus influenzae. Prevotella intermedia is a small-sized, Gram-negative short rod in sputum, resembling Haemophilus influenzae. Capnocytophaga sp. is a Gram-negative fusiform (thin needle shape) rod present in clinical specimens, resembling Fusobacterium nucleatum. PMID:20560458

  4. STRUCTURE OF ISOLATED PLANT GOLGI APPARATUS REVEALED BY NEGATIVE STAINING

    PubMed Central

    Cunningham, William P.; Morré, D. James; Mollenhauer, H. H.

    1966-01-01

    Sucrose-gradient-purified dictyosomes of plant Golgi apparatus appear, after glutaraldehyde stabilization, as stacks of highly fenestrate and tubate cisternae when negatively stained with phosphotungstic acid, shadowed with heavy metal, or OsO4-stained in thin section. The tubular proliferations (diameter 200 to 400 A) extend for several microns from the central region and are united at intervals into an anastomosing network. Associated with the tubules are two kinds of vesicles which are distinguishable on the basis of texture, size, shape, and staining characteristics. One vesicle type is rough-surfaced, nearly spherical, and of uniform dimensions (diameter approximately 600 A). Metal shadowing shows that these vesicles remain spherical after drying. The other vesicle type is smooth-surfaced and varies in both size and shape. Intercisternal elements are revealed, by negative staining, on the surface of internal cisternae after fragmentation of the dictyosome. The progressive differentiation of cisternae from the forming face to the maturing face is observed in thin sections of these isolated preparations. The morphological characteristics observed in negatively stained dictyosomes indicate regions of functional specialization within the dictyosome cisternae and reveal a dictyosome structure more extensive than that envisioned from sections. PMID:4161888

  5. 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.

  6. 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.

  7. Evaluation of forensic examination of extremely aged seminal stains.

    PubMed

    Nakanishi, Hiroaki; Hara, Masaaki; Takahashi, Shirushi; Takada, Aya; Saito, Kazuyuki

    2014-09-01

    The results of forensic tests, such as semen identification and short tandem repeat (STR) analysis of extremely aged seminal stains from unsolved sex crimes can provide important evidence. In this study we evaluated whether current forensic methods could be applied to seminal stains that were stored at room temperature for 33-56years (n=2, 33years old; n=1, 41years old; n=1, 44years old; n=1, 56years old). The prostatic acid phosphatase (SM-test reagent), microscopic (Baecchi stain method) and semenogelin (RSID™ Semen Laboratory Kit) tests were performed as discriminative tests for semen. In addition, the mRNA levels of the semen-specific proteins semenogelin 1 (SEMG1) and protamine 2 (PRM2) were investigated. STRs were analyzed using the AmpFlSTR® Identifiler™ PCR Amplification Kit. All samples were positive in the prostatic acid phosphatase and semenogelin tests, and sperm heads were identified in all samples. The staining degree of the aged sperm heads was similar to that of fresh sperm. Although SEMG1 mRNA was not detected in any sample, PRM2 mRNA was detected in three samples. In the STR analysis, all loci were detected in the 33-years-old sample and five loci were detected in the 56-years-old sample. We confirmed that current forensic examinations - including STR analysis - could be applied to extremely aged seminal stains. These results could be useful for forensic practice. PMID:24844186

  8. Evaluation of a fluorescent lectin-based staining technique for some acidophilic mining bacteria

    SciTech Connect

    Fife, D.J.; Bruhn, D.F.; Miller, K.S.; Stoner, D.L.

    2000-05-01

    A fluorescence-labeled wheat germ agglutinin staining technique was modified and found to be effective for staining gram-positive, acidophilic mining bacteria. Bacteria identified by others as being gram positive through 16S rRNA sequence analyses, yet clustering near the divergence of that group, stained weakly. Gram-negative bacteria did not stain. Background staining of environmental samples was negligible, and pyrite and soil particles in the samples did not interfere with the staining procedure.

  9. A staining protocol for identifying secondary compounds in Myrtaceae1

    PubMed Central

    Retamales, Hernan A.; Scharaschkin, Tanya

    2014-01-01

    • Premise of the study: Here we propose a staining protocol using toluidine blue (TBO) and ruthenium red to reliably identify secondary compounds in the leaves of some species of Myrtaceae. • Methods and Results: Leaves of 10 species representing 10 different genera of Myrtaceae were processed and stained using five different combinations of ruthenium red and TBO. Optimal staining conditions were determined as 1 min of ruthenium red (0.05% aqueous) and 45 s of TBO (0.1% aqueous). Secondary compounds clearly identified under this treatment include mucilage in the mesophyll, polyphenols in the cuticle, lignin in fibers and xylem, tannins and carboxylated polysaccharides in the epidermis, and pectic substances in the primary cell walls. • Conclusions: Potential applications of this protocol include systematic, phytochemical, and ecological investigations in Myrtaceae. It might be applicable to other plant families rich in secondary compounds and could be used as a preliminary screening method for extraction of these elements. PMID:25309840

  10. Feasibility of digitally stained multimodal confocal mosaics to simulate histopathology

    NASA Astrophysics Data System (ADS)

    Gareau, Daniel S.

    2009-05-01

    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.

  11. 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.

  12. When one plus one equals more than two--a novel stain for renal biopsies is a combination of two classical stains.

    PubMed

    Brodsky, Sergey V; Albawardi, Alia; Satoskar, Anjali A; Nadasdy, Gyongyi; Nadasdy, Tibor

    2010-11-01

    Histologic evaluation of renal biopsies includes multiple ancillary stains, including Periodic acid-Schiff's (PAS) and Masson's trichrome (Trichrome). Herein we report an innovative double-stain, derived from two standard stains (PAS and Trichrome). This novel stain not only has advantages of both ancestor stains, but became more distinguishable and colorful, when basement membranes stain dark-violet, whereas the interstitial collagen remains blue. This allows the pathologist immediate estimation of the amount of collagen, tubular atrophy and the degree of interstitial fibrosis in one section. Using computer-based analysis, we confirmed that our innovative double-stain highlights interstitial collagen better than Trichrome stain alone. We strongly recommend renal pathologists to try this innovative stain in their practice. PMID:20865661

  13. Spectroscopic studies on H2O2 damaging BSA induced by 1,2-dihydroxy-9, 10-anthraquinone-3-aminomethyl-N, N-diacetate-Ferrous(III)

    NASA Astrophysics Data System (ADS)

    Zou, Mingming; Li, Ying; Wang, Jun; Gao, Jingqun; Wang, Qi; Wang, Baoxin; Fan, Ping

    2013-08-01

    The interaction between 1,2-dihydroxy-9, 10-anthraquinone-3-aminomethyl-N, N-diacetate-Ferrous(III) (Alizarin-DA-Fe(III)) and bovine serum albumin (BSA) was studied by using UV-vis and fluorescence spectra. And then, the H2O2 damage of BSA induced by Alizarin-DA-Fe(III) was examined. The results show that due to the interaction the fluorescence of BSA solution can be obviously quenched by Alizarin-DA-Fe(III) and that the quenching process belongs to the static quenching. In addition, in the presence of Alizarin-DA-Fe(III) the BSA molecules were markedly damaged by H2O2. Meanwhile, the effects of the standing time, Alizarin-DA-Fe(III) concentration and H2O2 concentration on the damage of BSA molecules were also researched. The experimental results demonstrate that the damage degree increase with the increase of standing time, Alizarin-DA-Fe(III) concentration and H2O2 concentration. Finally, the generation of reactive oxygen species (ROS) from H2O2 induced by Alizarin-DA-Fe(III) as Fenton-like reagent was estimated by some quenchers. Because the Iminodiacetic-Ferrous(III) (IDA-Fe(III)) and Nitrilotriacetic-Ferrous(III) (NTA-Fe(III)) can be thought of as the active part of Alizarin-DA-Fe(III), they were used to compare the catalytic activity with Alizarin-DA-Fe(III). Owing to the special plane structure, the experiment results showed that the Alizarin-DA-Fe(III) exhibited higher damage ability than IDA-Fe(III) and NTA-Fe(III). Perhaps, the Alizarin-DA-Fe(III) may be used as a new antitumor compound to induce peroxides in body to kill cancer cells.

  14. News from the biological stain commission, no. 16.

    PubMed

    Lyon, H O

    2015-04-01

    In the 16(th) issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission's International Affairs Committee presents information from the 28(th) meeting of CEN/TC 140 In vitro diagnostic medical devices held on October 23, 2013 in Berlin, Germany. Information is also presented from the 19(th) meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on October 19 - 21, 2013 in Singapore. PMID:25747046

  15. Toward Digital Staining using Imaging Mass Spectrometry and Random Forests

    PubMed Central

    Hanselmann, Michael; Köthe, Ullrich; Kirchner, Marc; Renard, Bernhard Y.; Amstalden, Erika R.; Glunde, Kristine; Heeren, Ron M. A.; Hamprecht, Fred A.

    2009-01-01

    We show on Imaging Mass Spectrometry (IMS) data that the Random Forest classifier can be used for automated tissue classification and that it results in predictions with high sensitivities and positive predictive values, even when inter-sample variability is present in the data. We further demonstrate how Markov Random Fields and vector-valued median filtering can be applied to reduce noise effects to further improve the classification results in a post-hoc smoothing step. Our study gives clear evidence that digital staining by means of IMS constitutes a promising complement to chemical staining techniques. PMID:19469555

  16. A sensitive cytochemical staining method for glucose-6-phosphate dehydrogenase activity in individual erythrocytes. I. Optimalization of the staining procedure.

    PubMed

    van Noorden, C J; Vogels, I M; James, J; Tas, J

    1982-01-01

    A sensitive cytochemical staining method for glucose-6-phosphate dehydrogenase activity in individual human erythrocytes is described. This staining method can be used for the rapid routine discrimination of patients with a deficiency of the enzyme in its homozygote or heterozygote form, but also for quantitative localization of its activity in individual erythrocytes. The staining procedure in its optimal form consists of a treatment of the erythrocytes with sodium nitrite, then a "fixation" in 0.025% glutaraldehyde (under NADP+ protection of the active site of the enzyme), followed by incubation of the cells in suspension in the presence of tetranitro BT, 1-methoxyphenazine methosulphate and polyvinyl alcohol. Using this new technique, a sharp localization is obtained of the glucose-6-phosphate dehydrogenase activity, which enables discrimination between red cells with different levels of enzyme activity, as a consequence of enzyme deficiencies or age changes. PMID:6184337

  17. Alcian Blue Alizarin Red Skeletal Staining October 2003

    E-print Network

    De Robertis, Eddy M.

    by removing the skin and organs completely. Remove the fat from the mice but be careful that damage damage. 3. Replace 95% ethanol with Alcian blue staining solution for 1-3 days slowly rocking at room. Older animals will require more time than younger ones. If you want to remove any remaining fat or skin

  18. 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...

  19. 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...

  20. Original article Double staining (CTC-DAPI) for detection and

    E-print Network

    Paris-Sud XI, Université de

    Original article Double staining (CTC-DAPI) for detection and enumeration of viable but non cultivable INTRODUCTION A viable but non-culturable (VNC) bacte- rial state was originally detected of human pathogens: Escherichia coli (Xu et al, 1982), Salmonella enteritidi.s (Roszak et al, 1984), Vibrio

  1. 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.

  2. Quantitation of catalase activity by microspectrophotometry after diaminobenzidine staining

    Microsoft Academic Search

    A. Geerts; F. Roels

    1981-01-01

    The absorbance of the reaction product of catalase staining with diaminobenzidine is linearly proportional to enzyme activity. This is shown in semithin Epon sections of model systems containing serum albumin and catalase from bovine or guinea pig liver. Absorbance measurements were also performed on semithin sections of guinea pig liver, and from these, the activity of cytoplasmic (extraperoxisomal) catalase has

  3. STAINING OF TISSUE SECTIONS FOR ELECTRON MICROSCOPY WITH HEAVY METALS

    Microsoft Academic Search

    M. L. Watson

    1958-01-01

    ABS>Heavy metals may be incorporated from solution into tissue sections ; for electron microscopy. The resulting increase in density of the tissue ; provides greatly enhanced contrast with minimal distortion. Relative densities ; of various structures are found to depend on the heavy metal ions present and on ; the conditions of staining. Certain hitherto unobserved details are revealed and

  4. Epiluminescence Microscopy for Port-Wine Stains: Pretreatment Evaluation

    Microsoft Academic Search

    Enrico Maria Procaccini; Giuseppe Argenziano; Stefania Staibano; Gerardo Ferrara; Giuseppe Monfrecola

    2001-01-01

    Background: Port-wine stains (PWSs) are characterized by an increased number of ectatic vessels. The treatment of choice is the use of some lasers such as pulsed dye lasers. However, some lesions are nonresponsive to laser treatment. Perhaps the vessels’ depth and diameter and the thickness of the vessel wall are important factors influencing the effectiveness of the laser treatment. Methods:

  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. Infrared fluorescence microscopy of stained tissues: Principles and technic

    Microsoft Academic Search

    Holde Puchtler; Susan N. Meloan; L. D. Paschal

    1980-01-01

    Infrared photomicrography was used extensively from 1927 to the 1940's, but received little attention during the last decades. However, studies of infrared fluorescence of stained sections could not be found in the accessible literature. Ramsley (1968) published quantitative data on infrared fluorescence of approximately 250 dyes bound to textile fibers. The intensity of infrared fluorescence of many dyes varied widely

  7. 0038-9153/80/5503-0173$02200/0 STAIN TECHNOLOGY

    E-print Network

    Murphy, Bob

    procedures.This substitution significantly reduces the cost of staining for adenylate kinase, creatine kinase, glucosephosphate isomerase, mannosephosphate isomerase: phosphoglucomutase, and pyruvate kinase activity by uti the formation of adenosine-5'-triphosphate (ATP) in the initial reaction. ATP can then act as a cofactor

  8. Staining of Internal Limiting Membrane in Macular Hole Surgery

    Microsoft Academic Search

    Kazuaki Kadonosono; Norihiko Itoh; Eiichi Uchio; Satoshi Nakamura; Shigeaki Ohno; Arch Ophthalmol

    2000-01-01

    emoval of internal limiting membranes (ILMs) is a potentially useful surgical approach to close an idiopathic macular hole. However, the removal of ILMs is diffi- cult to perform because of poor visibility of the ILMs. We have developed a technique for staining the ILM with a solution of indocyanine green to facilitate the removal of ILMs in eyes with an

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

    SciTech Connect

    Chin, I.R.; Gorrell, T.A.; Scheffler, M.J. [Wiss, Janney, Elstner Associates, Inc., Chicago, IL (United States)

    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.

  10. Histochemistry of the Gram-staining Reaction for Microorganisms

    Microsoft Academic Search

    H. Henry; M. Stacey

    1943-01-01

    IN 1884 a Dane, Christian Gram, while working in Berlin, discovered a method of staining micro-organisms which has gone under his name ever since that date1. Although the method is in daily use by bacteriologists of various nationalities all over the world, and has by reason of its importance in ordinary routine diagnosis been the subject of extensive research, it

  11. Silver-Stained accessory structures on human sex chromosomes

    Microsoft Academic Search

    S. Pathak; F. F. B. Elder

    1980-01-01

    Using a combination of silver-staining and light microscopic techniques on human male meiotic preparations, it is feasible to study the morphology and behavior of both autosomal synaptonemal complexes and sex chromosome axes. During leptotene and early zygotene, the X and Y chromosomes are separate; their axes appearing as thin, filamentous structures. During late zygotene\\/early pachytene, the sex chromosomes come close

  12. Analytical and microbiological characterization of paper samples exhibiting foxing stains.

    PubMed

    Nunes, Margarida; Relvas, Cátia; Figueira, Francisca; Campelo, Joana; Candeias, António; Caldeira, Ana T; Ferreira, Teresa

    2015-02-01

    This work comprises the use of a multi-analytical approach combined with microbiological studies to characterize six paper samples, containing foxing stains, from the 20th century, regarding their cellulose matrix, fillers, and sizing materials, and to evaluate possible paper degradation that might have occurred during the foxing stains. Photography under different illuminations and optical microscopy were used for morphological characterization of the paper samples and foxing stains. Scanning electron microscopy coupled energy dispersive spectroscopy (SEM-EDS) was of particular importance for defining the presence of fiber disorder and disruption on the surface of some of the stains, and localized accumulations of mineral-like particles on the surface of others. SEM-EDS, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FT-IR), and energy dispersive X-ray fluorescence (EDXRF) were used for the identification of mineral fillers, whereas sizing agents were analyzed using ATR-FT-IR. EDXRF results showed that no differences, within the standard deviation, were found in iron and copper contents between the foxed and unfoxed areas. Fungi belonging to the genus Penicillium spp. were found in all the paper samples. Unfoxed areas presented lower contamination than the foxed areas. PMID:25787782

  13. Pathologic evaluation of inguinal sentinel lymph nodes in vulvar cancer patients: a comparison of immunohistochemical staining versus ultrastaging with hematoxylin and eosin staining

    Microsoft Academic Search

    Richard G Moore; C. O Granai; Walter Gajewski; Mary Gordinier; Margaret M Steinhoff

    2003-01-01

    ObjectiveTo evaluate the value of immunohistochemical (IHC) staining of inguinal sentinel lymph nodes (SLN) found to be negative for metastatic disease by ultrastaging with hematoxylin and eosin (H&E) staining.

  14. Fluorescent labeling of cranberry proanthocyanidins with 5-([4,6-dichlorotriazin-2-yl]amino)fluorescein (DTAF).

    PubMed

    Feliciano, Rodrigo P; Heintz, Joseph A; Krueger, Christian G; Vestling, Martha M; Reed, Jess D

    2015-01-01

    A novel methodology was developed to elucidate proanthocyanidins (PAC) interaction with extra-intestinal pathogenic Escherichia coli (ExPEC). PAC inhibit ExPEC invasion of epithelial cells and, therefore, may prevent transient gut colonization, conferring protection against subsequent extra-intestinal infections, such as urinary tract infections. Until now PAC have not been chemically labeled with fluorophores. In this work, cranberry PAC were labeled with 5-([4,6-dichlorotriazin-2-yl]amino) fluorescein (DTAF), detected by high-performance liquid chromatography with diode-array detection and characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). We report single and double fluorescent-labeled PAC with one or two chlorine atoms displaced from DTAF in alkaline pH via nucleophilic substitution. Fluorescent labeling was confirmed by fragmentation experiments using MALDI-TOF/TOF MS. Fluorescent labeled PAC were able to promote ExPEC agglutination when observed with fluorescence microscopy. DTAF tagged PAC may be used to trace the fate of PAC after they agglutinate ExPEC and follow PAC-ExPEC complexes in cell culture assays. PMID:25053065

  15. Quantitative observation of focused-ultrasound-induced vascular leakage and deformation via fluorescein angiography and optical coherence tomography.

    PubMed

    Tsai, Meng-Tsan; Lee, Cheng-Kuang; Lin, Kung-Min; Lin, Yu-Xiang; Lin, Tzu-Han; Chang, Ting-Chia; Lee, Jiann-Der; Liu, Hao-Li

    2013-10-01

    Focused ultrasound (FUS) is a recently discovered noninvasive technique for local and temporal enhancement of vascular permeability, which facilitates drug delivery from the vessels into the surrounding tissue. However, exposure to FUS at a high intensity may cause permanent damage. To investigate the effects of the FUS treatment on blood vessels, we propose to use fluorescein angiography (FA) and optical coherence tomography (OCT) for real-time observation of the diffusion of fluorescence dye from blood vessels and to evaluate the morphological changes of the vessels in vivo. With time-resolved FA imaging, the relationship between the exposed power and the improved permeability of the vessels can be assessed according to the enhancement of the fluorescent intensity due to the dye leakage. Furthermore, the variation of the time-resolved fluorescent intensities can be used to identify the occurrence of dye leakage. In contrast, OCT can be implemented for the reconstruction of tissue microstructures. To quantitatively evaluate the morphological changes of the vessels after the FUS exposure with OCT, a new algorithm was proposed to estimate the vessel area based on the comparison of backscattering properties resulting from the tissue and vascular structures. Results showed that the vessel area increased as the exposed power increased, and the area became significantly larger at a higher FUS exposure power of 10 W. In conclusion, integrated FA and OCT observation can be potentially effective for monitoring the outcome and investigating the effects of FUS treatment. PMID:23812607

  16. The fluorescein isothiocyanate-binding site of the plasma-membrane H+-ATPase of Neurospora crassa.

    PubMed

    Pardo, J P; Slayman, C W

    1988-12-15

    The mammalian (Na+,K+), Ca2+-, and (H+,K+)-ATPases contain a well-characterized lysine residue that reacts with fluorescein 5'-isothiocyanate (FITC); enzymatic activity is protected by ATP, suggesting that the residue is located in or near the nucleotide-binding domain. In this study, the plasma-membrane H+-ATPase of Neurospora crassa is also shown to be sensitive to FITC. The reaction occurs with pseudo first-order kinetics, has a pKa of 8.0, and is stimulated by Mg2+. Enzymatic activity is protected by MgADP with a Kd of 0.2-0.3 mM, close to the Ki with which MgADP serves as a competitive inhibitor of ATP hydrolysis. A tryptic peptide labeled with FITC in the absence, but not in the presence, of MgADP has been isolated and sequenced. The FITC-sensitive residue is Lys474, located in a region that exhibits significant homology with the mammalian cation-transporting ATPases. PMID:2904434

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

    E-print Network

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

  18. Silver stain for proteins in polyacrylamide gels: A modified procedure with enhanced uniform sensitivity

    Microsoft Academic Search

    JAMES H. MORRISSEY

    1981-01-01

    The rapid, ultrasensitive silver stains that have been developed recently for detecting proteins in polyacrylamide gels show variation in staining from gel to gel and do not stain certain proteins at all. It was found that treatment of gels with dithiothreitol prior to impregnation with silver nitrate results in more reproducible staining patterns that are also qualitatively similar to those

  19. NMR Structure and Dynamics of the Engineered Fluorescein-Binding Lipocalin FluA Reveals Rigidification of ?-Barrel and Variable Loops upon Enthalpy-Driven Ligand Binding

    PubMed Central

    Mills, Jeffrey L.; Liu, Gaohua; Skerra, Arne; Szyperski, Thomas

    2010-01-01

    The NMR structure of the 21 kDa lipocalin FluA, which was previously obtained by combinatorial design, elucidates a reshaped binding site specific for the dye fluorescein resulting from 21 side chain replacements with respect to the parental lipocalin, the naturally occurring bilin-binding protein (BBP). As expected, FluA exhibits the lipocalin fold of BBP, comprising eight antiparallel ?-strands forming a ?-barrel with an ?-helix attached to its side. Comparison of the NMR structure of the free FluA with the X-ray structures of BBP•biliverdin IX? and FluA•fluorescein complexes revealed significant conformational changes in the binding pocket, which is formed by four loops at the open end of the ?-barrel as well as adjoining ?-strand segments. An ‘induced fit’ became apparent for the side-chain conformations of Arg 88 and Phe 99, which contact the bound fluorescein in the complex and undergo concerted rearrangement upon ligand binding. Moreover, slower internal motional modes of the polypeptide backbone were identified by measuring transverse 15N backbone spin relaxation times in the rotating frame for the free FluA and also the FluA•fluorescein complex. A reduction of such motions was detected upon complex formation, indicating rigidification of the protein structure and loss of conformational entropy. This hypothesis was confirmed by isothermal titration calorimetry, showing that ligand binding is enthalpy driven, thus overcompensating negative entropy associated with both ligand binding per se and rigidification of the protein. Our investigation of the solution structure and dynamics as well as thermodynamics of lipocalin-ligand interaction does not only provide insight into the general mechanism of small molecule accommodation in the deep and narrow cavity of this abundant class of proteins but will also support the future design of corresponding binding proteins with novel specificities, so-called “anticalins”. PMID:19603796

  20. Fluorescein and eosin as sensitizing chromophores in near-infrared luminescent ytterbium(III), neodymium(III) and erbium(III) chelates

    Microsoft Academic Search

    Martinus H. V. Werts; Johannes W. Hofstraat; Frank A. J. Geurts; Jan W. Verhoeven

    1997-01-01

    Near-infrared luminescent ytterbium(III), neodymium(III) and erbium(III) chelates containing organic chromophores derived from fluorescein and eosin have been synthesized and studied spectroscopically. The complexes can be efficiently excited with visible light and show intense lanthanide luminescence at low concentrations (2? 10?6 mol l?1) in D2O as a result of energy transfer from the dye moiety to the rare earth ion. Quenching

  1. The effect of acetazolamide on passive and active transport of fluorescein across the blood-retina barrier in retinitis pigmentosa complicated by macular oedema

    Microsoft Academic Search

    Birgitte Moldow; Birgit Sander; Michael Larsen; Claus Engler; Bin Li; Thomas Rosenberg; Henrik Lund-Andersen

    1998-01-01

    · Background: The carbonic anhydrase inhibitor acetazolamide (AZM) reduces macular oedema in some patients with retinitis\\u000a pigmentosa. To better understand the oedema-reducing effect of AZM, the effect of AZM on passive permeability and active transport\\u000a of fluorescein across the blood-retina barrier was studied in patients with retinitis pigmentosa and varying degrees of macular\\u000a oedema.?· Method: The selection of patients was

  2. Detection of Rubella Virus-Specific Immunoglobulin G in Saliva by an Amplification-Based Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibody to Fluorescein Isothiocyanate

    Microsoft Academic Search

    A. J. VYSE; D. W. G. BROWN; B. J. COHEN; R. SAMUEL; D. J. NOKES; Coventry CV

    1999-01-01

    An immunoglobulin G (IgG)-capture enzyme-linked immunosorbent assay (ELISA) for rubella virus is described. The assay uses a fluorescein isothiocyanate (FITC)-anti-FITC amplification system. The detection limit of the ELISA was approximately 7 IU of rubella virus-specific IgG per ml of serum sample. For saliva samples the performances of the capture ELISA and previously described radioimmunoassay were assessed, and the results of

  3. Hypertrophic lupus erythematosus: the diagnostic utility of CD123 staining

    PubMed Central

    Ko, Christine J.; Srivastava, Bhaskar; Braverman, Irwin; Antaya, Richard J.; McNiff, Jennifer M.

    2014-01-01

    CD123-positive plasmacytoid dendrocytes are prominent in the infiltrate of discoid lupus erythematosus (LE). We hypothesized that these cells would also be present in hypertrophic LE and would aid in the histopathologic distinction from squamous cell carcinoma (SCC) and hypertrophic actinic keratosis (AK). Five cases of hypertrophic LE and 10 cases each of SCC and hypertrophic AK were stained with CD123. A heavy band of CD123-positive cells was present at the epidermal–dermal junction in all cases of hypertrophic LE, and only single or rare scattered clusters of CD123-positive cells were seen in SCC and actinic keratoses. The pattern of CD123 staining can be a useful feature to distinguish hypertrophic LE from SCC and hypertrophic AK. PMID:21955314

  4. Identification of active fluorescence stained bacteria by Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Krause, Mario; Beyer, Beatrice; Pietsch, Christian; Radt, Benno; Harz, Michaela; Rösch, Petra; Popp, Jürgen

    2008-04-01

    Microorganisms can be found everywhere e.g. in food both as useful ingredients or harmful contaminations causing food spoilage. Therefore, a fast and easy to handle analysis method is needed to detect bacteria in different kinds of samples like meat, juice or air to decide if the sample is contaminated by harmful microorganisms. Conventional identification methods in microbiology require always cultivation and therefore are time consuming. In this contribution we present an analysis approach to identify fluorescence stained bacteria on strain level by means of Raman spectroscopy. The stained bacteria are highlighted and can be localized easier against a complex sample environment e.g. in food. The use of Raman spectroscopy in combination with chemometrical methods allows the identification of single bacteria within minutes.

  5. Selection of Ovine Oocytes by Brilliant Cresyl Blue Staining

    PubMed Central

    Wang, Liqin; Lin, Jiapeng; Huang, Juncheng; Wang, Jing; Zhao, Yuncheng; Chen, Tong

    2012-01-01

    Sheep oocytes derived from the ovaries collected from the slaughterhouse are often used for research on in vitro embryo production, animal cloning, transgenesis, embryonic stem cells, and other embryo biotechnology aspects. Improving the in vitro culture efficiency of oocytes can provide more materials for similar studies. Generally, determination of oocyte quality is mostly based on the layers of cumulus cells and cytoplasm or cytoplasm uniformity and colors. This requires considerable experience to better identify oocyte quality because of the intense subjectivity involved (Gordon (2003), Madison et al. (1992) and De Loos et al. (1992)). BCB staining is a function of glucose-6-phosphate dehydrogenase (G6PD) activity, an enzyme synthesized in developing oocytes, which decreases in activity with maturation. Therefore, unstained oocytes (BCB?) are high in G6PD activity, while the less mature oocytes stains are deep blue (BCB+) due to insuffcient G6PD activity to decolorize the BCB dye. PMID:22675245

  6. Selection of ovine oocytes by brilliant cresyl blue staining.

    PubMed

    Wang, Liqin; Lin, Jiapeng; Huang, Juncheng; Wang, Jing; Zhao, Yuncheng; Chen, Tong

    2012-01-01

    Sheep oocytes derived from the ovaries collected from the slaughterhouse are often used for research on in vitro embryo production, animal cloning, transgenesis, embryonic stem cells, and other embryo biotechnology aspects. Improving the in vitro culture efficiency of oocytes can provide more materials for similar studies. Generally, determination of oocyte quality is mostly based on the layers of cumulus cells and cytoplasm or cytoplasm uniformity and colors. This requires considerable experience to better identify oocyte quality because of the intense subjectivity involved (Gordon (2003), Madison et al. (1992) and De Loos et al. (1992)). BCB staining is a function of glucose-6-phosphate dehydrogenase (G6PD) activity, an enzyme synthesized in developing oocytes, which decreases in activity with maturation. Therefore, unstained oocytes (BCB-) are high in G6PD activity, while the less mature oocytes stains are deep blue (BCB+) due to insuffcient G6PD activity to decolorize the BCB dye. PMID:22675245

  7. 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.

  8. Cytological detection of spermatozoa: comparison of three staining methods.

    PubMed

    Allery, J P; Telmon, N; Mieusset, R; Blanc, A; Rougé, D

    2001-03-01

    Sperm detection can be an important factor in confirming sexual assault in cases of rape. This paper compares three of the most commonly used staining methods cited in the scientific literature: Christmas tree. hematoxylin-eosin, and alkaline fuchsin. The population studied was composed of 174 consenting women seen at the Male Infertility Center in Toulouse. France. The date of their last sexual intercourse was accurately known. Alkaline fuchsin did not seem effective in detecting spermatozoa in vaginal samples. Compared with hematoxylin-eosin, Christmas tree stain appeared to be the most useful test in the first 72 h. Two external factors were associated with decreased detection of spermatozoa: time since in tercourse and sperm volume. PMID:11305439

  9. 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.

  10. MoMA: Paper: Pressed, Stained, Slashed, Folded

    NSDL National Science Digital Library

    Visitors can view 31 works created by about two dozen artists, both on or built from paper and paper pulp, at this exhibition website from the Museum of Modern Art (MoMA). The art in the show dates from the 1960s to the early 2000s, with many of the artists featured coming to prominence in the '60s. Much of the work challenges strict definitions of art, such as the selection from Ed Ruscha's portfolio of stains. These are sheets of paper stained with everyday substances, including nail polish, wine, and castor oil. Ruscha says he did not want the work to look like art, so he hired assistants to apply the substances to the paper with eyedroppers. Also employing unusual materials are Dieter Roth's pieces; sausage and cheese pressed into paper with a printing press. A piece by John Cage, titled "Wild edible Drawing #8" includes milkweed, cattail, saffron, and hijiki seaweed.

  11. 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. [Institute of Physics, Academia Sinica, Nankang, Taipei 11529, Taiwan (China); Liang, K. S.; Yin, G. C.; Chen, F. R. [National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan (China); Je, J. H. [Dept. Mater. Sci. Eng., Pohang University of Science and Technology, Pohang (Korea, Republic of); Margaritondo, G. [Ecole Polytechnique Federale, CH-1015 Lausanne (Switzerland); Hwu, Y. [Institute of Physics, Academia Sinica, Nankang, Taipei 11529, Taiwan (China); National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan (China); Institute of Optoelectronic Sciences, National Taiwan Ocean University, Keelong, Taiwan (China)

    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.

  12. [NBI magnifying endoscopic classification using crystal violet staining].

    PubMed

    Inoue, Haruhiro; Kodama, Kenta; Minami, Hitomi; Wada, Yoshiki; Kaga, Makoto; Sato, Yoshitaka; Sugaya, Satoshi; Kudo, Sinei

    2008-05-01

    NBI magnifying imaging with crystal violet (CV-NBI magnifying imaging) makes recognition of micro-vascular pattern and grandular structure in the gastric mucosa better. NBI image emphasizes micro-vascular structure in mucosal surface. Magnification endoscopy with crystal violet staining delineates surface grandular structure better than without it. Crystal violet stained epithelium is clearly observed as cobalt green with NBI imaging. In the classification of CV-NBI magnification findings, 71% of differentiated type lesion was classified into ILL (intralobular loop pattern), and the rest (29%) was diagnosed as FNP (fine network pattern) which was originally advocated by Nakayoshi, et al. ILL is the new category of magnifying endoscopy. ILL corresponded mainly to differentiated-type adenocarcinoma, but it also includes undifferentiated-type adenocarcinoma. Corkscrew pattern is corresponding well to undifferentiated-type adnocarcinoma (Nakayoshi, et al). CV-NBI magnifying classification is considered to be related to tissue characterization in gastric cancer. PMID:18464526

  13. Improved avidin—biotin—peroxidase complex (ABC) staining

    Microsoft Academic Search

    Giorgio Cattoretti; Emilio Berti; Raffaella Schiró; Lucia D'Amato; Costante Valeggio; Franco Rilke

    1988-01-01

    Summary  A considerable intensification of the avidin—biotin—peroxidase complex staining system (ABC) was obtained by sequentially overlaying the sections to be immunostained with an avidin-rich and a biotin-rich complex. Each sequential addition contributed to the deposition of horseradish peroxidase on the immunostained site and allowed the subsequent binding of a complementary complex. With this technique a higher dilution of the antisera could

  14. CMA staining analysis of chromosomes in several species of Aurantioideae

    Microsoft Academic Search

    Masashi Yamamoto; Asad Asadi Abkenar; Ryoji Matsumoto; Tatsuya Kubo; Shigeto Tominaga

    2008-01-01

    Fluorochrome staining with chromomycin A3 (CMA) was used to characterize and compare the CMA banding patterns of chromosomes of 17 species from 13 genera of Aurantioideae,\\u000a which is one of the seven subfamilies of Rutaceae. All species used in this study had 2n = 18 chromosomes. These chromosomes\\u000a were classified into five types based on the number and position of CMA-positive bands;

  15. An improved method for staining cell colonies in clonogenic assays

    Microsoft Academic Search

    Kishore Guda; Leanna Natale; Sanford D. Markowitz

    2007-01-01

    Clonogenic assay is a widely used experimental approach to test for the effects of drugs\\/genes on the growth and proliferative\\u000a characteristics of cells in vitro. Accurate quantitation of treatment effects in clonogeneic assays depends on the ability\\u000a to visualize and count cell colonies precisely. We report a novel method (referred as ETeB) for staining cell colonies grown\\u000a on plastic and

  16. Nile red: a selective fluorescent stain for intracellular lipid droplets

    Microsoft Academic Search

    PHILLIP GREENSPAN; EUGENE P. MAYER; STANLEY D. FOWLER

    1985-01-01

    We report that the dye nile red, 9-diethylamino-5H-benzo(a)phenoxazine-5-one, is an excellent vital stain for the detection of intracellular lipid droplets by fluorescence microscopy and flow cytofluorometry. The specificity of the dye for lipid droplets was assessed on cultured aortic smooth muscle cells and on cultured peritoneal macrophages that were incubated with acetylated low density lipoprotein to induce cytoplasmic lipid overloading.

  17. Method and apparatus for staining immobilized nucleic acids

    SciTech Connect

    Ramsey, J.M.; Foote, R.S.; Jacobson, S.C.

    2000-05-02

    A method for staining immobilized nucleic acids includes the following steps: 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.

  18. 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.

  19. The clinical measurement of tooth colour and stain.

    PubMed

    Brook, A H; Smith, R N; Lath, D J

    2007-10-01

    There are many contributory factors to tooth colour and different techniques for its measurement. The aim of this paper is to evaluate methods of tooth colour and stain measurement, with an emphasis on recent advances in objective clinical measurement techniques. The overall colour effect of natural teeth is created by a combination of light which is reflected and scattered by tooth enamel and the underlying dentine. Developmental defects of the dentition can affect the intrinsic discolouration of teeth, for example, amelogenesis imperfecta and dentinogenesis imperfecta. Extrinsic discolouration is predominantly caused by stain build up on a tooth surface from bacteria, foodstuffs or metalic compounds. Tooth colour and stain measurement are currently assessed using a wide range of measurement methods divided into subjective (visual shade matching) and objective instrumental assessment such as by colourimetry, spectrophotometry and digital image analysis. The most popular method of assessing tooth colour clinically is visual shade matching, as this approach is quick and simple to use. However, variation in results can occur as a consequence of the subjective nature of this method. The instrumental approaches including quantitative light-induced fluorescence remove or significantly reduce the subjective component. Image analysis appears to be the most suitable method for tooth colour measurement and further work is being carried out to establish this approach. PMID:17992918

  20. Application of stains-all staining to the analysis of axonemal tubulins: identification of beta-tubulin and beta-isotubulins.

    PubMed

    Nakamura, K; Masuyama, E; Wada, S; Okuno, M

    1990-01-01

    The cationic dye, Stains-all, is known to stain brain beta-tubulin blue and alpha-tubulin red (Serrano, L. et al. (1986) J. Biochem. Biophys. Methods 12, 281-287; Serrano, L. et al. (1989) Biochem. Int., 19, 235-246). The present experiments show that this stain can also be applied to detect beta-tubulin in axonemal tubulins from various sources such as cilia of protozoa, sperm flagella of echinoderm, and sperm flagella of mollusc. Furthermore, these experiments showed that it selectively stains isoforms of axonemal beta-tubulin blue following isoelectric focusing, whereas those of alpha-tubulin are stained red. These results indicate that Stains-all staining is a useful tool for electrophoretic analysis of axonemal tubulins. PMID:1704028

  1. Monoclonal antibody-targeted fluorescein-5-isothiocyanate-labeled biomimetic nanoapatites: a promising fluorescent probe for imaging applications.

    PubMed

    Oltolina, Francesca; Gregoletto, Luca; Colangelo, Donato; Gómez-Morales, Jaime; Delgado-López, José Manuel; Prat, Maria

    2015-02-10

    Multifunctional biomimetic nanoparticles (NPs) are acquiring increasing interest as carriers in medicine and basic research since they can efficiently combine labels for subsequent tracking, moieties for specific cell targeting, and bioactive molecules, e.g., drugs. In particular, because of their easy synthesis, low cost, good biocompatibility, high resorbability, easy surface functionalization, and pH-dependent solubility, nanocrystalline apatites are promising candidates as nanocarriers. This work describes the synthesis and characterization of bioinspired apatite nanoparticles to be used as fluorescent nanocarriers targeted against the Met/hepatocyte growth factor receptor, which is considered a tumor associated cell surface marker of many cancers. To this aim the nanoparticles have been labeled with Fluorescein-5-isothiocyanate (FITC) by simple isothermal adsorption, in the absence of organic, possibly toxic, molecules, and then functionalized with a monoclonal antibody (mAb) directed against such a receptor. Direct labeling of the nanoparticles allowed tracking the moieties with spatiotemporal resolution and thus following their interaction with cells, expressing or not the targeted receptor, as well as their fate in vitro. Cytofluorometry and confocal microscopy experiments showed that the functionalized nanocarriers, which emitted a strong fluorescent signal, were rapidly and specifically internalized in cells expressing the receptor. Indeed, we found that, once inside the cells expressing the receptor, mAb-functionalized FITC nanoparticles partially dissociated in their two components, with some mAbs being recycled to the cell surface and the FITC-labeled nanoparticles remaining in the cytosol. This work thus shows that FITC-labeled nanoapatites are very promising probes for targeted cell imaging applications. PMID:25602940

  2. Protection against oxidative damage by iron chelators: effect of lipophilic analogues and prodrugs of N,N'-bis(3,4,5-trimethoxybenzyl)ethylenediamine- N,N'-diacetic acid (OR10141).

    PubMed

    Galey, J B; Destrée, O; Dumats, J; Génard, S; Tachon, P

    2000-04-01

    N,N'-Bis(3,4,5-trimethoxybenzyl)ethylenediamine-N,N'-diacetic acid (1) was recently described as a new type of iron chelator for protection against oxidative damage. It has a low affinity for iron, but the corresponding iron complex undergoes a site-specific oxidation by hydrogen peroxide through intramolecular aromatic hydroxylation into a highly stable iron phenolato complex, which does not catalyze hydroxyl radical formation. The purpose of this local activation process is to minimize toxicity compared to strong iron chelators, which may interfere with normal iron metabolism. 1 efficiently protects biological molecules against oxidative damage in vitro but not intact cells because of poor membrane permeability. We show here that, among a series of prodrug esters and lipophilic analogues, membrane-permeant N,N'-bis(3,4,5-trimethoxybenzyl)ethylenediamine-N,N'-diacetic acid diacetoxymethyl ester (7) protects human skin fibroblasts against hydrogen peroxide toxicity with an IC(50) of 3 microM. These results thus demonstrate that, providing sufficient intracellular chelator concentration is reached, 1 efficiently protects cells against the deleterious effects of hydrogen peroxide. This strategy of oxidative activation should help the design of new chelators with better safety margins, which may be useful against oxidative damage under conditions where a prolonged administration is needed. PMID:10753479

  3. Stains and smears: resident guide to bedside diagnostic testing.

    PubMed

    Bronfenbrener, Roman

    2014-12-01

    Dermatology residents often are on the front line when it comes to treating patients with complicated skin disorders, frequently seeing these cases first before discussing their findings and plan with an attending physician. Bedside testing modalities can help facilitate arriving at a diagnosis quickly, allowing for rapid initiation of treatment. The potassium hydroxide (KOH) preparation, Tzanck smear, mineral oil preparation, and Gram stain are easy to perform quickly and provide valuable diagnostic information. The purpose of this article is to consolidate these frequently used tests into a useful reference for the training and practicing dermatologist. PMID:25566582

  4. DNA extraction and amplification from Giemsa-stained blood smears.

    PubMed

    Yokota, M; Tatsumi, N; Tsuda, I; Yano, I

    1995-01-01

    DNA extraction was attempted from Giemsa-stained blood smears on glass slides that had been stored for several years. High molecular weight DNA bands were clearly visible after electrophoresis when DNA was extracted from specimens stored up to 2 years. Specimens more than 4 years old demonstrated a smeared pattern, suggesting degeneration of the DNA, but they could be rescued by PCR amplification using primers of HLA-DQA1 genes. The recovery could be pursued even in 11-year-old specimens. PMID:8587007

  5. Some effects of salts on staining: Use of the Donnan Equilibrium to describe staining of tissue sections with acid and basic dyes

    Microsoft Academic Search

    P. J. Bennion; R. W. Horobin

    1974-01-01

    Summary  Using a wide variety of acid and basic dyes, and dyebaths of various pH’s and salt contents, it was shown that various effects\\u000a of neutral inorganic salts on the staining of tissue sections could be explained by the Donnan Membrane Equilibrium. Thus\\u000a the simultaneous increases in staining of some tissue components and decreases in staining of others, which occur on

  6. 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.

  7. Ring stains in the presence of electromagnetohydrodynamic interactions

    NASA Astrophysics Data System (ADS)

    Das, Siddhartha; Mitra, Sushanta K.; Chakraborty, Suman

    2012-11-01

    In a recent paper [Das , Phys. Rev. EPLEEE81539-375510.1103/PhysRevE.85.046311 85, 046311 (2012)], we delineated the role of electrokinetic transport in modifying the classical “coffee stain” effect. In this study, we extend this calculation to incorporate the consequences of a generalized electromagnetohydrodynamic transport in the coffee stain phenomenon. The magnetohydrodynamic (MHD) effect enhances the velocities at the beginning of the drop life, whereas the electrokinetic effect increases the “disordering” effect in particle deposition at the end of the drop, triggered by a velocity divergence. For a suitable combination of the strength of the MHD and electrokinetic transport, however, this disordering effect is substantially enhanced, and, most nonintuitively, such velocity divergence and the disordering effect may occur at a time that is much earlier than the end of the drop life, or may occur even instantaneously after the start of the drop evaporation. This work will provide useful insight in the understanding of the dynamics of mesoscopic patterns formed as the magnetic nanocrystals deposit in the presence of a combined transport driven by evaporation and magnetic field effects.

  8. Visualizing Peripheral Nerve Regeneration by Whole Mount Staining

    PubMed Central

    Dun, Xin-peng; Parkinson, David B.

    2015-01-01

    Peripheral nerve trauma triggers a well characterised sequence of events both proximal and distal to the site of injury. Axons distal to the injury degenerate, Schwann cells convert to a repair supportive phenotype and macrophages enter the nerve to clear myelin and axonal debris. Following these events, axons must regrow through the distal part of the nerve, re-innervate and finally are re-myelinated by Schwann cells. For nerve crush injuries (axonotmesis), in which the integrity of the nerve is maintained, repair may be relatively effective whereas for nerve transection (neurotmesis) repair will likely be very poor as few axons may be able to cross between the two parts of the severed nerve, across the newly generated nerve bridge, to enter the distal stump and regenerate. Analysing axon growth and the cell-cell interactions that occur following both nerve crush and cut injuries has largely been carried out by staining sections of nerve tissue, but this has the obvious disadvantage that it is not possible to follow the paths of regenerating axons in three dimensions within the nerve trunk or nerve bridge. To try and solve this problem, we describe the development and use of a novel whole mount staining protocol that allows the analysis of axonal regeneration, Schwann cell-axon interaction and re-vascularisation of the repairing nerve following nerve cut and crush injuries. PMID:25738874

  9. Vestibular schwannoma or tanycytic ependymoma: Immunohistologic staining reveals

    PubMed Central

    Divito, Anthony; Keller, Jeffrey T.; Hagen, Matthew; Zuccarello, Mario

    2014-01-01

    Background: The cerebellopontine angle (CPA) is a common location for primary tumors, most often vestibular schwannomas, and also meningiomas, dermoids, and a host of other neoplasms. Our case report illustrates how radiologic and histopathologic presentations of an unusual variant of ependymal neoplasm can be diagnostically challenging and how accurate diagnosis can affect treatment protocols. Case History: Our patient had a CPA mass that was a variant of ependymoma known as tanycytic ependymoma that mimicked vestibular schwannoma radiologically and during intraoperative pathologic examination. Diagnosis as a World Health Organization (WHO) grade II tanycytic ependymoma was supported by its appearance on evaluation of the permanent sections, its diffuse immunoreactivity for glial fibrillary acidic protein (GFAP), and the perinuclear dot-and-ring-like staining for epithelial membrane antigen (EMA). Conclusions: Our patient's CPA mass initially believed to be a vestibular schwannoma on preoperative evaluation, surgical appearance, and intraoperative pathologic consultation was then correctly diagnosed as a WHO grade II tanycytic ependymoma on permanent histologic sections with the assistance of immunohistochemical stains, including EMA. After this definitive diagnosis, our patient's adjuvant treatment was adjusted. Earlier diagnosis could have provided guidance for goals of resection and prompt initiation of adjuvant treatment. PMID:25506503

  10. Visualizing peripheral nerve regeneration by whole mount staining.

    PubMed

    Dun, Xin-Peng; Parkinson, David B

    2015-01-01

    Peripheral nerve trauma triggers a well characterised sequence of events both proximal and distal to the site of injury. Axons distal to the injury degenerate, Schwann cells convert to a repair supportive phenotype and macrophages enter the nerve to clear myelin and axonal debris. Following these events, axons must regrow through the distal part of the nerve, re-innervate and finally are re-myelinated by Schwann cells. For nerve crush injuries (axonotmesis), in which the integrity of the nerve is maintained, repair may be relatively effective whereas for nerve transection (neurotmesis) repair will likely be very poor as few axons may be able to cross between the two parts of the severed nerve, across the newly generated nerve bridge, to enter the distal stump and regenerate. Analysing axon growth and the cell-cell interactions that occur following both nerve crush and cut injuries has largely been carried out by staining sections of nerve tissue, but this has the obvious disadvantage that it is not possible to follow the paths of regenerating axons in three dimensions within the nerve trunk or nerve bridge. To try and solve this problem, we describe the development and use of a novel whole mount staining protocol that allows the analysis of axonal regeneration, Schwann cell-axon interaction and re-vascularisation of the repairing nerve following nerve cut and crush injuries. PMID:25738874

  11. Diff-Quik Stain as a Simplified Alternative to Papanicolaou Stain for Determination of Quality of Endocervical Specimens Submitted for PCR Detection ofChlamydia trachomatis

    Microsoft Academic Search

    JAMES A. KELLOGG; JOHN W. SEIPLE; JANET L. KLINEDINST; ANDERICA STROLL

    1996-01-01

    The simple, rapid, two-step Diff-Quik stain procedure (Baxter Diagnostics) was compared with the Pap- anicolaou stain for microscopic determination of endocervical specimen quality. Results from 230 (98.7%) of 233 specimens stained by both methods indicated agreement between the two staining methods for detection of the endocervical cells or erythrocytes indicating specimen adequacy. By using the Amplicor Chlamydia trachomatisTest(RocheDiagnosticSystems)todetectC.trachomatisandtheDiff-Quikstaintoassessspecimen adequacy,PCR-positiveresultswereobtainedfrom147(9.1%)of1,615microscopicallyadequatespecimensbut from

  12. X-34, A Fluorescent Derivative of Congo Red: A Novel Histochemical Stain for Alzheimer's Disease Pathology

    Microsoft Academic Search

    Scot D. Styren; Ronald L. Hamilton; Gisele C. Styren; William E. Klunk

    2000-01-01

    SUMMARY X-34, a lipophilic, highly fluorescent derivative of Congo red, was examined as a histochemical stain for pathological changes in Alzheimer's disease (AD). X-34 in- tensely stained neuritic and diffuse plaques, neurofibrillary tangles (NFTs), neuropil threads, and cerebrovascular amyloid. Comparison to standard methods of demonstrating AD pathology showed that X-34 correlated well with Bielschowsky and thioflavin-S stain- ing. X-34 staining

  13. Post-embedding staining of rat gastric mucous cells with lectins

    Microsoft Academic Search

    S. Suzuki; S. Tsuyama; F. Murata

    1982-01-01

    The mucous cells of the rat stomach were stained with lectins by two post-embedding staining methods for electron microscopy. The mucous granules of surface mucous cells and foveolar mucous cells were stained weakly by Ricinus communis agglutinin-ferritin and wheat germ agglutinin-ferritin. The mucous granules of mucous neck cells were stained by concanavalin A-ferritin, Ricinus communis agglutinin-ferritin and wheat germ agglutinin-ferritin.

  14. Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining

    Microsoft Academic Search

    Carll Goodpasture; Stephen E. Bloom

    1975-01-01

    A simple ammoniacal silver staining procedure, designated Ag-AS, differentially stains the chromosomal locations of ribosomal DNA in certain mammalian species. This was critically demonstrated by Ag-AS staining of the nucleolus organizer regions in karyotypes of the same species and cell lines used for locating the ribosomal cistrons by DNA\\/RNA in situ hybridization. With Ag-AS, silver stained NORs (Ag-NORs) are visualized

  15. Bacterial Viability and Antibiotic Susceptibility Testing with SYTOX Green Nucleic Acid Stain

    Microsoft Academic Search

    BRUCE L. ROTH; MARTIN POOT; STEPHEN T. YUE; PAUL J. MILLARD

    1997-01-01

    A fluorescent nucleic acid stain that does not penetrate living cells was used to assess the integrity of the plasma membranes of bacteria. SYTOX Green nucleic acid stain is an unsymmetrical cyanine dye with three positive charges that is completely excluded from live eukaryotic and prokaryotic cells. Binding of SYTOX Green stain to nucleic acids resulted in a >500-fold enhancement

  16. An improved technique for selective silver staining of nucleolar organizer regions in human chromosomes

    Microsoft Academic Search

    S. E. Bloom; C. Goodpasture

    1976-01-01

    A reliable technique for staining human chromosomal nucleolar organizers (NOR's) with silver solutions is described. The NOR's can be selectively stained dark brown by silver solutions leaving the chromosome arms unstained and available for counterstaining with orcein or Giemsa dyes. Unequivocal identification of chromosome pairs bearing NOR's can be achieved using fluorescent banding techniques followed by silver staining. The silver

  17. Increasing DNA extraction yield from saliva stains with a modified Chelex method

    Microsoft Academic Search

    David Sweet; Miguel Lorente; Aurora Valenzuela; José A. Lorente; J. Carlos Alvarez

    1996-01-01

    Recovery, preservation and analysis of body fluid stains is an important aspect of forensic science. PCR-based typing of DNA extracted from recovered stains is often a crucial method to identify a perpetrator or exclude an innocent suspect. This paper reports an improved method of extracting genomic DNA from saliva stains deposited on human skin in simulated bite mark situations. Results

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

    E-print Network

    Lebendiker, Mario

    in humans or animals. Contains DMSO. See safety data sheet supplied. Harmful Components RPN 5801 SYPRO Orange gel stain 5000x concentrate in DMSO, 500µl RPN 5802 SYPRO Orange gel stain 5000x concentrate in DMSO, 10x50µl RPN 5803 SYPRO Red gel stain 5000x concentrate in DMSO, 500µl RPN 5804 SYPRO Red gel

  19. 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

  20. 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...

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

    E-print Network

    Løw, Erik

    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

  2. Psychosocial Stress of Patients with Port Wine Stains and Expectations of Dye Laser Treatment

    Microsoft Academic Search

    M. Augustin; I. Zschocke; K. Wiek; M. Peschen; W. Vanscheidt

    1998-01-01

    Background: Port wine stains can be treated successfully with the dye laser in most cases. The indication for treatment is made on the basis of the emotional stress arising from the port wine stain. Studies of the extent of psychosocial stress to date have led to contradictory results. Objective: To address the question how many patients with port wine stain

  3. Phos-toolsTM Phos-tagTM 540 Phosphoprotein Blot Stain

    E-print Network

    Lebendiker, Mario

    FOR PVDF BLOTS 11 A. Fixing the Membrane 12 B. Blocking the Membrane 12 C. Staining the Membrane 12 D of phosphorylated proteins transferred to PVDF electroblot membrane. For Laboratory Use Caution: Research Chemicals (PVDF) membrane. Selective staining of phosphoproteins #12;5 with Phos-tag blot stain permits

  4. Coupling Between Precipitation and Contact-Line Dynamics: Multiring Stains and Stick-Slip Motion

    E-print Network

    Chang, Hsueh-Chia

    Coupling Between Precipitation and Contact-Line Dynamics: Multiring Stains and Stick-Slip Motion to be an important small scale fabrication process [5,6]. The formation of the well-known coffee-stain pattern the formation of the ringlike stain at the outer regions of the drop. At the same time, sustained evapora- tion

  5. Silver Staining of 2D Electrophoresis Gels Ccile Lelong, Mireille Chevallet, Sylvie Luche, Thierry Rabilloud

    E-print Network

    Paris-Sud XI, Université de

    1 Silver Staining of 2D Electrophoresis Gels Cécile Lelong, Mireille Chevallet, Sylvie Luche Cedex 9, France 1. Introduction Silver staining of polyacrylamide gels was introduced in 1979 by Switzer staining protocols for proteins in polyacrylamide gels can be found in the literature. However, all of them

  6. Color correction of pathological images for different staining-condition slides

    Microsoft Academic Search

    T. Abe; M. Yamaguchi; Y. Murakami; N. Ohyama; Y. Yagi

    2004-01-01

    The color of hematoxylin & eosin (H&E) stained tissue image varies with the staining conditions and with the characteristics of the microscope and imaging devices. This color variation affects the diagnostic examination. This paper proposes a color correction method for images of pathological slides prepared under inappropriate staining-condition. In the proposed method, the spectral transmittance obtained by a multispectral digital

  7. Specific In Vivo Staining of Astrocytes in the Whole Brain after Intravenous Injection of Sulforhodamine Dyes

    E-print Network

    Paris-Sud XI, Université de

    become a reference for in vivo staining of the whole astrocytes population in animal modelsSpecific In Vivo Staining of Astrocytes in the Whole Brain after Intravenous Injection or electroencephalographic recordings in vivo. The high contrast of the staining facilitates the image processing and allows

  8. 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…

  9. 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.

  10. Ring stains in the presence of electrokinetic interactions

    NASA Astrophysics Data System (ADS)

    Das, Siddhartha; Chakraborty, Suman; Mitra, Sushanta K.

    2012-04-01

    In this paper, we delineate the consequences of electrokinetic interactions on the “coffee stain” effect, induced by the deposition of particles during drop evaporation. We consider evaporation of an electrolytic drop in contact with a charged substrate and probe the effects of electrical double layer formation at the drop-substrate interface on the dynamics of particles suspended inside the drop. We show that the simultaneous considerations of streaming potential and flow-actuation-mechanism-independent description of the evaporation flux and the depth average velocities result in an enhanced induced radial pressure gradient. As a result, the deposition speed of the particles in the disordered packing regime, occurring at the end of the lifetime of the drop [Marin , Phys. Rev. Lett.PRLTAO0031-900710.1103/PhysRevLett.107.085502 107, 085502 (2011)], is greatly enhanced. This, in turn, is likely to signify an augmented degree of disordering in the evaporation-induced particle deposition.

  11. 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

  12. Optimized Negative-Staining Electron Microscopy for Lipoprotein Studies

    PubMed Central

    Zhang, Lei; Tong, Huimin; Garewal, Mark; Ren, Gang

    2012-01-01

    Background Negative-staining (NS), a rapid, simple and conventional technique of electron microscopy (EM), has been commonly used to initially study the morphology and structure of proteins for half a century. Certain NS protocols however can cause artifacts, especially for structurally flexible or lipid-related proteins, such as lipoproteins. Lipoproteins were often observed in the form of rouleau as lipoprotein particles appeared to be stacked together by conventional NS protocols. The flexible components of lipoproteins, i.e. lipids and amphipathic apolipoproteins, resulted in the lipoprotein structure being sensitive to the NS sample preparation parameters, such as operational procedures, salt concentrations, and the staining reagents. Scope of review The most popular NS protocols that have been used to examine lipoprotein morphology and structure were reviewed. Major conclusions The comparisons show that an optimized NS (OpNS) protocol can eliminate the rouleau artifacts of lipoproteins, and that the lipoproteins are similar in size and shape as statistically measured from two EM methods, OpNS and cryo-electron microscopy (cryo-EM). OpNS is a high-throughput, high-contrast and high-resolution (near 1 nm, but rarely better than 1 nm) method which has been used to discover the mechanics of a small protein, 53 kDa cholesterol ester transfer protein (CETP), and the structure of an individual particle of a single protein by individual-particle electron tomography (IPET), i.e. a 14 Å-resolution IgG antibody three-dimensional map. General significance It is suggested that OpNS can be used as a general protocol to study the structure of proteins, especially highly dynamic proteins with equilibrium-fluctuating structures. PMID:23032862

  13. The Prognostic Significance of Lymphovascular Space Invasion in Endometrial Cancer When Conventional Hemotoxylin and Eosin Staining Is Compared to Immunohistochemical Staining

    Microsoft Academic Search

    N. Tsuruchi; T. Kaku; T. Kamura; N. Tsukamoto; M. Tsuneyoshi; K. Akazawa; H. Nakano

    1995-01-01

    The current study was undertaken to compare the usefulness of hemotoxylin and eosin (H&E) staining and immunohistochemical staining to identify lymphovascular space invasion (LVSI) in endometrial cancer and to evaluate the presence of LVSI detected by either technique as an independent prognostic factor. Histologic sections from 92 patients with clinical stage I-II endometrial cancer were reviewed, and representative sections were

  14. Ionic and non-ionic bonds in staining, with special reference to the action of urea and sodium chloride on the staining of elastic fibres and glycogen

    Microsoft Academic Search

    S. Africa

    Summary 1. A powerful hydrogen bonding agent such as urea may affect staining in several ways: (a) by competing for hydrogen bonding sites in the tissue or on the dye particle, it may inhibit staining in which the dye-substrate link is a hydrogen bond; (6) because it is a dipole, urea may have some affinity for charged sites in the

  15. Tunable filter-based multispectral imaging for detection of blood stains on construction material substrates part 2: realization of rapid blood stain detection.

    PubMed

    Janchaysang, Suwatwong; Sumriddetchkajorn, Sarun; Buranasiri, Prathan

    2013-07-10

    Based on the blood stain detection method and criteria established in part 1 of this article, we combine and organize all necessary tasks to realize the multispectral imaging-based rapid blood stain detection system. To rapidly detect blood stains on the test surface, the developed system automatically captures the spectral images, extracts their spectral data, determines the positions of blood stains, and accurately highlights the positions of blood stains on the display. To achieve such a system, several tasks are newly introduced, including adjustment of camera exposure times to prevent image saturation or excessive darkness, the search for the sampled clean positions of the substrate to determine the substrate reflectance spectrum, and suitable detection procedures and proper arrangement of criteria to eliminate unnecessary calculations. Parallel processes between image capturing and blood stain identification help shorten the time for blood stain identifications despite a large amount of spectral data to be processed. The developed system can identify blood against several other reddish brown stains on several substrates. The measured average identification times on different test surfaces range from only 23.3 to 28.7 s, including the image capturing process. PMID:23852205

  16. Van Gieson's picrofuchsin. The staining mechanisms for collagen and cytoplasm, and an examination of the dye diffusion rate model of differential staining

    Microsoft Academic Search

    Poul Prentø

    1993-01-01

    The staining mechanism of van Gieson's picrofuchsin was studied by use of simple protein model systems and tissue sections, and by spectrophotometry and dialysis experiments. At the endpoint of the staining reaction (equilibrium) cytoplasm is yellow. Dye dilution experiments demonstrated that the highest affinity in the tissue section — picrofuchsin system is between binding sites in cytoplasmic protein and acid

  17. X-34, a fluorescent derivative of Congo red: a novel histochemical stain for Alzheimer's disease pathology.

    PubMed

    Styren, S D; Hamilton, R L; Styren, G C; Klunk, W E

    2000-09-01

    X-34, a lipophilic, highly fluorescent derivative of Congo red, was examined as a histochemical stain for pathological changes in Alzheimer's disease (AD). X-34 intensely stained neuritic and diffuse plaques, neurofibrillary tangles (NFTs), neuropil threads, and cerebrovascular amyloid. Comparison to standard methods of demonstrating AD pathology showed that X-34 correlated well with Bielschowsky and thioflavin-S staining. X-34 staining of NFTs correlated closely with anti-TAU antibody staining. A 1:1 correspondence of X-34 and anti-A beta antibody staining of plaques and cerebrovascular amyloid was observed. Both X-34 and thioflavin-S staining were eliminated by formic acid pretreatment, suggesting that beta-sheet secondary protein structure is a necessary determinant of staining. X-34 may be a general amyloid stain, like Congo red, because it also stains systemic amyloid deposits due to lambda-light chain monoclonal gammopathy. In conclusion, X-34 is a highly fluorescent marker for beta-sheet structures and intensely labels amyloid plaques, NFTs, neuropil threads, and vascular amyloid in AD brains. It can be used with both paraffin-embedded and frozen tissues as well as in combination with immunohistochemistry for double labeling. The intensity of staining and the simplicity and reproducibility of the technique suggest that it may be a useful addition to the standard techniques for evaluation of AD neuropathology. (J Histochem Cytochem 48:1223-1232, 2000) PMID:10950879

  18. Diff-Quik stain as a simplified alternative to Papanicolaou stain for determination of quality of endocervical specimens submitted for PCR detection of Chlamydia trachomatis.

    PubMed

    Kellogg, J A; Seiple, J W; Klinedinst, J L; Stroll, E

    1996-10-01

    The simple, rapid, two-step Diff-Quik stain procedure (Baxter Diagnostics) was compared with the Papanicolaou stain for microscopic determination of endocervical specimen quality. Results from 230 (98.7%) of 233 specimens stained by both methods indicated agreement between the two staining methods for detection of the endocervical cells or erythrocytes indicating specimen adequacy. By using the Amplicor Chlamydia trachomatis Test (Roche Diagnostic Systems) to detect C.trachomatis and the Diff-Quik stain to assess specimen adequacy, PCR-positive results were obtained from 147 (9.1%) of 1,615 microscopically adequate specimens but from only 13 (2.2%) of the 583 inadequate specimens (P < 0.001). PMID:8880526

  19. 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. (Univ. of Alberta, Edmonton (Canada)); Schaefer, F.W. III (Environmental Protection Agency, Cincinnati, OH (United States))

    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.

  20. On the barrier properties of the cornea: a microscopy study of the penetration of fluorescently labeled nanoparticles, polymers, and sodium fluorescein.

    PubMed

    Mun, Ellina A; Morrison, Peter W J; Williams, Adrian C; Khutoryanskiy, Vitaliy V

    2014-10-01

    Overcoming the natural defensive barrier functions of the eye remains one of the greatest challenges of ocular drug delivery. Cornea is a chemical and mechanical barrier preventing the passage of any foreign bodies including drugs into the eye, but the factors limiting penetration of permeants and nanoparticulate drug delivery systems through the cornea are still not fully understood. In this study, we investigate these barrier properties of the cornea using thiolated and PEGylated (750 and 5000 Da) nanoparticles, sodium fluorescein, and two linear polymers (dextran and polyethylene glycol). Experiments used intact bovine cornea in addition to bovine cornea de-epithelialized or tissues pretreated with cyclodextrin. It was shown that corneal epithelium is the major barrier for permeation; pretreatment of the cornea with ?-cyclodextrin provides higher permeation of low molecular weight compounds, such as sodium fluorescein, but does not enhance penetration of nanoparticles and larger molecules. Studying penetration of thiolated and PEGylated (750 and 5000 Da) nanoparticles into the de-epithelialized ocular tissue revealed that interactions between corneal surface and thiol groups of nanoparticles were more significant determinants of penetration than particle size (for the sizes used here). PEGylation with polyethylene glycol of a higher molecular weight (5000 Da) allows penetration of nanoparticles into the stroma, which proceeds gradually, after an initial 1 h lag phase. PMID:25165886

  1. Hydrogen peroxide staining to visualize intracellular bacterial infections of seedling root cells.

    PubMed

    White, James F; Torres, Mónica S; Somu, Mohini P; Johnson, Holly; Irizarry, Ivelisse; Chen, Qiang; Zhang, Ning; Walsh, Emily; Tadych, Mariusz; Bergen, Marshall

    2014-08-01

    Visualization of bacteria in living plant cells and tissues is often problematic due to lack of stains that pass through living plant cell membranes and selectively stain bacterial cells. In this article, we report the use of 3,3'-diaminobenzidine tetrachloride (DAB) to stain hydrogen peroxide associated with bacterial invasion of eukaryotic cells. Tissues were counterstained with aniline blue/lactophenol to stain protein in bacterial cells. Using this staining method to visualize intracellular bacterial (Burkholderia gladioli) colonization of seedling roots of switch grass (Panicum virgatum), we compared bacterial free seedling roots and those inoculated with the bacterium. To further assess application of the technique in multiple species of vascular plants, we examined vascular plants for seedling root colonization by naturally occurring seed-transmitted bacteria. Colonization by bacteria was only observed to occur within epidermal (including root hairs) and cortical cells of root tissues, suggesting that bacteria may not be penetrating deeply into root tissues. DAB/peroxidase with counter stain aniline blue/lactophenol was effective in penetration of root cells to selectively stain bacteria. Furthermore, this stain combination permitted the visualization of the bacterial lysis process. Before any evidence of H2 O2 staining, intracellular bacteria were seen to stain blue for protein content with aniline blue/lactophenol. After H2 O2 staining became evident, bacteria were often swollen, without internal staining by aniline blue/lactophenol; this suggests loss of protein content. This staining method was effective for seedling root tissues; however, it was not effective at staining bacteria in shoot tissues due to poor penetration. PMID:24825573

  2. Discrimination of p53 immunohistochemistry-positive tumors by its staining pattern in gastric cancer

    PubMed Central

    Ando, Koji; Oki, Eiji; Saeki, Hiroshi; Yan, Zhao; Tsuda, Yasuo; Hidaka, Gen; Kasagi, Yuta; Otsu, Hajime; Kawano, Hiroyuki; Kitao, Hiroyuki; Morita, Masaru; Maehara, Yoshihiko

    2015-01-01

    Immunohistochemistry staining of p53 is a cheap and simple method to detect aberrant function of p53. However, there are some discrepancies between the result of immunohistochemistry staining and mutation analysis. This study attempted to find a new definition of p53 staining by its staining pattern. Immunohistochemistry staining of p53 and TP53 gene mutation analysis were performed in 148 gastric cancer patients. Also SNP-CGH array analysis was conducted to four cases. Positive staining of p53 was observed in 88 (59.5%) tumors. Tumors with positive p53 staining showed malignant features compared to negative tumors. Mutation of TP53 gene was observed in 29 (19.6%) tumors with higher age and differentiated type. In positive p53 tumors, two types could be distinguished; aberrant type and scattered type. With comparison to TP53 gene mutation analysis, all the scattered type had wild-type TP53 gene (P = 0.0003). SNP-CGH array showed that scattered-type tumors had no change in the structure of chromosome 17. P53-scattered-type staining tumors may reflect a functionally active nonmutated TP53 gene. In interpretation of p53 immunohistochemistry staining, distinguishing p53-positive tumors by their staining pattern may be important in gastric cancer. PMID:25354498

  3. Hyperspectral imaging of the crime scene for detection and identification of blood stains

    NASA Astrophysics Data System (ADS)

    Edelman, G. J.; van Leeuwen, T. G.; Aalders, M. C. G.

    2013-05-01

    Blood stains are an important source of information in forensic investigations. Extraction of DNA may lead to the identification of victims or suspects, while the blood stain pattern may reveal useful information for the reconstruction of a crime. Consequently, techniques for the detection and identification of blood stains are ideally non-destructive in order not to hamper both DNA and the blood stain pattern analysis. Currently, forensic investigators mainly detect and identify blood stains using chemical or optical methods, which are often either destructive or subject to human interpretation. We demonstrated the feasibility of hyperspectral imaging of the crime scene to detect and identify blood stains remotely. Blood stains outside the human body comprise the main chromophores oxy-hemoglobin, methemoglobin and hemichrome. Consequently, the reflectance spectra of blood stains are influenced by the composite of the optical properties of the individual chromophores and the substrate. Using the coefficient of determination between a non-linear least squares multi-component fit and the measured spectra blood stains were successfully distinguished from other substances visually resembling blood (e.g. ketchup, red wine and lip stick) with a sensitivity of 100 % and a specificity of 85 %. The practical applicability of this technique was demonstrated at a mock crime scene, where blood stains were successfully identified automatically.

  4. 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

  5. Hyperspectral imaging for the age estimation of blood stains at the crime scene.

    PubMed

    Edelman, Gerda; van Leeuwen, Ton G; Aalders, Maurice C G

    2012-11-30

    The age estimation of blood stains can provide important information on the temporal aspects of a crime. As previously shown, visible spectroscopy of blood stains can successfully be used for their age estimation. In the present study we evaluated the feasibility to use hyperspectral imaging for this purpose. Visible reflectance spectra of blood stains were recorded using a pushbroom hyperspectral imaging system. From these spectra, the relative amounts of oxyhemoglobin, methemoglobin and hemichrome within the blood stains were derived. By comparison of the hemoglobin derivative fractions with a reference dataset, the age of blood stains up to 200 days old was estimated. The absolute error of the age estimation task increased with age, with a median relative error of 13.4% of the actual age. To test the practical applicability of this method, a simulated crime scene was analyzed, in which blood stains of several ages were deposited. Hyperspectral imaging combined with the proposed analysis provided insight in the absolute age of the blood stains. Additionally, the blood stains were clustered based on their hemoglobin derivative fractions, without the use of a reference dataset. Results demonstrated that the order of formation of blood stains can be determined, even under unknown environmental circumstances, when no proficient reference dataset is available. These findings are an important step toward the practical implementation of blood stain age estimation in forensic casework. PMID:22938693

  6. Modified fields' stain: ideal to differentiate Dientamoeba fragilis and Blastocystis sp.

    PubMed

    Ragavan, Anitamalar Devi; Govind, Suresh Kumar

    2015-03-01

    Dientamoeba fragilis, a trichomonad parasite is usually found in the gastrointestinal tract of human, and it is known to be the cause for gastrointestinal disease. The parasite is globally distributed and mostly found in rural and urban areas. The parasite is found in humans and nonhuman primates such as the macaques, baboons, and gorillas. Often, the parasite is confused with another largely found organism in stools called Blastocystis sp. especially when seen directly under light microscopy on culture samples containing both parasites. Both sometimes are seen with two nuclei with sizes tending to be similar which complicates identification. Stools were collected fresh from nine previously diagnosed persons infected with D. fragilis who also were found to be positive for Blastocystis sp. Samples were then cultured in Loeffler's medium and were stained with Giemsa, iron hematoxylin, and modified Fields' (MF) stain, respectively. D. fragilis was differentiated from Blastocystis sp. when stained with MF stain by the presence of a thinner outer membrane with clearly demarcated nuclei in the center of the cell whilst Blastocystis sp. had a darker and thicker stained outer membrane with the presence of two nuclei. The staining contrast was more evident with modified Fields' stain when compared with the other two. The simplicity in preparing the stain as well as the speed of the staining procedure make MF stain an ideal alternate. The modified Fields' stain is faster and easier to prepare when compared to the other two stains. MF stain provides a better contrast differentiating the two organisms and therefore provides a more reliable diagnostic method to precisely identify one from the other especially when cultures show mixed infections. PMID:25614298

  7. Digital separation of diaminobenzidine-stained tissues via an automatic color-filtering for immunohistochemical quantification

    PubMed Central

    Fu, Rong; Ma, Xiaomian; Bian, Zhaoying; Ma, Jianhua

    2015-01-01

    The digital separation of diaminobenzidine (DAB)-stained tissues from hematoxylin background is an important pre-processing step to analyze immunostains. In most stain separation methods, specific color channels (for example: RGB, HSI, CMYK) or color deconvolution matrices are used to obtain different tissue contrasts between DAB- and hematoxylin-stained areas. However, these methods could produce incomplete separation or color changes because the color spectra of stains and co-localized stains overlap in histological images. Therefore, we proposed an automatic color-filtering to separate hematoxylin- and DAB-stained tissues. In implantation, the RGB images of DAB-labeled immunostains are first converted to 8-bit BN images by a mathematical translation to produce the largest contrast between brown DAB-stained tissues and blue hematoxylin-stained tissues. The first valley in the histogram revised by nonuniform quantization is set as the cut-off point to obtain a brown filter. DAB-stained tissues are accurately delineated from the background counterstain, resulting in DAB-only-image and De-DAB-image. Subsequently, a blue filter is designed in the CIE-Lab color space to further delineate the hematoxylin-stained tissues from the De-DAB-image. Finally, the average values of the remaining pixels of the De-DAB-image are set as the background color of the DAB-only-image to manage uneven dyeing and provide DAB-stained-image for adaptive immunohistochemistry quantitation. Extensive experimental results demonstrated that the proposed method has significant advantages compared with existing methods in terms of complete stain separation without changing the color in DAB-stained areas. PMID:25780744

  8. 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.

  9. Diffuse reflectance FTIR of stains on grit blasted metals

    SciTech Connect

    Powell, G.L.; Hallman, R.L. Jr.; Cox, R.L. [Oak Ridge Centers for Manufacturing Technologies, TN (United States)

    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.

  10. 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.

  11. Modelling multiple laser pulses for port wine stain treatment.

    PubMed

    Verkruysse, W; van Gemert, M J; Smithies, D J; Nelson, J S

    2000-12-01

    Many port wine stains (PWS) are still resistant to pulsed dye laser treatment. However, anecdotal information suggests that multiple-pulse laser irradiation improves patient outcome. Our aims in this note are to explain the underlying mechanism and estimate the possible thermal effects of multiple pulses in vascular structures typical of PWS. Based on linear response theory, the linear combination of two thermal contributions is responsible for the total increase in temperature in laser irradiated blood vessels: direct light absorption by blood and direct bilateral thermal heat conduction from adjacent blood vessels. The latter contribution to the increase in temperature in the targeted vessel can be significant, particularly if some adjacent vessels are in close proximity, such as in cases of optical shielding of the targeted vessel, or if the vessels are relatively distant but many in number. We present evidence that multiple-pulse laser irradiation targets blood vessels that are optically shielded by other vessels. Therefore, it may be a means of enhancing PWS therapy for lesions that fail to respond to single-pulse dye laser treatment. PMID:11131209

  12. Immunofluorescence staining with frozen mouse or chick embryonic tissue sections.

    PubMed

    Wang, Hui; Matise, Michael P

    2013-01-01

    Immunofluorescence (IF), a form of immunohistochemistry (IHC) with specific applications, is commonly used for both basic research and clinical studies, including diagnostics, and involves visualizing the cellular distribution of target molecules (e.g., proteins, DNA, and small molecules) using a microscope capable of exciting and detecting fluorochrome compounds that emit light at specific, largely nonoverlapping wavelengths. The procedure for carrying out IF varies according to the tissue type and methods for processing and preparing tissue (e.g., fixative used to preserve tissue morphology and antigenicity). The protocol presented here provides a general guideline for multichannel IF staining using frozen embryonic mouse or chicken tissue sectioned on a cryostat. In general, the procedure involves the following: (1) fixing freshly dissected tissues in a 4 % paraformaldehyde solution buffered in the physiological pH range, (2) cryopreservation of tissue in a 30 % sucrose solution, (3) embedding and sectioning tissue in Optimal Cutting Temperature (OCT) matrix compound, (4) direct or indirect detection of the target antigen/s using fluorochrome-conjugated antibodies. PMID:23681628

  13. Diffuse reflectance FTIR of stains on grit blasted metals

    SciTech Connect

    Powell, G.L.; Hallman, R.L. Jr.; Cox, R.L. [Oak Ridge Centers for Manufacturing Technologies, P.O. Box 2009, Oak Ridge, Tennessee 37831-8096 (United States)

    1998-06-01

    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 {minus}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. {copyright} {ital 1998 American Institute of Physics.}

  14. Novel genetic mutations in a sporadic port-wine stain.

    PubMed

    Lian, Christine Guo; Sholl, Lynette M; Zakka, Labib R; O, Teresa M; Liu, Cynthia; Xu, Shuyun; Stanek, Ewelina; Garcia, Elizabeth; Jia, Yonghui; MacConaill, Laura E; Murphy, George F; Waner, Milton; Mihm, Martin C

    2014-12-01

    IMPORTANCE Port-wine stains (PWSs) are common congenital cutaneous capillary malformations. A somatic GNAQ mutation was recently identified in patients with sporadic PWSs and Sturge-Weber syndrome. However, subsequent studies to confirm or extend this observation are lacking.OBSERVATIONS We report a long-standing, unilateral facial PWS of a man in his early 70s confirmed by histopathological analysis. Staged surgical excision of the vascular malformation was performed, and genomic DNA was extracted from the vascular malformation specimen and normal skin. Targeted next-generation sequencing of the coding sequence of 275 known cancer genes including GNAQ was performed in both specimens. A single-nucleotide variant(c.548G>A, p.Arg183Gln) in GNAQ was identified in the PWS-affected tissue but not in the normal skin sample. In addition, this sequencing approach uncovered several additional novel somatic mutations in the genes SMARCA4, EPHA3, MYB, PDGFR-?, and PIK3CA.CONCLUSIONS AND RELEVANCE Our findings confirm the presence of somatic mutations inGNAQ in the affected skin of a patient with congenital PWS, as well as alterations in several other novel genes of possible importance in the pathogenesis of PWS that may also offer substantial therapeutic targets. PMID:25188413

  15. Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester (CFSE)

    PubMed Central

    Deleyrolle, Loic P.; Rohaus, Mark R.; Fortin, Jeff M.; Reynolds, Brent A.; Azari, Hassan

    2012-01-01

    Tumor heterogeneity represents a fundamental feature supporting tumor robustness and presents a central obstacle to the development of therapeutic strategies1. To overcome the issue of tumor heterogeneity, it is essential to develop assays and tools enabling phenotypic, (epi)genetic and functional identification and characterization of tumor subpopulations that drive specific disease pathologies and represent clinically relevant targets. It is now well established that tumors exhibit distinct sub-fractions of cells with different frequencies of cell division, and that the functional criteria of being slow cycling is positively associated with tumor formation ability in several cancers including those of the brain, breast, skin and pancreas as well as leukemia2-8. The fluorescent dye carboxyfluorescein succinimidyl ester (CFSE) has been used for tracking the division frequency of cells in vitro and in vivo in blood-borne tumors and solid tumors such as glioblastoma2,7,8. The cell-permeant non-fluorescent pro-drug of CFSE is converted by intracellular esterases into a fluorescent compound, which is retained within cells by covalently binding to proteins through reaction of its succinimidyl moiety with intracellular amine groups to form stable amide bonds9. The fluorescent dye is equally distributed between daughter cells upon divisions, leading to the halving of the fluorescence intensity with every cell division. This enables tracking of cell cycle frequency up to eight to ten rounds of division10. CFSE retention capacity was used with brain tumor cells to identify and isolate a slow cycling subpopulation (top 5% dye-retaining cells) demonstrated to be enriched in cancer stem cell activity2. This protocol describes the technique of staining cells with CFSE and the isolation of individual populations within a culture of human glioblastoma (GBM)-derived cells possessing differing division rates using flow cytometry2. The technique has served to identify and isolate a brain tumor slow-cycling population of cells by virtue of their ability to retain the CFSE labeling. PMID:22565048

  16. 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.

  17. A comparison of the iodine and fluorescent antibody methods for staining trachoma inclusions in the conjunctiva*

    PubMed Central

    Sowa, J.; Collier, L. H.; Sowa, Shiona

    1971-01-01

    In terms of the rate of positive diagnoses the indirect fluorescent antibody (FA) test was rather more effective than iodine for demonstrating trachoma (TRIC) inclusions in conjunctival scrapings, but the degree of advantage was not statistically significant. In duplicate scrapings stained at random by one or the other method, FA staining yielded the higher inclusion count significantly more often than did iodine. Some inclusions that failed to stain with FA were found on subsequent staining with Giemsa. A method is described for improving the post-FA Giemsa staining of conjunctival smears stored at subzero temperatures. Given adequate facilities, the FA stain is preferable to iodine for demonstrating TRIC inclusions in the conjunctiva; but the iodine method, properly used, holds advantages for field use. ImagesPlate 1Plate 1Plate 1Plate 1 PMID:4109203

  18. Analysis of Formation of Pad Stains in Copper Chemical Mechanical Planarization

    NASA Astrophysics Data System (ADS)

    Lee, Hyosang; Borucki, Leonard; Zhuang, Yun; Joh, Sooyun; O'Moore, Fergal; Philipossian, Ara

    2009-12-01

    A stain model was developed to simulate stain formation on the pad surface in copper chemical and mechanical planarization (CMP). The model consisted of the incompressible Navier-Stokes equations, the heat equation with advection, material removal rate model, a model for generation, transport and deposition of the polishing by-product that produces the stain. Slurry velocity simulations showed shear flow on the land areas and wafer-driven circulation in the grooves. The simulated temperature on the pad and the wafer surface increased gradually in the radial direction; furthermore, temperature simulations showed a 12 °C rise in the reaction temperature on the copper wafer surface. The simulated pad stains deposited on the copper land areas were darker in the direction of wafer rotation, suggesting that the generated staining agents were advected downstream by the slurry flow and deposited on the pad surface in the direction of the wafer rotation. Simulated stain images were in qualitative agreement with experimental results.

  19. Juvenile Localized Scleroderma with Port Wine Stain: Coincidental or Possible Common Pathogenetic Association

    PubMed Central

    Kacar, Seval Dogruk; Ozuguz, Pinar; Polat, Serap; Kacar, Emre; Polat, Onur; Tokyol, Cigdem

    2015-01-01

    Port wine stain and juvenile localized scleroderma are two different dermatoses usually encountered in pediatric age group. Up to now, there are reports of morphea patients initially diagnosed and treated as port wine stain. Coexistence of both diseases is not found yet. We herein present a case of juvenile localized scleroderma on the left side of trunk, with congenital port wine stain located on the ipsilateral face at V1-V2 distribution.

  20. Restoring tetracycline-stained teeth with a conservative preparation for porcelain veneers: case presentation.

    PubMed

    Bassett, Joyce; Patrick, Brad

    2004-08-01

    Tetracycline exposure in utero and in early childhood often results in intrinsic tooth staining that varies in severity based upon timing, duration, and form of tetracycline administered. Traditionally, dental aesthetics compromised by tetracycline staining have been restored with modalities requiring aggressive tooth preparation. In this case involving a patient with extremely severe staining of healthy and aesthetically shaped dentition, a conservative tooth preparation strategy and porcelain veneers were utilized to preserve tooth shape and arch form while restoring natural color. PMID:15485160

  1. Intrapartum amnioinfusion for meconium-stained fluid: meta-analysis of prospective clinical trials

    Microsoft Academic Search

    John Pierce; Francisco L Gaudier; Luis Sanchez-Ramos

    2000-01-01

    Objective: To evaluate the effectiveness of intrapartum prophylactic amnioinfusion in pregnancies complicated by meconium-stained amniotic fluid.Data Sources: We identified prospective clinical trials of amnioinfusion in pregnancies complicated by meconium-stained amniotic fluid (AF) published in English by using computerized databases, references in published studies, and index reviews.Methods of Study Selection: We analyzed prospective studies of intrapartum amnioinfusion for meconium-stained AF. In

  2. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with oil red O

    Microsoft Academic Search

    J. L. Ramírez-Zacarías; F. Castro-Muñozledo; W. Kuri-Harcuch

    1992-01-01

    Cultured 3T3-F442A cells differentiate into adipocytes and accumulate lipid droplets in the cytoplasm. When fat cells are stained with Oil red O, the degree of staining seems to be proportional to the extent of cell differentiation. We report here a fast and simple method to quantitate the extent of adipose conversion by staining the accumulated lipid with Oil red O

  3. 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

  4. 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.

  5. 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.

  6. Electrode potential-dependent colorimetric response of fluorescein-modified layer-by-layer films in the presence of hydrogen peroxide.

    PubMed

    Nagasaka, Munenari; Yoshida, Kentaro; Sato, Katsuhiko; Hoshi, Tomonori; Anzai, Jun-ichi

    2010-08-15

    Layer-by-layer (LbL) thin films composed of fluorescein-modified poly(allylamine) (F-PAH) and poly(styrenesulfonic acid) (PSS) were prepared on the surface of an indium-tin oxide (ITO) electrode and the electrode potential-dependent colorimetric response of the LbL films was studied in the presence of hydrogen peroxide (H(2)O(2)). The LbL films were prepared by an alternate deposition of F-PAH and PSS on the surface through an electrostatic force of attraction. The LbL films exhibited a UV-visible absorption band around 500 nm originating from fluorescein residues in the film and the intensity of the absorption band depended on the pH of the solution to which the LbL film is exposed. The absorbance of the film was higher at neutral pH than that in weakly acidic solutions. The intensity of the absorption band decreased when an electrode potential higher than 0.6 V was applied in the presence of H(2)O(2), while virtually no response was observed at lower electrode potential. The colorimetric response was suppressed in solutions with higher buffer capacity. The results were rationalized on the basis of the changes in local pH at the vicinity of the electrode surface, which in turn was induced by electrolysis of H(2)O(2) on the electrode surface. A possible application of the system for colorimetric sensing of H(2)O(2) was discussed. PMID:20621819

  7. Photoluminescence from stain-etched polycrystalline Si thin films A. J. Steckl, J. Xu, and H. C. Mogul

    E-print Network

    Steckl, Andrew J.

    Photoluminescence from stain-etched polycrystalline Si thin films A. J. Steckl, J. Xu, and H. C 1992) Visible room-temperature photoluminescence (PL) has been observed from stainSi) after stain-etching in a 1:3:5 solution of HF:HN03:H,0. Under UV excitation, the stain-etched doped

  8. Evaluation of confirmatory stains used for direct microscopic somatic cell counting of sheep milk.

    PubMed

    Petersson, K H; Connor, L A; Petersson-Wolfe, C S; Rego, K A

    2011-04-01

    Current FDA regulatory screening and confirmatory methods, electronic cell counting and the direct microscopic somatic cell count (DMSCC), differ for the detection of abnormal milk in sheep and goats. The DNA-specific electronic SCC screening methods such as Fossomatic (Foss, Hillerød, Denmark) can be used for both sheep and goat milk; however, the nonspecific methylene blue-based stains used for DMSCC in sheep cannot be used for goats as they nonspecifically stain cytoplasmic particles naturally present in goat milk. The DNA-specific stain pyronin Y-methyl green (PMG) is currently used for DMSCC in goats. Sheep also shed cytoplasmic particles during the milk secretory process, but in fewer numbers than goats. The objective of this study was to determine whether the nonspecific, methylene blue-based Levowitz-Weber (L-W) stain is the appropriate regulatory stain to use for DMSCC in sheep milk. Composite milk samples from 42 commercial dairy ewes were collected every 4 wk for the duration of each ewe's lactation for a total of 10 sample days. Milk samples were subjected to 3 methods of SCC determination: automated Fossomatic counting, DMSCC with L-W stain, and DMSCC with PMG stain conducted according to FDA regulatory procedures (2400 series forms). The DMSCC from milk smears stained with L-W were greater than those from smears stained with PMG and those from the Fossomatic analysis on 6 of the 10 sampling days. Milk smears stained with PMG did not differ from Fossomatic analysis at any sampling. The average milk SCC for L-W, PMG, and Fossomatic were (mean±SE) 275±36×10(3), 174±24×10(3), and 164±24×10(3) cells/mL, respectively. The DMSCC for milk stained with L-W was 58% greater than that with PMG and 68% greater than that obtained with the Fossomatic analysis. In conclusion, DMSCC of sheep milk stained with the nonspecific, methylene blue L-W stain resulted in a marked increase in SCC over that of the DNA-specific stain PMG and Fossomatic SCC analysis. The findings of this study support the continued use of Fossomatic SCC but recommend the replacement of the methylene blue-based stains with DNA-specific PMG for determination of DMSCC in sheep milk. PMID:21426980

  9. 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

  10. A new flow cytometric method for discrimination of apoptotic cells and detection of their cell cycle specificity through staining of F-actin and DNA.

    PubMed

    Endresen, P C; Prytz, P S; Aarbakke, J

    1995-06-01

    Drug-initiated apoptosis of human leukemia HL-60, THP-1, and U-937 cells was studied via multiparameter flow cytometry and cell sorting. A new flow cytometric method that allows both identification and quantitation of apoptotic cells and estimation of their cell cycle specificity is presented. The method is based on paraformaldehyde fixation followed by staining of F-actin and DNA with fluorescein isothiocyanate (FITC)-phalloidin and propidium iodide (PI), respectively. Bivariate green fluorescence (F-actin) vs. side scatterplots of HL-60 cells treated with 10 microM etoposide for 4 h showed two cell populations, one with high green fluorescence and low side scatter and one with low green fluorescence and high side scatter. Sorting revealed cells with intact nuclei in the high green fluorescence/low side scatter population and cells with fragmented nuclei in the low green fluorescence/high side scatter population, demonstrating that the cells in the latter population were apoptotic. Exposure of HL-60 cells to 10 microM etoposide for 4 h resulted in S-phase selective apoptosis, whereas 5 micrograms/ml cycloheximide initiated apoptosis mainly in G0/G1-phase and S-phase cells. The apoptotic response of HL-60 cells to 20 GY gamma-irradiation was selective for S-phase and G2 + M-phase cells. The present method offers the opportunity to estimate the cell cycle distributions of both the apoptotic and the nonapoptotic cell populations, which is especially valuable when apoptosis occurs in association with cell cycle perturbations. A similar shift from one to two cell populations in green fluorescence vs. side scatter-plots, similar to that observed for HL-60 cells, was observed in the THP-1 and U-937 cell lines secondary to etoposide treatment. PMID:7545098

  11. 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

  12. An evaluation of Retaine™ ophthalmic emulsion in the management of tear film stability and ocular surface staining in patients diagnosed with dry eye

    PubMed Central

    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

  13. Silver staining in transcriptionally active NORs of meiotic and mitotic cells in Acheta domesticus (L.) (Orthoptera)

    Microsoft Academic Search

    R. Czaker; Universitfit Wien

    1978-01-01

    The behaviour of the NOR material in mitotic and meiotic cells of Acheta domesticus was studied by silver staining. — In mitotic chromosomes black silver staining is observed in the centromeric region of 2 pairs of acrocentric chromosomes. Additionally a polymorphic silver positive region is found at the telomere of a large submetacentric chromosome. — The Ag-pattern of the amplified

  14. Flow cytometric discrimination of bacterial populations in seawater based on SYTO 13 staining of nucleic acids

    Microsoft Academic Search

    Marc Troussellier; Claude Courties; Philippe Lebaron; Pierre Servais

    1999-01-01

    Flow cytometric (FCM) counts of bacteria stained with SYTO 13, a cyanine dye, were highly correlated with DAPI epifluorescence microscopic counts in coastal seawater samples. Fluorescence intensity of stained cells appeared to depend on nucleic acid content and on the polarization of cell membranes. Right angle light scatter values of bacterial populations were clearly related to cell size. By FCM

  15. Characterization of six textile dyes as fluorescent stains for flow cytometry.

    PubMed

    Simmons, D M; Mercer, A V; Hallas, G; Dyson, J E

    1990-01-01

    Few fluorescent stains specific for cell constituents other than DNA are available. To assess their potential use as fluorescent stains for flow cytometry, the cell staining specificity of 55 compounds, originally synthesized for use as textile dyes and fluorescent brighteners, was explored and their excitation and emission wavebands determined. From these, six dyes were chosen for more detailed analysis. All six are vital stains, with excitation wavelengths allowing their use with an argon ion laser, and specific for a range of cell structures including mitochondria, Golgi bodies, lipid droplets, nuclear membrane, and endoplasmic reticulum. Concentrations as low as 0.01-0.25 microM were found to be adequate for most purposes, and high background fluorescence was not a problem. Their specificity allows differentiation between non-cycling and cycling cells. The properties of two of the stains allows their combination with propidium iodide or ethidium bromide for simultaneous determination of DNA content profiles. Being vital stains, usable at very low concentrations, and specific for a range of cell organelles, these six stains may be of considerable utility in flow cytofluorometry. We suggest that other textile dyes may be of use in flow cytofluorometry, or that their structures may form a starting point for the synthesis of further fluorescent stains of enhanced specificity. PMID:1688448

  16. ABOUT THE MECHANISM OF INTERFERENCE OF SILVER STAINING WITH PEPTIDE MASS SPECTROMETRY

    E-print Network

    Paris-Sud XI, Université de

    1 ABOUT THE MECHANISM OF INTERFERENCE OF SILVER STAINING WITH PEPTIDE MASS SPECTROMETRY Sophie staining interference with mass spectrometry *: to whom correspondence should be addressed Correspondence; MALDI: Matrix-Assisted Laser Desorption and Ionization; MS: mass spectrometry; RuBPS: ruthenium II tris

  17. A berberine-aniline blue fluorescent staining procedure for suberin, lignin, and callose in plant tissue

    Microsoft Academic Search

    Mark C. Brundrett; Daryl E. Enstone; Carol A. Peterson

    1988-01-01

    Summary A fluorescent staining procedure to detect suberin, lignin and callose in plants has been developed. This procedure greatly improves on previous methods for visualizing Casparian bands in root exodermal and endodermal cells, and performs equally well on a variety of other plant tissues. Berberine was selected as the most suitable replacement forChelidonium majus root extract after comparing the staining

  18. Thioflavin S fluorescent and congo red anisotropic stainings in the histologic demonstration of amyloid

    Microsoft Academic Search

    G. Kelényi

    1967-01-01

    Two methods employed for the histological detection of amyloid, the recently developed Thioflavin S fluorescent microscopic procedure and the Congo red anisotropic staining were compared. It was observed that in senile alterations of the brain, heart, and pancreas and in secondary amyloidosis both methods demonstrate the same structures. Dichroism of Congo red stained structures was also studied and it was

  19. Indocyanine green staining and removal of internal limiting membrane in macular hole surgery: histology and outcome

    Microsoft Academic Search

    Alvin K. H Kwok; Winnie W. Y Li; C. P Pang; Timothy Y. Y Lai; Gary H. F Yam; Nongnart R Chan; Dennis S. C Lam

    2001-01-01

    PURPOSE: To report the surgical technique, outcome, and histologic findings involving indocyanine green staining and removal of internal limiting membrane in primary macular hole surgery.METHODS: Prospectively, consecutive patients with idiopathic macular hole or myopic macular hole with retinal detachment were recruited. After pars plana vitrectomy and epiretinal membrane removal, the internal limiting membrane was stained and removed. The specimens were

  20. A rapid silver staining and destaining technique for the nucleolus organizer region.

    PubMed

    Dhar, P K; Kumar, M R; Nayak, S; Rao, T R; Joseph, A; Devi, S; Kumari, U; Bhat, S M; Bhat, K R

    1995-11-01

    Silver staining of nucleolar organizing regions (NOR) is common, but a standard protocol is lacking. A modification of a rapid silver nitrate staining technique for NORs is presented here. Advantages of the modified technique include reliability, speed, cost and the fact that it can be carried out in the light. PMID:9044659

  1. METHODS FOR THE USE OF INDIUM AS AN ELECTRON STAIN FOR NUCLEIC ACIDS

    Microsoft Academic Search

    MICHAEL L. WATSON; WILLIAM G. ALDRIDGE

    1961-01-01

    Methods are presented for the staining of blocks of tissue with trivalent indium so that good contrast and good specificity for nucleic acids is achieved for the electron microscope. The tissue is fixed in organic fixative, dehydrated, subjected to reduction by lithium borohydride, acetylated by acetic anhydride, stained with trivalent indium dissolved in organic solvent, and embedded. The embedding material

  2. Staining of the Lens Capsule for Circular Continuous Capsulorrhexis in Eyes With White Cataract

    Microsoft Academic Search

    Masayuki Horiguchi; Kensaku Miyake; Ichiro Ohta; Yasuki Ito

    e have developed a technique of staining the anterior capsule with a solution of indocyanine green that facilitates performance of the circular continuous cap- sulorrhexis in eyes with a mature cataract. We compared the results of phaco- emulsification and intraocular lens implantation in 10 eyes with the capsule stained with results of 10 eyes having the same procedure with standard

  3. Differential giemsa staining of sister chromatids and the study of sister chromatid exchanges without autoradiography

    Microsoft Academic Search

    Sheldon Wolff; Paul Perry

    1974-01-01

    Chinese hamster ovary cells grown for two rounds of DNA replication in the presence of BrdUrd contain sister chromatids that fluoresce differentially when stained with Hoechst 33258. If such fluorescent treatments are followed by incubation in 2 X SSC or water at 62° C and staining in 3% Giemsa, the chromosomes now contain one dark (unifilarly substituted) chromatid and one

  4. 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.

  5. Selection of optimum tetrazolium salts for use in histochemistry: The value of structure-staining correlations

    Microsoft Academic Search

    Richard W. Horobin

    1982-01-01

    Summary  A structure-staining correlation study was made of a number of tetrazolium salts and formazans used in histochemistry. Numerical parameters describing molecular structure were calculated from structural formulae. Staining data were from previously published work. It was found that several significant practical variables (namely, uptake and reducibility of tetrazolium salts; substantivity, lipid solubility, and crystal size of formazans) could be predicted

  6. Evidence from dithizone and selenium zinc histochemistry that perivascular mossy fiber boutons stain preferentially “in vivo”

    Microsoft Academic Search

    C. A. Howell; C. J. Frederickson; G. Danscher

    1989-01-01

    This paper describes a perivascular staining pattern that is obtained when dithizone or sodium selenite are used to label zinc intravitally. Our observations indicate that the perivascular staining is a result of zinc labeling in mossy fiber boutons adjacent to capillaries and suggest that there might be a special blood brain barrier in the mossy fiber regions.

  7. Histopathological Findings in Immunohistological Staining of the Granulomatous Tissue Reaction Associated with Tuberculosis

    PubMed Central

    Karimi, Shirin; Pourabdollah, Mihan; Sadr, Makan; Karbasi, Mehrdad; Bahadori, Moslem

    2014-01-01

    Purpose. The histological diagnosis of Mycobacterium tuberculosis (MTB) remains a diagnostic challenge despite different methods. Immunohistochemistry (IHC) not only could confirm granulomatous tissue involvement but also can demonstrate MTB antigen immunolocalization. This study tries to clarify the details of immunohistochemical staining for MTB with pAbBCG. Materials/Methods. Twenty-three confirmed TB granulomatous tissue samples were studied by Ziehl-Neelsen and immunohistochemistry (IHC) staining with pAbBCG. Samples were selected from the archive of the Department of Pathology, National Research Institute of Tuberculosis and Lung Disease, Tehran, Iran. Results. IHC staining was positive in all samples, whereas Ziehl-Neelsen was positive in 9 cases out of 23 (39.1%). Tissue types used were pleural tissue, lymph nodes, and lung tissue. IHC showed positive coarse granular cytoplasmic and round, fragmented bacillary staining. In this study, epithelioid cells clearly showed more positive staining at the periphery of the granuloma rather than the center of granuloma. There is also positive staining in endothelial cells, fibroblasts, plasma cells, lymphocytes, and macrophages outside the granuloma. Conclusion. Considering the criteria of positive immunohistochemical staining of TB granulomatous reactions, this stain not only highlights the presence of mycobacterial antigens for tissue diagnosis, but also could morphologically localize its distribution in different cells. PMID:24511393

  8. 9 CFR 147.3 - The stained-antigen, rapid, whole-blood test. 3

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...The stained-antigen, rapid, whole-blood test. 3 147.3 Section 147.3...ON NATIONAL POULTRY IMPROVEMENT PLAN Blood Testing Procedures § 147.3 The stained-antigen, rapid, whole-blood test. 3 3 The procedure...

  9. DNA cytofluorometry on large and small cell nuclei stained with pararosaniline Feulgen

    Microsoft Academic Search

    Setsuya Fujita

    1973-01-01

    To examine reliability of DNA cytofluorometry with conventional Feulgen staining, measurement was carried out on smears of small and large neurons of human cerebellum, using a Digital-Microfluorometer with incident green light excitation. Nuclear fluorescence of the inner granule neuron and the Purkinje cell came to good agreement with each other when the staining was reduced so that the maximal extinction

  10. Inhibitory effect of a 2,4-bis(4-hydroxyphenyl)-2-butenal diacetate on neuro-inflammatory reactions via inhibition of STAT1 and STAT3 activation in cultured astrocytes and microglial BV-2 cells.

    PubMed

    Kim, Jin A; Yun, Hyung-Mun; Jin, Peng; Lee, Hee Pom; Han, Jin Yi; Udumula, Venkatareddy; Moon, Dong Cheul; Han, Sang Bae; Oh, Ki Wan; Ham, Young Wan; Jung, Heon-Sang; Song, Ho Sueb; Hong, Jin Tae

    2014-04-01

    2,4-Bis(p-hydroxyphenyl)-2-butenal (Butenal), a tyrosine-fructose Maillard reaction product has been demonstrated as an effective compound for prevention of neuroinflammatory diseases. However, this compound was vulnerable to environmental factors. Our research has been continuously made to improve druggability of Butenal and identified 2,4-bis(4-hydroxyphenyl)but-2-enal diacetate (HPBD) as an alternative. Herein, to investigate potential anti-neuroinflammatory and anti-amyloidogenic effects of HPBD, we treated HPBD (0.5, 1, and 2 ?g/ml) on the lipopolysaccharides (LPS) (1 ?g/ml) stimulated astrocytes and microglial BV-2 cell. HPBD inhibited LPS-induced NO and ROS production, and LPS-elevated expression of iNOS, COX2, ?-site APP-cleaving enzyme 1 (BACE1), C99, and A?1-42 levels as well as attenuation of ?-secretase activities. The activation of nuclear factor-kappaB (NF-?B), signal transducer and activator of transcription1 (STAT1), and STAT3 was concomitantly inhibited by HPBD. Moreover, siRNA targeting STAT3 abolished HPBD-induced inhibitory effects on neuro-inflammation and amyloidogenesis. In addition, pull down assay and docking model showed interaction of HPBD with STAT3. These findings suggest that HPBD may be useful and potentially therapeutic choices for the treatment of neuroinflammatory diseases. PMID:23891616

  11. Uncovering the origin of the black stains in Lascaux Cave in France.

    PubMed

    Saiz-Jimenez, Cesareo; Miller, Ana Z; Martin-Sanchez, Pedro M; Hernandez-Marine, Mariona

    2012-12-01

    Lascaux Cave in France was discovered in 1940. Since being opened to visitors the cave has suffered three major microbial outbreaks. The current problem is the fast dissemination of black stains which are threatening the Palaeolithic paintings. Previous data pointed to the involvement of new fungal species in the formation of black stains on the rock walls and ceiling. However, it appears that there could be other reasons for the formation of different and extensive black stains coating the surface of the clayey sediments. Our analyses reveal that black stains on clayey sediments are mainly produced by Acremonium nepalense, a manganese oxide-depositing fungus, widely distributed in the cave. Thus, in Lascaux Cave, the black stains have a dual origin: on limestone rocks they are mainly produced by the accumulation of fungal melanins, and on clayey sediments by the biogenic deposition of black manganese oxides. PMID:23106913

  12. Picrosirius red staining: a useful tool to appraise collagen networks in normal and pathological tissues.

    PubMed

    Lattouf, Raed; Younes, Ronald; Lutomski, Didier; Naaman, Nada; Godeau, Gaston; Senni, Karim; Changotade, Sylvie

    2014-10-01

    Specific staining of the extracellular matrix components is especially helpful in studying tissue remodeling, particularly in the case of connective tissue pathologies. As developed by Junqueira and colleagues in 1979, specific staining by Picrosirius red is one of the most important stains to study collagen networks in different tissues. Under polarized light, collagen bundles appear green, red or yellow, and are easily differentiated from the black background, thus allowing for quantitative morphometric analysis. As Junqueira and colleagues point out, many studies use color staining to differentiate collagen bundles and to specify collagen types, yet other studies report that polarized colors only reflect fiber thickness and packing. Using a simple histological example, our study illustrates the inability of Picrosirius red staining to differentiate collagen types, since the absorbed amount of polarized light by this dye strictly depends on the orientation of the collagen bundles. PMID:25023614

  13. An automatic stain removal algorithm of series aerial photograph based on flat-field correction

    NASA Astrophysics Data System (ADS)

    Wang, Gang; Yan, Dongmei; Yang, Yang

    2010-10-01

    The dust on the camera's lens will leave dark stains on the image. Calibrating and compensating the intensity of the stained pixels play an important role in the airborne image processing. This article introduces an automatic compensation algorithm for the dark stains. It's based on the theory of flat-field correction. We produced a whiteboard reference image by aggregating hundreds of images recorded in one flight and use their average pixel values to simulate the uniform white light irradiation. Then we constructed a look-up table function based on this whiteboard image to calibrate the stained image. The experiment result shows that the proposed procedure can remove lens stains effectively and automatically.

  14. 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.

  15. Porcine Intestinal Mast Cells. Evaluation of Different Fixatives for Histochemical Staining Techniques Considering Tissue Shrinkage

    PubMed Central

    Rieger, J.; Twardziok, S.; Huenigen, H.; Hirschberg, R.M.; Plendl, J.

    2013-01-01

    Staining of mast cells (MCs), including porcine ones, is critically dependent upon the fixation and staining technique. In the pig, mucosal and submucosal MCs do not stain or stain only faintly after formalin fixation. Some fixation methods are particularly recommended for MC staining, for example the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF) has been reported to work excellently for the preservation of fixation-sensitive antigens. The aim of this study was to establish a reliable histological method for counting of MCs in the porcine intestinum. For this purpose, different tissue fixation and staining methods that also allow potential subsequent immunohistochemical investigations were evaluated in the porcine mucosa, as well as submucosa of small and large intestine. Tissues were fixed in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with either polychromatic methylene blue, alcian blue or toluidine blue. For the first time our study reveals that ZSF, a heavy metal fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not suitable for histochemical visualization of MCs in the pig intestine. All other tested fixatives were suitable. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs difficult. The polychromatic methylene blue proved to be the optimal staining. In order to compare MC counting results of the different fixation methods, tissue shrinkage was taken into account. As even the same fixation caused shrinkagedifferences between tissue from small and large intestine, different factors for each single fixation and intestinal localization had to be calculated. Tissue shrinkage varied between 19% and 57%, the highest tissue shrinkage was found after fixation with ZSF in the large intestine, the lowest one in the small intestine after lead acetate fixation. Our study emphasizes that MC counting results from data using different fixation techniques can only be compared if the respective studyimmanent shrinkage factor has been determined and quantification results are adjusted accordingly. PMID:24085270

  16. NMDAR-1 staining in the lateral geniculate nucleus of normal and visually deprived cats.

    PubMed

    Ziburkus, J; Bickford, M E; Guido, W

    2000-01-01

    In normal adult cats, a monoclonal antibody directed toward the NR-1 subunit of the N-methyl-D-aspartate (NMDA) receptor (Pharmingen, clone 54.1) produced dense cellular and neuropil labeling throughout all layers of the lateral geniculate nucleus (LGN) and adjacent thalamic nuclei, including the thalamic reticular, perigeniculate, medial intralaminar, and ventral lateral geniculate nuclei. Cellular staining revealed well-defined somata, and in some cases proximal dendrites. NMDAR-1 cell labeling was also evident in the LGN of early postnatal kittens, suggesting that developing LGN cells possess this receptor subunit at or before eye opening. Within the A-layers of the adult LGN, staining encompassed a wide range of soma sizes. Soma size comparisons of NMDAR-1 stained cells with those stained with an antibody directed toward a nonphosphorylated neurofilament protein (SMI-32), which selectively stains Y-relay cells (Bickford et al., 1998), or an antibody to glutamic acid decarboxylase (GAD), which stains for GABAergic interneurons, suggested that NMDA receptors are utilized by relay cells and interneurons. NMDAR-1 staining was also observed in the LGN of cats with early monocular lid suture. Although labeling was apparent in both deprived and nondeprived A-layers of LGN, the distribution of soma sizes was significantly different. In the deprived A-layers of LGN, staining was limited to small- and medium-sized cells. Cells with relatively large soma were lacking. However, cell density measurements as well as soma size comparisons with cells stained for Nissl substance suggested these differences were due to deprivation-induced cell shrinkage and not to a loss of NMDAR-1 staining in Y-cells. Taken together, these results suggest that NMDA receptors are utilized by both relay cells and interneurons in LGN and that alterations in early visual experience do not necessarily affect the expression of NMDA receptors in the LGN. PMID:10824673

  17. Degradation of Giardia lamblia cysts in mixed human and swine wastes.

    PubMed

    Deng, M Y; Cliver, D O

    1992-08-01

    This study was conducted to determine the persistence of Giardia lamblia cysts in mixed septic tank effluent and swine manure slurry and to correlate fluorescein diacetate-propidium iodide staining of G. lamblia cysts with their morphology under low-voltage scanning electron microscopy. Under field conditions, G. lamblia cysts were degraded more rapidly in the mixed waste than in the control Dulbecco's phosphate-buffered saline (PBS). For total and viable cysts, the mixed waste had D values (time for a 90% reduction in number of cysts) of 18.3 and 15.5 days, and the Dulbecco's PBS control had D values of 41.6 and 26.8 days. The rates of cyst degradation in septic tank effluent and in Dulbecco's PBS were similar. Increasing the proportion of swine manure slurry in the mixed waste favored degradation of the parasite. These results indicate that the mixed waste treatment was the predominant factor affecting the cyst persistence and that it was swine manure slurry that played the role of degrading the parasite. Visualization of viable and nonviable Giardia cysts with low-voltage scanning electron microscopy revealed an excellent correlation between the viability of the cysts determined by fluorescein diacetate-propidium iodide staining and their electron microscopic morphology. PMID:1381171

  18. Subjective and objective measures of corneal staining related to multipurpose care systems.

    PubMed

    Pritchard, Nicola; Young, Graeme; Coleman, Sarah; Hunt, Chris

    2003-03-01

    An objective, digital-imaging method of measuring corneal staining was evaluated in 24 subjects wearing soft contact lenses. The method was used to compare the clinical performance of common multipurpose care systems (MPS) for soft contact lens care. Subjects used three different MPS, one containing polyquaternium-1 (PQ) and two containing polyhexanide (PX1 and PX2), for 2 weeks in a randomised, single-masked (investigator) crossover study. Corneal staining induced with the three MPS was analysed using an image-processing program (ImageTool, UTHSCSA Version 2, University of Texas, USA). Conjunctival hyperaemia and papillae were also evaluated. The intraclass correlation coefficient was similar with image analysis to that of investigator grading (0.876, 0.879, respectively). Significant differences in staining response were detected using the objective method. There was significantly less staining area with polyquaternium-1 (PQ) than polyhexanide (PQ: 0.12 mm(2), PX2: 0.91 mm(2)). Inferior palpebral papillae were significantly greater with PX2 than with PQ (1.0, 0.7 (0-4), respectively). The technique was shown to be an effective method of evaluating different corneal staining responses. Bilateral corneal staining in three or more quadrants is useful in the diagnosis of MPS-related staining. PMID:16303491

  19. 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

  20. Effects of histological staining on the analysis of human DNA from archived slides.

    PubMed

    Simons, Joanne L; Vintiner, Sue K

    2011-01-01

    Archived slides of cell smears treated with histological stains for sperm detection are often the only source of DNA available when cold cases are reopened. There have been conflicting reports as to the negative effects of particular histological stains on DNA recovery and quality from human cells, making stain selection an important consideration for forensic laboratories. This study investigates the effect of several staining systems on DNA recovery from histological slide samples stored from 0 to 10 weeks. DNA profiles obtained after analysis of these samples with AmpFlSTR(®) Identifiler™ and increased cycle AmpFlSTR(®) SGM Plus™ short tandem repeat (STR) profiling systems and the effects that these stains have on DNA quantity and quality over time are described. Results indicate that Christmas Tree and Hematoxylin and Eosin stains do not have significantly different effects on DNA quality after 10-week storage of slides. This research will assist scientists to select staining systems that have minimal deleterious effects on the DNA recovered. PMID:21198622

  1. Gram Stains: A Resource for Retrospective Analysis of Bacterial Pathogens in Clinical Studies

    PubMed Central

    Srinivasan, Usha; Ponnaluri, Sreelatha; Villareal, Lisa; Gillespie, Brenda; Wen, Ai; Miles, Arianna; Bucholz, Brigette; Marrs, Carl F.; Iyer, Ram K.; Misra, Dawn; Foxman, Betsy

    2012-01-01

    We demonstrate the feasibility of using qPCR on DNA extracted from vaginal Gram stain slides to estimate the presence and relative abundance of specific bacterial pathogens. We first tested Gram stained slides spiked with a mix of 108 cfu/ml of Escherichia coli and 105 cfu/ml of Lactobacillus acidophilus. Primers were designed for amplification of total and species-specific bacterial DNA based on 16S ribosomal gene regions. Sample DNA was pre-amplified with nearly full length 16S rDNA ribosomal gene fragment, followed by quantitative PCR with genera and species-specific 16S rDNA primers. Pre-amplification PCR increased the bacterial amounts; relative proportions of Escherichia coli and Lactobacillus recovered from spiked slides remained unchanged. We applied this method to forty two archived Gram stained slides available from a clinical trial of cerclage in pregnant women at high risk of preterm birth. We found a high correlation between Nugent scores based on bacterial morphology of Lactobacillus, Gardenerella and Mobiluncus and amounts of quantitative PCR estimated genus specific DNA (rrn copies) from Gram stained slides. Testing of a convenience sample of eight paired vaginal swabs and Gram stains freshly collected from healthy women found similar qPCR generated estimates of Lactobacillus proportions from Gram stained slides and vaginal swabs. Archived Gram stained slides collected from large scale epidemiologic and clinical studies represent a valuable, untapped resource for research on the composition of bacterial communities that colonize human mucosal surfaces. PMID:23071487

  2. 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

  3. CdS/TiO2-fluorescein isothiocyanate nanoparticles as fluorescence resonance energy transfer probe for the determination of trace alkaline phosphatase based on affinity adsorption assay.

    PubMed

    Liu, Jia-Ming; Lin, Li-ping; Jiao, Li; Cui, Ma-Lin; Wang, Xin-Xing; Zhang, Li-Hong; Zheng, Zhi-Yong

    2012-08-30

    The CdS/TiO(2)-fluorescein isothiocyanate (FITC) luminescent nanoparticles (CdS/TiO(2)-FITC) with the particle size of 20 nm have been synthesized by sol-gel method. CdS/TiO(2)-FITC could emit the fluorescence of both FITC and CdS/TiO(2). The fluorescence resonance energy transfer (FRET) occurred between the donor CdS/TiO(2) and the acceptor FITC in the CdS/TiO(2)-FITC. Taking advantages of the excellent characteristics of FRET, a new CdS/TiO(2)-FITC FRET labeling reagent and a CdS/TiO(2)-FITC-wheat germ agglutinin (CdS/TiO(2)-FITC-WGA) fluorescent probe have been developed. The FRET occurring between the donor CdS/TiO(2) and the acceptor FITC in the labelled product CdS/TiO(2)-FITC-WGA-AP, formed in the affinity adsorption reaction between the WGA in this CdS/TiO(2)-FITC-WGA fluorescent probe and alkaline phosphatase (AP), sharply enhanced the fluorescence signal of FITC and quench the fluorescence signal of CdS/TiO(2). Moreover, the ?F (the change of the fluorescence signal) of FITC and CdS/TiO(2) were proportional to the content of AP, respectively. Thus, a new method that CdS/TiO(2)-fluorescein isothiocyanate nanoparticles for the determination of trace AP based on FRET-affinity adsorption assay has been established. The limit of quantification (LOQ) of the method was 1.3×10(-17) g AP mL(-1) for CdS/TiO(2) and 1.1×10(-17) g AP mL(-1) for FITC, respectively. This sensitive, rapid, high selective and precise method has been applied to the determination of AP in human serum and the prediction of human disease with the results agreed well with enzyme-linked immunosorbent assay (ELISA) in Zhangzhou Municipal Hospital of Fujian Province. Simultaneously, the reaction mechanism for the determination of AP was also discussed. PMID:22939139

  4. 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.

  5. Lack of clinical utility of urine gram stain for suspected urinary tract infection in pediatric patients.

    PubMed

    Cantey, Joseph B; Gaviria-Agudelo, Claudia; McElvania TeKippe, Erin; Doern, Christopher D

    2015-04-01

    Urinary tract infection (UTI) is one of the most common infections in children. Urine culture remains the gold standard for diagnosis, but the utility of urine Gram stain relative to urinalysis (UA) is unclear. We reviewed 312 pediatric patients with suspected UTI who had urine culture, UA, and urine Gram stain performed from a single urine specimen. UA was considered positive if ?10 leukocytes per oil immersion field were seen or if either nitrates or leukocyte esterase testing was positive. Urine Gram stain was considered positive if any organisms were seen. Sensitivity, specificity, and positive and negative predictive values were calculated using urine culture as the gold standard. Thirty-seven (12%) patients had a culture-proven UTI. Compared to urine Gram stain, UA had equal sensitivity (97.3% versus 97.5%) and higher specificity (85% versus 74%). Empirical therapy was prescribed before the Gram stain result was known in 40 (49%) patients and after in 42 (51%) patients. The antibiotics chosen did not differ between the two groups (P = 0.81), nor did they differ for patients with Gram-negative rods on urine Gram stain compared to those with Gram-positive cocci (P = 0.67). From these data, we conclude that UA has excellent negative predictive value that is not enhanced by urine Gram stain and that antibiotic selection did not vary based on the urine Gram stain result. In conclusion, the clinical utility of urine Gram stain does not warrant the time or cost it requires. PMID:25653411

  6. Utility of ancillary stains for Helicobacter pylori in near-normal gastric biopsies.

    PubMed

    Panarelli, Nicole C; Ross, Dara S; Bernheim, Oren E; Landzberg, Zachary B; Schuetz, Audrey N; Jenkins, Stephen G; Landzberg, Brian R; Jessurun, Jose; Yantiss, Rhonda K

    2015-03-01

    Documentation of Helicobacter pylori infection and eradication is important, prompting some clinicians and pathologists to request ancillary stains on all gastric samples that do not demonstrate H. pylori on initial histologic review. Studies evaluating the utility of ancillary stains in patients with minimal inflammation are lacking. We used Giemsa, Warthin-Starry, acridine orange, and immunohistochemical stains to search for organisms in 56 patients with biochemical evidence of H. pylori infection (positive Campylobacter-like organism test) and gastric mucosal samples interpreted to be H pylori negative by hematoxylin and eosin (H&E). We correlated the findings with severity of inflammation and patients' histories of medication use. Nineteen (34%) patients had histologically normal mucosae, 22 (39%) had chronic inflammation with or without focal activity, and 15 (27%) had chemical gastropathy. Fifty (89%) cases were negative for H. pylori with additional stains, and 6 contained bacteria that were detected with all 4 ancillary stains and on retrospective review of H&E-stained sections that also showed chronic inflammation. Eleven (20%) patients were taking proton pump inhibitors, and 4 (7%) had previously received H. pylori eradication therapy. We conclude that H&E stains demonstrate H. pylori in most infected patients, so preemptive stain requests are largely unnecessary. Failure to identify bacteria by H&E evaluation generally reflects their absence in biopsy material, even among Campylobacter-like organism test--positive patients. However, organisms may be overlooked in patients with mild inflammation and in those receiving proton pump inhibitor or antibiotic therapy, so one should consider ordering ancillary stains to enhance detection of bacteria in these settings. PMID:25582501

  7. Mallory stain may indicate differential rates of RNA synthesis: II. Comparative observations in vertebrate nuclei.

    PubMed

    Chieffi Baccari, G; Marmorino, C; Minucci, S; Di Matteo, L; d'Istria, M

    1992-01-01

    The differential staining of nuclei by the use of the Mallory trichrome method was investigated in a variety of tissues of representative vertebrates. By this method nuclei stained orange or blue; erythrocyte nuclei stained red. Since the higher affinity for aniline blue is due to an increased RNA synthesis, it was possible to reveal not only the changing metabolic status of a cell type, as shown for instance in the liver parenchyma and other glandular tissues, and nervous tissue, but also in different cell populations in the same tissue, such as the spleen. PMID:1380852

  8. Development of a Whole Blood Staining Device for use During Space Shuttle Flights

    NASA Technical Reports Server (NTRS)

    Sams, Clarence F.; Crucian, Brian E.; Clift, Vaughan L.; Meinelt, Ellen M.

    1999-01-01

    Exposure to microgravity during space flight results in profound physiologic changes. Numerous studies have shown changes in circulating populations of peripheral blood immune cells immediately after space flight. It is currently unknown if these changes result from exposure to microgravity or are caused by the stress of reentry and readaptation to gravity. We have developed the whole blood staining device as a system for the staining of whole blood collected during space flight for subsequent flow cytometric analysis, This device contains all liquids to address safety issues concerned with space flight and also moves the cells through the staining, lyse/fixation and dilution steps.

  9. Reduced Fluorescein Angiography and Fundus Photography Use in the Management of Neovascular Macular Degeneration and Macular Edema During the Past Decade

    PubMed Central

    Schneider, Eric W.; Mruthyunjaya, Prithvi; Talwar, Nidhi; Harris Nwanyanwu, Kristen; Nan, Bin; Stein, Joshua D.

    2014-01-01

    Purpose. We assessed recent trends in the use of diagnostic testing for neovascular age-related macular degeneration (NVAMD) and macular edema (ME). Methods. Claims data from a managed-care network were analyzed on patients with NVAMD (n = 22,954) or ME (n = 31,810) to assess the use of fluorescein angiography (FA), fundus photography (FP), and optical coherence tomography (OCT) from 2001 to 2009. Repeated-measures logistic regression was performed to compare patients' odds of undergoing these procedures in 2001, 2005, and 2009. In addition, the proportions of patients with an incident NVAMD or ME diagnosis in 2003 or 2008 who underwent FA, FP, and OCT were compared. Results. From 2001 to 2009, among patients with NVAMD, the odds of undergoing OCT increased 23-fold, whereas the odds of receiving FA and FP decreased by 68% and 79%, respectively. Similar trends were observed for ME. From 2003 to 2008, the proportion of patients undergoing OCT within 1 year of initial diagnosis increased by 315% for NVAMD and by 143% for ME; the proportion undergoing OCT without FA within 1 year increased by 463% for NVAMD and by 216% for ME. Conclusions. Use of OCT increased dramatically during the past decade, whereas use of FA and FP declined considerably, suggesting that OCT may be replacing more traditional diagnostic testing in patients with NVAMD or ME. Future studies should evaluate whether this increased reliance on OCT instead of FA and FP affects patient outcomes. PMID:24346174

  10. Photodetection of early cancer in the upper aerodigestive tract and the bronchi using photofrin II and colorectal adenocarcinoma with fluoresceinated monoclonal antibodies

    NASA Astrophysics Data System (ADS)

    Wagnieres, Georges A.; Braichotte, Daniel; Chatelain, Andre; Depeursinge, Christian D.; Monnier, Philippe; Savary, Jean-Francois; Fontolliet, Charlotte; Calmes, J.-M.; Givel, Jean-Claude; Chapuis, G.; Folli, S.; Pelegrin, A.; Buchegger, F.; Mach, J.-P.; van den Bergh, Hubert

    1991-11-01

    The performance of a fluorescence endoscope for the detection of early cancer is clinically evaluated. the apparatus is based on the imaging of the laser-induced fluorescence (LIF) of a dye which localizes in the tumor after IV injection with a higher concentration than in the surrounding normal tissue. The tests are carried out in several of the hollow organs, such as the upper aerodigestive tract, the bronchi, and the colon. In the two former cases the dye used is photofrin II, whereas in the latter case conjugates between monoclonal antibodies (Mab) directed against carcinoembrionic antigen (CEA) and fluorescein molecules are injected. The fluorescence contrast between tumor and surrounding tissue is enhanced by real-time image processing which eliminates most of the tissue autofluorescence as well as the fluorescence due to the relatively small amount of dye localized in the normal tissue. This is done by recording the fluorescence image in two spectral domains, after which these two images are digitized and manipulated with a mathematical operator (lookup-table). The sources of false positives and false negatives are evaluated in terms of the fluorescent dye and tissue optical properties.

  11. Design of highly sensitive phosphorescence sensor for determination of procaterol hydrochloride based on inhibition of KClO3 oxidation fluorescein isothiocyanate.

    PubMed

    Liu, Jia-Ming; Huang, Qitong; Liu, Zhen-Bo; Zhang, Li-Hong; Zheng, Zhi-Yong; Lin, Chang-Qing

    2015-06-01

    Procaterol hydrochloride (Prh) can inhibit KClO3 oxidation of fluorescein isothiocyanate (FITC) to form a non-phosphorescent compound, which causes room temperature phosphorescence (RTP) of FITC in the system to enhance sharply the linear relationship between ?Ip and the Prh content. Thus, a rapid response and highly sensitive phosphorescence sensor for the determination of Prh has been developed based on the inhibiting effect of Prh on KClO3 oxidation of FITC. This simple, high sensitivity (detection limit (LD) calculated by 3Sb /k was 0.019 fg/spot, sample volume 0.40 µl, corresponding concentration 4.8?×?10(-14) g ml(-1) ) and selective sensor with a wide linear range (0.080-11.20 g/spot) has been applied to detect Prh in blood samples, and the results were consistent with those obtained by high-performance liquid chromatography (HPLC). Simultaneously, the mechanism of the phosphorescence sensor for the detection of Prh was also investigated using infrared spectroscopy. Copyright © 2014 John Wiley & Sons, Ltd. PMID:25044504

  12. 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.

  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. Detection of Rubella Virus-Specific Immunoglobulin G in Saliva by an Amplification-Based Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibody to Fluorescein Isothiocyanate

    PubMed Central

    Vyse, A. J.; Brown, D. W. G.; Cohen, B. J.; Samuel, R.; Nokes, D. J.

    1999-01-01

    An immunoglobulin G (IgG)–capture enzyme-linked immunosorbent assay (ELISA) for rubella virus is described. The assay uses a fluorescein isothiocyanate (FITC)–anti-FITC amplification system. The detection limit of the ELISA was approximately 7 IU of rubella virus-specific IgG per ml of serum sample. For saliva samples the performances of the capture ELISA and previously described radioimmunoassay were assessed, and the results of those two assays were compared to the rubella virus-specific IgG result obtained by a commercial ELISA (Behring Enzygnost) with a panel of paired serum and saliva samples. This comparison showed that the capture ELISA with saliva was more sensitive than the radioimmunoassay and that the results correlated better with the serum IgG result than the results of the radioimmunoassay did, with an overall sensitivity of 82% and a rank correlation of 0.68, whereas the sensitivity and rank correlation for the radioimmunoassay were 74% and 0.45, respectively. For subjects of 10 years of age or younger, the ELISA with saliva had a sensitivity of 94% and a specificity of 100% compared to the results of the ELISA (Behring Enzygnost) for rubella virus-specific IgG with corresponding serum samples. The sensitivity was much lower for subjects ages 17 years or older. The assay may have wider epidemiological use with saliva specimens, particularly those from children. PMID:9889225

  15. 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....

  16. 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

  17. Oil red-O stains non-adipogenic cells: a precautionary note.

    PubMed

    Kinkel, Adrienne D; Fernyhough, Melinda E; Helterline, Deri L; Vierck, Janet L; Oberg, Karen S; Vance, Tyler J; Hausman, Gary J; Hill, Rodney A; Dodson, Michael V

    2004-09-01

    Bovine adipofibroblasts, 3T3-L1 cells, L-6 myogenic cells, and sheep satellite cells were allowed to proliferate for 48 h. Oil red-O (ORO) was dissolved in three different solvents isopropanol, propylene glycol and triethyl phosphate. At 48 h, the proliferative cultures were stained with the three stains. ORO stain prepared in both propylene glycol and triethyl phosphate resulted in bright red droplets appearing in all cultures, whereas ORO dissolved in isopropanol was not taken up by any of the cells. These data suggest that certain preparations of ORO may stain cells in non-adipogenic lineages as well as undifferentiated pre-adipocytes. Caution must be exercised when choosing solvents for ORO in differentiation studies using cells of the fat/adipose lineage. PMID:19003258

  18. Pollen viability of Polygala paniculata L. (Polygalaceae) using different staining methods.

    PubMed

    Frescura, Viviane Dal-Souto; Laughinghouse, Haywood Dail; do Canto-Dorow, Thais Scotti; Tedesco, Solange Bosio

    2012-12-01

    Polygala paniculata L. is a medicinal plant that grows in the Brazilian Atlantic coast, known as 'barba-de-São-João', 'barba-de-bode', 'vassourinha branca', and 'mimosa'. In this study, pollen viability was estimated by three different staining methods: 2% acetic orcein, 2% acetic carmine, and Alexander's stain. The young inflorescences of twenty accessions were collected and fixed in a solution of ethanol: acetic acid (3:1) for 24 hours, then stored in ethanol 70% under refrigeration. Six slides per plant, two for each stain, were prepared by squashing, and 300 pollen grains per slide were analyzed. Pollen viability was high (> 70%) for most accessions of P. paniculata using the Alexander's stain, which proved the most adequate method to estimate pollen viability. PMID:23682430

  19. Embedding, Serial Sectioning and Staining of Zebrafish Embryos Using JB-4™ Resin

    PubMed Central

    Sullivan-Brown, Jessica; Bisher, Margaret E.; Burdine, Rebecca D.

    2011-01-01

    Histological techniques are critical for observing tissue and cellular morphology. Here, we outline our protocol for embedding, serial sectioning, staining, and visualizing zebrafish embryos embedded in JB-4™ plastic resin – a glycol methacrylate-based medium that results in excellent preservation of tissue morphology. In addition we describe our procedures for staining plastic sections with Toluidine Blue or Hematoxylin and Eosin (H&E), and show how to couple these stains with whole-mount RNA in situ hybridization. We also describe how to maintain and visualize immunofluorescence and GFP signals in JB-4™ resin. The protocol we outline – from embryo preparation, embedding, sectioning and staining to visualization - can be accomplished in three days. Overall, we reinforce that plastic embedding can provide higher resolution of cellular details and is a valuable tool for cellular and morphological studies in zebrafish. PMID:21212782

  20. Efficacy of evaluation of rooster sperm morphology using different staining methods.

    PubMed

    Lukaszewicz, E; Jerysz, A; Partyka, A; Siudzi?ska, A

    2008-12-01

    This work focused on inexpensive methods of evaluation fowl sperm morphology, based on eosin-nigrosin smears, which can determine disorders in spermatogenesis and can be recommended for evaluating the fertilising potency and selecting males in flocks reproduced by artificial insemination. Four fowl breeds (Black Minorca, Italian Partridge, Forwerk and Greenleg Partridge) were used to determine the efficacy of sperm morphology evaluation using four eosin-nigrosin staining methods (according to Blom, Bakst and Cecil, Morisson, Ja?kowski) and three examiners of different experience (high, medium, novice). There were significant (P< or = 0.01) differences in sperm morphology between Blom's staining method and those of Bakst and Cecil, Morisson or Ja?kowski, irrespective of fowl breed and examiners experience. Blom stain caused sperm head swelling and showed a drastic reduction in the proportion of live spermatozoa with normal morphology. The staining method had a greater influence on sperm morphology evaluation than the experience of the examiners. PMID:18486956

  1. Extrinsic Iron Staining in Infant Teeth from Iron-Fortified Formula and Rice Cereal

    PubMed Central

    Adcock, Kim G.; Hogan, Shirley M.

    2008-01-01

    Extrinsic staining of teeth due to excessive iron intake has been reported previously in the literature. We describe a 7-month-old infant who presented with extrinsic teeth staining due to inadvertent over consumption of dietary iron. The infant was fed iron-fortified formula and rice cereal. Rice cereal, fortified with iron, was being used as part of a normal infant diet and as a thickening agent when added to the formula for treatment of gastroesophageal reflux. After several months of administration, “blackening” of the infant's teeth was noted by the mother. The stain was removed by the pediatric dentist who simply scraped the affected teeth. No further staining occurred after the amount of dietary iron was reduced. PMID:23055877

  2. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...DEPARTMENT OF THE TREASURY ARTICLES CONDITIONALLY FREE, SUBJECT TO A REDUCED RATE, ETC. General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted,...

  3. 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

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

    E-print Network

    Ottino, Julio M.

    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

  5. The art of stained glass: metaphor for the art of nursing.

    PubMed

    Hess, J D

    1995-12-01

    The aesthetic is a way of knowing the meaning of and the meaning in the art of nursing. The art of creating stained glass offers a personal metaphor for nursing's essence; the art of caring. Both arts aim to fulfil the potential of their subjects to achieve a harmony that goes beyond their individual components. Stained glass artistry and caring in nursing require technical expertise, yet technical skill and knowledge are not the substance of either art. Both transcend space and time, and the art of stained glass and the art of nursing are influenced by the artist's/nurse's personal, social and cultural history. Just as the artisan transforms the glass and lead and is transformed in the creative moment, so does the caring transaction transform both patient and nurse. This personal reflection explores the nature of caring in nursing as mirrored by the author's work with stained glass. PMID:8705607

  6. Application of Prussian blue staining in the diagnosis of ocular siderosis

    PubMed Central

    Yang, Zhen; Yang, Xiao-Li; Xu, Li-Shuai; Dai, Le; Yi, Mei-Chao

    2014-01-01

    AIM To explore the value of Prussian blue staining in the diagnosis of ocular siderosis. METHODS Between January 2012 and January 2013, the Prussian blue stain used in anterior lens capsule and vitreous liquid after centrifugation from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis. At the same time, give a negative control. RESULTS Anterior lens capsule membrane and liquid of vitreous cavity from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis revealed ferric ions that stained positively with Prussian blue. In the control group, there is no positive reaction. CONCLUSION Prussian blue staining in the diagnosis of ocular siderosis has a very significant worth, suspected cases can be definitive diagnosed. PMID:25349794

  7. Transient partial permeabilization with saponin enables cellular barcoding prior to surface marker staining1

    PubMed Central

    Behbehani, Gregory K.; Thom, Colin; Zunder, Eli R.; Finck, Rachel; Gaudilliere, Brice; Fragiadakis, Gabriela K.; Fantl, Wendy J.; Nolan, Garry P.

    2015-01-01

    Fluorescent cellular barcoding and mass-tag cellular barcoding are cytometric methods that enable high sample throughput, minimize inter-sample variation, and reduce reagent consumption. Previously employed barcoding protocols require that barcoding be performed after surface marker staining, complicating combining the technique with measurement of alcohol-sensitive surface epitopes. This report describes a method of barcoding fixed cells after a transient partial permeabilization with 0.02% saponin that results in efficient and consistent barcode staining with fluorescent or mass-tagged reagents while preserving surface marker staining. This approach simplifies barcoding protocols and allows direct comparison of surface marker staining of multiple samples without concern for variations in the antibody cocktail volume, antigen-antibody ratio, or machine sensitivity. Using this protocol, cellular barcoding can be used to reliably detect subtle differences in surface marker expression. PMID:25274027

  8. Staining Pattern Classification of Antinuclear Autoantibodies Based on Block Segmentation in Indirect Immunofluorescence Images

    PubMed Central

    Li, Jiaqian; Tseng, Kuo-Kun; Hsieh, Zu Yi; Yang, Ching Wen; Huang, Huang-Nan

    2014-01-01

    Indirect immunofluorescence based on HEp-2 cell substrate is the most commonly used staining method for antinuclear autoantibodies associated with different types of autoimmune pathologies. The aim of this paper is to design an automatic system to identify the staining patterns based on block segmentation compared to the cell segmentation most used in previous research. Various feature descriptors and classifiers are tested and compared in the classification of the staining pattern of blocks and it is found that the technique of the combination of the local binary pattern and the k-nearest neighbor algorithm achieve the best performance. Relying on the results of block pattern classification, experiments on the whole images show that classifier fusion rules are able to identify the staining patterns of the whole well (specimen image) with a total accuracy of about 94.62%. PMID:25474260

  9. Reversible MHC multimer staining for functional isolation of T-cell populations and effective adoptive transfer.

    PubMed

    Knabel, Michael; Franz, Tobias J; Schiemann, Matthias; Wulf, Anna; Villmow, Brigitte; Schmidt, Burkhard; Bernhard, Helga; Wagner, Hermann; Busch, Dirk H

    2002-06-01

    Recently developed major histocompatibility complex (MHC) multimer technologies allow visualization and isolation of antigen-specific T cells. However, functional analysis and in vivo transfer of MHC multimer-stained cells is hampered by the persistence of T-cell receptor (TCR) MHC interactions and subsequently induced signaling events. As MHC monomers do not stably bind to TCRs, we postulated that targeted disassembly of multimers into MHC monomers would result in dissociation of surface-bound TCR ligands. We generated a new type of MHC multimers, which can be monomerized in the presence of a competitor, resulting in rapid loss of the staining reagent. Following dissociation, the T cells are phenotypically and functionally indistinguishable from untreated cells. This 'reversible' T-cell staining procedure, which maintains the specificity and sensitivity of MHC multimer staining while preserving the functional status of T lymphocytes, may be of broad benefit for ex vivo investigation of T-cell functions and clinical applications. PMID:12042816

  10. 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

  11. The effect of staining on the monotonic tensile mechanical properties of human cortical bone

    PubMed Central

    Kayacan, Ramazan

    2007-01-01

    Microdamage in the form of microcracks has been observed in cortical bone following in vivo and in vitro fatigue loading. It has been suggested that bone has an inherent ability to repair microdamage at physiological activity levels. If the biological remodelling and repair process cannot keep up with the rate of damage accumulation, as in ageing bone and in individuals such as athletes and military recruits, microdamage may accumulate even at physiological activity levels. Such microdamage accumulation is thought to contribute to stress and fragility fractures. It is therefore important to obtain quantitative data on the rate of damage accumulation so as to understand the etiology of skeletal fractures. Sequential labelling of microdamage using fluorochrome stains at different stages of mechanical loading is becoming standard for assessing damage evolution. Although verification of this staining technique is provided in the literature, it has not yet been reported if the stains change the mechanical properties of cortical bone. In this study, monotonic tensile tests were performed to investigate the effect of the staining on the monotonic tensile mechanical properties of cortical bone. Forty-eight specimens were machined from human femora obtained from three male subjects, aged 52–55 years, and all 48 specimens were systematically divided into one control and three treatment groups. Specimens in the first (n = 12) and second treatment groups (n = 12) were stained with alizarin complexone and calcein (0.0005 m), respectively, for 16 h under 50 mmHg vacuum. Specimens in the third treatment group (n = 12) were kept in calcium-supplemented saline solution under the same conditions of the first and second treatment groups. Specimens in the control group (n = 12) were removed from the freezer prior to testing and allowed to thaw at room temperature in saline solution. Differences among the mean values of the mechanical properties for four testing groups were determined by the Mann–Whitney test at a significance level of P < 0.05. The statistical results indicated that the chelating stains and the staining conditions have no significant effect on the mechanical properties of the cortical bone under monotonic tensile loading. This study demonstrated that microcrack labelling with the chelating stains under aforementioned conditions (stain concentration, staining time, etc.) is a reliable method in that staining cortical bone with alizarin complexone and calcein prior to testing does not affect tensile properties. PMID:17894797

  12. Weathering patinas on the medieval (S. XIV) stained glass windows of the Pedralbes Monastery (Barcelona, Spain)

    Microsoft Academic Search

    Meritxell Aulinas; Maite Garcia-Valles; Domingo Gimeno; Jose Luis Fernandez-Turiel; Flavia Ruggieri; Montserrat Pugès

    2009-01-01

    Background, aim, and scope  The first step in the restoration of a medieval stained glass window is the evaluation of its degree of degradation. This\\u000a implies the study of the chemical composition of the stained glass as well as the new mineral phases developed on its surface\\u000a (patinas). Patinas are clearly related to glass composition, time, environmental conditions, microenvironments developed in

  13. Use of Romanowsky type (Diff3) stain for detecting Helicobacter pylori in smears and tissue sections

    Microsoft Academic Search

    A M Zaitoun

    1992-01-01

    A Romanowsky type (Diff-3) stain was used for identifying Helicobacter pylori in gastric biopsy specimens from 50 patients with ulcer and non-ulcer dyspepsia. Air dried smears were prepared from fresh biopsy tissue and histological sections were prepared from paraffin wax processed tissue. The Diff-3 technique is accomplished in five steps and takes about 30 seconds. Results using the Diff-3 stain

  14. MITOCHONDRIAL LOCALIZATION OF OXIDATIVE ENZYMES: STAINING RESULTS WITH TWO TETRAZOLIUM SALTS

    Microsoft Academic Search

    ALEX B. NOVIKOFF; WOO-YUNG SHIN; JOAN DRUCKER

    1961-01-01

    A comparison is made of the staining results obtained with Nitro-BT and MTT-Co ++ as acceptors when DPNH is the substrate in frozen sections of cold formol-calcium-fixed rat kidney (normal and following ligation of the blood vessels) and human liver containing lipofuscin granules. The kidney results are evaluated in terms of mitoehondrial morphology seen after classical mitochondrial stains and in

  15. 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.

  16. 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.

  17. In vitro capacitation of dog spermatozoa as assessed by chlortetracycline staining

    Microsoft Academic Search

    P. Guérin; M. Ferrer; A. Fontbonne; L. Bénigni; M. Jacquet; Y. Ménézo

    1999-01-01

    We developed an assay for detecting capacitation and acrosome status in dog spermatozoa using chlortetracycline (CTC) as a fluorescent probe. Sperm cells were stained after incubation in modified canine capacitation medium (mCCM). Calcium ionophore A23187 permitted the induction of acrosomal exocytosis of capacitated sperm cells. Spermac® staining and transmission electron microscopy were used as control tests to detect acrosome-reacted spermatozoa.Three

  18. Fluorescence and confocal laser scanning microscopy imaging of elastic fibers in hematoxylin-eosin stained sections

    Microsoft Academic Search

    Hernandes Faustino de Carvalho; Sebastião Roberto Taboga

    1996-01-01

    We have studied the possibility of associating fluorescence microscopy and hematoxylin-eosin staining for the identification\\u000a of elastic fibers in elastin-rich tissues. Elastic fibers and elastic laminae were consistently identified by the proposed\\u000a procedure, which revealed it-self to be easy and useful for the determination of such structures and their distribution. The\\u000a fluorescence properties of stained elastic fibers are due to

  19. Evaluation of the one-step eosin-nigrosin staining technique for human sperm vitality assessment

    Microsoft Academic Search

    L. Bjorndahl; I. Soderlund; U. Kvist

    2003-01-01

    BACKGROUND: The one-step eosin-nigrosin staining technique for assessment of sperm vitality was developed in the 1950s for various mammalian species. Although commonly used on human sperm in semen, a validation for this use has not previously been published. METHODS: The technique was evaluated on 1235 consecutive semen samples. RESULTS: The one-step eosin-nigrosin staining technique gave valid results when evaluated with

  20. Should Gram Stains Have a Role in Diagnosing Hip Arthroplasty Infections?

    Microsoft Academic Search

    Aaron J. Johnson; Michael G. Zywiel; D. Alex Stroh; David R. Marker; Michael A. Mont

    2010-01-01

    Background  The utility of Gram stains in diagnosing periprosthetic infections following total hip arthroplasty has recently been questioned.\\u000a Several studies report low sensitivity of the test, and its poor ability to either confirm or rule out infection in patients\\u000a undergoing revision total hip arthroplasty. Despite this, many institutions including that of the senior author continue to\\u000a perform Gram stains during revision